Nutritional Evaluation and Glycaemic Response of Food Products Prepared Using Stevia Rebaudiana

Transcription

1 Nutritional Evaluation and Glycaemic Response of Food Products Prepared Using Stevia Rebaudiana By SUHANI ARORA 2009HS69D Thesis submitted to CCS Haryana Agricultural University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY IN FOODS AND NUTRITION Department of Foods and Nutrition I.C. College of Home Science CCS Haryana Agricultural University Hisar

2 CERTIFICATE I This is to certify that this thesis entitled, Nutritional Evaluation and Glycaemic Response of Food Products Prepared Using Stevia Rebaudiana, submitted for the degree of Doctor of Philosophy, in the subject of Foods and Nutrition to the CCS Haryana Agricultural University, is a bonafide research work carried out by Ms. Suhani Arora (Admn. No. 2009HS69D) under my supervision and that no part of this dissertation has been submitted for any other degree. The assistance and help received during the course of investigation have been fully acknowledged. Dr. (Mrs.) Sudesh Jood Major Advisor Professor Deptt. of Foods and Nutrition CCS HAU, Hisar

3 CERTIFICATE II This is to certify that this thesis entitled, Nutritional Evaluation and Glycaemic Response of Food Products Prepared Using Stevia Rebaudiana, submitted by Ms. Suhani Arora (Admn. No. 2009HS69D) to the CCS Haryana Agricultural University in partial fulfillment of the requirements for the degree of Doctor of Philosophy, in the subject of Foods and Nutrition, has been approved by the Student s Advisory Committee after an oral examination on the same, in collaboration with an External Examiner. MAJOR ADVISOR EXTERNAL EXAMINER HEAD OF THE DEPARTMENT DEAN, POSTGRADUATE STUDIES

4 CERTIFICATE III FORMAT FOR P. G. THESIS It is certified that the thesis submitted by Ms. Suhani Arora, Adm. No. 2009HS69D, Ph.D. student of this department has been checked and found as per specification of the format circulated by the Dean, PGS vide his Memo No. PGS/A-1/09/ dated MAJOR ADVISOR PROFESSOR AND HEAD

5 ACKNOWLEDGEMENT Gratitude cannot be seen or expressed, it can only be felt deep in heart and is beyond description. Although thanks are poor expression of debt of gratitude one feels, yet there is no better way to express it. With the grace of Baba Ji Maharaj I am able to carve another milestone in my academic journey. I allide this present opportunity to express my deep sense of gratitude and obligation to my esteemed major advisor Dr. (Mrs.) Sudesh Jood, Professor, Department of Foods and Nutrition, CCS HAU, Hisar for her valuable and sustaining guidance, continous encouragement, constructive and valuable suggestions throughout the course of this investigation. I wish to convey my heartiest appreciation for patience, understanding and round the clock help extended to me by my co-major advisor Dr. (Mrs.) Darshan Punia, Sr. Scientist, Foods and Nutrition. I also honour and thanks this same sparke to the members of my advisory committee Dr. R.K Sharma, Sr. Scientist, Department of Horticulture; Dr. R.S. Dabur, Professor, Department of Animal Products Technology and Dr. Sheela Sangwan, Professor, Department of HDFS for their timely help and valuable guidance rendered during the course of investigation and for valuable suggestions for the improvement of manuscript. I am also greatly thankful to Dr. (Mrs.) Pinky Boora, Professor and Head, Department of Foods and Nutrition for timely providing all the required facilities. With all sincerely, care, love and affection I express my heartfelt thanks to my loveable Mom, Dad, Akash, Angel, Karan, Gulshan Chachu, Komal Chachi, My cute little bittle AAnya, Mamta, Shweta, Kiran Di, Priyanka, Alka and Mani for precious praise, worthy suggestions and cooperation which went a long way for their patience, moral support and love extended during the entire course of study. Last but not the least days come and days goes with every passing day the memories may fade away. But for me its not just the matter of 24 hrs but for the life which I m living just because of you. I still can t forget the days and memories when no one came for my help but you gave your hand and let all my worries goes off. I don t need any occasion to show my gratefulness towards you because every moment of my life is a blessing of yours. Because emotions, feelings come direct from heart only for those people which we love truly and these should be describe straightforwardly. These are my pure emotions just for you. You will always be my angel in heaven who will always be loved. Love you so so much and miss you Prateek. They say memories are golden, well maybe that is true, I never wanted memories, I only wanted you. A million times I needed you, a million times I cried. In life I loved you dearly, In death I love you still, In my heart you hold a place no one else could ever fill. If love could build a stairway and heartache build a lane, I'd walk the path to heaven and bring you back again. Our family chain is broken, and nothing seems the same. But as God calls us one by one, the chain will link again. Place : Hisar Dated : March, 2015 Suhani Arora

6 CONTENTS CHAPTER DESCRIPTION PAGE(S) I INTRODUCTION 1-3 II REVIEW OF LITERATURE 4-16 III MATERIALS AND METHODS IV RESULTS V DISCUSSION VI SUMMARY AND CONCLUSION REFERENCES i-vii ANNEXURES I-IV

7 Table No. LIST OF TABLES Description 2.1 Proximate analysis of dried Stevia rebandiana leaves (g/100g on dry weight basis) 2.2 Mineral contents of dried Stevia rebandiana leaves (mg/100g on dry weight basis) 4.1 Moisture, ash and sugars contents of Stevia leaves and Stevia powder g/100g, on dry matter basis) 4.2 Mineral contents of Stevia leaves and Stevia powder g/100g, on dry matter basis) 4.3 Mean scores of organoleptic characteristics of tea Mean scores of organoleptic characteristics of coffee Mean scores of organoleptic characteristics of milk shake Mean scores of organoleptic characteristics of RTS Mean scores of organoleptic characteristics of biscuit Mean scores of organoleptic characteristics of cake Mean scores of organoleptic characteristics of bun Mean scores of organoleptic characteristics of sweet porridge Mean scores of organoleptic characteristics of carrot halwa Mean scores of organoleptic characteristics of kheer Proximate composition of tea (% as is basis) Sugar and starch content of tea (% as is basis) Mineral contents of tea (% as is basis) Proximate composition of coffee (% as is basis) Sugar and starch content of coffee (% as is basis) Mineral contents of coffee (% as is basis) Proximate composition of milk shake (% as is basis) Sugar and starch content of milk shake (% as is basis) Mineral contents of milk shake (% as is basis) Proximate composition of RTS (% as is basis) Sugar and starch content of RTS (% as is basis) Mineral contents of RTS (% as is basis) Proximate composition of biscuit (g/100g, on dry matter basis) Sugar and starch content of biscuit (g/100g, on dry matter basis) Mineral contents of biscuit (g/100g, on dry matter basis) Proximate composition of cake (g/100g, on dry matter basis) 62 Page No

8 4.29 Sugar and starch content of cake (g/100g, on dry matter basis) Mineral contents of cake (g/100g, on dry matter basis) Proximate composition of bun (g/100g, on dry matter basis) Sugar and starch content of bun (g/100g, on dry matter basis) Mineral contents of bun (g/100g, on dry matter basis) Proximate composition of sweet porridge (g/100g, on dry matter basis) Sugar and starch content of sweet porridge (g/100g, on dry matter basis) Mineral contents of sweet porridge (g/100g, on dry matter basis) Proximate composition of carrot halwa (g/100g, on dry matter basis) Sugar and starch contents of carrot halwa (g/100g, on dry matter basis) Mineral contents of carrot halwa (g/100g, on dry matter basis) Proximate composition of kheer (g/100g, on dry matter basis) Sugar and starch contents of kheer (g/100g, on dry matter basis) Mineral contents of kheer (g/100g, on dry matter basis) Anthropometric measurements of subjects Amount of beverages ingested 25 g of equicarbohydrate portion for glycaemic index / person 4.45 Amount of food products ingested 50g of equicarbohydrate portion for glycaemic index / person 4.46 Area under blood glucose curve (mg/dl) of milk shake prepared using sugar, sucralose and Stevia powder for normal subjects Area under blood glucose curve (mg/dl) of milk shake prepared using sugar, sucralose and Stevia powder for obese subjects 4.48 Area under blood glucose curve (mg/dl) of milk shake prepared using sugar, sucralose and Stevia powder for diabetic subjects 4.49 Glycaemic Index of milk shake in normal, obese and diabetic subjects Area under blood glucose curve (mg/dl) of RTS prepared using sugar, sucralose and Stevia powder for normal subjects Area under blood glucose curve (mg/dl) of RTS prepared using sugar, sucralose and Stevia powder for obese subjects 4.52 Area under blood glucose curve (mg/ dl) of RTS prepared using sugar, sucralose and Stevia powder for diabetic subjects 4.53 Glycaemic Index of RTS in normal, obese and diabetic subjects Area under blood glucose response curve (mg/dl) of biscuits prepared using sugar, sucralose and Stevia powder in normal subjects Area under blood glucose response curve (mg/dl) of biscuits prepared using sugar, sucralose and Stevia powder in obese subjects 4.56 Area under blood glucose response curve (mg/dl) of biscuits prepared using sugar, sucralose and Stevia powder in diabetic subjects 88 90

9 4.57 Glycaemic Index of biscuits in normal, obese and diabetic subjects Area under blood glucose curve (mg/dl) of cake using sugar, sucralose and Stevia powder in normal subjects 4.59 Area under blood glucose curve (mg/dl) of cake using sugar, sucralose and Stevia powder in obese subjects 4.60 Area under blood glucose curve (mg/dl) of cake using sugar, sucralose and Stevia powder in diabetic subjects 4.61 Glycaemic Index of cake in normal, obese and diabetic subjects Area under blood glucose curve (mg/dl) of bun using sugar, sucralose and Stevia powder in normal subjects 4.63 Area under blood glucose curve (mg/dl) of bun using sugar, sucralose and Stevia powder in obese subjects 4.64 Area under blood glucose curve (mg/dl) of bun using sugar, sucralose and Stevia powder in diabetic subjects 4.65 Glycaemic Index of bun in normal, obese and diabetic subjects Area under blood glucose curve (mg/dl) of sweet porridge using sugar, sucralose and Stevia powder in normal subjects 4.67 Area under blood glucose curve (mg/dl) of sweet porridge using sugar, sucralose and Stevia powder in obese subjects 4.68 Area under blood glucose curve (mg/dl) of sweet porridge using sugar, sucralose and Stevia powder in diabetic subjects 4.69 Glycaemic Index of sweet porridge in normal, obese and diabetic subjects Area under blood glucose curve (mg/dl) of carrot halwa using sugar, sucralose and Stevia powder in normal subjects 4.71 Area under blood glucose curve (mg/dl) of carrot halwa using sugar, sucralose and Stevia powder in obese subjects 4.72 Area under blood glucose curve (mg/dl) of carrot halwa using sugar, sucralose and Stevia powder in diabetic subjects 4.73 Glycaemic Index of carrot halwa in normal, obese and diabetic subjects Area under blood glucose curve (mg/dl) of kheer using sugar, sucralose and Stevia powder in normal subjects 4.75 Area under blood glucose curve (mg/dl) of kheer using sugar, sucralose and Stevia powder in obese subjects 4.76 Area under blood glucose curve (mg/dl) of kheer using sugar, sucralose and Stevia powder in diabetic subjects 4.77 Glycaemic Index of kheer in normal, obese and diabetic subjects Cost (Rs./100g) of the food products prepared using sugar, sucralose and acceptable level of Stevia powder 116

10 LIST OF FIGURES Fig. No. Description Page No. 4.1 Blood glucose curve of milk shake in normal subjects Blood glucose curve of milk shake in obese subjects Blood glucose curve of milk shake in diabetic subjects Blood glucose curve of RTS in normal subjects Blood glucose curve of RTS in obese subjects Blood glucose curve of RTS in diabetic subjects Blood glucose curve of biscuits in normal subjects Blood glucose curve of biscuits in obese subjects Blood glucose curve of biscuits in diabetic subjects Blood glucose curve of cake in normal subjects Blood glucose curve of cake in obese subjects Blood glucose curve of cake in diabetic subjects Blood glucose curve of bun in normal subjects Blood glucose curve of bun in obese subjects Blood glucose curve of bun in diabetic subjects Blood glucose curve of sweet porridge in normal subjects Blood glucose curve of sweet porridge in obese subjects Blood glucose curve of sweet porridge in diabetic subjects Blood glucose curve of carrot halwa in normal subjects Blood glucose curve of carrot halwa in obese subjects Blood glucose curve of carrot halwa in diabetic subjects Blood glucose curve of kheer in normal subjects Blood glucose curve of kheer in obese subjects Blood glucose curve of kheer in diabetic subjects 115

11 LIST OF PLATES Fig. No. Description Page No. 1 Stevia leaves and powder 18 2 Tea 19 3 Coffee 20 4 Milk shake 20 5 RTS 21 6 Biscuits 22 7 Cake 22 8 Bun 23 9 Sweet porridge Carrot halwa Kheer 25

12 CHAPTER-I INTRODUCTION Stevia (Stevia rebaudiana) is a perennial herb that belongs to the Asteracae family. There are around 150 species within the Stevia family including Stevia dianathoidea, Stevia phlebophylla, Stevia anisostemma, Stevia bertholdii, Stevia crenata, Stevia enigmatica, Stevia eupatoria, Stevia lemmonii, Stevia micranatha, Stevia ovata, Stevia plummerae, Stevia rebaudiana, Stevia salicifolia, Stevia serrata and Stevia viscida with all plants being sweet but Stevia rebaudiana having the highest sweetness levels. It is widely used in many parts of the world as sweetener and grown commercially in Central America, Korea, Paraguay, Brazil, Thailand and China (Haroslav et al., 2006; Gupta et al., 2014). In India cultivation of Stevia as a crop is still restricted to the research level. However, Department of Ayurveda, Yoga and Naturopathy, Unani, Sidha and Homeopathy (AYUSH), (Government of India) has sanctioned proposals for the prospects of S. rebaudiana cultivation of various states like West Bengal, Uttranchal, Haryana and Punjab. The isolated diterpene glycoside which is secondary metabolites responsible for the sweet taste of Stevia (Ahmed et al., 2011). The leaves of Stevia contain a natural complex mixture of eight sweet diterpene glycosides including isosteviol, stevioside, rebaudioside (A, B, C, D, E, F), steviolbioside and dulcoside A (Rajasekaran et al., 2008; Goyal et al., 2010). Out of various steviol glycosides (SGs), stevioside and rebaudioside A are the major metabolites and these compounds are 250 to 300 times as sweet as sucrose (Debnath, 2008; Gupta et al., 2013), ph-stable, heat stable, not fermentable (Abdullateef and Osman, 2012) and posses healthy promoting potential. Along with sweetness, Stevia has some bitter affect due to the presence of some essential oils, tannins and flavonoids (Parsons, 2001). Stevia commonly known in Sanskrit as madhu patra, meaning sweet leaf is natural and healthy alternative to sugar and artificial sweeteners. It is also known as honey yerba and honey leaf and by some other variations of these names (Ahmed et al., 2011). It has also been reported that Stevia rebaudiana, as a non-calorie first natural sweetener used in medicinal green teas for treating heart burn and other ailments (Vanek et al., 2001). Even though there are more than 200 species of the genus Stevia, only S.rebaudiana gives the sweetest essence (Savita et al., 2004). It has been reported that Stevia and its products in cooked or baked foods, processed foods, beverages and juices, sherbets,etc. have been used since a long time in Japan, China, Russia and USA ( Goyal et al., 2010; Balaswamy et al., 2014). In 1970 s while cyclamate and saccharin were suspected as carcinogens, Japan began cultivating Stevia as an alternative which soon gained popularity and got commercialized, hence today it is cultivated in many countries (Chatsudthipong and Manprasat, 2009; Gupta et al., 2014). 1

13 Stevia leaves have sensory and functional properties superior to those of many other high-potency sweeteners and is likely to become a major source of natural sweetener for the growing food market. Leaves of Stevia rebanudiana has many medical applications like antimicrobial, anti-viral, anti-fungal, anti-hypertensive, anti-hyperglycaemic, anti-tumour, antiinflammatory, anti-diarrhoeal, diuretic and anti-human rota-virus activities (Mohan and Robert, 2009; Chatsudthipong and Muanprasat, 2009). The toxicological studies have shown that secondary metabolites present in Stevia does not have teratogenic, mutagenic or carcinogenic effects and no allergic reactions have been observed after consuming it as an sweetener (Pol et al., 2007). The major compounds of Stevia as steviol glycosides are metabolized and dominated through similar pathways in both human and animals (Genus et al., 2007). Rebaudioside A in the digestive tract is first metabolized by microbes in the colon to stevioside which is further concerted into glucose molecule and steviol. The released glucose molecule is used by bacteria in the colon and is not absorbed into the blood stream. The metabolized components essentially leave the body and there is no accumulation (Genus et al., 2007; Pol et al., 2007). Obesity and its related health problems like diabetes, hyperlipidemia, heart disease and hypertension are the results of modern lifestyle and changed diets with balance tilted towards refined foods specially sugar and fat. The over consumption of refined sugars, especially sucrose promotes inappropriate positive caloric balance, loss of body weight control, excessive weight gain, which can lead to obesity, a risk factor for some chronic diseases such as diabetes (Thomas and Glade, 2010; Agarwal et al., 2010; Pande and Khetmalas, 2012; Gupta et al., 2014). There is an alarming increase in the incidence of diabetes in India and with world s largest population being noted, India is labeled as Diabetic capital of world. According to International Diabetes Federations, diabetes currently affects 382 million people worldwide and India has the second largest number of people with diabetes (IDF, 2013; Gupta et al., 2014). Therefore, substituting sugar with low calorie sweeteners may be efficacious in reducing the weight and its related health problems. So sugar substitutes like saccharin, sucralose and aspartame gained importance in reducing calorie intake. However, these are artificial sweeteners can cause more health problems than they cure. These artificial sweeteners attack vital organs that could lead to serious complications after prolonged use (Thomas and Glade, 2010; Balaswamy et al., 2014). Globally, scientists have concluded that Stevia sweeteners are safe for people of all ages. Stevia leaf or extracted forms like stevioside, rebaudioside A and steviol glycosides was approved by US FDA as a dietary supplement considered (Generally recognized as safe). Currently, the joint FAO/WHO Expert Committee on Food Additives reviewed the safety of 2

14 steviol glycosides and an acceptable daily intake (ADI) for steviol glycosides (expressed as steviol equivalents) of 4 mg/kg body weight/day was recommended (European Food Safety Authority, 2010). Nowadays, food industry is searching for solutions to reduce the levels of certain ingredients in their products, such as fat, salt and sugars in order to produce healthier foods that meet the consumer s expectations. As consumers are becoming more cautions regarding their health in terms of calories intake. The market potential for Stevia sweetener is still untapped. It is estimated that about 65.1 million Indian people are presently suffering from diabetes. With such a huge share of the population being diabetic, the new ventures in the food industry are focused entirely on low calorie food products for diabetics (IDF, 2013; Gupta et al., 2014). Keeping in mind the sweetening property and other medicinal uses of Stevia rebaudiana, an attempt has been made to explore the use of Stevia rebaudiana as a sweetener in comparison with the other artificial sweetener available in the market, so that the diabetic and calorie conscious people can add variety to the diet and relish the food. Keeping these facts in mind, the present study was planned with the following objectives: 1. To determine nutrient composition of Stevia powder 2. To standardize and develop food products using Stevia powder and to study their organoleptic acceptability 3. To study the nutrient composition and glycaemic response of most acceptable developed food products 3

15 CHAPTER II REVIEW OF LITERATURE The literature regarding nutrient composition of Stevia leaves and Stevia powder and the developed food products using Stevia powder and low calorie artificial sweeteners for their nutrient composition and glycaemic index has been suitably reviewed under the following heads and subheads: 2.1 Nutritional composition of Stevia leaves and Stevia powder 2.2 Organoleptic characteristics and nutritional composition of food products using Stevia and artificial sweetners 2.3 Therapeutic implications and glycaemic response of food products 2.1 Nutritional composition of Stevia leaves and Stevia powder Stevia planted in experimental plot in India contained 4 per cent steviosides and 0.2 per cent rebaudiosides (Rosalizon, 2010). The chemical contents were lower compared with plants grown in other countries. Leaves of Paraguan Stevia contains 9 to 13 per cent of the sweet steviosides/ rebaudiosides molecules, Chinese Stevia contains only 5 to 6 per cent and Indian Stevia contains about 9.08 per cent steviosides based on dry weight of leaves (Yadav et al., 2011). Measurements Stevia is the new emerging alternative source of calorie free sweetener having no carbohydrate and fat. It is 20 to 30 times sweet then cane sugar and beet sugar, highly nutritious, delicious, non toxic and non additive sugar. It enhances the flavour, helpful in digestion, weight reduction, antioxidant, prevents dental caries and having antimicrobial and anti-plaque properties. It increases mental alertness, energy levels but does not affect the blood sugar level and can also be used in hypertension and hypoglycaemic. It can also be used as alternative source of food confectionaries, bakeries, fruit juices, jams, biscuits, chocolates, vegetables and other food stuffs (Arora et al., 2008; Agarwal et al., Proximate composition and mineral contents of Stevia rebaudiana leaves have been reported by various workers are given in Table 2.1 and 2.2 Table 2.1: Proximate analysis of dried Stevia rebandiana leaves (g/100g on dry weight basis) Parameters Tadhani and Subhash (2006) Goyal et al. (2010) Kaushik et al. (2010) Mishra et al. (2010) Moisture ND Protein Fat Ash Fibre ND 15.2 ND 18.0 Carbohydrate 35.2 ND ND

16 Table 2.2: Mineral contents of dried Stevia rebandiana leaves (g/100g on dry weight basis) Parameters Serio (2010) Abon-Arab et al. (2010) Atteh et al. (2011) Calcium Phosphorus 318 ND 2.6 Sodium ND Potassium Iron Magnesium Zinc ND Savita et al. (2004 a) also analysed nutritional composition of Stevia on dry weight basis and it was reported Stevia only produce 2.7 Kcal/g energy which may be entitled as the low calorie sweeteners. Other major nutrients like protein, fat, carbohydrate, ash and crude fibre were 9.8, 2.5, 52.0, 10.5 and 18.5 g/100g, respectively. Stevia also contained good amount of minerals such as calcium (464.6), phosphorus (55.3), iron (11.4), sodium (19.0) and potassium (1800.0) mg/100g, respectively. Stevia was found to have higher percentage of anti-nutritional factor (oxalic acid) which may hinder the bioavailability of minerals. Manish and Rema (2006) reported that Stevia leaves contained g protein, 4.34 g fat, g carbohydrate and g/100g, respectively ash content on dry weight basis. They also reported high content of K, Ca, Na, P, Mg and Mo in Stevia leaves. Mishra et al. (2010) determined nutritional and mineral composition of Stevia leaves on dry weight basis and observed that Stevia powder contained moisture, protein, fat, total carbohydrate, ash and crude fibre to be 7.0, 10.0, 3.0, 52.0, 11.0, and 18.0 g/100g, respectively. They reported 11 per cent of ash in leaves of S. rebaudiana. A significant content of Ca (464 mg/100g), P (11 mg/100g), Na (190 mg/100g), K (1800 mg/100g), Mg (349 mg/100g), Fe (55 mg/100g) and Zn (1.5 mg/100g) have also been reported in Stevia leaves. Abou Arab et al. (2010) analysed dried Stevia leaves for chemical composition, amino acids content and mineral contents. The moisture, protein, fat, fibre, ash, and carbohydrates were found to be 5.37, 11.41, 3.73, 15.52, 7.41 and per cent, respectively. Stevia leaves also contained 5.88, 9.77 and per cent for reducing sugars, non- reducing sugars and total soluble carbohydrates, respectively. The mean concentrations of macro minerals (potassium, calcium, sodium and magnesium) and micro elements (copper, manganese, iron and zinc) were also determined in dried Stevia leaves. Potassium (21.15 mg/100 g ) followed by calcium (17.70 mg/100 g), sodium (14.93 mg/100 g) and magnesium (3.26 mg/100 g) were detected. On the other hand, the leaves contained lower amount of 5

17 copper, manganese, iron and zinc which have been recorded as 0.73, 2.89, 5.89 and 1.26 mg/100 g, respectively. It was observed that sucrose had higher acceptability scores of sweet taste followed by syrup stevioside. They also reported that stevioside had slight bitterness, some astringency, after taste and showed fairly low general acceptability among people. Similarly, Atteh et al. (2011) also analyzed Stevia leaves and Stevia stem for nutritional composition. Protein, fat, crude fibre and ash in Stevia leaves were 16.0, 2.6, 6.8 and 15.5 per cent and in Stevia stem were found to be 6.7, 1.1, 45.1 and 5.6 per cent, respectively. The Ca, P, Na, K, Mg, Zn, Mn, Fe and Cu content in leaves and stem were reported to be 800, 200, 70, 1700, 240, 200, 30.03, 36.6, 600 mg/100g, respectively and 40, 120, 50, 1850, 90, 800, 60, 120 and 200 mg/100 g, respectively. A significant variation was observed in fatty acid profiles of Stevia leaves and Stevia stem. Stevia leaves had higher content of saturated fatty acids as compared to Stevia stems. Pande and Khetmalas (2011) analyzed Stevia extract and stevioside powder and reported 31 per cent carbohydrates, 25 per cent proteins and 9 per cent reducing sugars, respectively. Whereas, the concentration of protein, carbohydrates and reducing sugar in stevioside powder were found to be 5, 28.2 and 0.8 g/100 g, respectively. Yadav and Guleria (2012) also evaluated Stevia leaves for their proximate and mineral composition on dry weight basis. Stevia leaves contained high amount of protein, carbohydrate and ash contents. The values were 20.45, and g/100 g, respectively. Stevia leaves contained low amount of fat i.e g/100 g. Potassium, calcium, magnesium, phosphorus, sodium, and sulphur were found in higher amounts whereas, copper, cobalt, manganese, zinc, selenium, and molybdenum were found in trace amounts. Samsudin and Aziz (2013) reported freshly harvested Stevia leaves contained 80 per cent moisture content and 1.6 per cent soluble sugars. Dried Stevia leaves contained protein, fat, carbohydrate, fibre and ash contents were in the range of 15.65, 1.0, 65.18, and 8.8 g/100g, respectively. The Stevia leaves were also found to contain K, Ca and P ( , and mg/100 g on dry weight basis) in higher amount and Fe and Na (24.62 and mg/100 g of dry weight basis) in lower amount, respectively. Similarly, Chaturvedi et al. (2013) estimated Stevia leaves for nutritional composition and mineral content. It was reported that Stevia leaves contained 9.5 ± 2.5 g protein, 3.6 ± 0.02 g fat and ± 1.5 g fibre /100 g, respectively. The ash content was found to be 10.4 ± 1.61 g/100 g which indicated that Stevia leaves are good source of minerals. Stevia leaves powder contained calcium (390.4 ± 4.5 mg/100 g), phosphorus (16.5 ± 0.57 mg/100 g) and iron (29.4 ± 3.5 mg/100 g), respectively. Stevia leaves also had high amount of glycosides, flavonoids, terpenoids, tannins and saponins. Campos et al. (2014) analysed the functional properties, proximate and dietary fibre composition of two varieties of Stevia Bertoni (S. Morita II and S. Criolla) and reported that 6

18 Morita II had higher (0.28 g water/g sample) water holding and organic molecule absorption capacity in comparison to Criolla variety (and 0.25 g water/g sample). Both varieties showed similar oil holding capacity. For proximate composition, it was reported that S. Morita II and S. Criolla contained carbohydrates (64.06 and g), protein (12.11 and g), crude fiber (5.92 and 9.52 g) and ash (7.82 and g) /100 g, respectively. Whereas Morita II and Criolla leaves were found to be poor source of fat (3.23 and 3.04g) /100 g. TDF content in the S. Morita II and S. Criolla leaves powder were and g/100 g, respectively, The major content represented by IDF per cent in S. Morita II and per cent in S. Criolla leave powder. The remainder was constituted by SDF. Gasmalla et al. (2014) determined the effect of three methods of drying, viz, sun, oven and microwave on Stevia (Stevia rebaudian Bertoni) leaves for their physiochemical properties, chemical composition and microbiological contamination and reported that Stevia leaves contained very high amount of ash (12.06 %) and total soluble substance (17.03 %), high carbohydrates (63.10 %) and protein (13.68 %), whereas, low amount of moisture (10.73 %), fiber (5.03 %), fat (6.13 %) and reducing sugar (4.50 %) contents. For sun, oven and microwave drying moisture content ranged from 4.45 to per cent; ash ranged from 4.65 to per cent; protein ranged from to per cent; fat ranged from 4.18 to 6.13 per cent; total dietary fiber ranged from 4.35 to 5.26 per cent and total carbohydrates ranged from to per cent, respectively. Its high ash content indicates that the Stevia leaves are good source of inorganic minerals. 2.2 Organoleptic characteristics and nutritional composition of food products using Stevia and artificial sweeteners Stevia rebaudiana is perennial herb with claimed medicinal and culinary characteristics. It is a plant of daisy family that grows naturally in South America (Geuns, 2003). Quality of Stevia sweetness is preferable to that of aspartame or saccharin. It serves as a flavour enhancer and remains stable when combined with acidic foods. High temperature does not destroy its sweetening properties and it does not discolour. Savita et al. (2004 b) incorporated Stevia leaf powder at different levels of substitution i.e., 50, 60, 75 and 100 per cent in ten products: biscuit, grape juice, sweet bun, tea, milk shake basen ladoo, wheat ladoo, sweet bun and chikki, etc. The recipes developed were also tested on organoleptic acceptability and glycaemic index. The mean scores of sensory evaluation for appearance, texture, flavour, taste and over all acceptability showed no significant variation between the products. All the products developed were equally accepted. They also reported that Stevia contained 9.8 g of protein, 10.5 g of ash, 18.4 g of crude fibre and 269 Kcal. Mineral composition indicated fairly high amounts of calcium, iron, sodium and potassium with high amount of oxalic acid also. Glycaemic index of products found to be lower in diabetics as well as in normal individuals. 7

19 Singh et al. (2005) prepared flavoured milk beverage with incorporation of 10, 20 and 30 per cent of carrot juice and evaluated for organoleptic characteristics. Results revealed that though all types of flavoured milk were found acceptable but 20 per cent carrot juice flavoured milk scored the significantly high scores. Similarly, Sharma (2006) prepared low calorie aonla-ginger, aonla-lime and lime-ginger RTS, sugar can be successfully substituted by Stevia up to the extent of 50, 50 and 75 per cent, respectively. Jagadeesh and Hegde (2008) developed Stevia leaf extract based Ready-to-Serve (RTS) beverages of mango and pomegranate in different combinations and studied for organoleptic scores. It was reported that RTS prepared with Stevia extract alone had significantly lower scores. The RTS prepared from cane sugar alone (control) as a source of sweetness obtained the highest scores in both the fruits individually and in combinations. However, pomegranate and mango (50:50) with 25 per cent Stevia extract and 75 per cent sugar ranked statistically at par with the control. Ahmed et al. (2008) prepared and tested fifteen beverages with sucrose, cyclamate, aspartame and acesulfame K for sensory characteristics. The drink with 100 per cent sucrose was used as a control. Control sample got higher scores of taste as compared to treated samples. It was noted that diet drink based on sweetener got less scores as compared to drinks in which combination of sweeteners were used. Drink prepared by incombination of cyclamate (%): aspartame (%): acesulfame-k 1:2:1 got the maximum score for flavour. While lower flavour score was reported for drink based on aspartame and acesulfame K (3:1) and non-significant difference was observed in rest of the drinks with control. Jagadeesh and Laxminarayan (2008) made mango and pomegranate RTS using Stevia extract and found that the RTS containing 15 per cent pomegranate juice and 2.13 per cent Stevia aqueous extract with 8 per cent sucrose was found to be comparable with control (sugar). Similarly, fruit dietetic beverages (5 % bittergourd juice) with Stevia aqueous extract (13 %) which were found equally comparable in sensory characteristics in comparison to the control beverage (sugar based) (Singh et al., 2012). Sujith and Chithra (2009) substituted Stevia powder in the place of cane sugar in rice flakes kheer, wheat halwa, fruit custard and bread pudding. The mean scores obtained by the recipes ranged between 3.0, 4.0, 4.0 and 4.4 out of 5.0 with respect to overall acceptability. The Stevia rice flakes kheer contained energy values ( Kcal), carbohydrate content ( g), fat content ( g), protein ( g), and fibre ( g), respectively. Similarly, Mogra and Dashora (2009) tested sweetness equivalence by comparing sweetness of Stevia extract with sugar and other sweeteners like sugar free, equals, natura, sugar free natura and saccharine Eleven recipes viz milk, coffee, tea, gajar halwa, milk shake, kheer, curd, lemon water, custard, halwa and lapsi were prepared using Stevia extract and other sweeteners. The mean scores of sensory evaluation for appearance, color, flavour, texture, 8

20 taste, overall acceptability showed perceivable variation between the products. All the products with Stevia extract were well accepted by the panel members ( ) and ranked first when compared with other artificial sweeteners ( ) and found statistically at par (p>0.001 %) with sugar ( ) in coffee, halwa, milk shake, kheer and lapsi while superior over the rest of recipes. They also reported proximate composition values for the 100g cooked product, The prepared recipe using 25 mg Stevia in custard, kulfi and sandesh had g of moisture, 3.91 g of protein, 1.35 g of fat, 0.18 g of fibre, 1.09 g of ash, g of carbohydrates content in custard, whereas 58.81g of moisture, 9.37 g of protein, 13.1 g of fat, 0.47 g of fibre, 2.3 g ash,15.95 g of carbohydrates were present in kulfi and recipe of sandesh had 25 mg protein, 1.77 g of fat, 0.05 g of fibre, 3.57 g of ash, 8.37 g of carbohydrates, respectively. Salem et al. (2009) developed regular yoghurt cake and yoghurt cake for diabetics using the replacement of sugar with hot water extract of Stevia leaves of equal sweetness. Yoghurt diabetic cake was found to have good sensory score and found acceptable by all the panelists. Agarwal et al. (2009) prepared sweet cereal products namely malpura, mithi roti, pinni and sevian using Stevia powder at three different levels i.e. 25, 50 and 75 mg, respectively. Developed products were also evaluated for organoleptic characteristics and nutritional composition. Malpura with 50 mg Stevia powder, mithi roti with 75 mg Stevia powder and pinni and sevian with 37.5 mg of Stevia powder were found to be acceptable in comparison to control (sugar based) recipe. The modified recipe of malpura had g of protein, 3.41 g of fat, g of carbohydrates and provided 287 Kcal of energy. The modified recipe of mithi roti had of protein, g of fat, g of carbohydrate and provided 374 Kcal of energy. The modified recipe of pinni had g of protein, g of fat, g of carbohydrate and provided 399 Kcal of energy. The modified recipe of sevian had 4.42 g of protein, 5.60 g of fat, g of carbohydrate and provided 119 Kcal of energy. The percent decrease in calories provided by modified recipe compared to the basic recipe for malpura, mithi roti, pinni and sevian were reported to be per cent, per cent, per cent and per cent, respectively. Sweet cereal products using Stevia powder were highly acceptable upto 75 mg and are low in calories as compared to the basic recipe which makes them suitable for consumption by diabetics to satisfy their craving for sweet foods. Arora et al. (2009) formulated sweetened burfi using sugar (6 %) and aspartame ( %) per cent aspartame burfi was analysed for sensory evaluation. The higher scores were reported by control burfi. Sucrose helps to form a network and create a binding effect in the product. Hence, the inclusion of sugar improved the body and texture of the product. Sugar also added to the caramelised colour of the control product. On the other hand, aspartame had insufficient water binding capacity, which resulted in a product with 9

21 a slightly lower score. Aspartame-sweetened burfi could not retain most textural attributes like hardness, adhesiveness, springiness and chewiness whereas chewiness was retained by the aspartame-sweetened burfi. The lightness was significantly less (p < 0.05) incontrol samples than aspartame-sweetened burfi. Agarwal et al. (2010) developed custard, kulfi and sandesh using Stevia powder and evaluated for organoleptic characteristics and nutritional composition. In custard, Stevia powder was incorporated at 12.5 mg, 25 mg and 50 mg and 6 g of sugar was used in control. In kulfi, the amount of Stevia powder used was 25 mg, 37.5 mg and 50 mg and 25 g of sugar was used in control. Whereas, 25 mg, 50 mg and 62.5 mg of Stevia powder was incorporated in sandesh where 10 g of sugar added was taken as control. All the products with three levels were organoleptically evaluated. The results revealed that custard, kulfi and sandesh with 25 mg Stevia powder was liked very much whereas 12.5 mg of Stevia powder in custard, 50 mg of Stevia powder in kulfi and 62.5 mg of Stevia powder in sandesh were neither liked nor disliked by the panelists. The modified recipe of custard had g of moisture, 3.91 g of protein, 1.35 g of fat, g of carbohydrates and provided 74 Kcal of energy. The modified recipe of kulfi had 58.81g of moisture, 9.37 g of protein, 13.1g of fat, g of carbohydrate and provided 219 Kcal of energy. The modified recipe of sandesh had g of moisture, g of protein, 1.77 g of fat, 8.37 g of carbohydrate and provided 125 Kcal of energy. The percent decrease in calories provided by modified recipe compared to the basic recipe for custard, kulfi and sandesh were reported to be 23.71, and per cent, respectively. Sweet milk products using Stevia powder were highly acceptable upto 25 mg and are low in calories as compared to the basic recipe which makes them suitable for consumption by diabetics. Bhattacharya (2010) developed rosogolla with alternative sweetener (commercial sucralose). Rosogolla with 7 per cent sucrose and 5 per cent alternative sweetener (sucralose) syrup was prepared by dissolving 50 g of sugar free natura (sucralose) in 1000 ml cold water. Textural, shelf life, cost effectiveness and acceptance by people were studied and reported that hardness of control rosogolla (5.56±1.19) was significantly lower than the experimental rosogolla (7.86±0.28) samples. The cohesiveness of control rosogolla was found to be at par with those of experimental rosogolla. Springiness of experimental sample was nonsignificantly differed from that of control. Gumminess of experimental rosogolla was higher than the control rosogolla. Chewiness of experimental samples was significantly higher than control rosogolla. Byanna and Gowda (2010) developed ready-to-serve beverages using sweet orange (Citrus sinensis), pomegranate and kokum with sugar and sugar substitutes (sucralose and fructose) in different combinations. The RTS were also evaluated for sensory attributes. Nectar with sugar and sucralose (50:50) and sucrose and fructose (50:50) were liked very 10

22 much by the panel of judges. Sweet orange and kokum (88 : 12) squash with sucrose and fructose (50:50) and sweet orange and pomegranate with sucrose and fructose (50:50) were rated as superior for over all acceptability scores in comparison to control for sensory evaluation. Mishra (2011) replaced Stevia leaf powder (0.5 g and 1.0 g) with sugar in tea/ coffee and milk shake. All the products were found to be acceptable by all the judges as compared to control. Similarly, Giri et al. (2012) prepared dietetic kulfi with 50, 60 and 70 per cent sugar replacement with 0.05, 0.06 and 0.07 per cent refined Stevia extract powder. All the products were evaluated for sensory characteristics. It was found that above 50 per cent sugar replacement resulted in bitterness, lack of brownish appearance and presence of icy texture. Paul et al. (2013) prepared fifteen experimental treatments of lassi by incorporation of whey and Stevia powder with five varying proportions of whey and curd in ratios viz. 70:30, 60:40, 50:50, 40:60 and 30:70, respectively and three variations of Stevia powder (viz.0.13, 0.15 and 0.17 %, respectively ). Sensory evaluation of the three products were conducted and reported that treatment with 70 per cent curd, 30 per cent whey and 0.15 per cent Stevia powder was found best in case of flavor, taste, consistency and overall acceptability. Whereas 70 per cent curd, 30 per cent whey and 0.13 per cent Stevia powder proved to be best in terms of colour and appearance. In all the combinations of 70 per cent curd, 30 per cent whey and 0.15 per cent Stevia powder was found to be the most acceptable combination. Rice kheer, fruit punch and papaya shake were developed by incorporating Stevia leaves powder with 25, 50, 75 and 100 per cent. The products developed by incorporating 25 per cent Stevia leaves powder was found to be liked very much where as with the increase in the quantity of Stevia leaves decreased the acceptability of the products and there were found in the category of neither liked nor disliked by the judges (Chaturvedi et al., 2013). Similarly, Alizadeh et al. (2014) developed a low calorie and low glycaemic index (GI) ice creams by using Stevia. Sucralose was used in control ice cream. Ice cream prepared using Stevia (100 %) had low glycaemic index as compared to control ice cream. Balaswamy et al. (2014) utilized Stevia aqueous extract and stevioside powder to prepare RTS beverage based on fresh juice of jamun, pomegranate, pineapple and purple grapes. Stevia aqueous extract (1.5, 2.0 and 1.5 %), stevioside powder (20, 25, 30 and 35 mg/100 ml) in RTS beverage was found to be palatable with 10, 15, 20 and 25 per cent juice concentration. The results of sensory evaluation revealed that jamun beverage with stevioside was found to be liked very much by the panelists. All the other RTS beverages with stevioside were liked moderately by the panelists. Mango beverage with Stevia aqueous extract was found to be unacceptable as Stevia aqueous extract imparted raw flavour and dull shade to the beverages. 11

23 Singh and Singh (2014) formulated aonla based low calorie blended nectar from aonla, mango, guava, jamun and jackfruit using Stevia for low calorie. Sugar and Stevia were used in ratios (100: 0, 75:25, 50:50, 25:75, 0:100) per cent, respectively. It was found that half of the sugar can be successfully substituted by Stevia in the preparation of aonla based low calorie blended nectar without impairing the quality of beverages. Over all acceptability of aonla pulp and mango pulp (25:75) incorporating sugar and Stevia (50:50) was found to be significantly superior. Whereas, low calorie nectar prepared from aonla pulp and guava pulp (75:25) with sugar and Stevia powder (75:25) were found to be liked very much by the panellists. Perera et al. (2014) developed a ready to serve herbal drink using a water extract of Syzygium cumini bark. Four combinations of ready to serve drink samples were prepared by S.cumini decoction and sucralose solution (0.01g/100 ml). Results indicated that colour, sweetness and overall acceptability of the herbal drinks were not significantly different. The selected sample with the highest overall acceptability contained 20 ml of S.cumini decoction and 0.01 per cent sucralose solution. Mehrotra et al. (2014) formulated shrikhand by incorporating Stevia powder and extract to replace sugar. Stevia powder and extract were added in different combination in the experimental products. Shrikhand by 100 per cent sugar was served as control. Combination of sugar and Stevia extract/stevia powder in combinations 100 per cent; 90:10; 80:20; 70:30 and 60:40 was incorporated and same proportions for Stevia powder was also prepared and tested for organoleptic characteristics. They reported that the mean sensory score for colour and appearance of shrikhand showed a slight decrease in colour and appearance as the amount of Stevia leaves powder was increasing. The variation in colour and appearance was not significant upto 30 per cent thereafter a significant (P 0.05) variation was observed. However the variation was not significant upto 30 per cent replacement of sugar content. The mean sensory score for flavour of shrikhand showed that 30 per cent replacement of sugar with Stevia leaf powder scored better, however the substitution had significantly no effect on the flavour and texture of shrikhand in comparison to the control sample. However 40 per cent replacement of sugar with Stevia extract decreased the sensory attributes of shrikhand significantly (P 0.05). Mean sensory score for overall acceptability of shrikhand showed that the replacement of sugar upto 30 per cent with Stevia leaf powder scored the highest. The replacement of sugar by the powder till 30 per cent did not differ significantly from the control sample. However, 40 per cent replacement of sugar with Stevia powder decreased significantly (P 0.05) the overall acceptability. Shrikhand prepared by replacing 30 per cent sugar with Stevia powder showed an increase in moisture, protein and fat. Increasing trend was observed in protein from 5.80 to 5.90, moisture to and fat from 8.75 to

24 per cent, respectively. On the other hand, decrease in energy value was observed on substitution of sugar with Stevia powder. 2.3 Therapeutic implications and glycaemic response of food products Stevia helps to treat many ailments like high blood pressure, hyperlipidemia, hyperglycemia, obesity, skin diseases and digestive disorders. It has no side effects and is safe for human consumption (Hore et al., 2002; Ferri et al., 2006; Anton et al., 2010).Studies on Stevia produces evidence that the natural sweetners stevioside and steviol, a product formed by enzymatic hydroxylation within the plant which is found in plant S. rebaudiana has been used for many years in the treatment of diabetics among Indians, Paraguay and Brazil (Akhtar et al., 2001). Stevioside and steviol stimulate insulin secretion by direct action on beta cells and these compounds have potential role as anti-hyperglycemia agent in the treatment of type 2 Diabetes Mellitus (Jeppesen et al., 2000; Atteh et al., 2011; Ali et al., 2013) Gregersen et al. (2004) conducted study on twelve type 2 diabetic patients (4 females/8 males) with a mean age of 65.8 yrs, a diabetes duration of 6.0 years, a mean body mass index of 28.5 m 2 /kg in the study. A standard test meal was served either with 1 g of stevioside encapsulated in gelatin and administered orally. The total energy content of the test meal was 412 Kcal. Stevioside suppresses the postprandial blood glucose level in type 2 diabetic subjects by an average 18 per cent. However, when calculated the insulinogenic index, a significant increase of 40 per cent was found. A blood glucose lowering effect of stevioside both when administered intravenously and when given orally was observed. Savita et al. (2004b) determined the effect of consumption of Stevia rebaudiana on diabetic and hypertensive group. Thus, eight hypertensives (2 men and 6 women) and 6 diabetics (4 men and 2 women) were selected between the age group of yrs. About 0.5 g and 1.0 g of Stevia leaf powder was substituted to sweet bun and chikki respectively per 100 g of the end product, where, bun without Stevia and glucose (75 g available carbohydrate) was served as reference food. The feeding trial was conducted for 30 days. The results revealed that the effect of consumption of Stevia products on blood sugar levels were in variance, but the mean values of serum insulin and urine sodium level increased and the mean values of blood pressure, lipid profile and weight of subjects decreased. It was also found to have anti-hypertensive effect. Manish and Rema (2006) incorporated and standardised Stevia leaf powder in selected recipes. Out of the 10 standardised recipes, sweet bun and chikki were selected and fed to groups of diabetic (N=6) and hypertensive (N=8) respectively belonging to the age group of years for a period of 30 days. Sweet bun had 100 per cent sugar replacement where as chikki had 50 per cent jaggery with Stevia leaf powder. Fasting and post prandial blood glucose levels, serum insulin levels and weights of the subject were assessed before and after the study period. The results revealed that mean initial serum insulin level was 8.2 IU/ml 13

25 and after 30 days the mean serum insulin level was 15.3 IU/ml. An increase was observed in mean serum insulin level after 30 days of consumption of Stevia but increase was nonsignificant. Freckmann et al. (2007) also conducted a study on 24 healthy volunteers [12 female, 12 male, age 27.1 ± 3.6 yrs, body mass index (BMI) 22.6 ± 1.7 kg/m 2 ]. Subjects received standardized meals for breakfast, lunch and free choice dinner from a buffet equicarbohydrate portion of 50 g. The mean 24-hour interstitial glucose concentration was 89.3 ± 6.2 mg/dl (range mg/dl), with a mean glucose concentration of 93.0 ± 7.0 mg/dl and 81.8 ± 6.3 mg/dl before all meals. The highest postprandial interstitial glucose concentrations were observed after ingestion of meals with fast absorption characteristics, rice pudding with sugar and cinnamon peak glucose concentrations were ± 14.4 and ± 21.1 mg/dl. Areas under the postprandial tissue glucose curve after ingestion of fast absorption characteristics showed the highest values in the first 2 hours and a rapid decline after 2 hours. In contrast, meal containing high fat and low sugar induced only a small, flat increase in tissue glucose concentrations. Meal consists of fruits or boiled vegetables induced a prolonged increase in tissue glucose concentrations with the highest areas under the curve in the first 2 hours and a slow decline thereafter. Aston et al. (2008) studied GI values of forty-two healthy adult volunteers. Subjects were served with 25 and 50 g portions of glucose on three occasions, followed by a selection of test foods providing an equal amount of available carbohydrate, in random order. Capillary blood glucose levels were measured in the fasted state, 30, 60, 90 and over the 120 min followed by commencement of consumption of the foods. Two of the breads were tested using portions containing both 25 and 50 g available carbohydrate, to compare the results obtained. A significant difference was not found between GI values obtained with different portion sizes for either of these breads, indicating that it is acceptable to use a smaller portion size where necessary when measuring GI. Kochar et al. (2008) conducted a study on 90 non-insulin dependent male subjects between 40 to 50 yrs, free from sickness complications were selected from Punjab Agriculture University (PAU) hospital, Ludhiana to see the effect of supplementation of Stevia leaves powder and nutrition counselling on anthropometric parameters and gain in knowledge. The subjects selected were divided into three groups namely group I, group II and group III having 30 subjects each. Group I subjects were only given medicine prescribed by the doctor, group II subjects were given medicine prescribed by the doctor and nutrition counselling, group III subjects were given medicine, nutrition counselling and Stevia leaves powder. One g of Stevia leaves powder was incorporated in 500 mg capsules and were given to the subjects daily during breakfast and lunch for three months. Nutrition education was given to the subjects of group II and group III for 3 months after fifteen days interval. It was observed that majority of 14

26 the subjects in all the three groups had craving for sweet foods, liked fried foods. Frequency of consumption of vegetables and fruits by the subjects was increased in group II and III after nutrition counselling. Physical activity pattern of the subjects of group II and group III was increased after nutrition counselling, significant decrease in the weight of the subjects of group II and III was observed. It was seen that there was a significant increase in knowledge, attitude and practices and hence gain in total knowledge of subjects in group II and III after three months of the study. While there was a non-significant increase in the knowledge of the subjects of group I. Hence, it was concluded that supplementation of Stevia leaves powder along with nutrition counselling significantly improved the nutritional status of diabetic patients. Sharma et al. (2009) determined the effect of consumption of Stevia extract on selected 20 hypercholesteromic women. Stevia extract was standardized in the laboratory; 20 ml of extract was used to intervene one subject in a glass of water (200 ml). The results revealed that the consumption of Stevia extract reduced the levels of cholesterol, triglyceride, LDL-C significantly while in an increase in HDL-C was noted. Thus concluded that Stevia extract have hypolipidaemic effect and can be used to reduce the risk of CVD in future. Sujith and Chithra (2009) substituted Stevia in the place of cane sugar in rice flakes kheer, wheat halwa, fruit custard and bread pudding. In the selected volunteers, the mean fasting blood glucose recorded was around 77 mg/dl. On administration of glucose, the mean blood glucose level rose to mg/dl in 30 min. But administration of Stevia incorporated rice flakes kheer recorded a mean blood glucose level of only 103 mg/dl after 30 min showing the difference of around 33 mg/dl. Similarly, after 60 min, the mean blood glucose levels registered were mg/dl and 96.3 mg/dl on administration of the reference food and the test food, respectively showing the difference of around 20 mg/dl. A similar trend was also observed after 120 min also. The glycaemic index of the Stevia incorporated rice flakes kheer was found to be 49.7 (low) and glycaemic load was found to be 12.4 (medium). Mishra (2011) interrogated fifteen patients were who were vegetarian, Out of fifteen, nine patients were women and six patients were men between the age group of yrs. Feeding trial was for 45 days. First 15 days, patient was given medicine and their fasting and post prandial diabetic level were measured, then for next 15 days, they were not given any medicine under normal diet and their prandial and fasting glucose level were measured. Then for further 15 days, they were given Stevia leaf three times a day with tea and their glucose level were measured. About 0.5 g and 1g of Stevia leaf powder was consumed with tea in place of sugar. Fasting blood sugar (FBS) and post prandial (PPBS) blood sugar were taken FBS was and PPBS was For next 15 days, patient were asked to stop taking medicine under normal diet and in the supervision of physician and their FBS and PPBS were measured as and 283 mg/dl, respectively. There was significant difference in both FBS 15

27 and PPBS which showed that while taking medicine their FBS and PPBS were under control. For further 15 days, patients were given Stevia leaf powder (0.1g to 5g) with tea (three times a day). FBS and PPBS were measured as and Significant decrease in the FBS and PPBS level while consuming Stevia leaf powder was observed. There was significant decrease in FBS with Stevia consumption. Parimalavalli and Radhaisri (2011) determined glycaemic index of chapati prepared using Stevia in six women having diabetes. The reference food (glucose), control food (chapati with 2 % sugar) and test food (2 % Stevia incorporated chapati) were given to each of the women on first, second and third day, respectively. The mean value of glucose for fasting 30, 60, 90 and 120 min were reported to be 105.2±20.1, 205.3±6.3, 229.5±74.9, 220.8±45.8 and ± 43.8 mg/dl, respectively. The mean glycaemic response of control chapati (multimix flour and 2% sugar) at fasting, 30, 60, 90 and 120 min were observed to be 105.8±8.4, 184.3±59.8, ± 41.2, ± 75.6 and ± 40.1 mg/dl, respectively. Whereas, chapati with Stevia showed the mean glycaemic response 95.3 ± 11.4, ± 6.1, ± 31.9, ± 30.9 and 132 ±24.4 mg/dl at fasting, 30, 60, 90 and 120 min, respectively. The peak value (229.5 mg/dl) was observed at 60 min for glucose, whereas the peak value 184 mg/dl and 176 mg/dl were recorded for control chapati and Stevia chapati at 30 min. The levels showed a reducing trend at 120 min. Glucose recorded the highest response (179.3 mg/dl) followed by control chappati (147.7 mg/dl) and lowest response (132 mg/dl) was recorded by the Stevia chapati. The glycaemic index of control chapati and Stevia chapati was 71.2 and 61.9 respectively. The GI of Stevia chapati was significantly lower than control chappati. Alizadeh et al. (2014) developed a low calorie and low glycaemic index (GI) of soft ice cream using mixture of sucrose and Stevia. Five formulations were developed using the combinations of sucrose and Stevia.Young, apparently healthy adults with average age of 23.3 ± 4.16 years and mean body mass index was 23 ± 4.37 were included for study. The mean glycaemic index of ice cream formulated with and without sucrose was reported as ± 4.01 and ± 5.27, respectively. Mean calorie values of two ice creams were recorded to be and The remarkable reduction in caloric value and GI of Stevia based ice creams, concluded that substitution of Stevia with sugars brings a new relatively healthy choice for food basket of families with high risk of life style related diseases including diabetes mellitus. Some studies have shown that Stevia have a re-vitalizing effect on β-cells of pancreas, improve insulin, sensitivity in rats, and possibly even to promote additional insulin production, helping to reverse diabetes and metabolic syndrome. Stevia consumed before meals significantly reduced postparandial insulin levels compared to both aspartame and sucrose. It was concluded that Stevia sweetners would likely have benefit for diabetic patients (Benford et al., 2006; Brusick, 2008). 16

28 CHAPTER III MATERIAL AND METHODS The present investigation was conducted in the Department of Foods and Nutrition, I.C. College of Home Science, Chaudhary Charan Singh Haryana Agricultural University, Hisar (Haryana). This study was designed to analyze the nutritient composition and glycaemic response of products developed by incorporating Stevia powder This chapter delineates information pertaining to the research design and methodological steps used for the present investigation. The research procedures have been distinctly described under the following heads and subheads: 3.1 Procurement and Preparation of Products Procurement Preparation 3.2 Standardization and development of food products Beverages Tea Coffee Milk shake RTS Baked Products Biscuits Cake Bun Traditional Products Sweet porridge Carrot halwa Kheer 3.3 Organoleptic evaluation of food products 3.4 Nutrient composition of Stevia leaf, Stevia powder and food products developed by incorporating Stevia powder Proximate composition Moisture Crude protein Fat Crude fibre Ash 17

29 Total carbohydrates Carbohydrate Total soluble sugars Reducing sugars Non-reducing sugars Starch Dietary fibre Total minerals Calcium Iron Zinc Magnesium Sodium Potassium 3.5 Glycaemic response of developed food products using sugar (control), sucralose and Stevia powder 3.6 Cost calculation of the developed food products 3.7 Statistical analysis 3.1 Procurement and preparation of products Procurement: The raw products namely milk, coffee, tea, sugar, rice, carrot, lemon, mint leaves, refined flour, yeast, oil and margarine were procured from local market of Hisar city. The Stevia powder was procured from IARI, Pusa, New Delhi. Plate 1 : Stevia leaves and powder 3.3 Standardization and development of food products Various products were standardized and developed by incorporating Stevia powder, sucralose and sugar. The products developed using sugar was served as control. The following products were developed: 18

30 3.3.1 Beverages Tea, Coffee, Milk shake & RTS Preparation schedule for Tea Ingredients: Replacement in sugar based tea (control) was made with two levels of Stevia powder and sucralose individually. Types of Tea Milk (ml) Water (ml) Tea (g) Sugar (g) Sucralose (mg) Stevia powder (mg) Control (Sugar) Sucralose Stevia powder T T Method Plate 2 : Tea Milk, water and tea leaves were heated on high flame. A pinch of cardamom powder was also added. Then sugar/sucralose/stevia powder was added. Tea was boiled for 3 min on low flame. Tea was strained, poured and served Preparation schedule for Coffee Ingredients: Replacement in sugar based coffee (control) was made with two levels of Stevia powder and sucralose individually. Types of Coffees Milk (ml) Water (ml) Coffee powder (g) Sugar (g) Sucralose (mg) Stevia powder (mg) Control (Sugar) Sucralose Stevia powder T 1 T

31 Plate 3: Coffee Method Milk was boiled. Coffee powder was dissolved in water and added to boiled milk. Then sugar/sucralose/stevia powder was added. Poured and served Preparation schedule for Milk shake Ingredients: Replacement in sugar based milk shake (control) was made with two levels of Stevia powder and sucralose individually. Milk (ml) Cocoa powder (g) Powdered Sugar (g) Sucralose (mg) Stevia powder (mg) Control (Sugar) Sucralose Types of Milk shakes Stevia powder T1 T2 Plate 4: Milk shake Method: Milk was boiled and cooled. Cocoa powder and powdered sugar/sucralose/stevia powder was added. The milk shake was blended nicely in the homogenizer/mixer. Milk shake was chilled and served. 20

32 Preparation schedule for RTS Ingredients: Replacement in sugar based RTS (control) was made with two levels of Stevia powder and sucralose individually. Types of RTS Mint leaves (g) Water (ml) Bengal Gram Flour (g) Lemon juice (ml) Sugar (g) Sucralose (mg) Stevia powder (mg) Control (Sugar) Sucralose Stevia powder T T Method Flour was roasted. Plate 5 : RTS Mint leaves and lemon juice was added. Powdered sugar/sucralose/stevia powder, salt, black pepper and cumin powder were mixed thoroughly. Water was added and blended in mixer. Serve chilled Baked products Preparation schedule for Biscuit Ingredients: Replacement in sugar based biscuit (control) was made with two levels of Stevia powder and sucralose individually Types of Biscuit Wheat flour (g) Ghee (g) Milk (g) Ammonia (g) Sodium bicarbonate (g) Sugar (g) Sucralose (mg) Stevia Powder (mg) Control (Sugar) Sucralose Stevia powder - - T T

33 Method Creamed ghee. Plate 6 : Biscuits Sugar/sucralose/Stevia powder was added and creamed it. Added sodium bicarbonate and ammonia powder and again creamed. Added milk and flour and mixed well. Placed the dough for conditioning for 1 hr in freezer. Kneaded the dough again and prepared biscuits by biscuit making machine. Baked at 160ºC in preheated oven for 15 min or till brown Preparation schedule for Cake Ingredients: Replacement in sugar based cake (control) was made with two levels of Stevia powder and sucralose individually. Types of Cakes Control (Sugar) Refined flour (g) Refined oil (ml) Curd (g) Baking powder (g) Sodium bicarbonate (g) Sugar (g) Sucralose (mg) Stevia powder (mg) Sucralose Stevia powder - - T T Plate 7 : Cake Method Sieved refined flour, sodium bicarbonate and baking powder together. Creamed oil and sugar /sucralose/stevia powder and whipped it. Added curd, water and mixed well. 22

34 Added sieved flour in it and added more water if required. Poured the mixture in greased tin and baked at 160oC for min or till brown Preparation schedule for Bun : Ingredients: Replacement in sugar based bun (control) was made with two levels of Stevia powder and sucralose individually. Refined Flour (g) Refined oil (ml) Yeast (g) Cherries (g) Mix fruit essence Salt (g) Sugar (g) Sucralose (mg) Stevia powder (mg) Control (Sugar) Sucralose Types of Buns Stevia powder TI T2 Plate 8 : Bun Method Sieved refined flour and added warm water and then mixed with yeast properly. Added sugar with yeast in control recipe. Then sucralose/stevia powder (T1 and T2) were added to the flour for other three modified recipes before adding the yeast. Mixed all the mixtures properly and dough was divided into equal balls and placed them in oven tray for initial proofing for 15 min. Take out the tray and cherries were added and the buns were remoulded and kept for 2nd proofing for 30 min. Finally buns were taken out from incubator. Baked at 160ºC for 5-10 min Traditional products Preparation schedule for Sweet porridge Ingredients: Replacement in sugar based sweet porridge (control) was made with two levels of Stevia powder and sucralose individually 23

35 Types of Sweet porridge Wheat Grits (g) Ghee (g) Water (ml) Salt (g) Sugar (g) Sucralose (mg) Stevia powder (mg) Control (Sugar) Sucralose Stevia powder T I T Method Plate 9 : Sweet porridge Wheat grits were roasted with ghee in pan. Sugar/sucralose/Stevia powder and salt were added to it. Required amount of water was added in all the recipes and boiled in pressure cooker. Cooked for 7-8 min on high flame in pressure cooker with closed lid Preparation schedule for Carrot halwa Ingredients: Replacement in control was made with different levels of Stevia powder and sucralose. Types of Carrot halwa Carrot (g) Ghee (g) Milk (ml) Cardamom powder (g) Sugar (g) Sucralose (mg) Stevia powder (mg) Control (Sugar) Sucralose Stevia powder T T Plate 10 : Carrot halwa 24

36 Method Washed the carrots and grated them. Added milk to the grated carrots and cooked them till milk evaporated. Added sugar (control) / sucralose / Stevia powder separately to the above mixture. Cooked the mixture for a while and served hot Preparation schedule for Kheer Ingredients: Replacement in sugar based kheer (control) was made with two levels of Stevia powder and sucralose individually Types of Kheer Rice (g) Milk (ml) Cardamom powder (g) Sugar (g) Sucralose (mg) Stevia powder (mg) Control ( Sugar) Sucralose Stevia powder T T Plate 11 : Kheer Method Rice was washed properly. Milk was added in washed rice and cooked on slow flame. Sugar (control) / sucralose/ Stevia powder was added separately in the cooked kheer. Removed it from fire after getting proper consistency. Then added cardamom powder and served hot. 3.3 Organoleptic evaluation of value-added products For selecting the acceptable products for further study, the above mentioned developed products were subjected to organoleptic evaluation by a panel of ten judges from the I.C. College of Home Science, Chaudhary Charan Singh Haryana Agricultural University, Hisar, Haryana. The judges were asked to record the quality characteristics i.e. colour, appearance, aroma, texture/consistency, taste and overall acceptability by employing a ninepoint Hedonic Rating Scale as given in Annexure-I. Judges were asked to rinse their mouth 25

37 before and after testing each product. Mean scores for all sensory characteristics were calculated. 3.4 Nutritional composition of Stevia leaf, Stevia powder and food products developed by incorporating Stevia powder The most acceptable products having desirable sensory properties were further analyzed for the following parameters Proximate composition Moisture (AOAC, 2000). Procedure Moisture content was estimated by employing the standard method of analysis Five ml of wet sample and 5 gram food sample was weighed in a petridish and dried in oven at 50 o C for 8 h for beverages and for 6 h for food products, respectively. The sample was reweighed after cooling it in a desiccator. Moisture was calculated by using the following equation Loss in weight (g) Moisture (%) = 100 Weight of sample (g) Crude protein Crude protein was estimated by standard method of analysis (AOAC, 2000), using KEL PLUS Automatic Nitrogen Estimation System. Reagents i) Hydrochloric acid (0.01N) ii) iii) iv) Boric acid solution (40%): Dissolved 400 g of boric acid in distilled water and diluted to one litre. Sodium hydroxide solution (40%): Dissolved 400 g of carbonate free sodium hydroxide (NaOH) in distilled water and diluted to one litre. Copper sulphate catalyst: Mixed copper sulphate (CuSO 4 :5H 2 O) and potassium sulphate in ratio of 1:9. v) Mixed indicator solution: Dissolved 2 g of methyl red and 1 g methylene blue or bromo-cresol green in one litre of ethanol. Stored in dark brown bottle. vi) Digestion mixture : K 2 SO 4 : CuSO 4 (5 : 1) Procedure Fifteen g K 2 SO 4, 1 ml CuSO 4, 5H 2 O catalyst solution and 8-10 boiling chips were added to digestion flask. Milk was warmed to C and mix thoroughly. Five ml sample for beverages and 200 mg of sample for food products were taken and immediately placed in digestion flask. 25 ml H 2 SO 4 was added, rinsing any milk on neck of flask down into bulb. Flask may be stopped and held for digestion at later time. 26

38 Digestion Temperature was set to C in the controller. The samples and chemicals were prepared. The tubes with samples + sulphuric acid + digestion mixture (3 g) in insert rack were loaded. Then the manifold was placed over the tubes. The rack was inserted + manifold were loaded in the digestion block. The water connection was opened. After one to two hour the rack was removed and it was noted whether all the samples got digested. If not, the tubes in the block were replaced and were left for another 15 min. The end point of digestion was the appearance of bluish green colour and flames got reduced. After digestion, the insert rack was removed from the block and was placed in the cooling stand. It was removed slowly after 15 min till the tubes got cooled. Then finally, the water connection was closed. Now, the samples were ready for distillation. Distillation First, the macro tube containing the digested samples was loaded in the space provided in apparatus. One empty conical flask was put on the receiver side and ran the programme of the equipment. Boric acid (20 ml) was automatically dropped into the conical flask. This was pink in colour initially. The 40 ml of 40 per cent NaOH in auto mode was added slowly in the order of 10 ml each time (till the colour in the test tube changed from bluish green to brown precipitate). Then the process was set. After 6 min, the colour in the conical flask changed from pink to green. This was the end point of distillation of the sample. The flask was taken out for titration. Titration: Titrated the above solution with 0.1 N HCl till colour changed from green to permanent pale pink colour. This was the end point of titration. Where, 14 Titrant value Normality of acid 100 Total N (%) = 1000 sample weight Titrate value = Volume of N/10 HCl used for titration Crude Fat For beverages: Procedure Crude fat was determined by AOAC (2000) method Ten ml of sample was taken and heated if necessary, then 1.5 ml of NH3-solution was added and heated in a water bath for 15 min at C. The mixture was shaken occasionally. Then the mixture was cooled, 35 ml ethyl alcohol was added and mixed. 25 ml of ethyl ether was added. The cylinder was closed tightly and the contents were mixed by turning the cylinder up and down for 1 minute. 25 ml of petroleum ether was added and the procedure was repeated again. The mixture was allowed to stand for at least 2 hrs. The ether 27

39 phase was quite clear and completely separated from the water phase. Ether phase was transferred to a flask by means of the pipette, which is then cleaned with a little amount of ether. This ether was also transferred to the flask. Take care that none of the water phase is introduced into the flask. The procedure was further repeated for two times, each time 25 ml of petroleum ether was added. Same flask was used each time. After the final extraction, the ether was evaporated on a hot plate or similar and flask was dried for about one hour in an oven at 102 C ± 2 C. Flask is cooled at room temperature in a desiccator and weighed. The procedure was repeated i.e oven dried and cooled until a constant weight was reached. For food products: Fat was estimated by employing the standard method of analysis (AOAC, 2000) using the Automatic SOCS plus Solvent Extraction System. Procedure: The beaker was washed thoroughly and dried in hot air oven at 60 0 C. The weight of empty beaker was recorded. Five gram of moisture free sample was weighed and transferred into cellulose thimble. The thimble holder along with the sample was kept into the beaker. Required quantity of solvent (petroleum ether, boiling point C) into the beaker was taken. The beakers were loaded into the system and set on required temperature (90 0 C according to boiling point of solvent) in the controller. The extraction was carried out for one hour at 90 0 C. After the completion of extraction period, the temperature was raised at C, the stopper was closed in order to collect the solvent in the solvent compartment. The beaker was removed along with the fat and kept in hot air oven at 60 0 C temperature, till a constant weight was obtained. The beaker was weighed after cooling it in a dessicator. Where, W 2 W 1 Fat (%) = 100 W W = Weight of sample (g) W 1 = Weight of empty beaker W 2 = Weight of beaker with fat Crude fibre For beverages samples were dried after extracting fat in petridish in oven and scraps were obtained and were further grounded to powder for further analysis. The crude fibre was estimated by standard method of analysis (AOAC, 2000) using Automatic Fibre plus system. 28

40 Reagents i) Sulphuric acid solution (1.25 %): Dissolved 12.5 ml of sulphuric acid in 200 ml ii) Procedure distilled water and made volume to one litre. Sodium hydroxide solution (1.25 %): Dissolved 12.5 g sodium hydroxide in distilled water and diluted to one litre. One g fat free dried sample was taken and put in weighed crucibles. The crucibles were fixed in apparatus. One fifty ml of 1.25 per cent H 2 SO 4 was filled in each of the beaker through funnels. The instrument was then connected to water supply. All the knobs were set on closed mode and the instrument was switched on. The temperature was fixed to 550 C by pushing the set button. When solution in beakers started boiling, the temperature was reduced to 400 C and the instrument was allowed to run for 45 min. After 45 min, the instrument was switched off. The water source was then changed from direct to via suction apparatus. All the knobs were put in open position and the big knob was changed from pressure to vaccum. The suction pump apparatus was also switched on. This whole process was then repeated twice distilled water followed by NaOH solution and again two times with distilled water. After that, the instrument was switched off. The crucibles were taken off and put in hot air oven till dry and then transferred to muffle furnace at 550 C for 2 h. After 2 h, the crucibles were taken out, put in dessicator and weighed again. Initial weight of crucible with sample final weight of crucible with sample Crude fiber (%) = Weight of sample Ash Procedure Ash in the sample was estimated by employing the standard method of AOAC (2000). Five ml wet sample and 5 gram of oven dried for food sample was weighed in the silica crucible. It was ignited till no charred particles remained in the crucible. The crucible was put in muffle furnace (550 O C) for 5-6 hours until a white ash was obtained. Then the crucible was cooled in a dessicator and weighed. The loss in weight represented the organic matter and residue being the ash content and it was calculated as follows: Loss in weight (g) Ash (%) = 100 Weight of sample (g) Total carbohydrates The total carbohydrate was calculated by difference method. Total carbohydrate (%) = 100 [Crude protein (%) + Crude fat (%) + Crude fibre (%) + Ash (%)] 29

41 3.4.2 Carbohydrates Total sugars For beverages: Total sugars for beverages were estimated according to Yemm and Willis (1954) Reagents i) Standard sugar solution: Dissolved 25 mg glucose in water and made to 100 ml. This ii) Estimation solution contained 250 µg glucose per ml. For obtaining the standard curve, 0.2 ml to 1.0 ml of this solution was added. Anthrone Reagent (0.2 % anthrone in 70 % H 2 SO 4 ): This reagent was prepared fresh daily and allowed to stand for 30 to 40 min before use. Twenty five ml of sample was diluted with distilled water and volume was made upto 100 ml. The volume was filtered and the filterate was used as sugar extract. For food products Total soluble sugars were extracted according to procedure of Cerning and Guilhot (1973) and estimated by Yemm and Willis (1954). Estimation Twenty five ml ethanol (80 %) was added to 0.5 g sample in a round bottomed flask. The flask was connected to a condenser and kept on a heating mantle for 30 min with occasional stirring. The extract was cooled, centrifuged at 8000 rpm for 15 min and supernatant collected. The above procedure was repeated twice, each time extracting the residue in 25 ml 80 per cent ethanol. The combined extract in the beaker was evaporated to dryness on a boiling water bath. The residue was dissolved in distilled water and made to 50 ml. The prepared sample was sugar free pellet. Procedure Freshly prepared 10 ml anthrone reagent was pipetted in a test tube, chilled and kept in ice cold water. One ml of the sugar extract was taken as such for beverages and diluted to 10 ml with water for food products. Out of the diluted sugar extract, one ml was taken for further analysis. The extract was layered on the anthrone reagent. After cooling for 3-5 min, the contents were thoroughly mixed, while still immersed in ice cold water. The contents in the tube were heated vigorously in a boiling water bath for 10 min and then immediately cooled in cold water. The absorbance was then read at 625 nm against blank. Amount of sugar was estimated by referring to standard curve prepared with glucose previously. The total soluble sugars were calculated by the formula: C x V x 100 Total Soluble Sugars (g/100g) = W x V 1 30

42 Where, C = concentration of glucose from curve (µg) V = volume of extract made W = weight of sample taken V 1 = volume of aliquot taken Reducing Sugars Reagents For beverages: reducing sugars were estimated by Lane and Ennoyn method (1996) Fehling's Solution: i) g of reagent grade crystalline copper sulfate pentahydrate (CuSO4 5H2O) was ii) Estimation dissolved in purified water and diluted to 500 ml volume. 173 g of reagent grade potassium sodium tartrate (KnaC4H4O6 4H2O) and 50 g of reagent grade sodium hydroxide (NaOH) were dissolved in purified water and diluted to 500 ml volume. Solution i&ii were added in equal quantities and mixed. Twenty five ml of sample was taken and transfered to 250 ml volumetric flask 10 ml of neutral lead acetate solution was added and volume was diluted with water and filtered. An aliquot of 25 ml of the clarified filtrate transfered to 500 ml volume flask contained 100 ml of water. Potassium oxalate was added in small amounts until there is no further precipitation. Volume was made. The solution was mixed well and filtered through Whatman No. 1 filter paper. The filtrate was transfered to a 50 ml burette. Preliminary Titration Five ml of Fehling A and B solution was pipeted into 250 ml conical flask. Ten ml water and a few boiling chips or glass beads were added. The flask was heated to boiling. 3 drops of methylene blue indicator was added. The solution was added dropwise until the blue colour disappears to a brick-red end point. (The concentration of the sample solution was such that the titre value is between 15 and 50 ml). The titre value was noted down. Final Titration Five ml each of Fehling A and B was pipetted. Sample solution 2 ml or less than titre value was added to the preliminary titration. The flask was heated and boiled within 3 min and the titration was completed. The titration was performed in duplicates and the average was recorded. the reducing sugars % was calculated as shown below. Dilution Factor of Fehling (in gm) *100 Reducing Sugars % = Weight of sample *titre For food products Reducing sugars were estimated by the method of Somogyi (1945). 31

43 Reagents i) Copper reagent (A): Dissolved 25 g anhydrous sodium carbonate, 25 g potassium ii) iii) iv) sodium tartarate, 20 g sodium bicarbonate and 200 g anhydrous sodium sulphate in about 800 ml distilled water and diluted to one litre. Copper reagent (B): Dissolved 15 g CuSo 4 in 100 ml distilled water containing two drops of HCl. Arsenomolybdate reagent: Dissolved 25 g ammonium molybdate in 450 ml distilled water by warming. Added 21 ml conc. H 2 SO 4 with stirring. Three g sodium hydrogen arsenate dissolved in 25 ml distilled water was added with stirring. The solution was kept in an incubator at 37 0 C for 24 h before use. The reagent was kept in a glass stoppered brown bottle and stored in refrigerator. Copper Reagent: A and B mixed in the ratio of 25:1 (V/V) before use. v) Standard sugar solution: Dissolved 25 mg glucose and made to 100 ml with water. Estimation This contained 250 µg glucose/ ml. One ml test extract was taken in blood sugar tube graduated at 25 ml. One ml mixed copper reagent (iv) was added and then heated for 20 min in boiling water bath. To this one ml arsenomolybdate reagent was added, mixed thoroughly and the contents diluted to 25 ml. A stable blue colour appeared quickly which was read at 520 nm against blank. The amount of reducing sugar was then determined by referring to the glucose standard curve and using the following formula: C x V x 100 Reducing sugar (g/100g) = W x V 1 x 1000 Where, C= concentration of glucose from curve (µg) V= volume of extract made W= weight of sample taken V 1 = volume of aliquot taken Non-Reducing sugars The amount of non-reducing sugars was calculated as the difference between total sugars and reducing sugars in beverages and food products Starch For beverages (2000) Determination of Starch in beverages was estimated by standard method of AOAC 32

44 Procedure Twenty five ml sample was weighed in a 250 ml beaker. Twenty ml of alcohol was added. The sample was filtered by a filter paper and the precipitate was collected and washed with 50% alcohol till the precipitate is free from sugar. The precipitate was transfered to a 500 ml flask with about 200 ml water and10 ml Conc. HCl was added to hydrolyse and refluxed the starch by boiling water bath for 2.5 hrs. Neutralized and cooled with 10 % Sodium Hydroxide and Sodium Carbonate. The volume was made upto 500 ml with water. Shaked well and filtered. Starch was calculated with the following formula : % starch = % reducing sugar x 0.9 for milk and milk products % starch = % glucose *0.9 for RTS For food products : Starch from the sugar free pellet obtained after centrifugation was estimated by the method of Clegg (1956). Extraction Five ml water was added to the aforesaid residue of test material and while stirring, 6.5 ml of 52 per cent perchloric acid was added. The contents were stirred continuously for five min and then occasionally for next 15 min. 20 ml water was added and centrifuged. The supernatant was collected in a 100 ml volumetric flask. Five ml water was added to the residue and repeated the extraction with 52 per cent perchloric acid stirring occasionally for next 30 min. The contents of the tube were washed into a volumetric flask containing the test extract and made it to 100 ml with distilled water. It was then filtered discarding first 5 ml of the filtrate. One ml of the extract was taken and diluted to 15 ml with water. Out of the diluted extract a suitable aliquot (1.0 ml) was used for glucose estimation using anthrone reagent by method of Yemm and Willis (1954) as described under Starch was calculated using the following formula: Starch= Glucose x Total dietary fibre Total dietary fibre constituents were determined by the enzymatic method given by Furda (1981). Reagents i) N HCl ii) Phosphate buffer (ph 10) iii) EDTA iv) Enzymes: Alpha amylase and protease enzymes were obtained from Sigma Chemical Company, USA. v) Ethanol (75% and absolute) vi) Acetone 33

45 Procedure i) Sample preparation: 5 g sample of less than 1 mm particle size food material was defatted on a Soxhlet or Goldfish apparatus. ii) Extraction of water-soluble material: The prepared sample weighing about 2.0 g was dispersed in 200 ml of N HCl and boiled for 20 min. The suspension was then cooled down to 60 C; 0.3 g of disodium EDTA was added and then adjusted to ph with 12 ml of phosphate buffer ph 10. The extraction was continued for an additional 40 min at 60 C to ensure the extraction with minimal degradation. iii) Starch and protein hydrolysis: Adjusted the ph to bring the solution closer to the ph optimum of amylase and protease. The suspension was cooled to C before incubation overnight with 10 mg of bacterial alpha-amylase and 10 mg of bacterial protease. The incubation was accompanied by slow stirring with a magnetic bar. iv) Isolation of insoluble dietary fibre: The suspension was filtered through a coarsetarred Gooch filtering crucible containing glass wool and the insoluble residue was washed with a small amount of water. The filtrate was saved for the next step. The insoluble residue was then washed with water, alcohol and acetone before being dried at 70 C in a vacuum oven overnight. The dry residue constitutes insoluble dietary fibre (IDF). v) Precipitation and isolation of soluble dietary fibre (SDF): The saved filtrate was acidified with a few drops of concentrated hydrochloric acid to ph 2-3; this ph tended to facilitate the rapid precipitation of polysaccharides. Slowly added four volumes of ethanol and left suspension to stand for about 1 hour. Filtered the precipitate on a tarred, coarse Gooch crucible containing glass wool, then washed with 75% ethanol, absolute ethanol, and acetone before drying at 70 C in a vacuum oven overnight. The residue was weighed in the crucible to give the soluble dietary fibre (SDF) content of the original material. The SDF fraction was corrected for ash and for-co-precipitated protein. vi) Total dietary fibre (TDF): The sum of insoluble dietary fibre and soluble dietary fibre contents were calculated. TDF = IDF + SDF Total minerals Procedure Acid digestion: Five ml sample for beverages and 5 g sample for food products were taken in a 150 ml conical flask, ml of diacid mixture (HNO 3 : HClO 4 :: 5:1, v/v) was added and kept overnight. The contents were digested by heating till clear white precipitates settled 34

46 down at the bottom. The volume was made to 50 ml with double distilled water The crystals were filtered through Whatman No. 42 filter paper and used for determination of total Calcium, Iron, Zinc, Sodium, Magnesium and Pottassium Estimation Calcium, Iron, zinc, sodium, magnesium and pottassium in acid digested samples were determined by Atomic Absorption Spectrophotometer according to the method of Lindsey and Norwell (1969). 3.5 Glycaemic response of developed food products using sugar (control), sucralose and Stevia powder On the basis of organoleptic and nutritional evaluation the food products having desirable quality were selected to assess the glycaemic response Selection of subjects To study the glycaemic response of value added products, normal healthy, obese and diabetic adult subjects (n=30) in the age group of 35 to 55 yrs were selected randomly. None of the subjects were under medication. All the subjects were also informed beforehand about the experiment and their voluntary consents were taken before conducting the experiment (Annexure II and III). Their anthropometric measurements i.e. height (m) and weight (kg) were recorded and BMI was calculated as per method of Garrow (1981) Meal tolerance test Assessment of glycaemic response was done by Finger prick method by taking the drop of blood on glucose test strip was checked through Glucometer. For glycaemic response, the feeding trials were carried out on the selected subjects. On first occasion, 25 and 50 g carbohydrates in the form of glucose (reference food) and on subsequent occasions, a 25 and 50 g carbohydrate test food was given to the subjects. The subjects were asked to come for blood glucose test after overnight fast. For the glycaemic index of reference food, blood glucose was measured before feeding the glucose i.e. at zero hour and then 25 and 50 g glucose dissolved in 250 ml water was given to the subjects. The blood samples were drawn and checked after 30, 60, 90 and 120 min for the postprandial glucose level. For test foods, the volunteers were asked to consume test food with 150 ml of potable water, within min. The blood samples were drawn and checked after 30, 60, 90 and 120 min for the postprandial glucose level. In the same manner, daily a freshly prepared new recipe was fed to the subjects and blood glucose levels at 0, 30, 60, 90 and 120 min were measured Evaluation of glycaemic index of value added products For estimating glycaemic index of each product, the portion of food product equivalent to 50 g carbohydrate was computed from analyzed value of recipes. The blood glucose response curves were plotted for both the oral glucose tolerance test and the test 35

47 carbohydrate meal. The glycaemic index was calculated using the formula given by Jenkins et al. (1981). Formula for calculating area under curve is given below: Area under glucose curve of test meal Glycaemic Index food products = 100 Area under glucose curve of reference meal 3.6 Cost calculation of the developed food products: Cost of the developed food products was calculated by adding the cost of each raw ingredient as procured on the retail price from market. 3.7 Statistical analysis The data were subjected to statistical analysis for analysis of variance in a complete randomized design using standard methods of Sheoran and Pannu (1999). Whereas, for nutritional evaluation of leaves and powder were analyzed using students t test by Panse and Sukhatme (1961). 36

48 CHAPTER-IV RESULTS In the present study, efforts were made to develop food products by using Stevia powder and sucralose by 100 per cent replacement of sugar. The developed food products were evaluated for organoleptic characteristics and nutritional composition. The most acceptable developed food products were also studied for their glycaemic response in normal, obese and diabetic subjects. The results pertaining to this study have been presented and discussed under the following heads and sub-heads: 4.1 Nutritional composition of Stevia leaves and Stevia powder Proximate composition Moisture Ash Sugars Total Sugars Reducing Sugars Non Reducing Sugars Total minerals Calcium Iron Zinc Sodiun Magnesium Potassium 4.2 Development and organoleptic evaluation of developed food products prepared using sucralose and Stevia powder Beverages Tea Coffee Milk shake RTS Baked products Biscuits Cake Bun Traditional products Sweet porridge Carrot halwa Kheer 37

49 4.3 Nutritional evaluation of developed food products prepared using sugar, sucralose and Stevia powder Proximate composition Moisture Crude protein Crude Fat Crude fibre Ash Total carbohydrates Available carbohydrates Total sugars Reducing sugars Non-reducing sugars Starch Total minerals Calcium Iron Zinc Magnesium Sodiun Potassium 4.4 Glycaemic response of developed food products using sucralose and Stevia powder 4.5 Cost calculation of the developed food products 4.1 Nutritional composition of Stevia leaves and Stevia powder Proximate composition The data pertaining to moisture, ash and sugars content in Stevialeaves and Stevia powder are presented in Table 4.1. Table 4.1: Moisture, ash and sugars contents of Stevia leaves and Stevia powder (g /100 g, on dry weight basis) Stevia Moisture Ash Total sugars Reducing sugars Non reducing sugars Leaves 6.21± ± ± ± ±0.11 Powder 2.92± ± ± ± ±0.06 t value 7.84** 5.98** 7.48** 8.22** 5.25** Values are mean± S.E of three independent observations. *Significant at 5%level NS-= non-significant **Significant at 1% level 38

50 Moisture content of Stevia leaves and Stevia powder was 6.21 and 2.92 per cent, respectively. Stevia leaves contained significantly (p<0.05) higher content of moisture as compared to Stevia white powder Ash content in Stevia leaves was significantly higher as compared to Stevia powder. The values were and 3.24 per cent, respectively. Stevia leaves contained total soluble, reducing and non reducing sugar 15.63, 5.81 and 9.82 per cent, respectively. Stevia powder were found to have significantly (p<0.01) lower total soluble (1.78 %), reducing (0.84 %) and non reducing sugar (0.93 %) contents. A significant (p<0.01) difference was observed between Stevia leaves and Stevia powder sugar contents. Table 4.2: Mineral contents of Stevia leaves and Stevia powder (mg/100 g, on dry matter basis) Stevia Calcium Iron Zinc Sodium Magnesium Potassium Leaves ± ± ± ± ± ±0.02 Powder ± ± ± ± ± ±0.03 t value 5.12** 6.59** 7.73** 5.63** 6.01** 5.91** Values are mean± S.E of three independent observations. NS= Non-significant *Significant at 5%level **Significant at 1% level Significantly higher calcium content was observed in Stevia leaves ( mg/100 g) and lower was observed in Stevia powder ( mg/100 g). Concentration of iron in Stevia leaves and Stevia powder was observed to be and 6.31 mg/100 g, respectively. Significantly higher iron content was observed in Stevia leaves whereas lower content was observed in Stevia powder. A significant (P<0.05) differences was observed in zinc content of both the samples. Higher value of zinc was observed in Stevia leaves (3.57 mg/100 g) and lower was observed in Stevia powder (0.12 mg/100 g). The sodium content of Stevia leaves and Stevia powder was observed as and mg/100 g, respectively. The sodium content of both the samples varied significantly. Among the Stevia leaves and Stevia powder significant differences were observed in magnesium content. Stevia leaves and Stevia powder contained and mg/100 g magnesium content, respectively. The values differed significantly for magnesium content. Stevia leaves exhibited significantly higher amount of potassium content as compared to Stevia powder. The potassium content in Stevia leaves was observed to be higher ( mg/100 g) whereas Stevia powder had comparatively lower mg/100 g potassium content. 39

51 4.2 Development and organoleptic evaluation of developed food products prepared using sucralose and Stevia powder Beverages, baked and traditional products were prepared using sugar, sucralose and Stevia powder (at two levels). In test recipes, Stevia powder was added at two levels and sucralose (artificial sweetner) was added individually for comparison. While in the control recipe, required amount of sugar was added in 100g of recipe. The organoleptic evaluation was done to select the most acceptable level of Stevia powder in all the recipes. The panel of judges were asked to score the test samples for their colour, appearance, flavour, texture/consistency, taste and overall acceptability compared to the control (sugar based) recipes and sucralose incorporated recipes Beverages Tea Mean scores of colour, appearance, aroma, texture, taste and overall acceptability of tea prepared using sugar (control), sucralose and Stevia powder at two levels T 1 (6 mg) and T 2 (12 mg) are presented in Table 4.3. Table 4.3: Mean scores of organoleptic characteristics of Tea Types of Tea Colour Appearance Aroma Texture/ Consistency Taste Overall acceptability Control (Sugar) 7.60± ± ± ± ± ±0.22 Sucralose 7.30± ± ± ± ± ±0.15 Stevia powder T ± ± ± ± ± ±0.09 T ± ± ± ± ± ±0.35 CD (P<0.05) NS NS 0.22 NS Values are mean ± SE of ten independent observations NS = Non-significant Control = Basic recipe with 6 g of sugar Sucralose = Recipe with 3 mg of sucralose T 1 = Recipe with 6 mg of Stevia powder T 2 = Recipe with 12 mg of Stevia powder The mean scores of colour ranged from 7.00 to 7.60, with the highest for tea prepared using sugar (control) and lowest for tea prepared using Stevia powder T 2 (12 mg) which revealed that the colour scores of all four types of teas were found in the category of liked moderately by the judges. Similarly mean scores of appearance of all the four types of tea were found in the range of 7.00 to 7.63, with the highest for control tea and lowest for Stevia powder (T 2 ) based tea. Mean scores of aroma of all four types of tea prepared using sugar, sucralose and Stevia powder (T 1 ) were 7.85, 7.00 and 7.40 which fell in the category of liked moderately. But on increasing the concentration of Stevia powder T 2 (12 mg) in tea caused significant decrease in mean score of aroma (5.90), which revealed that aroma score of tea neither liked nor disliked by the judges. Similarly, mean scores of texture/consistency of tea 40

52 prepared using sugar (control), sucralose and Stevia powder (T 1 ) did not differ significantly and ranged from 7.00 to However, mean score of texture/consistency of tea prepared using Stevia powder (T 2 ) was 6.90 which fell in the category of liked slightly. The taste scores ranged from 5.90 to 7.80, with the highest for control tea and lower for tea prepared using Stevia powder (T 2 ). Taste score of tea prepared by using Stevia powder T 2 (12 mg) was neither liked nor disliked by the judges due to bitter after taste. The overall acceptability was highest for control tea followed by sucralose and Stevia powder T 1 (6 mg) based tea. However, tea prepared using Stevia powder T 2 (12 mg) was not acceptable in terms of aroma and taste and rated as neither liked nor disliked by the panelists Coffee Mean scores of colour, appearance, aroma, texture, taste and overall acceptability of coffee prepared using sugar (control), sucralose and Stevia powder at two levels T 1 (8 mg) and T 2 (16 mg) are presented in Table 4.4. Table 4.4: Mean scores of organoleptic characteristics of Coffee Types of Coffee Colour Appearance Aroma Texture/ Consistency Taste Overall acceptability Control (Sugar) 7.40± ± ± ± ± ±0.08 Sucralose 7.30± ± ± ± ± ±0.12 Stevia powder T ± ± ± ± ± ±0.23 T ± ± ± ± ± ±0.57 CD (P<0.05) NS NS 0.22 NS Values are mean ± SE of ten independent observations Sucralose = Recipe with 4 mg of sucralose NS =Non-significant T 1 = Recipe with 8 mg of Stevia powder Control = Basic recipe with 8 g of sugar T 2 = Recipe with 16 mg of Stevia powder Mean scores of colour ranged from 7.00 to 7.40, with the highest for coffee prepared using sugar (control) and lowest for coffee prepared using Stevia powder T 2 (16 mg) which indicated that the colour scores of all four types of coffees were found in the category of liked moderately by the judges. Similarly, mean scores of appearance of all the four types of coffees were found in the range of 7.10 to 7.70, with the highest for control coffee and lowest for Stevia powder (T 2 ) based coffee. Mean scores for aroma of all four types of coffee prepared using sugar, sucralose and Stevia powder (T 1 ) were 7.30, 6.90 and 7.10, respectively. Coffee prepared using sugar and Stevia powder T 1 fell in the category of liked moderately whereas, coffee prepared using sucralose liked slightly by the panelist. On increasing the concentration, Stevia powder T 2 (16 mg) in coffee significant decrease was observed in mean score of aroma (5.60), which revealed that aroma score of coffee was neither liked nor disliked by the judges. Similarly mean scores for consistency of coffee prepared using sugar 41

53 (control), sucralose and Stevia powder (T 1 ) and (T 2 ) did not differ significantly and ranged from 7.00 to However, a significant decrease in the mean score for test was observed in coffees. Coffee prepared using sugar (control) and Stevia powder T 1 (8 mg) showed a nonsignificant difference and scored similarly mean scores i.e for taste. Mean scores for taste of coffee prepared using sugar and Stevia powder (T 1 ) based coffee was rated as liked moderately by the panelist. The overall acceptability was highest for control coffee followed by Stevia powder (T 1 ) based and sucralose based coffee. However, Stevia powder T 2 (16 mg) based coffee was not acceptable in terms of aroma and rated neither liked nor disliked whereas the overall acceptability was liked slightly by the judges Milk shake Mean score of colour, appearance, aroma, taste, texture and overall acceptability of milkshake prepared using sugar (control), sucralose, Stevia powder in two concentrations T 1 (12 mg) and T 1 (24 mg) is depicted in Table 4.5. Table 4.5: Mean scores of organoleptic characteristics of Milk shake Types of Milk shake Colour Appearance Aroma Texture/ Consistency Taste Overall acceptability Control (Sugar) 7.90± ± ± ± ± ±0.22 Sucralose 7.60± ± ± ± ± ±0.11 Stevia powder T ± ± ± ± ± ±0.10 T ± ± ± ± ± ±0.24 CD (P<0.05) NS NS Values are mean ± SE of ten independent observations Sucralose = Recipe with 6 mg of sucralose NS =Non-significant T 1 = Recipe with 12 mg of Stevia powder Control = Basic recipe with 8 g of sugar T 2 = Recipe with 24 mg of Stevia powder Mean scores for colour of all types of milk shakes ranged from 7.60 to 7.90, with the highest for milk shake prepared using sugar followed by milk shake prepared using Stevia powder T 1 (12 mg). Similar but lower mean scores were observed for milk shake prepared using sucralose and Stevia powder T 1 (24 mg) which revealed that all four types of milk shakes were found in the category of liked moderately by the judges. Whereas, mean scores for appearance of all four types of milk shake ranged from 7.10 to 7.80, with the highest for sucralose based and Stevia powder T 1 (12 mg) based milk shake and lowest for Stevia powder T 2 (24 mg) based milk shakes. Mean scores for aroma of all four types of milk shakes prepared using sugar and Stevia powder T 2 (12 mg) were 8.10 and 5.70, respectively which fell in the categories of liked very much for control and neither liked nor disliked for Stevia powder T 2 (24 mg) based milk shake. Milk shake prepared using sucralose and Stevia powder T 1 (12 mg) scored similar mean scores i.e for aroma, which fell in the category of liked moderately. Similarly, the mean scores of milk shakes for consistency varied 42

54 significantly. Highest mean score was observed for milk shake prepared using sugar (control) and lowest score for consistency was found to be in Stevia powder (T 2 ) based milkshake. The mean score of milk shake prepared using sucralose, Stevia powder (T 1 ) and (T 2 ) was found to have a non-significant difference for texture (consistency) and fell in the category of liked moderately whereas milk shake prepared using sugar (control) was liked very much by the panelists. The taste score ranged from 6.60 to 8.40, with the highest of control milkshake and lowest for milk shake prepared using Stevia powder T 2 (24 mg). The mean scores of taste of milk shake prepared using Stevia powder T 1 (12 mg) and sucralose was liked very much by the panelists. Whereas, milk shake prepared using Stevia powder T 2 was liked slightly by the judges. Increase in the amount of Stevia powder lead to licorice type after taste making it non likeable for judges. The overall acceptability was highest for control milk shake followed by Stevia powder T 1 (12 mg) based and sucralose based milk shake. However milk shake prepared using Stevia powder T 2 (24 mg) was acceptable but not at par in terms of aroma and was liked slightly in terms of taste and overall acceptability RTS Mean scores of colour, appearance, aroma, texture, taste and overall acceptability of RTS prepared using sugar (control), sucralose and Stevia powder at two levels T 1 (10 mg) and T 2 (20 mg) are presented in Table 4.6. Table 4.6: Mean scores of organoleptic characteristics of RTS Types of RTS Colour Appearance Aroma Texture/ Consistency Taste Overall acceptability Control (Sugar) 7.90± ± ± ± ± ±0.12 Sucralose 7.60± ± ± ± ± ±0.13 Stevia powder T ± ± ± ± ± ±0.12 T ± ± ± ± ± ±0.34 CD (P<0.05) NS NS 0.74 NS Values are mean ± SE of ten independent observations Sucralose = Recipe with 5 mg of sucralose NS =Non-significant Control = Basic recipe with 10 g of sugar T 1 = Recipe with 10 mg of Stevia powder T 2 = Recipe with 20 mg of Stevia powder Mean score of colour, ranged from 7.30 to 7.90, with the highest for RTS prepared using sugar (control) and lowest for Stevia powder T 2 (20 mg) based RTS, which revealed that the mean score for colour of all types of RTS were found in the category of liked moderately. Similarly mean score for appearance of all the four types RTS were found in the range of 7.00 to 7.20, with the highest for sucralose based RTS and lowest for Stevia powder T 2 (20 mg) based RTS. Mean score for aroma of all four types of RTS prepared using sugar were 7.30 which fell in the category of liked moderately. But on replacing sugar with sucralose and Stevia powder in high concentration in RTS caused significant decrease in the 43

55 mean scores of aroma, which revealed that aroma score of RTS fell in the category of liked slightly by the judge. Whereas, RTS prepared using sugar (control) and Stevia powder T 1 (10 mg) was liked moderately by the panelist. However, the mean score for consistency of RTS prepared using sugar, sucralose, Stevia powder T 1 (10 mg) and Stevia powder T 2 (20 mg) did not differ significantly and ranged from 7.00 to The mean score for taste ranged from 5.50 to 7.10, with the highest for control RTS and lowest for RTS prepared using Stevia powder T 2. Taste score for RTS prepared using sucralose and Stevia powder T 1 was liked slightly. Whereas, RTS prepared using Stevia powder in higher concentration i.e. Stevia powder based T 2 (20 mg) was neither liked nor disliked by the judges due to bitter and prominent after taste of Stevia powder. The overall acceptability was highest for control RTS followed by sucralose based and Stevia powder T 1 based RTS. However, RTS prepared using Stevia powder T 2 (20 mg) was not acceptable in terms of taste and rated neither liked nor disliked whereas for aroma and overall acceptability the RTS prepared using Stevia powder T 2 was liked slightly by the panel of judges Baked products Biscuits Mean score of colour, appearance, aroma, texture, taste and overall acceptability of biscuits prepared using sugar (control), sucralose and Stevia powder at two levels T 1 (60 mg) and T 2 (120 mg) are presented in Table 4.7 Table 4.7: Mean scores of organoleptic characteristics of Biscuits Types Biscuits of Colour Appearance Aroma Texture Taste Overall Acceptability Control (Sugar) 7.70± ± ± ± ± ±0.09 Sucralose 7.30± ± ± ± ± ±0.13 Stevia powder T ± ± ± ± ± ±0.17 T ± ± ± ± ± ±0.25 CD (P<0.05) NS NS Values are mean ± SE of ten independent observations NS =Non-significant Control = Basic recipe with 60 g of sugar Sucralose = Recipe with 30 mg of sucralose T 1 = Recipe with 60 mg of Stevia powder T 2 = Recipe with 120 mg of Stevia powder Mean scores of colour ranged from 7.30 to 7.70, with the highest for biscuits prepared using sugar (control) and Stevia powder T 1 (60 mg) and lowest for biscuits prepared using sucralose which revealed that all four types of biscuits were found in the category of liked moderately by the judges. Similarly, mean scores of appearance of all the four types of biscuits were found in the range of 7.20 to 7.90, with the highest for Stevia powder T 1 based biscuits and lowest for sucralose based biscuits. Mean scores of aroma of control and Stevia 44

56 powder T 1 based biscuits were 7.70 and 7.40 which fell in the category of liked moderately. however, biscuits prepared using sucralose and Stevia powder T 2 (120 mg) obtained mean scores 6.10 and 6.70 for aroma which fell in the category of liked slightly. On increasing the concentration of Stevia powder (T mg) in biscuits caused significant decrease in mean score of aroma. Similarly, mean scores of texture of biscuits prepared using sugar (control) and Stevia powder (T 1 ) did not differed significantly and were 7.60 and However, mean score of texture of biscuits prepared using sucralose and Stevia powder (T 2 ) differed significantly from control biscuits. The mean score for texture of sucralose based and Stevia powder (T 2 ) based biscuits were 6.20 and 6.80 which fell in the category of liked slightly. The taste scores ranged from 6.40 to 7.80, with the highest for control biscuits and lowest for Stevia powder T 2 (120 mg) based biscuits. Taste score of biscuits prepared by using Stevia powder T 2 (120 mg) was liked slightly by the judges. The overall acceptability was highest for control biscuits followed by Stevia powder (T 1 ) and Stevia powder (T 2 ) based biscuits. However, biscuits prepared using sucralose was neither liked nor disliked in terms of aroma, texture and overall acceptability Cake Mean score of colour, appearance, aroma, texture, taste and overall acceptability of cake prepared using sugar (control), sucralose and Stevia powder in two concentrations T 1 (60 mg) and T 2 (120 mg) are presented in Table 4.8. Table 4.8: Mean scores of organoleptic characteristics of Cake Types of Cake Colour Appearance Aroma Texture Taste Overall acceptability Control (Sugar) 7.30± ± ± ± ± ±0.22 Sucralose 7.20± ± ± ± ± ±0.13 Stevia Powder T ± ± ± ± ± ±0.23 T ± ± ± ± ± ±0.41 CD (P<0.05) Values are mean ± SE of ten independent observations NS =Non-significant Control = Basic recipe with 60 g of sugar Sucralose = Recipe with 30 mg of sucralose T 1 = Recipe with 60 mg of Stevia powder T 2 = Recipe with 120 mg of Stevia powder Mean scores of colour ranged from 7.20 to 8.00, with the highest for Stevia powder T 1 (60 mg) incorporated cake and lowest for sucralose incorporated and Stevia powder T 2 (120 mg) incorporated cake, which revealed that the colour score of three types of cake control, sucralose incorporated and Stevia powder T 2 (120 mg) incorporated were found in the category of liked moderately. Similarly, mean score of appearance of all four types of cake ranged from 7.10 to 8.00 with the highest for Stevia powder T 1 (60 mg) incorporated and lowest for sucralose incorporated and Stevia powder T 2 (120 mg) incorporated cake. Mean score for aroma of all types of cake prepared using sugar, sucralose, Stevia powder (T 1 ) and 45

57 Stevia powder (T 2 ) were 7.40, 7.40, 8.10 and Stevia powder (T 1 ) incorporated cake was liked very much whereas cake prepared using sugar and sucralose were fell in the category of liked moderately. But an increasing the concentrations of Stevia powder T 2 (120 mg) in cake caused significant decrease in mean score of aroma (6.30), which revealed that aroma score of cake was liked slightly by the judges. Similarly, mean score of texture of cake prepared using Stevia powder (T 1 ) differed significantly from control, sucralose incorporated and Stevia powder T 2 incorporated cake. The mean score of Stevia powder T 1 cake was 7.60 which fell in the category of liked moderately. The taste scores ranged from 6.40 to 8.00, with the highest for Stevia powder (T 1 ) incorporated cake and lowest for Stevia powder T 2 incorporated cake. Taste score of cake prepared using sucralose and Stevia powder T 2 was liked slightly as both imparted after taste to the cake. Overall acceptability was highest for Stevia powder T 1 incorporated cake followed by control and sucralose cake. Stevia powder T 2 incorporated cake was liked slightly in terms of aroma, texture, taste and overall acceptability Buns Mean score of colour, appearance, aroma, texture, taste and overall acceptability of bun prepared using sugar (control), sucralose and Stevia powder at two levels T 1 (30 mg) and T 2 (60 mg) are presented in Table 4.9. Table 4.9: Mean scores of organoleptic characteristics of Buns Types of Buns Colour Appearance Aroma Texture Taste Overall acceptability Control (Sugar) 7.40± ± ± ± ± ±0.06 Sucralose 7.30± ± ± ± ± ±0.08 Stevia powder T ± ± ± ± ± ±0.08 T ± ± ± ± ± ±0.04 CD (P 0.05) NS NS Values are mean ± SE of ten independent observations NS =Non-significant Control = Basic recipe with 30 g of sugar Sucralose = Recipe with 15 mg of sucralose T 1 = Recipe with 30 mg of Stevia powder T 2 = Recipe with 60 mg of Stevia powder The mean scores of colour ranged from 7.20 to 7.80, with the highest for bun prepared using Stevia powder T 1 (30 mg) and lowest for Stevia powder T 2 (60 mg) incorporated bun which indicated that the colour score of all the four types of bun were found in the category of liked moderately by the judges. Similarly, mean score for appearance of all four types of bun and in the range of 7.40 to 7.90, with the highest for Stevia powder T 1 (30 mg) incorporated bun and lowest for Stevia powder T 2 (60 mg) incorporated bun. Mean scores of aroma of all four types of bun prepared using sugar, sucralose, Stevia powder T 1 (30 mg) and Stevia powder T 2 (60 mg) were 7.30, 7.30, 7.90 and 6.00, respectively. Mean scores 46

58 for aroma of buns prepared using sugar, sucralose and Stevia powder (T 1 ) fell in the category of liked moderately. However, mean score for aroma of bun prepared using the higher concentration of Stevia powder i.e. T 2 (60 mg) bun was liked slightly by the panelist. Similarly, mean score of texture of bun prepared using sugar, sucralose and Stevia powder (T 1 & T 2 ) did not differed significantly and fell in the category of liked moderately with the mean score of 7.90, 7.20 and 7.30, respectively. However, mean score for texture of buns prepared using Stevia powder T 2 differed significantly from control, sucralose and Stevia powder (T 1 ) based buns. The taste scores ranged from 6.10 to 7.60, with the highest for control bun and lowest for bun prepared using Stevia powder (T 2 ). Taste score of bun prepared by using Stevia powder T 2 was 6.10 which fell in the category of liked slightly. The overall acceptability score was highest for bun prepared using Stevia powder (T 1 ) followed by control and sucralose based buns. Bun prepared using Stevia powder (T 2 ) was liked slightly in terms of aroma, texture, taste and overall acceptability by the panelists Traditional Products Sweet porridge Mean score of colour, appearance, aroma, texture, taste and overall acceptability of sweet porridge prepared using sugar (control), sucralose and Stevia powder at two levels T 1 (30 mg) and T 2 (60 mg) are presented in Table Table 4.10: Mean scores of organoleptic characteristics of Sweet porridge Types of Sweet porridge Colour Appearance Aroma Texture Taste Overall acceptability Control (Sugar) 8.10± ± ± ± ± ±0.12 Sucralose 7.90± ± ± ± ± ±0.09 Stevia powder T ± ± ± ± ± ±0.21 T ± ± ± ± ± ±0.35 CD(P<0.05) NS NS 0.80 NS 0.50 NS Values are mean ± SE of ten independent observations NS =Non-significant Control = Basic recipe with 30 g of sugar Sucralose = Recipe with 15 mg of sucralose T 1 = Recipe with 30 mg of Stevia powder T 2 = Recipe with 60 mg of Stevia powder Mean scores of colour ranged from 7.70 to 8.10, with the highest for sweet porridge prepared using sugar (control) and lowest for sweet porridge prepared using Stevia powder T 2 (60 mg), which exhibited that the colour score of control was found in the category of liked very much. However, the colour scores of sweet porridge prepared using sucralose, Stevia powder T 1 (30 mg) and T 2 (60 mg) were found in the category of liked moderately by the judges. Similarly, the mean score of appearance of all the four types of sweet porridge were found in the range of 7.40 to 7.80, with the highest for control and lowest for Stevia powder (T 2 ) based sweet porridge. Mean score of aroma of all four types of aroma of all types of sweet porridge using sugar, sucralose and Stevia powder T 1 and T 2 were 8.50, 7.30, 7.70 and 47

59 6.60, respectively. Sweet porridge prepared using sucralose and Stevia powder T 1 fell in the category of liked moderately. Whereas sweet porridge prepared using sugar was liked very much and sweet porridge prepared using Stevia powder T 2 was liked slightly for aroma by the judges. Similarly, mean scores of texture for all types of sweet porridge prepared using sugar, sucralose and Stevia powder T 1 and T 2 did not differ significantly and ranged between 7.50 to 7.90, respectively. However, similar texture score i.e was observed for sweet porridge prepared using sucralose and Stevia powder T 2 and which fell in the category of liked moderately by the judges. The taste scores ranged from 6.10 to 8.00, with the highest for control and lowest for sweet porridge prepared using Stevia powder T 2 (60 mg). Taste score of sweet porridge prepared by using Stevia powder T 2 (60 mg) was liked slightly by the judges due to the high amount of sweetness and its impacts remains for longer period, which caused decrease in the taste score. The overall acceptability score was highest for control followed by Stevia powder (T 1 ) and sucralose based sweet porridge. However, sweet porridge prepared using Stevia powder (T 2 ) scored 6.60 and 6.10 and rated as liked slightly for aroma and taste, whereas, liked moderately for overall acceptability by the judges Carrot halwa Mean score of colour, appearance, aroma, texture, taste and overall acceptability of carrot halwa prepared using sugar (control), sucralose and Stevia powder at two level T 1 (30 mg) and T 2 (60 mg) are presented in Table Table 4.11: Mean scores of organoleptic characteristics of Carrot halwa Types of Carrot Colour Appearance Aroma Texture Taste Overall halwa acceptability Control (Sugar) 8.70± ± ± ± ± ±0.12 Sucralose 8.60± ± ± ± ± ±0.09 Stevia powder T I 8.80± ± ± ± ± ±0.11 T ± ± ± ± ± ±0.12 CD (P 0.05) NS NS 0.14 NS Values are mean ± SE of ten independent observations NS =Non-significant Control = Basic recipe with 30 g of sugar Sucralose = Recipe with 15 mg of sucralose T 1 = Recipe with 30 mg of Stevia powder T 2 = Recipe with 60 mg of Stevia powder Mean scores of colour ranged from 8.30 to 8.70, with the highest was observed for sucralose and lowest for Stevia powder T 2 (60 mg) based carrot halwa, which revealed that all four types of carrot halwa were found in the category of liked very much by the judges. Similarly, the mean scores of appearance of all four types of carrot halwa were found in the category of liked very much by the judges. The mean scores for appearance of carrot halwa prepared using sugar, sucralose and Stevia powder T 1 (30 mg) and T 2 (60 mg) were in the range of 8.40 to 8.70 with the highest for Stevia powder T 1 and lowest for Stevia powder T 2 based carrot halwa. Mean scores for aroma of carrot halwa prepared using sugar and sucralose were similar i.e However, 8.60 and 7.00 aroma score were obtained by carrot 48

60 halwa prepared using Stevia powder T 1 (30 mg) and T 2 (60 mg), respectively which fell in the category of liked very much for carrot halwa prepared using Stevia powder (T 1 ) and liked moderately for carrot halwa prepared using Stevia powder (T 2 ). Similarly, mean scores of texture of carrot halwa prepared using sugar, sucralose, Stevia powder (T 1 ) and (T 2 ) did not differ significantly and ranged from 8.20 to All four types of carrot halwa for texture fell in the category of liked very much by the panelists. The taste scores ranged 7.50 to 8.60, with the highest for control and lowest for Stevia powder (T 2 ) based carrot halwa. Carrot halwa prepared using Stevia powder (T 2 ) differed significantly and scored 7.50 for taste in comparison to control, sucralose and Stevia powder (T 1 ) and rated as liked moderately by the judges. The overall acceptability was highest for control carrot halwa followed by Stevia powder (T 1 ) and sucralose based carrot halwa. However, carrot halwa prepared using Stevia powder T 2 (60 mg) was liked moderately in terms of aroma, taste and overall acceptability Kheer Mean score of colour, appearance, aroma, texture, taste and overall acceptability of kheer prepared using sugar (control), sucralose and Stevia powder at two levels T 1 (20 mg) and T 2 (40 mg) are presented in Table 4.12 Table 4.12: Mean scores of organoleptic characteristics of Kheer Types of Kheer Colour Appearance Aroma Texture Taste Overall acceptability Control 7.60± ± ± ± ± ±0.18 Sucralose 7.10± ± ± ± ± ±0.16 Stevia powder T ± ± ± ± ± ±0.15 T ± ± ± ± ± ±0.28 CD (P 0.05) NS NS 0.45 NS Values are mean ± SE of ten independent observations NS =Non-significant Control = Basic recipe with 20 g of sugar Sucralose = Recipe with 10 mg of sucralose T 1 = Recipe with 20 mg of Stevia powder T 2 = Recipe with 40 mg of Stevia powder Mean scores of colour ranged from 7.10 to 7.60, with the highest for kheer prepared using sugar (control) and lowest for kheer prepared using sucralose which revealed that the colour scores of all four types of kheer were found in category of liked moderately by the judges. Similarly, mean scores of appearance of all the four types of kheer were found in the range of 7.40 to 7.70, with the highest for control and Stevia powder T 1 (20 mg) based and lowest for Stevia powder T 2 (40 mg) based kheer. Mean scores of aroma differed significantly for all types of kheer. However, kheer prepared using control and Stevia powder T 1 (20 mg) were liked moderately and kheer prepared using sucralose and Stevia powder T 2 (40 mg) were liked slightly by the panelists. Mean scores of aroma for all four types of kheer prepared using sugar, sucralose, Stevia powder T 1 and Stevia powder T 2 were 7.90, 6.90, 7.30 and 6.40, respectively. Similarly, mean score for texture of kheer prepared using sugar 49

61 (control), sucralose, Stevia powder T 1 and Stevia powder T 2 did not differ significantly and ranged from 7.20 to However, mean score for texture of all types kheer prepared using sugar, sucralose, Stevia powder T 1 and T 2 fell in the category of liked moderately by the judges. The taste scores ranged from 6.10 to 7.90, with the highest for control kheer and lowest for kheer prepared using Stevia powder (T 2 ). Taste scores for kheer prepared using Stevia powder (T 2 ) was liked slightly by the judges due to licorice after taste was more prominent and remained in mouth for long. The overall acceptability was highest for control kheer followed by Stevia powder (T 1 ) based kheer and sucralose based kheer. However, kheer prepared using Stevia powder T 2 (40 mg) was liked slightly in terms of aroma, taste and overall acceptability by the judges. 4.3 Nutritional evaluation of developed products prepared using sugar, sucralose and Stevia powder Each recipe with sugar i.e control and corresponding recipes with acceptable level of Stevia powder and sucralose were analyzed for nutritional composition Proximate compositions of Beverages Tea The data pertaining to proximate composition of tea prepared using sugar (control), sucralose and Stevia powder individually are presented in Table Table 4.13: Proximate composition of Tea (% as is basis) Types of Tea Moisture Protein Fat Fibre Ash Total carbohydrates Control (Sugar) 85.84± ± ± ± ± ±0.02 Sucralose 87.42± ± ± ± ± ±0.04 Stevia powder 88.79± ± ± ± ± ±0.08 CD (P<0.05) 0.37 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 6gm of sugar Sucralose = Recipe with 3 mg of sucralose Stevia powder = Recipe with 6 mg of Stevia powder Significant differences were observed in moisture content of control, sucralose and Stevia based tea. In control tea, moisture content was observed as per cent which increased to per cent in sucralose based and per cent in Stevia based tea. The protein and fat content ranged from content of control, sucralose and Stevia based tea were 1.40 to 1.52 per cent and 0.63 to 0.64 per cent, respectively. A non-significant difference was observed in protein and fat content in all types of teas. On the other hand, almost negligible amount of fibre was found in all types of teas which ranged from 0.04 to 0.09 per cent. The ash content ranged from 0.62 to 0.85 per cent, with the highest amount was observed in tea prepared with Stevia powder and lowest amount was found in tea prepared with sugar (control). Whereas, significant difference was noted in total carbohydrates content 50

62 of all three types of tea.there were per cent in sugar based tea, 9.69 per cent in sucralose based tea and 8.17 per cent in Stevia based tea Sugars and starch The data in respect to total soluble sugar, reducing sugar, non-reducing sugar and starch content of tea prepared using sugar, sucralose and Stevia powder are presented in Table Table 4.14: Sugars and starch contents of Tea (% as is basis) Types of Tea Sugars Total Reducing Non reducing Starch Control (sugar) 14.13± ± ± ±0.02 Sucralose 9.82± ± ± ±0.15 Stevia powder 9.73± ± ± ±0.02 CD (P<0.05) 1.80 NS 0.47 NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 6 gm of sugar Sucralose = Recipe with 3 mg of sucralose Stevia powder = Recipe with 6 mg of Stevia powder Control, sucralose and Stevia based tea exhibited significant differences for the sugars content. The total soluble, reducing and non-reducing sugar content of control tea were 14.13, 4.62 and 9.51 g/100 ml. Tea prepared using Stevia showed significantly (P<0.05) lowest amounts of total soluble (9.73 g/100 ml), reducing (4.68 g/100 ml) and non-reducing (5.05 g/100 ml) sugar content. While, sucralose based tea was found to have 9.82, 4.66 and 5.15 g/100 ml total soluble, reducing and non-reducing sugar content, respectively which were significantly (P<0.05) lower than control tea but, higher than Stevia based tea. Starch content of all types of tea differed non- significantly. The values were ranged from 1.01 to 1.04 per cent, with the lowest starch content (1.01 %) in control tea and highest starch content (1.04 %) in tea prepared with Stevia powder Minerals The data in respect to total calcium, iron, zinc, sodium, magnesium and potassium content of three types of tea are presented in Table 4.15 Table 4.15: Mineral contents of Tea (mg/100 ml as is basis) Types of Tea Calcium Iron Zinc Sodium Potassium Magnesium Control (Sugar) 89.93± ± ± ± ± ±0.09 Sucralose 89.95± ± ± ± ± ±0.08 Stevia powder 90.06± ± ± ± ± ±0.05 CD (P<0.05) 0.05 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Sucralose = Recipe with 3 mg of sucralose Stevia powder = Recipe with 6 mg of Stevia powder Control = Basic recipe with 6gm of sugar Stevia based tea differed significantly in calcium content in comparison to control and sucralose based tea. Maximum calcium content was observed in Stevia based tea (

63 mg/100 ml) followed by sucralose based tea (89.95 mg/100 ml) and control tea (89.93 mg/100 ml). A non-significant difference was observed in control and sucralose based tea. Iron concentration in different types of tea ranged from 0.51 to 0.53 mg/100 ml. Although there was a non-significant difference in all types of tea but still highest iron content was observed in sucralose based tea and lowest iron content was observed in control tea. Zinc content in three types of tea ranged from 0.22 to 0.25 mg/100 ml. A nonsignificant difference was observed among all types of tea. Control, sucralose based and Stevia based tea had 0.22, 0.24 and 0.25 mg/100 ml of zinc, respectively. Highest amount of zinc was found in Stevia powder based tea followed by sucralose and control tea. The sodium content of control, sucralose and Stevia based was observed as 19.32, and mg/100 ml, respectively. Sucralose based tea had higher sodium content followed by Stevia based tea and control tea. However, non-significant difference was observed among the sodium content of all the three types of tea. The results showed that incorporation of sucralose or Stevia in place of sugar in tea leads to increase in the potassium content of tea. Control, sucralose and Stevia based tea contained 51.12, mg and /100ml total potassium content, respectively. Maximum amount of potassium was observed in Stevia based tea and minimum in control tea. Sucralose based and control tea did not vary significantly in potassium content. Stevia based tea had significantly (P<0.05) higher value for magnesium (5.13 mg/100 ml) followed by sucralose based tea (4.58 mg/100 ml) and the lowest (4.51mg/100 ml) was observed in control tea. A non-significant difference was observed in sucralose based and control tea. Stevia based tea differed significantly in magnesium content in comparison to control and sucralose based tea Coffee Proximate composition Data depicting the results of the proximate composition of coffee prepared using sugar, sucralose and Stevia powder are given in Table Table 4.16: Proximate composition of Coffee (% as is basis) Types of Coffee Moisture Protein Fat Fibre Ash Total carbohydrate Control (Sugar) 82.60± ± ± ± ± ±0.04 Sucralose 84.82± ± ± ± ± ±0.06 Stevia powder 85.88± ± ± ± ± ±0.02 CD (P<0.05) 1.04 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Sucralose = Recipe with 4 mg of sucralose Stevia powder = Recipe with 8 mg of Stevia powder Control = Basic recipe with 8 g of sugar Significant differences were observed in moisture content of control, sucralose and Stevia based coffee. In control coffee, moisture content was observed as per cent which increased to per cent in sucralose based and per cent in Stevia based. The protein 52

64 and fat content ranged from content of control, sucralose and Stevia based were 4.68 to 4.65 per cent and 0.86 to 0.84 per cent, respectively. A Non-significant difference was observed in protein and fat content in all types of coffee. On the other hand, amount of fibre was found in all types of coffees which ranged from 0.10 to 0.12 per cent. The ash content ranged from 0.95 to 1.31 per cent, with the highest amount was observed in coffee prepared with Stevia powder and lowest amount was found in coffee prepared with sugar (control). Whereas, significant difference was noted in total carbohydrates content of all three types of coffees.there were per cent of total carbohydrates in sugar based coffee, 8.55 per cent in sucralose based coffee and 7.31 per cent in Stevia based coffee Sugars and starch The data regarding sugars (total sugar, reducing sugar and non-reducing sugars) and starch contents of coffee prepared using sugar, sucralose and Stevia based are presented in Table Table 4.17 : Sugars and starch contents of Coffee (% as is basis) Types of Coffee Sugars Starch Total Reducing Non reducing Control (Sugar ) 17.38± ± ± ±0.03 Sucralose 10.64± ± ± ±0.02 Stevia powder 10.44± ± ± ±0.07 CD (P<0.05) NS Values are mean ± SE of three independent observations Sucralose = Recipe with 4 mg of sucralose NS = Non-significant Stevia powder = Recipe with 8 mg of Stevia powder Control = Basic recipe with 8 g of sugar All the coffee was statistically different for their sugar contents. Significantly (P<0.05) higher amount of total soluble (17.38 %), reducing (6.94%) and non-reducing sugars (11.44%) were observed in sugar based coffee. The total soluble, reducing and non-reducing sugar content in sucralose based coffee were observed to be 10.64, 5.89 and 4.75 per cent, respectively. While, Stevia based coffee had 10.44, 5.85, and 4.59 per cent total soluble, reducing and non-reducing sugar content, respectively which were significantly (P<0.05) lower in total soluble sugars and non reducing sugars content among control, sucralose and Stevia based coffees. Whereas, reducing sugar content in sucralose based and Stevia based coffee differed non-significantly. Although a non-significant difference was observed among starch content of sugar based, sucralose based and Stevia based coffee. The starch content were observed to be highest (1.96%) in sucralose based coffee, followed by (1.93%) in Stevia based and (1.92%) in control coffee. 53

65 Minerals Data in respect to total iron, calcium, zinc, sodium, magnesium and potassium content of sugar, sucralose and Stevia based coffee are presented in the Table Table 4.18: Mineral contents of Coffee (mg/100 ml as is basis) Types of Coffee Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar) 97.34± ± ± ± ± ±0.11 Sucralose 97.37± ± ± ± ± ±0.02 Stevia powder 97.42± ± ± ± ± ±0.07 CD (P<0.05) NS NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 4 mg of sucralose NS = Non-significant Stevia powder = Recipe with 8 mg of Stevia powder Control = Basic recipe with 8 g of sugar All the coffees were non-statistically different for their calcium content. Maximum calcium content was observed in Stevia based (97.42 mg/100 ml) followed by sucralose based (97.37mg/100 ml), and sugar based (97.34 mg/100 ml) coffee, respectively. Concentration of total iron in different coffees ranged from 0.72 to 0.83 mg/100 ml. Highest iron content was observed in Stevia based coffee (0.83 mg/100 ml) and lowest iron content was observed in sugar based coffee (0.72 mg/100 ml). All the values for iron content of coffees differed non-significantly. Zinc content of all the three types of coffees differed non-significantly. Stevia based and sucralose based coffees had similar zinc (0.33 mg/100 ml) content. Whereas, sugar based coffee (control) had lowest amount of zinc i.e 0.31 mg / 100 ml. The sodium content in control, Stevia based and sucralose based coffees were observed as 23.68, and mg/100ml, respectively. Although all types of coffee differed non-significantly for the sodium content but sucralose based coffee had the highest amount of total sodium whereas control had the lowest Stevia powder based coffee was found to have a significant difference in magnesium content in comparison to control and sucralose based coffee. Stevia based coffee was found to have higher (7.79 mg/100 ml) magnesium content followed by sucralose based coffee (6.55 mg/100 ml) and sugar based coffee (6.47 mg/100 ml). Incorporation of coffee with sucralose and Stevia increased the potassium content. Higher potassium content was observed in Stevia powder based (74.72 mg/100 ml) followed by sucralose based (73.23 mg/100 ml) and sugar based coffee (73.21mg/100 ml). The potassium content of Stevia powder based coffee was observed to be significantly higher than other two types of coffees whereas control coffee and sucralose based coffee did not differ significantly among each other. 54

66 Milk shake Proximate composition Results of proximate composition of three types of milk shake prepared using sugar (control), sucralose and Stevia powder are given in Table Table 4.19 : Proximate composition of Milk Shake (% as is basis) Types of Milk shake Moisture Protein Fat Fibre Ash Total carbohydrates Control (Sugar ) 80.31± ± ± ± ± ±0.17 Sucralose 82.17± ± ± ± ± ±0.12 Stevia powder 84.84± ± ± ± ± ±0.14 CD (P<0.05) 2.04 NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 6 mg of sucralose NS = Non-significant Stevia powder = Recipe with 12 mg of Stevia powder Control = Basic recipe with 12 g of sugar Significant differences were observed in moisture content of control, sucralose and Stevia based milk shake. In control milk shake, moisture content was observed as per cent which increased to per cent in sucralose based and per cent in Stevia based milk shake. The protein and fat content ranged from content of control, sucralose and Stevia based milk shakes were 5.42 to 5.44 per cent and 1.29 to 1.31 per cent, respectively. A Nonsignificant difference was observed in protein and fat content in all types of milk shakes. On the other hand, lower amount of fibre was found in all types of milk shakes which ranged from 0.61 to 0.66 per cent. The ash content ranged from 1.92 to 2.07 per cent, with the highest amount was observed in milk shake prepared with Stevia powder and lowest amount was found in milk shake prepared with sugar (control) whereas, significant difference was noted in total carbohydrates content of all three types of milk shakes. There were per cent in sugar based milk shake, 8.42 per cent in sucralose based tea and 5.73 per cent in Stevia based milk shake Sugars and Starch The data in respect to sugars (total soluble sugar, reducing sugar and non-reducing sugars) and starch contents of milk shakesprepared using sugar (control), sucralose and Stevia powder are presented in Table Table 4.20 : Sugars and starch contents of Milk Shake (% as is basis) Types of Milk Sugars shake Total Reducing Non reducing Starch Control (sugar) 21.12± ± ± ±0.08 Sucralose 12.72± ± ± ±0.02 Stevia powder 12.70± ± ± ±0.04 CD (P<0.05) NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 12 g of sugar Sucralose = Recipe with 6 mg of sucralose Stevia powder = Recipe with 12 mg of Stevia powder 55

67 The total, reducing and non-reducing sugars content in control milk shakes were observed to be 21.12, 8.76 and per cent, respectively. Stevia based milk shake showed significantly lowest amounts of total soluble (12.70 %), reducing (6.51 %) and non-reducing sugar (6.19 %) content. Sucralose based milk shake was found to have 12.72, 6.56 and 6.16 per cent total, reducing and non-reducing sugar content, respectively. Sucralose based and Stevia based milk shake exhibited a non-significant difference in total, reducing sugars and non reducing sugars content. Stevia based milk shake exhibited significantly lower carbohydrate profile in comparison to control. Starch content of control milk shake was 4.02 per cent which increased slightly by addition of sucralose and Stevia. The values of starch content in sucralose based and Stevia based were 4.10 and 4.06 per cent, respectively. Maximum amount of starch was found to be in sucralose based milk shake followed by Stevia powder based milk shake. Non-significant difference was observed in the starch content of all types of milk shakes Minerals Results regarding iron, calcium, sodium, magnesium, zinc and potassium content of milk shake incorporating sugar (control), sucralose and Stevia are given in the Table Table 4.21: Mineral contents of Milk Shake (mg/ 100 ml on as is basis) Types of Milk shake Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar ) ± ± ± ± ± ±5.24 Sucralose ± ± ± ± ± ±4.88 Stevia powder ± ± ± ± ± ±2.24 CD (P<0.05) NS NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 6 mg of sucralose NS = Non-significant Stevia powder = Recipe with 12 mg of Stevia powder Control = Basic recipe with 12 g of sugar Milk shake prepared incorporating Stevia powder was found to vary in calcium content. Maximum calcium content was observed in Stevia based milkshake ( mg/100 ml) followed by sucralose based ( mg/100 ml), and sugar based ( mg/100 ml) milk shakes. All values for calcium content differed non-significantly among each other for control, sucralose based and Stevia powder based milk shakes. Concentration of iron in different types of milk shakes ranged from 0.59 to 0.61 mg/100ml, respectively. Highest iron content was observed in control milk shake (0.61 mg/100 ml) and lowest iron content was observed in sucralose based milk shake (0.59 mg/100 ml). A non-significant difference in iron content was observed among all the developed milk shakes. Non-significant differences were observed in all three types of milk shakes for zinc content. Similar zinc content (0.41mg/100 ml) was found in Stevia powder based and control 56

68 milk shake. Milk shake prepared using sucralose was found to have lowest zinc content (0.39 mg/100 ml). The sodium content of control, sucralose based and Stevia based milk shakes were observed as 42.22, and mg/100 ml, respectively. Sucralose milk shake contained significantly (P<0.05) highest amount of total sodium content and control had the lowest. Non-significant difference was observed among all types of milk shake. Among the milk shakes Stevia powder based milk shake was found to have significantly higher magnesium content in comparison to control and sucralose based milk shake. Stevia based milk shake with a value of mg/100 ml had significantly (P<0.05) higher magnesium content followed by sucralose based (11.26 mg/100 ml) and sugar based (11.23 mg/100 ml) milkshake. Total potassium content of Stevia powder based milk shake varied significantly (P<0.05) in comparison to control and sucralose based milk shakes. Maximum potassium ( mg/100 ml) was observed in Stevia based milk shake followed by ( mg/100 ml) sucralose based and sugar based ( mg/100 ml) milk shakes, respectively. Sucralose based and control milk shake did not differ significantly RTS Proximate composition Data regarding the proximate composition of RTS prepared with sugar, sucralose and Stevia powder are given in Table Table 4.22: Proximate composition of RTS (% as is basis) Types of RTS Moisture Protein Fat Fibre Ash Total carbohydrates Control (Sugar) 87.46± ± ± ± ± ±0.72 Sucralose 90.52± ± ± ± ± ±0.43 Stevia powder 90.97± ± ± ± ± ±0.86 CD (P<0.05) 0.33 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 10 g of sugar Sucralose = Recipe with 5 mg of sucralose Stevia powder = Recipe with 10 mg of Stevia powder On mean basis, significant difference was found in moisture content of control and incorporated RTS. Moisture content of both RTS incorporated sucralose and Stevia was higher than that of control RTS. In control RTS, the moisture was found to be per cent which increased to per cent in sucralose based RTS and per cent in Stevia powder incorporated RTS. Protein content indicated that, the control RTS had 0.94 per cent protein content. Sucralose and Stevia powder based RTS differed non-significantly for protein content. Maximum protein content i.e per cent was found in Stevia powder based RTS in comparison to the RTS prepared using sugar and sucralose. Fat content of control, sucralose 57

69 and Stevia based RTS was found to be 0.23, 0.24 and 0.26 per cent, respectively. A nonsignificant difference was observed in fat content of all types of RTS. Control RTS exhibited 0.41 per cent crude fibre. A non-significant difference was observed in protein values for sucralose based and Stevia based RTS. Maximum amount of crude fibre was found to be in sucralose based RTS i.e 0.43 per cent. Stevia based RTS showed significantly higher ash content. The ash content of control RTS was 0.11 per cent, whereas, sucralose and Stevia powder based RTS were found to have 0.18 and 0.33 per cent, respectively. Ash content of sucralose and Stevia based RTS was more than the control RTS. Total carbohydrates content of Stevia based RTS was significantly (P<0.05) lower than control RTS. Maximum amount of total carbohydrates was exhibited by control RTS (10.85 %) followed by sucralose based RTS (7.67 %) and Stevia based RTS (7.18 %) Sugars and starch The data in respect to sugar and starch contents of RTS prepared using sugar (control), sucralose and Stevia powder are presented in Table Table 4.23 : Sugars and starch contents of RTS (% as is basis) Types of RTS Sugars Total Reducing Non reducing Starch Control (Sugar ) 10.42± ± ± ±0.07 Sucralose 6.46± ± ± ±0.02 Stevia powder 6.45± ± ± ±0.05 CD (P<0.05) NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 10 g of sugar Sucralose = Recipe with 5 mg of sucralose Stevia powder = Recipe with 10 mg of Stevia powder Control, sucralose and Stevia based RTS exhibited significant differences for the sugars content. The total, reducing and non-reducing sugar content of control RTS were 10.42, 0.52 and 9.89 per cent Stevia RTS showed significantly (P<0.05) lowest amounts of total (6.45 %), reducing (0.44 %) and non-reducing (6.01 %) sugar content. Sucralose based RTS had 6.46, 0.41 and 6.05 per cent total, reducing and non-reducing sugar content, respectively which was significantly (P<0.05) lower than control RTS but higher than Stevia RTS. Starch content of control RTS was found to be 6.92 per cent, which was observed to be higher when sucralose and Stevia powder incorporated individually. Non-significant difference in starch content was observed for all types of RTS. The values for starch content was found to be 6.92 and 6.94 per cent, respectively for sucralose and Stevia based RTS. 58

70 Mineral The result regarding the mineral content of sugar, sucralose and Stevia based RTS is presented in Table Table 4.24 : Mineral contents of RTS (mg /100 ml as is basis) Types of RTS Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar) 3.01± ± ± ± ± ±0.07 Sucralose 2.97± ± ± ± ± ±0.03 Stevia powder 3.14± ± ± ± ± ±0.04 CD (P<0.05) NS NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 5 mg of sucralose NS = Non-significant Stevia powder = Recipe with 10 mg of Stevia powder Control = Basic recipe with 10 g of sugar Stevia powder based RTS had 3.14 mg/100 ml of total calcium, which was found to be higher among all the RTS. Sucralose and control RTS was found to have 2.97 and 3.01 mg/100 ml total calcium content, respectively. Least amount of calcium was found in sucralose incorporated RTS. Total iron content in sucralose, control and Stevia RTS were found to be 0.40, 0.41 and 0.42 mg/100 ml, respectively. A non-significant difference was observed in total iron content of RTS, whereas highest total iron content was found in RTS prepared using Stevia powder. Total zinc content of control, sucralose and Stevia based RTS were 0.21, 0.22 and 0.24 mg/100 ml, respectively. All three types of RTS did not vary significantly. Stevia powder based RTS exhibited the maximum zinc content followed by sucralose and control RTS. Total sodium content was higher in sucralose based RTS (5.17 mg/100 ml) followed by Stevia powder based RTS (5.13 mg/100 ml) and control RTS (5.12 mg/100 ml). Minimum amount of total sodium was observed in control RTS. Significantly higher amount (15.39 mg/100 ml) of total magnesium was found in Stevia powder based RTS followed by (14.17 mg/100 ml) in sucralose based and (14.14 mg/100 ml) in sugar based RTS. A non-significant difference was observed in magnesium content of control and sucralose based RTS. Stevia based RTS depicted highest amount (9.99 mg/100 ml) of potassium. The potassium content in control and sucralose based RTS were found to be 8.64 and 8.66 mg/100 ml respectively. A non-significant (P<0.05) difference was observed in the potassium content of sucralose based and control RTS. 59

71 4.3.2 Nutritional composition of baked products Biscuits Proximate composition The data regarding proximate composition of biscuits prepared using sugar, sucralose and Stevia powder based are depicted in the Table Table 4.25: Proximate composition of Biscuits (g /100 g on dry matter basis) Types of Biscuits Moisture Protein Fat Fibre Ash Total carbohydrate Control (Sugar) 2.30± ± ± ± ± ±0.01 Sucralose 2.83± ± ± ± ± ±0.03 Stevia powder 3.11± ± ± ± ± ±0.08 CD (P<0.05) 0.15 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 60 g of sugar Sucralose = Recipe with 30 mg of sucralose Stevia powder = Recipe with 60 mg of Stevia powder Control, sucralose based and Stevia powder based biscuits differ significantly (P<0.05) for their moisture content. The moisture content in sugar, sucralose and Stevia powder based biscuits were found to be 2.30, 2.83 and 3.11 g/100 g, respectively. The significantly highest amount of moisture was found in Stevia powder based biscuits and lowest moisture content was found in sugar based biscuits. The protein content of sucralose, control and Stevia powder based biscuits was found to be 7.81, 7.83 and 7.86 per cent, respectively. All developed samples did not vary significantly for their protein content Control and sucralose biscuit were found to have and per cent fat content, respectively. Fat content in Stevia based biscuits was found to be lower (15.97%) than control and sucralose based biscuits. The control biscuit exhibited a value 2.85 per cent for crude fibre which increased with the incorporation of sucralose and Stevia powder. The crude fibre content in sucralose and Stevia powder based biscuits was found to be 2.86 and 2.88 per cent, respectively. Nonsignificant difference was observed among control, sucralose and Stevia based biscuits for crude fibre content. Stevia based biscuits showed significantly (P<0.05) higher (1.94 %) ash content than control and sucralose based biscuits. The ash content in control and sucralose based biscuits were found to be 1.70 and 1.74 per cent, respectively. Maximum amount of total carbohydrates was found to be in control (69.31 %) followed by sucralose based (68.69 %) and the minimum observed in Stevia powder based biscuits (67.17 %). Total carbohydrates content in sugar based biscuits was found to be significantly (P<0.01) highest and Stevia powder based biscuits being the lowest. 60

72 Sugars and starch The data in respect to sugars and starch content of biscuits prepared using sugar, sucralose and Stevia powder are presented in Table Table 4.26: Sugars and starch contents of Biscuits (g/100 gm on dry matter basis) Types of Biscuits Sugars Starch Total Reducing Non reducing Control (Sugar) 20.23± ± ± ±0.01 Sucralose 15.03± ± ± ±0.08 Stevia powder 15.01± ± ± ±0.01 CD (P<0.05) NS Values are mean ± SE of three independent observations Sucralose = Recipe with 30 mg of sucralose NS = Non-significant Stevia powder = Recipe with 60 mg of Stevia powder Control = Basic recipe with 60 g of sugar The total, reducing and non-reducing sugar content of control biscuits were 20.23, 3.76 and g/100 g, respectively. Stevia powder based biscuits were found to have significantly (P<0.05) lower amounts of total (15.01 g/100 g), reducing (1.56 g/100 g) and non-reducing (13.45 g/100 g) sugars, respectively. Sucralose based biscuits were found to have 15.03, 1.52 and g/100 g total soluble, reducing and non-reducing sugar content, respectively. Significantly (P<0.05) highest amount was found in control biscuits whereas, lowest amount was found in Stevia powder biscuits for total, reducing and non-reducing sugars content. Starch content of control biscuits was found to be g/100 g. The values for starch content were observed to be and g/100 g, respectively for biscuits incorporating sucralose and Stevia powder. Control, sucralose and Stevia powder biscuits were found to have a non-significant (P<0.05) difference in starch content Minerals The data in respect to calcium, iron, sodium, magnesium, zinc and potassium content of biscuits prepared using, sugar (control), sucralose and Stevia powder are depicted in Table Table 4.27: Mineral contents of Biscuit (mg /100 gm on dry matter basis) Types of Biscuit Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar ) 74.31± ± ± ± ± ±0.20 Sucralose 74.34± ± ± ± ± ±0.14 Stevia Powder 75.27± ± ± ± ± ±0.23 CD (P<0.05) 0.71 NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 30 mg of sucralose NS = Non-significant Stevia powder = Recipe with 60 mg of Stevia powder Control = Basic recipe with 60 g of sugar 61

73 Stevia powder biscuits were found to have a significant difference in calcium content in comparison to sucralose based and sugar based biscuits. Maximum calcium content was observed in Stevia based (75.27 mg/100 g) followed by sucralose based (74.34 mg/100 g) and control (74.31 mg/100 g) biscuits. mg/100 g. Total iron concentration in different types of biscuits ranged from 2.61 to 2.63 Non-significant differences were found among all types of biscuits. Highest (2.63mg/100 g) iron content was observed in sucralose based biscuits and similar (2.61 mg/100 g) iron content was observed in sugar and Stevia powder based biscuits. Non-significant differences were found among developed biscuits for zinc content. Maximum amount of zinc was found to be in sucralose based biscuits with a value of 0.44 mg / 100 g and similar zinc content was found in control and Stevia powder (0.43 mg/100 g) based biscuits. The amount of total sodium content of control, sucralose and Stevia powder based biscuits were found to be 99.21, and mg/100 g, respectively. A significant difference was observed in Stevia powder based biscuits and other two types of biscuits for magnesium content. The highest magnesium content was observed in Stevia powder based biscuits (20.69 mg/100 g) followed by sucralose based (19.28 mg /100 g) and control (19.26 mg/100 g) biscuits. Significantly higher total potassium content was found in Stevia powder based biscuits in comparison to sucralose and control biscuits. Stevia powder based biscuits found to have mg/100 g of potassium content. While, control and sucralose based biscuits were found to have significantly lower potassium content i.e and mg /100 g Cake Proximate composition The data in respect to proximate composition of cake prepared using, sugar (control), sucralose and Stevia powder are depicted in Table Table 4.28: Proximate composition of Cake (g/100 g on dry matter basis) Types of Cake Moisture Protein Fat Fibre Ash Total carbohydrate Control (sugar) 27.28± ± ± ± ± ±2.03 Sucralose 31.79± ± ± ± ± ±2.01 Stevia powder 31.81± ± ± ± ± ±3.06 CD (P<0.05) 1.06 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 60 g of sugar Sucralose = Recipe with 30 mg of sucralose Stevia powder = Recipe with 60 mg of Stevia powder 62

74 Stevia powder based and sucralose based cakes did not differ significantly (P<0.05) for their moisture content. The highest moisture content was found to be per cent in Stevia powder based cake followed by per cent in sucralose based and per cent in sugar based cake. Control cake was found to have significantly (p<0.05) lowest moisture content in comparison to cakes prepared using sucralose and Stevia powder. Almost similar protein content was found in control and sucralose based cake i.e 8.44 and 8.45 per cent, respectively. A non-significant but higher protein content (8.48 %) was found in Stevia based cake. Cake prepared using sugar was served as control. Similar fat content (19.65 %) was found in sucralose based and control cake. Stevia based cake was found to have slightly lower (19.64 %) fat content in comparison to sucralose based and control cake. The cake exhibited a value of 1.44 to 1.48 per cent crude fibre content. Replacement of sugar with sucralose and Stevia powder increased the fibre content among cakes whereas, the difference was found to be non-significant. Stevia powder based cake were found to have significantly (P<0.01) higher ash content than control cake. Control cake, sucralose cake and Stevia powder cake were found to have 1.38, 1.40 and 1.89 per cent, respectively ash content. Sucralose based cake differed non-significantly for ash content in comparison to control whereas, cake prepared using Stevia powder differed significantly in comparison to sugar and sucralose. Significantly higher total carbohydrate content was found in control (41.76 %) followed by sucralose (34.24 %) and Stevia powder (34.14 %) based cake. Stevia powder cake differed significantly for total carbohydrate content in comparison to control Sugars and starch The data in respect to total, reducing and non-reducing sugar and starch contents of cake prepared using sugar, sucralose and Stevia powder are presented in Table Table 4.29 :Sugars and starch contents of cake (g/100 g on dry matter basis) Types of Cake Sugars Starch Total Reducing Non reducing Control (sugar) 35.45± ± ± ±0.12 Sucralose 24.79± ± ± ±0.13 Stevia powder 24.71± ± ± ±0.24 CD (P<0.05) NS Values are mean ± SE of three independent observations Sucralose = Recipe with 30 mg of sucralose NS = Non-significant Stevia powder = Recipe with 60 mg of Stevia powder Control = Basic recipe with 60 g of sugar The total, reducing and non-reducing sugar content of control cake were found to be 35.45, 3.80 and g/100 g, respectively. Sucralose based cake was found to have 63

75 significantly (P<0.05) lower amounts of total soluble (24.79 g/100 g), reducing (3.66 g/100 g) and non-reducing (21.13 g/100 g) sugars, respectively in comparison to control cake. Stevia powder based cake was found to have 24.71, 3.71 and g/100 g total, reducing and nonreducing sugar content, respectively. Significantly (P<0.05) higher amount of total, reducing and non-reducing sugars was observed in control cake whereas, Stevia powder based cake was found to have the lower total soluble and non reducing sugars. Control and Stevia powder based cake was found to have similar starch content (39.85g/100 g). Starch content of sucralose based cake was found to be g/100g, respectively. All types of cakes differed non-significantly (P<0.05) for starch content Minerals The data in respect to total iron, calcium, sodium, magnesium, zinc and potassium content in cake prepared using sugar, sucralose and Stevia powder are depicted in Table Table 4.30 : Mineral contents of Cake (mg/100 g on dry matter basis) Types of cake Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar) 75.23± ± ± ± ± ±5.18 Sucralose 75.27± ± ± ± ± ±4.24 Stevia powder 77.11± ± ± ± ± ±2.21 CD (P<0.05) 0.97 NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 30 mg of sucralose NS = Non-significant Stevia powder = Recipe with 60 mg of Stevia powder Control = Basic recipe with 60 g of sugar Stevia powder based cake was found to have a significant difference in calcium content. Maximum calcium content was observed in Stevia powder based cake (77.11 mg/100 g) followed by sucralose cake (75.27 mg/100 g) and control (75.23 mg/100 g). Iron concentration in different types of cake prepared using sugar, sucralose and Stevia powder were found to be 1.52, 1.53 and 1.52 mg/100 g. Similar value for total iron content was observed in control and Stevia powder based cake. A higher value for iron content was observed in sucralose based cake, whereas difference among different types of cake was observed to be non-significant. Zinc content of control and sucralose based cake was found to be similar i.e 0.54 mg/100 g. Stevia powder based cake had 0.53 mg/100 g of total zinc content. All types of cakes differed non-significantly for zinc content among each other. Total sodium content of Stevia powder, sugar (control) and sucralose based cake were found to be , and mg/100 g, respectively. A non-significant (P<0.05) difference was observed among all types of cake. Sucralose based cake was found to have the highest sodium content and Stevia powder based cake exhibited the lowest sodium content. 64

76 Stevia powder based cake differed statistically for the magnesium content. Maximum magnesium content was observed in Stevia powder based cake (23.85 mg/100 g) followed by sucralose based (22.34 mg/100 g) and control (22.23 mg/100 g) cake. Maximum potassium was observed in Stevia powder based cake with a value of mg/100 g and minimum was found in control (154.65mg/100 g). Potassium content was found to be mg/100 g in sucralose based cake. Cake prepared using sugar and sucralose were not found to be statistically different for their potassium content Buns Proximate composition The data regarding proximate composition of buns prepared using sugar, sucralose and Stevia powder are tabulated in the Table Table 4.31: Proximate composition of Buns (g/100 g on dry matter basis) Types of Buns Moisture Protein Fat Fibre Ash Total carbohydrate Control (Sugar) 36.27± ± ± ± ± ±0.07 Sucralose 39.12± ± ± ± ± ±0.02 Stevia powder 39.34± ± ± ± ± ±0.06 CD (P<0.05) 0.07 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 30 g of sugar Sucralose = Recipe with 15 mg of sucralose Stevia powder = Recipe with 30 mg of Stevia powder Mean basis, significant differences were observed in moisture content of control and buns prepared using sucralose and Stevia powder. Control bun had per cent which increased to per cent in sucralose and per cent in Stevia powder based bun. Protein content of bun was also observed to have no significant differences. Higher amount of protein was found in Stevia powder based bun (6.25 %) followed by sucralose based bun and control. A similar value (6.23 %) for protein content was observed in sucralose based bun and control. Fat content of control bun was per cent. Incorporation of sucralose and Stevia powder showed no significant difference in the fat content in sucralose and Stevia powder based bun. The control bun exhibited 1.36 per cent crude fibre which increased by incorporation of sucralose and Stevia powder. The similar crude fibre content i.e 1.37 per cent was found in sucralose and Stevia powder based buns. All three types of buns differ nonsignificantly among each other for crude fibre content. Ash content of incorporated buns was more than the control bun. The ash content of three types of buns was 1.59 per cent (control), 1.62 per cent (sucralose based bun) and 1.92 per cent (Stevia powder based buns). Stevia powder based buns showed significantly (P<0.05) higher ash content as compared to control and sucralose based buns. Total carbohydrates content of sucralose and Stevia powder based bun was significantly (P<0.05) lower than control buns. Maximum amount of total 65

77 carbohydrates was found in control buns (41.41%) followed by sucralose based bun (38.56 %) and Stevia powder based bun (38.45%) Sugars and starch The data in respect to total sugar, reducing sugar, non-reducing sugar and starch contents are presented in Table The total, reducing and non-reducing sugar content of control bun were found to be 21.12, 8.76 and g/100 g. Stevia powder incorporated bun had significantly (P<0.05) lower amounts of total sugars (13.70 g/100 g), reducing (4.58 g/100 g) and non-reducing sugar content (8.42 g/100 g) and sucralose incorporated bun was found to have 13.72, 4.42 and 9.36 g/100 g total, reducing and non-reducing sugar content, respectively. A significantly (P<0.05) higher amount of total and non-reducing sugars was observed in control bun whereas, bun prepared using sucralose and Stevia powder differed non-significantly for reducing sugars content. Table 4.32 : Total sugars and starch content of Bun (g/100 gm on dry matter basis) Types of Bun Sugars Starch Total Reducing Non reducing Control (Sugar) 21.12± ± ± ±2.02 Sucralose 13.72± ± ± ±2.06 Stevia powder 13.70± ± ± ±1.08 CD (P<0.05) NS Values are mean ± SE of three independent observations Sucralose = Recipe with 15 mg of sucralose NS = Non-significant Stevia powder = Recipe with 30 mg of Stevia powder Control = Basic recipe with 30 g of sugar Starch content of control bun was g/100 g. The starch content was found to be and g/100 g for Stevia powder and sucralose incorporated bun, respectively. All types of buns did not differ significantly (P<0.05) for starch content Minerals Data in respect to total calcium, iron, sodium, magnesium, zinc and potassium content of bun prepared using sugar, sucralose and Stevia powder is presented in the Table Table 4.33 : Mineral contents of Bun (g /100 g on dry matter basis) Type of bun Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar) 74.28± ± ± ± ± ±3.08 Sucralose 74.19± ± ± ± ± ±4.14 Stevia powder 74.47± ± ± ± ± ±5.15 CD (P<0.05) NS NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 15 mg of sucralose NS = Non-significant Stevia powder = Recipe with 30 mg of Stevia powder Control = Basic recipe with 30 g of sugar 66

78 All three types of bun did not differ statistically for their calcium content. Maximum calcium content was observed in Stevia powder incorporated bun (74.47 mg/100 g) followed by control (74.28 mg/100 g), and sucralose incorporated bun (74.19 mg/100 g). Concentration of total iron in different buns prepared using sugar, sucralose and Stevia powder ranged from 1.57 to 1.60 mg/100g, respectively. Iron content did not differ significantly but highest (1.60 mg/100 g) iron content was observed in control bun and lower (1.57 and 1.58 mg/100 g) was found to be in sucralose and Stevia powder based bun. Zinc content in buns did not vary significantly (P<0.05). Similar zinc content (0.71 mg/100 g) was found to be in control and Stevia powder based bun. Sucralose based bun was found to have 0.70 mg/100 g zinc content. The sodium content of control, sucralose and Stevia powder were observed as , and mg/100 g, respectively. Sodium content of buns did not differ significantly (P<0.05). Stevia powder based bun was found to have a significant difference in magnesium content in comparison to control and sucralose based buns. Stevia powder bun with a value of mg/ 100 g had significantly (P<0.05) higher magnesium followed by control (20.23 mg/100 g) and sucralose based (20.21 mg/100 g) buns. Stevia powder based bun was found to have significantly higher total potassium content in comparison to control and sucralose based bun. Total potassium were found to be , and mg/100 g, respectively in control, sucralose based and Stevia powder based bun Traditional products Sweet porridge Proximate composition The data regarding proximate composition of sweet porridge prepared using sugar (control), sucralose and Stevia powder are presented in the Table Table 4.34 : Proximate composition of Sweet porridge (g /100 g on dry matter basis) Types of Sweet porridge Moisture Protein Fat Fibre Ash Total carbohydrate Control (Sugar ) 61.22± ± ± ± ± ±2.08 Sucralose 65.54± ± ± ± ± ±2.04 Stevia powder 65.90± ± ± ± ± ±2.02 CD (P<0.05) 3.65 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 30 g of sugar Sucralose = Recipe with 15 mg of sucralose Stevia powder = Recipe with 30 mg of Stevia powder 67

79 Significant differences were observed in moisture content of control and sucralose and Stevia powder incorporated sweet porridge. Moisture content of sweet porridge prepared using Stevia powder and sucralose were found to be higher than that of control sweet porridge. In control sweet porridge, moisture was found to be per cent which increased to per cent in sucralose and per cent in Stevia powder incorporated sweet porridge. Protein content of sucralose, control and Stevia powder incorporated sweet porridge was found to be 9.05, 9.08 and 9.10 per cent, respectively. Protein content increased with the addition of Stevia powder, where as the difference was found to be non-significant in protein content of sweet porridge with the incorporation of sucralose. Fat content in all types of sweet porridge was found to have non-significant difference. The maximum fat content was observed in sucralose sweet porridge (2.09%) followed by control (2.07%) and Stevia powder (2.06%) sweet porridge. The control sweet porridge was found to have 1.29 per cent crude fibre which increased but not significantly (P<0.05) upon incorporation of sucralose and Stevia powder. The crude fibre content in sucralose and Stevia powder incorporated sweet porridge was observed to be 1.30 and 1.32 per cent, respectively. Stevia powder incorporated sweet porridge showed significantly (P<0.05) higher ash content. The ash content of control sweet porridge was 1.98 per cent, sucralose was 2.00 per cent and Stevia powder 2.39 per cent, respectively. Sweet porridge incorporated sucralose and Stevia powder ash content was found to be more than the control sweet porridge. Total carbohydrates content of Stevia powder incorporated sweet porridge was significantly (P<0.05) lower than control sweet porridge. Maximum amount of total carbohydrates was exhibited by control sweet porridge (24.43 %) followed by sucralose incorporated sweet porridge (20.09 %) and Stevia powder incorporated sweet porridge (19.60%) Sugars and starch The data in respect to total soluble sugar, reducing sugar, non-reducing sugar and starch content of sweet porridge prepared using sugar (control), sucralose and Stevia powder are presented in Table Table 4.35: Sugars and starch contents of Sweet Porridge (g/100 gm on dry matter basis) Types of sweet porridge Sugars Starch Total Reducing Non reducing Control (Sugar) 19.24± ± ± ±2.02 Sucralose 16.10± ± ± ±3.01 Stevia powder 15.09± ± ± ±2.01 CD (P<0.05) NS Values are mean ± SE of three independent observations Sucralose = Recipe with 15 mg of sucralose NS = Non-significant Stevia powder = Recipe with 30 mg of Stevia powder Control = Basic recipe with 30 g of sugar 68

80 The total, reducing and non-reducing sugar content of control sweet porridge were 19.24, 3.62 and g/100 g. Stevia powder incorporated sweet porridge showed significantly (P<0.05) lower amounts of total (15.09 g/100 g), reducing sugars (4.30 g/100 g) and non-reducing sugar (10.79 g/100 g) content, respectively. Sweet porridge prepared using sucralose was found to have 16.10, 3.32 and g/100 g total, reducing and non-reducing sugar content, respectively. Significantly (P<0.05) higher amount of total, reducing and nonreducing sugars was found to be in control sweet porridge followed by porridge prepared using sucralose and Stevia powder incorporated sweet porridge. Starch content of control sweet porridge was g/100 g which increased but not significantly (P<0.05) in Stevia powder incorporated sweet porridge. The values of starch content were found to be and g/100 g, respectively for Stevia powder and sucralose incorporated sweet porridge Minerals The data in respect to total iron, calcium, sodium, magnesium, zinc and potassium content of sweet porridge incorporated with sugar, sucralose and Stevia powder are depicted in Table Table 4.36: Mineral contents of Sweet porridge (g /100 g on dry matter basis) Type of Sweet porridge Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar) 42.54± ± ± ± ± ±3.20 Sucralose 42.57± ± ± ± ± ±3.14 Stevia powder 43.06± ± ± ± ± ±3.20 CD (P<0.05) 0.31 NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 15 mg of sucralose NS = Non-significant Stevia powder = Recipe with 30 mg of Stevia powder Control = Basic recipe with 30 g of sugar All the sweet porridge was found to be statistically different for their calcium content. Maximum calcium content was observed in Stevia powder incorporated sweet porridge (43.06 mg/100 g) followed by sucralose incorporated sweet porridge (42.57 mg/100 g) and control sweet porridge (42.54 mg/100 g). Iron concentration in different sweet porridge ranged from 2.18 to 2.21 mg/100 g. Higher iron content was found to be in control sweet porridge (2.21 mg/100 g) and similar value for iron content was found to be in sucralose and Stevia powder incorporated sweet porridge (2.18 mg/100 g). A non-significant difference was found in iron content of all the developed sweet porridge. Zinc content of all types of sweet porridge did not vary significantly (P<0.05). Similar zinc content was found in sugar (control) and sucralose incorporated sweet porridge (1.23mg/100 g) followed by Stevia powder incorporated sweet porridge (1.21 mg/100 g). 69

81 A non-significant difference was found to be among all sweet porridge for total sodium content. The sodium content in Stevia powder, control and sucralose incorporated sweet porridge were found to be , and mg/100 g, respectively. Maximum magnesium content was observed in Stevia powder incorporated sweet porridge with a value of mg/100 g and minimum was found in control sweet porridge (57.73 mg/100 g). Significant difference was observed in total magnesium content of Stevia powder incorporated sweet porridge in comparison to control and sucralose incorporated sweet porridge. Significant difference in total potassium content was found to be in Stevia powder incorporated sweet porridge in comparison to sucralose incorporated sweet porridge and control. The potassium content in control, sucralose and Stevia powder incorporated sweet porridge were exhibited to be , and mg/100 g, respectively Carrot halwa Proximate composition The data regarding proximate composition of carrot halwa prepared using sugar, sucralose and Stevia powder are depicted in Table Table 4.37: Proximate composition of Carrot halwa (g /100 g on dry matter basis) Types of Carrot halwa Moisture Protein Fat Fibre Ash Total carbohydrate Control (Sugar) 52.18± ± ± ± ± ±2.03 Sucralose 57.31± ± ± ± ± ±3.04 Stevia powder 57.26± ± ± ± ± ±3.05 CD (P<0.05) 2.27 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 30 g of sugar Sucralose = Recipe with 15 mg of sucralose Stevia powder = Recipe with 30 mg of Stevia powder Sucralose and Stevia powder based carrot halwa did not differ significantly (P<0.01) for their moisture content. In control carrot halwa moisture content was found to be per cent, in sucralose based carrot halwa it was found to be per cent and in Stevia powder based carrot carrot halwa it was found to be per cent, respectively. Protein content of control and Stevia powder based carrot halwa was 6.21 and 6.17 per cent, respectively. Non-significant difference in protein content was observed in control, sucralose and Stevia powder based carrot halwa. Similar fat content 5.43 per cent was found to be in control and Stevia powder based halwa. Fat content in sucralose based carrot halwa was observed to be 5.44 per cent. All the values for fat content in all types of halwa did not differed significantly. The control and Stevia powder based carrot halwa exhibited 1.31 per cent crude fibre which increased but non-significantly (P<0.01) upon replacement of sugar with sucralose. The crude fibre content in sucralose based carrot halwa was found to be 1.32 per cent. 70

82 Stevia powder based carrot halwa showed significantly (P<0.01) higher ash content (2.58%) than control carrot halwa. The ash content in control carrot halwa was 2.32 per cent and in sucralose based carrot halwa was 2.37 per cent. Maximum total carbohydrates was found to be in control carrot halwa (32.51 %) followed by sucralose based carrot halwa (27.21%). Stevia powder based carrot halwa was found to have significantly (P<0.05) lower (27.06%) total carbohydrates content Sugars and Starch The data regarding sugars and starch contents of carrot halwa prepared using sugar (control), sucralose and Stevia powder are depicted in Table Table 4.38 : Sugars and starch contents of Carrot halwa (g/100 g on dry matter basis) Types of Carrot halwa Sugars Total Reducing Non reducing Starch Control (Sugar) 31.73± ± ± ±0.07 Sucralose 22.60± ± ± ±0.01 Stevia powder 22.54± ± ± ±0.01 CD (P<0.05) NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 30 g of sugar Sucralose = Recipe with 15 mg of sucralose Stevia powder = Recipe with 30 mg of Stevia powder The total, reducing and non-reducing sugar content of control 31.73, and g/100 g. Stevia powder based carrot halwa showed significantly (P<0.05) lower amounts of total (22.54 g/100 g), reducing (7.35 g/100 g) and non-reducing (15.19 mg/100 g) sugars, respectively. Sucralose based carrot halwa was found to have 22.60, 6.64 and g/100 g total, reducing and non-reducing sugar content, respectively. Significantly (P<0.05) higher amount of total, reducing and non-reducing sugars was found to be in control carrot halwa while, lowest was exhibited in Stevia powder based carrot halwa. Starch content of control carrot halwa was found to be 1.73 g/100 g. Starch content for sucralose and Stevia powder based carrot halwa were found to be 1.74 and 1.76 g/100 g, respectively. Starch content in all types of carrot halwa did not differed significantly Minerals The data in respect to total iron, calcium, sodium, magnesium, zinc and potassium content of carrot halwa prepared by incorporating sugar, sucralose and Stevia powder are depicted in Table

83 Table 4.39 : Mineral contents of Carrot halwa (mg /100 g on dry weight basis) Types of Carrot halwa Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar) ± ± ± ± ± ±1.02 Sucralose ± ± ± ± ± ±1.01 Stevia powder ± ± ± ± ± ±1.05 CD (P<0.05) 0.46 NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 15 mg of sucralose NS = Non-significant Stevia powder = Recipe with 30 mg of Stevia powder Control = Basic recipe with 30 g of sugar Stevia powder based carrot halwa were found to have a significant difference in calcium content in comparison to control and sucralose. Maximum calcium content was observed in Stevia powder based carrot halwa ( mg/100 g) followed by sucralose based carrot halwa ( mg/100 g) and control carrot carrot halwa ( mg/100 g). Iron concentration in different types of carrot halwa ranged from 0.29 to 0.31 mg/100 g. Higher iron content was found to be in control carrot halwa followed by Stevia powder based carrot halwa and lowest was found to be in sucralose based halwa. A non-significant difference in iron content was exhibited in all developed carrot halwa. Non-significant differences were observed among developed carrot halwa for total zinc content. Maximum amount of total zinc was observed in Stevia powder based carrot halwa i.e 0.91 mg/100 g and minimum was found to be in sucralose based carrot halwa (0.86 mg/100 g). Zinc content was found to be higher in control carrot halwa than sucralose based carrot halwa. The sodium content of control, Stevia powder and sucralose based carrot halwa were found to be 51.47, and mg /100 g, respectively. Sucralose based carrot halwa were found to have higher amount of total sodium content and control the lowest. Sodium content in all types of carrot halwa differed non-significantly. A significant difference was found to be in Stevia powder based carrot halwa for total magnesium content. Stevia powder based carrot halwa was found to have higher magnesium content (32.21 mg/100 g) followed by control carrot halwa (31.62mg/100 g) and sucralose based carrot halwa (31.57 mg/100 g). Significant differences in potassium content were exhibited among all developed carrot halwa. Stevia powder based carrot halwa had significantly (P<0.05) higher value for potassium content ( mg/100 g) while, control and sucralose based carrot halwa were found to have lower potassium content ( and mg /100 g). A non-significant difference was observed among control and sucralose based carrot halwa. 72

84 Kheer Proximate composition Data in respect to proximate composition of kheer prepared using sugar (control), sucralose and Stevia power are presented in Table Table 4.40 : Proximate composition of Kheer (g /100 g on dry matter basis) Types of Kheer Moisture Protein Fat Fibre Ash Total carbohydrates Control (Sugar) 59.94± ± ± ± ± ±1.28 Sucralose 68.03± ± ± ± ± ±1.18 Stevia powder 68.20± ± ± ± ± ±1.05 CD (P<0.05) 1.38 NS NS NS Values are mean ± SE of three independent observations NS = Non-significant Control = Basic recipe with 20 g of sugar Sucralose = Recipe with 10 mg of sucralose Stevia powder = Recipe with 20 mg of Stevia powder Significant differences were observed in moisture content of control, sucralose and Stevia powder based kheer. Control kheer had per cent which increased to per cent in sucralose and per cent in Stevia powder kheer. Protein content of kheer was found to have non-significant difference. Higher amount of protein was found in Stevia powder based kheer (8.15 %) followed by control (8.12%) and sucralose based kheer. Similar fat content was found to be in control and Stevia powder based kheer i.e 6.30 per cent, which differed non-significantly (P<0.05). Sucralose based kheer had 6.31 per cent fat content. The control kheer exhibited 0.81 per cent crude fibre (P<0.05). Incorporation of sucralose and Stevia powder in kheer increase the crude fibre content in kheer. The difference was found to be non-significant. Ash content of sucralose and Stevia powder incorporated kheer was more than the control kheer. The ash content of three types of kheer was found to be 2.82 per cent (control), 2.84 per cent (sucralose) and 3.05 per cent (Stevia powder). Stevia powder based kheer showed significantly (P<0.05) higher ash content as compared to control and sucralose based kheer. Total carbohydrates content of incorporated kheer was significantly (P<0.05) lower than control kheer. Maximum amount of total carbohydrates was found in control kheer (22.01 %) followed by sucralose based kheer (14.78 %) and Stevia powder based kheer (13.46%) Sugars and starch The data in respect to total soluble sugar, reducing sugar and non-reducing sugar content kheer incorporated with sugar, sucralose and Stevia powder are presented in Table

85 Table 4.41: Sugars and starch contents of Kheer (g/100 g on dry matter basis) Types of Kheer Sugars Total Reducing Non reducing Starch Control (Sugar) 32.21± ± ± ±1.02 Sucralose 24.26± ± ± ±1.07 Stevia powder 24.24± ± ± ±1.02 CD (P<0.05) NS Values are mean ± SE of three independent observations Sucralose = Recipe with 10 mg of sucralose NS = Non-significant Stevia powder = Recipe with 20 mg of Stevia powder Control = Basic recipe with 20 g of sugar The total, reducing and non-reducing sugar content of control kheer were found to be 32.21, and g/100 g, respectively. Sucralose based kheer was found to have significantly (P<0.05) lower total (24.26 g/100 g) and non reducing sugars (16.42 g/100 g) than control whereas, reducing sugar (7.84 g/100 g) content was found to be significantly (P<0.05) lower than Stevia powder based kheer. Stevia powder based kheer was found to have 24.24, 8.55 and g/100 g total, reducing and non-reducing sugar content, respectively. Total, reducing and non-reducing sugars were found to be in significantly highest in control kheer and Stevia powder based kheer the lowest. Starch content of control kheer was found to be g/100 g. Starch content for Stevia powder and sucralose based kheer were found to be and g/100 g, respectively. Starch content in all types of kheer did not differ significantly Minerals The data pertaining to calcium, iron, zinc, sodium, magnesium and potassium content in sugar (control), sucralose and Stevia powder kheer is presented in Table Table 4.42: Mineral contents of Kheer (mg /100 g, on dry weight basis) Types of Kheer Calcium Iron Zinc Sodium Magnesium Potassium Control (Sugar) ± ± ± ± ± ±0.88 Sucralose ± ± ± ± ± ±0.88 Stevia powder ± ± ± ± ± ±0.20 CD (P<0.05) 0.73 NS NS NS Values are mean ± SE of three independent observations Sucralose = Recipe with 10 mg of sucralose NS = Non-significant Stevia powder = Recipe with 20 mg of Stevia powder Control = Basic recipe with 20 g of sugar Stevia powder based kheer were found to have a significant difference in calcium content in comparison to control and sucralose based kheer. Maximum calcium content was found to be in Stevia powder based carrot kheer ( mg/100 g) followed by sucralose based kheer ( mg/100 g) and control kheer ( mg/100 g). 74

86 Iron concentration in different types of kheer ranged from 0.10 to 0.12 mg/100 g. Higher iron content was found to be in control kheer followed by Stevia powder and sucralose based kheer. A non-significant difference in iron content was exhibited in all developed kheer. Non-significant differences were observed among developed kheer for total zinc content. Maximum amount of total zinc was found to be in Stevia powder based kheer i.e 0.45 mg/100 g and minimum was found to be in sucralose based kheer (0.41 mg/100 g). Zinc content did not vary significantly for control, sucralose and Stevia powder based kheer. The sodium content of Stevia powder, control and sucralose based kheer were found to be 42.50, and mg/100 g, respectively. Sucralose based kheer were found to have higher total sodium content and Stevia powder based kheer the lowest. Sodium content in all types of kheer differed non-significantly. A significant difference was found to be in Stevia powder based kheer for total magnesium content. Stevia powder based kheer was found to have higher magnesium content (15.82 mg/100 g) followed by control kheer (14.64 mg/100 g) and sucralose based kheer (14.62 mg/100 g). Significant differences in potassium content were exhibited among all developed kheer. Stevia powder based kheer had significantly (P<0.05) higher value for potassium content ( mg/100 g) while, control and sucralose based kheer were found to have lower potassium content ( and mg /100 g). A non-significant difference was observed among control and sucralose based kheer. 4.4 Glycaemic response of selected food products Glycaemic index of selected food products were assessed in normal and healthy (n=10), obese (n=10) and diabetic subjects (n=10) free from any kind of ailments. The experimental subjects were in the age group of 40 to 51 yrs. The mean for age, height, weight and body mass index (BMI) of the normal, obese and diabetic subjects are presented in Table Table 4.43 :Anthropometric measurements of subjects Subjects Age Sex Height (cm) Weight (Kg) BMI Normal (n=10) 40.4 F Obese (n=10) 45.6 F Diabetic (n=10) 50.9 F The mean height of the subjects ranged from to cm and weight ranged between 62.9 to 84.8 kg, respectively, with an average height were found to be 162.0,

87 and cm and weight were found to be 62.9, 84.8 and 69.7 kg, respectively for normal, obese and diabetic subjects. Mean value of BMI to be 24.00, and kg/m 2 for normal, obese and diabetic subjects, respectively. Glycaemic response of selected food products On the basis of organoleptic acceptability, the most acceptable products (milk shake, RTS, biscuit, cake, bun, sweet porridge, carrot halwa and kheer) were selected for studying their glycaemic response in normal, obese and diabetic subjects. Available carbohydrate content of developed food products Data regarding the available carbohydrate contents of organoleptically acceptable products among beverages, baked and traditional products are presented in Tables 4.44 and As beverages were low in available carbohydrates and in order to provide 50 g of equicarbohydrate portion the amount was exceeding the portion size so, amount given to subjects was adjusted in order to provide 25 g of equicarbohydrates/ Serving for beverages and 50 g of equicarbohydrates/ Serving for baked and traditional developed food products as mentioned in tables 4.44 and Table 4.44 : Amount of beverages ingested 25 g of equicarbohydrate/serving portion for glycaemic index / person S.No. Control (Sugar) Sucralose Stevia powder Beverages Available carbohydrates (g) Amt/ serving (ml) Available carbohydrate (g) Amt/ serving (ml) Available carbohydrate (g) Amt/ serving (ml) Milk shake RTS Table 4.45: Amount of food products ingested 50g of equicarbohydrate/ Serving portion for glycaemic index / person S.No. Control (Sugar) Sucralose Stevia powder Products Baked products Available carbohydrates (g) Amt/ serving (g) Available carbohydrate (g) Amt/ serving (g) Available carbohydrate (g) Amt/ serving Biscuits Cake Bun Traditional products Sweet porridge Carrot halwa Kheer (g) 76

88 4.4.1 Glycaemic response of beverages Milk shake Glycaemic index of mix shake in normal subjects Data in respect to area under glucose curve and glycaemic index of Milk shake prepared using sugar (control), sucralose and Stevia powder are presented in Table Table 4.46 : Area under blood glucose curve (mg/dl) of milk shake prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean The area under blood glucose response curve for milk shake for normal ranged from 1574 to 1920, 1070 to1357 and 1067 to 1353 mg/dl for milk shakes based on sugar (control), sucralose and Stevia powder, respectively in normal subjects. The mean values of glycaemic index of control, sucralose and Stevia powder milk shake in normal subjects were observed as 46.35, and 33.06, respectively. Milk shake prepared using sugar was found to have maximum glycaemic index followed by sucralose and Stevia powder based milk shakes. Mean GI value of milk shake prepared using sucralose and Stevia powder were comparable in normal subjects. Blood glucose response to all types of milk shakes in normal subjects in comparison to glucose load of 25 g is presented in Fig

89 Mg/dl Time (Min.) Fig. 4.1: Blood glucose curve of milk shake in normal subjects The peak rise in blood glucose curve occurred at 30 minutes after the consumption of glucose as well as control, sucralose and Stevia powder based milk shakes. After the ingestion of glucose load blood glucose was 162 mg/dl which was higher than the peak rise in blood glucose with control milk shake (132 mg/dl). Whereas, after the consumption of sucralose and Stevia powder based milk shakes blood glucose was found to be 118 and 112 mg/dl, respectively. Stevia powder based milk shake the blood glucose dropped to 83 mg/dl at 90 min to the baseline and were found to maintain the plateau in normal subjects with very less fluctuations in blood glucose level. Whereas consumption of glucose load, control and sucralose based milk shake continuously resulted in dropping of blood glucose level at 120 min Glycaemic index of mix shake in obese subjects. Data in respect to area under blood glucose response curve to all types of milk shake in obese subjects in comparison to glucose load of 25 g is presented in Table Table 4.47 : Area under blood glucose curve (mg/dl) of milk shake prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean

90 The mean area under blood glucose response curve for obese was found to be 1988, 1340 and 1235 mg/dl in case of milk shake prepared using sugar (control), sucralose and Stevia powder, respectively in obese subjects. Mean GI of control, sucralose and Stevia powder based milk shake in obese subjects were observed to be 52.80, and 32.80, respectively. The glycaemic index of milk shake prepared using sugar was found to have maximum glycaemic index followed by sucralose and Stevia powder based milk shake. Whereas, the mean value of glycaemic index of milk shake prepared using sucralose and Stevia powder showed slight difference in obese subjects. Blood glucose curve of obese subjects in response to milk shake in comparison to glucose load of 25 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.2 : Blood glucose response curve for milk shake in obese subjects The peak rise in blood glucose curve occurred at 30 minutes after the ingestion of glucose as well as control, sucralose and Stevia powder based milk shakes. After the feeding of glucose load blood glucose was 184 mg/dl which was higher than the peak rise in blood glucose after ingestion of control milk shake (158 mg/dl). Whereas consumption of sucralose and Stevia powder resulted in the rise of blood glucose upto 128 and 110 mg/dl, respectively. After the maximum peak in blood glucose was observed in obese subjects, blood glucose level dropped at 60 and 90 min and reached the baseline value at 120 min for sugar and sucralose in obese subjects. Stevia based milk shake the blood glucose dropped to 97 mg/dl at 60 min to the baseline and were found to maintain the plateau in obese subjects with very less fluctuations in blood glucose level. Whereas ingestion of glucose load, control and sucralose based milk shake continuous dropping in blood glucose level was observed till 120 min. 79

91 Glycaemic index of mix shake in diabetic subjects Data in respect to area under glucose curve with glucose load of 25 g and glycaemic index of control, sucralose and Stevia powder based milk shake for diabetic subjects are presented in table Table 4.48 : Area under blood glucose curve (mg/dl) of milk shake prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia G.I Mean Mean area under blood glucose response curve for glucose was 4653 mg/dl followed by control milk shake (2752 mg/dl) sucralose based milk shake (2293 mg/dl) and Stevia powder based milk shake (2190 mg/dl). Mean values of glycaemic index of control, sucralose and Stevia powder based milk shake were found to be 59.15, and 47.07, respectively. Glycaemic index value of Stevia powder based milk shake was found to be lower than the control and sucralose based milk shake in diabetic subjects. Blood glucose curve of diabetic subjects in response to milk shake in comparison to glucose load of 25 g exhibited in Fig Mg/dl Time (Min.) Fig 4.3 : Blood glucose response curve for milk shake in diabetic subjects 80

92 The peak rise in blood glucose curve occurred at 30 minutes after the ingestion of glucose as well as control, sucralose and Stevia powder based milk shakes. After the consumption of glucose load blood glucose was found to be 210 mg/dl which was higher than the peak rise in blood glucose with control milk shake (170 mg/dl). Whereas consumption of sucralose and Stevia powder based milk shake resulted in the rise of blood glucose upto 154 and 148 mg/dl, respectively in diabetic subjects. After the maximum peak in blood glucose was observed in diabetic subjects, blood glucose level dropped after 60 and 90 min and reached the baseline value (137 and 133 mg min /100 ml) at 120 min for sugar and sucralose based milk shake. Consumption of Stevia powder based milk shake dropped the blood glucose to 130 mg/dl at 60 min and continue to maintain the plateau in diabetic subjects with very less fluctuations in blood glucose level. Whereas consumption of glucose load, control and sucralose based milk shake resulted in continuous dropping of blood glucose level till 120 min. Glycaemic index of milk shake in normal, obese and diabetic subjects A comparative data regarding glycaemic index of milk shake prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table Table 4.49 : Glycaemic Index of Milk shake in normal, obese and diabetic subjects Milk shakes Normal Obese Diabetic CD (P<0.05) Control (Sugar) ± ± ± Sucralose 34.35± ± ± Stevia powder 33.06± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations Glycaemic index of milk shake prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 48.35, and 33.06, respectively. Milk shake prepared using sugar had significantly higher (48.35) GI value, whereas milk shake prepared using sucralose and Stevia powder did not vary significantly in normal subjects. Similarly, in case of obese and diabetic subjects, GI values of milk shakes based on sucralose and Stevia powder did not vary significantly. However, GI values of milk shake (control) in obese (52.80) and in diabetic (59.15 ) subjects differed significantly, whereas GI values of milk shake based on sucralose and Stevia powder differed non-significantly in normal and obese subjects but differed significantly in normal and diabetic subjects. GI values of all the three 81

93 types of milk shakes were found higher in diabetic subjects followed by obese and normal subjects RTS Glycaemic index of RTS in normal subjects Data in respect to area under blood glucose response curve to all types of RTS in normal subjects in comparison to glucose load of 25 g is presented in Table 4.50 Table 4.50 : Area under blood glucose curve (mg/dl) of RTS prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose Sugar GI Sucralose GI Stevia GI (Control) Mean The mean area under blood glucose response curve for glucose was 3688 mg/dl followed by control RTS (1378 mg/dl), sucralose based RTS (1029 mg/dl) and Stevia powder (947 mg/dl) based RTS. The area under curve depicted that glycaemic index was found to be in the range of to for control, to for sucralose based and to for Stevia powder based RTS in normal subjects. Blood glucose curve of normal subjects in response to RTS in comparison to glucose load of 25 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.4 : Blood glucose response curve for RTS in normal subjects 82

94 The peak rise in blood glucose curve occurred at 30 minutes after the consumption of glucose as well as control, sucralose and Stevia powder based RTS. Peak was found to be 114, 106 and 102 mg/dl for glucose, control, sucralose and Stevia powder RTS. After the ingestion of glucose load blood glucose was found to be 142 mg/dl, which was higher than the peak rise in blood glucose in comparison to control milk shake. After the maximum peak in blood glucose was observed in normal subjects, blood glucose level dropped at 60 and 90 min and reached the baseline value at 120 min. for sugar and sucralose in normal subjects. After the consumption of milk shake (Stevia powder) the blood glucose dropped to 97 mg/dl and 93 mg min /100 ml at 60 and 90 min to the baseline and was found to maintain the plateau at 120 min in normal subjects. Whereas, glucose, control and sucralose based RTS at 120 min resulted in continuous dropping of blood glucose level Glycaemic index of RTS in obese subjects Data in respect to area under blood glucose response curve to all types of RTS in obese subjects in comparison to glucose load of 25 g is presented in Table 4.51 Table 4.51: Area under blood glucose curve (mg/dl) of milk shake prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean The table exhibited the comparison in area under curve on ingesting RTS prepared using sugar, sucralose and Stevia powder where RTS prepared using sugar was served as a control for obese subjects. The mean for area under curve for glucose was found to be 3766 mg min /100ml. The data in respect to area under glucose curve for RTS was found to be 83

95 maximum for sugar based RTS i.e mg/dl and Stevia powder based RTS i.e 812 mg/dl ml the minimum. The area under curve for RTS prepared using sucralose was found to be in the range of 818 to 1362 mg min / 100 ml for obese subjects. The mean glycaemic index for contol, sucralose and Stevia powder based RTS in obese subjects were found to be 38.43, and 21.56, respectively. Blood glucose curve of obese subjects in response to RTS in comparison to glucose load of 25 g exhibited in Fig Mg/dl Time (Min.) Fig 4.5. Blood glucose response curve for RTS in obese subject The peak rise in blood glucose curve occurred at 30 minutes after the consumption of glucose as well as control, sucralose and Stevia powder based RTS. After the ingestion of glucose load blood glucose was found to be 144 mg/dl which was higher than the peak rise in blood glucose with control RTS (118 mg/dl) whereas, ingestion of sucralose and Stevia powder resulted in the rise of blood glucose to 104 and 100 mg/dl, respectively. After the maximum peak in blood glucose was observed in obese subjects, blood glucose level dropped after 60 and 90 min and reached the baseline value after 120 min for glucose, sugar and sucralose in obese subjects. Stevia based RTS the blood glucose dropped to 92 mg/dl after 60 min to the baseline and were found to maintain same blood glucose level in obese subjects. Whereas consumption of glucose load and control RTS showed a sharp fall from 60 to 90 min after consumption i.e 128 to 119, 110 to 104 mg/dl, respectively Glycaemic index of RTS in diabetic subjects Data in respect to area under blood glucose response curve to all types of RTS in diabetic subjects in comparison to glucose load of 25 g is presented in Table

96 Table 4.52 : Area under blood glucose curve (mg/dl) of RTS prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean Area under blood glucose curve was found to be between the range of 4176 to 5208 mg/dl. The mean value for area under curve for RTS prepared using sugar, sucralose and Stevia powder were observed to be 2205, 1764 and 1613 mg/dl, respectively for diabetic patients. The highest mean glycaemic index was found to be for RTS prepared using sugar (49.53) followed by RTS prepared using sucralose (41.80). Stevia powder based RTS (35.88) were found to be the lowest. Blood glucose curve of diabetic subjects in response to RTS in comparison to glucose load of 25 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.6 : Blood glucose response curve for RTS in diabetic subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based RTS. After the consumption of glucose load blood glucose rised to 210 mg/dl which was higher than the peak rise in blood glucose after ingesting control RTS (173 mg/dl) whereas, consumption of sucralose and 85

97 Stevia powder based RTS resulted in the rise of blood glucose to 161 and 154 mg/dl, respectively after 30 min of the ingestion of RTS. Blood glucose level after the consumption of sucralose and Stevia powder based RTS dropped after 60 and 90 min and reached the baseline value (134 and 139 mg/dl) at 120 min for sugar and sucralose based RTS in diabetic subjects. Stevia based RTS the blood glucose was found to be 151, 145 and 139 mg/dl at 60, 90 min and 120 min, respectively. Consumption of glucose load, control, sucralose and Stevia powder based RTS resulted in continuous dropping of blood glucose level till 120 min. Glycaemic index of RTS in normal, obese and diabetic subjects A comparative data regarding glycaemic index of RTS prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table Table 4.53 : Glycaemic index of RTS in normal, obese and diabetic subjects RTS Normal Obese Diabetics CD(P<0.5) Control ± ± ± Sucralose 27.91± ± ± Stevia powder 25.68± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations Glycaemic index of RTS prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 37.38, and 25.68, respectively. RTS prepared using sugar had significantly higher (37.38) GI value, whereas RTS prepared using sucralose and Stevia powder did not vary significantly in normal subjects. Similarly, in case of obese and diabetic subjects, GI values of RTS based on sucralose and Stevia powder did not vary significantly. However, GI values of RTS (control) in obese (38.43) and in diabetic (47.38) subjects differed significantly, whereas GI values of RTS based on sucralose and Stevia powder differed non-significantly in normal subjects but differed significantly in obese and diabetic subjects. GI values of all the three types of RTS were found higher in diabetic subjects followed by obese and normal subjects Glycaemic response of baked products in normal, obese and diabetic subjects Biscuits Glycaemic index of biscuits in normal subjects Data in respect to area under blood glucose response curve to all types of biscuits in normal subjects in comparison to glucose load of 50 g is presented in Table

98 Table 4.54 : Area under blood glucose response curve (mg/dl) of biscuits prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean Mean area under blood glucose curve for glucose was found to 5810 mg/dl followed by control 3775 mg/dl, sucralose based biscuits 3100 mg/dl and Stevia powder based biscuits exhibited lowest area under blood glucose curve i.e mg/dl. The area under curve ranged between 2587 to 3886 mg min /100ml, 2669 to 3514 mg/dl and 2818 to 4577 mg/dl for Stevia powder based, sucralose based and control biscuits in normal subjects. Blood glucose curve of normal subjects in response to biscuits in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.7 : Blood glucose response curve for biscuits in normal subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based biscuits. After the consumption 87

99 of glucose load blood glucose raised to 178 mg/dl which was higher than the peak rise in blood glucose with control biscuits (142 mg/dl). Whereas, consumption of sucralose and Stevia powder based biscuits resulted in the rise of blood glucose to 128 and 120 mg min /100 ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based biscuits dropped at 60 (113 and 109 mg/dl) and 90 min (105 and 97 mg/dl), respectively. Almost similar baseline value (89 and 91 mg min /100 ml) was observed for sucralose based biscuits and Stevia powder based biscuits in normal subjects. After consuming Stevia based biscuits the blood glucose was found to be 109, 97 and 91 mg/dl after 60, 90 min and at 120 min, respectively. Consumption of glucose load, control and sucralose based biscuits resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of Stevia powder based biscuits showed a slight decline in blood glucose level at 60 min and reached to the baseline at 90 and 120 min Glycaemic index of biscuits in obese subjects Data in respect to area under glucose curve and glycaemic index of biscuits incorporating sucralose and Stevia powder in comparison to glucose load of 50 g in obese subjects are presented in table Table 4.55 : Area under blood glucose response curve (mg/dl) of biscuits prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean Area under blood glucose response curve in obese subjects ranged between 2510 to 4859, 2462 to 4400 and 2457 to 3647 mg/dl for control, sucralose and Stevia powder based biscuits, respectively. The mean values of glycaemic index of control, sucralose and Stevia powder based biscuits were observed as 68.09, and 51.31, respectively. The glycaemic 88

100 index of sucralose and Stevia powder based biscuits had lower glycaemic index in comparison to control in obese subjects. Blood glucose curve of normal subjects in response to biscuits in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.8 : Blood glucose response curve for biscuits in obese subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based biscuits, peak was found to be at 30 minutes. After the consumption of glucose load blood glucose raised to 183 mg/dl which was higher than the peak rise in blood glucose with control biscuits (142 mg/dl). Whereas, consumption of sucralose and Stevia powder based biscuits resulted in the rise of blood glucose to 130 and 126 mg min /100 ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based biscuits dropped at 60 (121 and 110 mg/dl) and 90 min (96 and 99 mg/dl), respectively. Consumption of sucralose and Stevia powder based biscuits resulted in rise of blood glucose level to 89 and 93 mg min /100 ml) in obese subjects. After consumption of Stevia powder based biscuits the blood glucose was found to be 110, 99 and 93 mg/dl at 60, 90 min and at 120 min, respectively. Consumption of glucose load, control and sucralose based biscuits resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of Stevia powder based biscuits showed a slight decline in blood glucose level at 60, 90 and 120 min Glycaemic index of biscuits in diabetic subjects Data in respect to area under glucose curve in comparison to glucose load of 50 g and glycaemic index of control, sucralose and Stevia powder based biscuits are depicted in Table

101 Table 4.56 : Area under blood glucose response curve (mg/dl ) of biscuits prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean The mean area under blood glucose response curve for glucose was mg/dl followed by control biscuits (5604 mg/dl) sucralose based biscuits (3974 mg/dl) and Stevia powder based biscuits (3798 mg/dl). The mean values of glycaemic index of control, sucralose and Stevia powder based biscuits were found to be 78.80, and respectively. The glycaemic index value of Stevia powder based biscuits was found to be lower than the control and sucralose based biscuits in diabetic subjects. Whereas sucralose based biscuits exhibited higher glycaemic index than Stevia powder based biscuits but exhibited lower glycaemic index than control biscuits in diabetic subjects. Blood glucose curve of diabetic subjects in response to biscuits in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.9 : Blood glucose response curve for biscuits in diabetic subjects 90

102 The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based biscuits, peak was found to be at 30 minutes. After the feeding of glucose load blood glucose raised to 296 mg/dl which was higher than the peak rise in blood glucose with control biscuits (219 mg/dl) whereas, consumption of sucralose and Stevia powder based biscuits resulted in the rise of blood glucose level to 197 and 185 mg/dl, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based biscuits dropped at 60 (156 and162 mg/dl).whereas sucralose based biscuit raised the blood glucose level again i.e 162 mg/dl and Stevia powder based biscuit decreased the blood glucose level i.e 151 mg/dl at 90 min and reached the baseline value at 120 min for control, sucralose and Stevia powder based biscuits in diabetic subjects. After consumption of Stevia based biscuits the blood glucose was found to be 162, 151 and 139 mg/dl at 60, 90 min and to the baseline, respectively. Consumption of glucose load, control, sucralose and Stevia powder based biscuits resulted in continuous dropping of blood glucose level till 120 min. Glycaemic Index of biscuits in normal, obese and diabetic subjects A comparative data regarding glycaemic index of biscuits prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table Table 4.57 : Glycaemic index of biscuits in normal, obese and diabetic subjects Normal Obese Diabetics CD(P<0.5) Control 64.99± ± ± Sucralose 53.37± ± ± Stevia powder 49.19± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations Glycaemic index of biscuits prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 64.99, and 49.19, respectively. Biscuits prepared using sugar had significantly higher (64.99) GI value, whereas biscuits prepared using sucralose and Stevia powder did not vary significantly in diabetic subjects. Whereas, in normal and obese subjects, GI values of biscuits based on sucralose and Stevia powder vary significantly. However, GI values of biscuits (control) in obese (68.09) and in diabetic (78.80) subjects differed significantly. GI values of biscuits based on sucralose and Stevia powder differed significantly in normal subjects as well as obese subjects. GI values of all the three types of biscuits were found higher in diabetic subjects followed by obese and normal subjects. 91

103 Cake Glycaemic index of cake in normal subjects Data in respect to area under glucose curve in comparison to glucose load of 50 g and glycaemic index of control, sucralose and Stevia powder cake are depicted in Table Table 4.58: Area under blood glucose curve (mg/dl) of cake prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean Mean area under blood glucose curve for glucose was 5810 mg/dl followed by control 3490 mg/dl, sucralose based cake 2868 mg/dl and Stevia powder based cake exhibited lowest area under blood glucose curve i.e mg/dl. The area under curve ranged between 1987 to 3396 mg min /100ml, 2043 to 3804 mg/dl and 2943 to 4048 mg/dl for Stevia powder based, sucralose based and control cake in normal subjects. The glycaemic index of control, sucralose and Stevia powder based cake was reported to be 60.07, and 47.25, respectively. The glycaemic index of Stevia powder based cake was found to be lower than the control and sucralose based cake in normal subjects. Whereas sucralose based cake exhibited higher glycaemic index than Stevia powder based cake but exhibited lower glycaemic index than control cake in normal subjects. Blood glucose curve of normal subjects in response to cake in comparison to glucose load of 50 g exhibited in Fig

104 Mg/dl Time (Min.) Fig. 4.10: Blood glucose response curve for cake in normal subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control cake. Whereas maximum peak in sucralose and Stevia powder based cake was observed to be at 60 min. After the consumption of glucose load blood glucose raised to 178 mg/dl which was higher than the peak rise in blood glucose with control cake (141 mg/dl). Whereas consumption of sucralose and Stevia powder based cake resulted in the rise of blood glucose to 120 and 113 mg min /100 ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based cake raised at 60 (123 and 115 mg/dl) and dropped slightly at 90 min (101 and 95 mg/dl), respectively. Almost similar baseline value (89 and 92 mg min /100ml) was observed for sucralose based and Stevia powder based cake in normal subjects. After consumption of Stevia powder based cake the blood glucose was found to be 115, 95 and 92 mg/dl at 60, 90 min and at 120 min, respectively. Consumption of glucose load and control cake resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of sucralose and Stevia powder based cake showed a rise at 60 min and slow decline at 90 min. Stevia powder based cake resulted in reduction in blood glucose level at 90 and 120 min where blood glucose level reached to the baseline Glycaemic index of cake in obese subjects Data in respect to area under glucose curve in comparison to glucose load of 50 g and glycaemic index of control, sucralose and Stevia powder based cake are depicted in Table

105 Table 4.59 : Area under blood glucose response curve (mg/dl) of cake prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Control GI Sucralose GI Stevia G.I Mean Values are mean ± SE of ten independent determinations Area under blood glucose response curve in obese subjects ranged between 3008 to 4742, 2385 to 3960 and 2358 to 3598 mg/dl for control, sucralose and Stevia powder based cake, respectively. The mean values of glycaemic index of control, sucralose and Stevia powder based cake were observed as 63.10, and 48.12, respectively. The glycaemic index of sucralose and Stevia powder based cake had lower glycaemic index in comparison to control in obese subjects. Blood glucose curve of obese subjects in response to cake in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.11: Blood glucose response curve for cake in obese subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control cake. After the consumption of glucose load blood glucose raised to 183 mg/dl which was higher than the peak raised in blood glucose level by control cake 94

106 (148 mg/dl). Whereas consumption of sucralose and Stevia powder based cake raised the blood glucose to 117 and 113 mg/ dl, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based cake was found to be 121and 115 mg/dl at 60 min and 96 and 101 mg/dl at 90 min, respectively. Consumption of sucralose and Stevia powder based cake resulted in 89 and 95 mg min /100 ml blood glucose level at 120 min in obese subjects. After consumption of Stevia powder based and sucralose based cake the blood glucose level raised at 60 min with a slow drop after 90 and 120 min, respectively. Whereas, consumption of glucose load and control cake exhibited a sharp increase after 30 min with a sharp decrease of blood glucose at 60 and slow drop at 90 and 120 min Glycaemic index of cake in diabetic subjects Data in respect to area under glucose curve in comparison to glucose load of 50 g and glycaemic index of control, sucralose and Stevia powder based cake in diabetic subjects are depicted in Table Table 4.60 : Area under blood glucose response curve (mg/ dl) of cake prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean The mean area under blood glucose response curve for glucose was 7112 mg/dl followed by control cake (5128 mg/dl) sucralose based cake (4145 mg/dl) and Stevia powder based cake (3894 mg/dl). The mean values of glycaemic index of control, sucralose and Stevia powder cake were found to be 72.10, and 54.75, respectively in diabetic subjects. The glycaemic index of Stevia powder based cake was found to be lower than the control and sucralose based cake in diabetic subjects. Whereas sucralose based cake exhibited higher glycaemic index than Stevia powder based cake but exhibited lower glycaemic index than control cake in diabetic subject. Blood glucose curve of diabetic subjects in response to cake in comparison to glucose load of 50 g exhibited in Fig

107 Mg/dl Time (Min.) Fig. 4.12: Blood glucose response curve for cake in diabetic subjects The peak rise in blood glucose curve occurred after 30 min after the consumption of glucose as well as control cake, whereas sucralose and Stevia powder based cake, peak was found to be at 60 minutes. After the feeding of glucose load blood glucose raised to 296 mg/dl which was higher than the peak rise in blood glucose with control cake (256 mg/dl) at 30 min. Whereas, consumption of sucralose and Stevia powder based cake resulted in the rise of blood glucose level to 189 and 165 mg/dl, respectively at 30 min. Blood glucose level after the consumption of sucralose and Stevia powder based cake raised at 60 min which was found to be 182 and 187 mg/dl. Blood glucose level dropped slightly at 90 min in Stevia powder based cake whereas increased slightly in case of sucralose based cake. Blood glucose level after consumption of sucralose and Stevia powder based cake after 90 and 120 min was found to be 192 and 181 and 157 and 173 mg/dl, respectively. After consumption of sucralose based cake the blood glucose was found to be 182, 192 and 157 mg/dl at 60, 90 min and 120 min, respectively. Data depicted states that although sucralose based cake dropped the blood glucose level but it does not protect diabetic subjects with fluctuations in blood glucose level whereas Stevia powder based cake was observed to be on the safer side. A comparative data regarding glycaemic index of cake prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table Table 4.61 : Glycaemic index of cake in normal, obese and diabetic subjects Cake Normal Obese Diabetics CD(P<0.5) Control ± ± ± Sucralose 49.37± ± ± Stevia powder 47.25± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations 96

108 Glycaemic index of cake prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 60.07, and 47.25, respectively. Cake prepared using sugar had significantly higher (60.07) GI value, whereas cake prepared using sucralose and Stevia powder did not vary significantly in normal, obese and diabetic subjects. However, GI values of cake (control) in obese (63.10) and in diabetic (72.10) subjects differed significantly. GI values of cake based on sucralose and Stevia powder differed significantly in normal, obese as well as diabetic subjects. GI values of all the three types of cake were found higher in diabetic subjects followed by obese and normal subjects Bun Glycaemic index of bun in normal subjects Data in respect to area under glucose curve in comparison to glucose load of 50 g and glycaemic index of control, sucralose and Stevia powder based bun in normal subjects are depicted in Table Table 4.62 : Area under blood glucose response curve (mg/dl) of bun prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose (Sugar) Control GI Sucralose GI Stevia GI Mean Mean area under blood glucose curve for glucose was 5810 mg/dl followed by control bun 3721 mg/dl, sucralose based bun 3003 mg/dl and Stevia powder based bun exhibited lowest area under curve i.e mg/dl. The area under curve ranged between 2124 to 4132 mg min /100 ml, 2167 to 4347 mg/dl and 3018 to 4856 mg/dl for Stevia powder based, sucralose based and control bun in normal subjects. Blood glucose curve of normal subjects in response to bun in comparison to glucose load of 50 g exhibited in Fig

109 Mg/dl Fig : Blood glucose response curve for bun in normal subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose, control, sucralose and Stevia powder based bun. After the consumption of glucose load blood glucose raised to 178 mg/dl which was higher than the peak rise in blood glucose with control bun (158 mg/dl) whereas, consumption of sucralose and Stevia powder based bun resulted in the rise of blood glucose to 146 and 134 mg min /100 ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based bun decreased (110 and 117 mg/dl) followed by further dropping in sucralose and Stevia powder based bun at 90 min (105 and 97 mg/dl), respectively. Lower baseline value (89 and 94 mg min /100 ml) was observed for sucralose based and Stevia powder based buns followed by control bun (99 mg/dl) in normal subjects. Consumption of glucose load, control and sucralose bun resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of Stevia powder based bun showed a rise at 30 min and slow decline at 60, 90 and 120 min and finally reaching baseline at 90 and 120 min in normal subjects Glycaemic index of bun in obese subjects Data in respect to area under glucose curve in comparison to glucose load of 50 g and glycaemic index of control, sucralose and Stevia powder based bun in obese subjects are depicted in Table Time (Min.) Table 4.63 : Area under blood glucose response curve (mg/dl) of bun prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Sugar GI Sucralose GI Stevia GI (Control) Mean

110 Area under blood glucose response curve for control, sucralose and Stevia powder based bun in obese subjects were found to be 4141, 3247 and 3049, respectively. The mean values of glycaemic index of control, sucralose and Stevia powder based bun were observed as 69.24, and 50.99, respectively. The glycaemic index of sucralose and Stevia powder based bun had lower glycaemic index in comparison to control in obese subjects. Blood glucose curve of obese subjects in response to bun in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.14: Blood glucose response curve for bun in obese subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose, control, sucralose and Stevia powder based buns. After the consumption of glucose load blood glucose raised to 183 mg/dl which was higher than the peak rise in blood glucose with control buns (163 mg/dl). Whereas, consumption of sucralose and Stevia powder based buns resulted in the rise of blood glucose to 151 and 137 mg min /100 ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based buns dropped at 60 (124 and 118 mg/dl) and 90 min (101 and 111 mg/dl), respectively. After 120 min of consumption of sucralose and Stevia powder based buns blood glucose level twas found to be 93 and 99 mg min /100ml) in obese subjects. After consumption of Stevia based buns the blood glucose was found to be 118, 111 and 99 mg/dl at 60, 90 min and at 120 min, respectively. Consumption of glucose load, control and sucralose based buns resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of Stevia powder based buns showed a slight and steady decline in blood glucose level at 60, 90 and 120 min Glycaemic index of bun in diabetic subjects Data in respect to area under glucose curve and glycaemic index of control, sucralose and Stevia powder based bun in comparison to glucose load of 50 g in diabetic subjects are depicted in Table

111 Table 4.64 : Area under blood glucose response curve (mg/dl) of bun prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean The mean area under blood glucose response curve for glucose was 7112 mg/dl followed by control bun (6003 mg/dl) sucralose bun (4748 mg/dl) and Stevia powder based bun (4322 mg/dl). The mean values of glycaemic index of control, sucralose and Stevia powder based bun were found to be 84.41, and respectively. The glycaemic index of Stevia powder based bun was found to be lower than the control and sucralose based buns in diabetic subjects. Whereas sucralose based buns exhibited higher glycaemic index than Stevia powder based bun but lower than control buns in diabetic subjects. Blood glucose curve of diabetic subjects in response to buns in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig : Blood glucose response curve for bun in diabetic subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based bun. After the ingestion of glucose load blood glucose raised to 296 mg/dl which was higher than the peak rise in blood 100

112 glucose with control buns (256 mg/dl) whereas, consumption of sucralose and Stevia powder resulted in the rise of blood glucose level to 225 and 200 mg/dl, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based buns dropped at 60 (174 and 165 mg/dl). Control, sucralose and Stevia powder based buns in diabetic subjects the blood glucose was observed to be 203, 182 and 155 mg/dl, respectively, after the consumption of buns at 90 min. and reached the baseline value at 120 min. Consumption of glucose load, control, sucralose based buns resulted in sharp and continuous dropping of blood glucose level whereas Stevia powder based buns was found to have a slow and steady effect on blood glucose level of the diabetic subjects till 120 min. Glycaemic index of bun in normal, obese and diabetic subjects A comparative data regarding glycaemic index of bun prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table Table 4.65 : Glycaemic index of bun in normal, obese and diabetic subjects Types of Buns Normal Obese Diabetics CD(P<0.5) Sugar (Control) ± ± ± Sucralose 51.70± ± ± Stevia powder 49.08± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations Glycaemic index of bun prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 64.05, and 49.08, respectively. Bun prepared using sugar had significantly higher (64.05) GI value, whereas bun prepared using sucralose and Stevia powder did not vary significantly in normal whereas, observed to decrease the glycaemic index significantly in obese and diabetic subjects. However, GI values of bun (control) in obese (69.24) and in diabetic (84.41) subjects differed significantly. GI values of buns based on sucralose and Stevia powder differed significantly in obese as well as diabetic subjects. Bun prepared using Stevia powder showed a non-significant difference in GI of normal subjects when compared to GI of obese subjects. GI values of all the three types of buns were found higher in diabetic subjects followed by obese and normal subjects Glycaemic response of traditional products in normal, obese and diabetic subjects Sweet porridge Glycaemic index of sweet porridge in normal subjects Data in respect to area under glucose curve and glycaemic index of control, sucralose and Stevia powder based sweet porridge in comparison to glucose load of 50 g in normal subjects are depicted in Table

113 Table 4.66 : Area under blood glucose response curve (mg/dl) of sweet porridge prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean Mean area under blood glucose curve for sweet porridge in normal subjects for glucose was 5810 mg/dl followed by control sweet porridge 3557 mg/dl, sucralose based sweet porridge 2893 mg/dl and Stevia powder based sweet porridge exhibited lowest area under curve i.e mg/dl. The area under curve ranged between 1806 to 3698 mg/dl, 2282 to 3437 mg/dl and 3240 to 3942 mg/dl for Stevia powder based, sucralose based and control sweet porridge in normal subjects. The glycaemic index value for control, sucralose and Stevia powder based sweet porridge was found to be 62.52, and 47.12, respectively. Blood glucose curve of normal subjects in response to sweet porridge in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig : Blood glucose response curve for sweet porridge in normal subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose, control, sucralose and Stevia powder based sweet porridge. After the consumption of glucose load blood glucose raised to 178 mg/dl which was higher than the peak rise in blood glucose with control sweet porridge (145 mg/dl). Whereas, consumption of sucralose and 102

114 Stevia powder based sweet porridge resulted in the rise of blood glucose to 124 and 117 mg min /100 ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based sweet porridge at 60 min were found to be 110 and 107 mg/dl and continue to drop in case of sucralose and Stevia powder based sweet porridge at 90 min (95 and 99 mg/dl), respectively. Lower baseline value (93 and 96 mg min /100 ml) was observed for sucralose based and Stevia powder based sweet porridge followed by control sweet porridge (99 mg/dl) in normal subjects. Consumption of glucose load, and control sweet porridge resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of sucralose and Stevia powder based sweet porridge showed a rise at 30 min and slow decline at 60, 90 and 120 min and finally reaching baseline at 90 and 120 min in normal subjects Glycaemic index of sweet porridge Data in respect to area under glucose curve and glycaemic index of control, sucralose and Stevia powder based sweet porridge in comparison to glucose load of 50 g in obese subjects are depicted in Table Table 4.67: Area under blood glucose response curve (mg/dl) of sweet porridge prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean Area under blood glucose response curve for control, sucralose and Stevia powder based sweet porridge in obese subjects were found to be 3902, 3084 and 2935 mg/dl, respectively. The mean values of glycaemic index of control, sucralose and Stevia powder based sweet porridge were observed as 65.24, and 49.07, respectively. The glycaemic index of sucralose and Stevia powder based sweet porridge had lower glycaemic index in comparison to control in obese subjects. Blood glucose curve of obese subjects in response to sweet porridge in comparison to glucose load of 50 g exhibited in Fig

115 Mg/dl Time (Min.) Fig : Blood glucose response curve for sweet porridge in obese subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose, control, sucralose and Stevia powder based sweet porridge. After the consumption of glucose load blood glucose raised to 183 mg/dl which was higher than the peak rise in blood glucose with control sweet porridge (150 mg/dl). Whereas, consumption of sucralose and Stevia powder based sweet porridge resulted in the rise of blood glucose to 138 and 126 mg min /100ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based sweet porridge dropped at 60 min (113 and 109 mg/dl) and 90 min (96 and 93 mg/dl), respectively. After 120 min of consumption of sucralose and Stevia powder based sweet porridge blood glucose level was found to be 93 and 99 mg min /100 ml) in obese subjects. After consumption of Stevia based sweet porridge the blood glucose was found to be 109, 93 and 99 mg/dl at 60, 90 min and at 120 min, respectively. Consumption of glucose load, control and sucralose based sweet porridge resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of Stevia powder based sweet porridge showed a slight and steady decline and then rise in blood glucose level at 60, 90 and 120 min, to maintain the blood glucose level of the subject to maintain a steady state of the subject Glycaemic index of sweet porridge in diabetic subjects Data in respect to area under glucose curve and glycaemic index of sweet porridge incorporating in comparison to glucose load of 50 g in diabetic subjects are presented in Table

116 Table 4.68 : Area under blood glucose response curve (mg/dl) and of sweet porridge prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean The mean area under blood glucose response curve for glucose was 7112 mg/dl followed by control sweet porridge (5233 mg/dl) sucralose based sweet porridge (4357 mg/dl) and Stevia powder based sweet porridge (3554 mg/dl). The mean values of glycaemic index of control, sucralose and Stevia powder based sweet porridge were found to be 73.57, and 55.85, respectively. The glycaemic index of Stevia powder based sweet porridge was found to be lower than the control and sucralose based sweet porridge in diabetic subjects. Whereas sucralose based sweet porridge exhibited higher glycaemic index than Stevia powder based sweet porridge but lower than control sweet porridge in diabetic subjects. Blood glucose curve of diabetic subjects in response to sweet porridge in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig : Blood glucose response curve for sweet porridge in diabetic subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based sweet porridge. After the 105

117 consumption of glucose load blood glucose raised to 296 mg/dl which was higher than the peak rise in blood glucose with control sweet porridge (238 mg/dl). Whereas, consumption of sucralose and Stevia powder sweet porridge resulted in the rise of blood glucose level to 198 and 184 mg/dl, respectively at 30 min. After the consumption of sucralose and Stevia powder based sweet porridge the blood glucose level dropped at 60 min (176 and163 mg/dl). Control, sucralose and Stevia powder based sweet porridge in diabetic subjects the blood glucose level was observed to be 209, 181 and 159 mg/dl, respectively, after the consumption of sweet porridge at 90 min. At 120 min, after the consumption of all types of sweet porridge blood glucose level reached the baseline value. Consumption of glucose load, control, sucralose based sweet porridge resulted in sharp and continuous dropping of blood glucose level whereas Stevia powder based sweet porridge was found to have a slow and steady effect on blood glucose level of the diabetic subjects. Glycaemic index of sweet porridge in normal, obese and diabetic subjects A comparative data regarding glycaemic index of sweet porridge prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table Table 4.69 : Glycaemic index of sweet porridge in normal, obese and diabetic subjects Normal Obese Diabetics CD(P<0.5) Control ± ± ± Sucralose 49.81± ± ± Stevia powder 47.12± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations Glycaemic index of sweet porridge prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 62.52, and 47.12, respectively. Sweet porridge prepared using sugar had significantly higher (62.52) GI value, whereas sweet porridge prepared using sucralose and Stevia powder did not vary significantly in normal whereas, observed to decrease the glycaemic index significantly in diabetic subjects. However, GI values of sweet porridge (control) in obese (65.24) and in diabetic (73.52) subjects differed significantly. GI values of sweet porridge based on sucralose and Stevia powder differed significantly in obese as well as diabetic subjects. Sweet porridge prepared using Stevia powder and sucralose showed a non-significant difference in GI of normal subjects when compared to GI of obese subjects. GI values of all the three types of sweet porridge were found higher in diabetic subjects followed by obese and normal subjects Carrot halwa Glycaemic index of carrot halwa in normal subjects Data in respect to area under glucose curve and glycaemic index of carrot halwa incorporating sucralose and Stevia powder in comparison to glucose load of 50 g in normal subjects are presented in Table

118 Table 4.70 : Area under blood glucose response curve (mg/dl) of carrot halwa prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose Sugar GI Sucralose GI Stevia GI (Control) Mean Mean area under blood glucose curve for carrot halwa in normal subjects for glucose was 5810 mg/dl followed by control carrot halwa 2971 mg/dl, sucralose based carrot halwa 2242 mg/dl and Stevia powder based carrot halwa exhibited lowest area under curve i.e mg/dl. The area under curve ranged between 1561 to 2972 mg/dl, 1821 to 2408 mg/dl and 2237 to 3684 mg/dl for Stevia powder based, sucralose based and control carrot halwa in normal subjects, respectively. The glycaemic index value for control, sucralose and Stevia powder based sweet carrot halwa was found to be 51.15, and 35.99, respectively. Blood glucose curve of normal subjects in response to carrot halwa in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.19: Blood glucose response curve for carrot halwa in normal subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose, control, sucralose and Stevia powder based carrot halwa. After the consumption of glucose load blood glucose raised to 178 mg/dl which was higher than the peak rise in blood glucose with control carrot halwa (134 mg/dl). Whereas, consumption of sucralose and Stevia 107

119 powder based carrot halwa resulted in the rise of blood glucose to 117 and 112 mg/dl, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based carrot halwa at 60 min were found to be 120 and 117 mg/dl and continue to drop in case of sucralose and Stevia powder based carrot halwa at 90 min (107 and 103 mg/dl), respectively. Lower baseline value (93 and 98 mg/dl) was observed for sucralose based and Stevia powder based carrot halwa followed by control carrot halwa (101 mg/dl) in normal subjects. Consumption of glucose load, and control carrot halwa resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of sucralose and Stevia powder based carrot halwa showed a rise at 30 min and slow decline at 60, 90 and 120 min and finally reaching baseline at 90 and 120 min in normal subjects Glycaemic index of carrot halwa in obese subjects Data in respect to area under glucose curve and glycaemic index of carrot halwa incorporating sucralose and Stevia powder in comparison to glucose load of 50 g in obese subjects are presented in Table Table 4.71 : Area under blood glucose response curve (mg/dl) of carrot halwa prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Sugar GI Sucralose GI Stevia GI (Control) Mean Area under blood glucose response curve for control, sucralose and Stevia powder based carrot halwa in obese subjects were found to be 3330, 2472 and 2354 mg/dl, respectively. The mean values of glycaemic index of control, sucralose and Stevia powder based carrot halwa were observed to be 55.67, and 39.37, respectively. The glycaemic index of sucralose and Stevia powder based carrot halwa had lower glycaemic index in comparison to control in obese subjects. Blood glucose curve of obese subjects in response to carrot halwa in comparison to glucose load of 50 g exhibited in Fig

120 Mg/dl Fig. 4.20: Blood glucose response curve for carrot halwa in obese subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control carrot halwa. After the consumption of glucose load blood glucose raised to 183 mg/dl which was higher than the peak raised in blood glucose level by control carrot halwa (142 mg/dl). Whereas, consumption of sucralose and Stevia powder based carrot halwa raised the blood glucose to 118 and 109 mg min /100 ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based carrot halwa was found to be 124 and 120 mg/dl at 60 min and 96 and 103 mg/dl at 90 min, respectively. Consumption of sucralose and Stevia powder based carrot halwa resulted in 93 and 95 mg min /100ml blood glucose level at 120 min in obese subjects. After consumption of Stevia based and sucralose based carrot halwa the blood glucose level raised at 60 min with a slow drop at 90 and 120 min, respectively. Whereas, consumption of glucose load and control carrot halwa exhibited a sharp increase at 30 min with a sharp decrease of blood glucose after 60, and slow drop after 90 and 120 min Glycaemic index of carrot halwa in diabetic subjects Data in respect to area under glucose curve and glycaemic index of carrot halwa incorporating sucralose and Stevia powder in comparison to glucose load of 50 g in diabetic subjects are presented in Table Time (Min.) Table 4.72 : Area under blood glucose response curve (mg/dl) of carrot halwa prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Sugar GI Sucralose GI Stevia GI (Control) Mean

121 The mean area under blood glucose response curve for glucose was 7112 mg/dl followed by control carrot halwa (4941 mg/dl) sucralose based carrot halwa (3832 mg/dl) and Stevia powder based carrot halwa (3812 mg/dl). The mean values of glycaemic index of control, sucralose and Stevia powder carrot halwa were found to be 69.46, and 51.58, respectively. The glycaemic index of Stevia powder based carrot halwa was found to be lower than the control and sucralose based carrot halwa in diabetic subjects. Whereas sucralose based carrot halwa exhibited higher glycaemic index than Stevia powder based carrot halwa but lower than control carrot halwa in diabetic subjects. Blood glucose curve of diabetic subjects in response to carrot halwa in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig : Blood glucose response curve for carrot halwa in diabetic subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control carrot halwa, whereas sucralose and Stevia powder based carrot halwa, peak was found to be at 60 minutes.after the feeding of glucose load blood glucose raised to 296 mg/dl which was higher than the peak rise in blood glucose with control carrot halwa (210 mg/dl) at 30 min. Whereas, consumption of sucralose and Stevia powder based carrot halwa resulted in the rise of blood glucose level to 170 and 164 mg/dl, respectively at 30 min. Blood glucose level after the consumption of sucralose and Stevia powder based carrot halwa raised at 60 min which was found to be almost similar i. e. 173 and 172 mg/dl. Blood glucose level dropped slightly at 90 min in Stevia powder based carrot halwa whereas increased slightly in case of sucralose based carrot halwa. Blood glucose level after consumption of sucralose and Stevia powder based carrot halwa at 90 and 120 min was found to be 158 and 154 and 140 and 148 mg/dl, respectively. After consumption of sucralose based carrot halwa the blood glucose was found to be 173, 158 and 140 mg/dl at 60, 90 min and 120 min, respectively. Data depicted states that although sucralose drops the blood glucose level but it does not protect diabetic subjects with fluctuations in blood glucose level whereas Stevia powder based carrot halwa was observed to be on the safer side. 110

122 Glycaemic index of carrot halwa in normal, obese and diabetic subjects A comparative data regarding glycaemic index of carrot halwa prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table 4.73 Table 4.73 : Glycaemic index of carrot halwa in normal, obese and diabetic subjects Types of Halwa Normal Obese Diabetics CD(P<0.5) Control ± ± ± Sucralose 38.60± ± ± Stevia powder 35.99± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations Glycaemic index of carrot halwa prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 51.15, and 35.99, respectively. Carrot halwa prepared using sugar had significantly higher (51.15) GI value, whereas carrot halwa prepared using sucralose and Stevia powder did not vary significantly in normal, obese and diabetic subjects. However, GI values of carrot halwa (control) in obese (54.02) and in diabetic (69.46) subjects differed significantly. GI values of carrot halwa based on sucralose and Stevia powder differed significantly in obese as well as diabetic subjects. Carrot halwa prepared using Stevia powder and sucralose showed a non-significant difference in GI of normal subjects when compared to GI of obese subjects. GI values of all the three types of carrot halwa were found higher in diabetic subjects followed by obese and normal subjects Kheer Glycaemic index of kheer in normal subjects Data in respect to area under glucose curve and glycaemic index of kheer incorporating sucralose and Stevia powder in comparison to glucose load of 50 g in normal subjects are presented in Table Table 4.74 : Area under blood glucose response curve (mg/dl) kheer prepared using sugar, sucralose and Stevia powder in normal subjects S.No. Glucose Sugar GI Sucralose GI Stevia GI (Control) Mean

123 Mean area under blood glucose curve for kheer in normal subjects for glucose was 5810 mg/dl followed by control kheer 3548 mg/dl, sucralose based kheer 2671 mg/dl and Stevia powder based kheer exhibited lowest area under curve i.e mg/dl. The area under curve ranged between 2011 to 3384 mg min /100 ml, 2089 to 3345 mg/dl and 2698 to 4982 mg/dl for Stevia powder based, sucralose based and control kheer in normal subjects. The glycaemic index value for control, sucralose and Stevia powder based kheer was found to be 66.71, and 44.37, respectively Blood glucose curve of normal subjects in response to kheer in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.22: Blood glucose response curve for kheer in normal subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose, control, sucralose and Stevia powder based kheer. After the consumption of glucose load blood glucose raised to 178 mg/dl which was higher than the peak rise in blood glucose with control kheer (159 mg/dl). Whereas, consumption of sucralose and Stevia powder based kheer raised blood glucose to 134 and 122 mg min /100ml, respectively. Blood glucose level after the consumption of sucralose and Stevia powder based kheer after 60 min were found to be 120 and 114 mg/dl and continue to drop in case of sucralose and Stevia powder based kheer at 90 min (99 and 107 mg/dl), respectively. Lower baseline value (93 and 96 mg min /100 ml) was observed for sucralose based and Stevia powder based kheer followed by control kheer (99 mg/dl) in normal subjects. Consumption of glucose load, and control kheer resulted in sharp dropping of blood glucose level till 120 min. Whereas, consumption of sucralose based kheer showed a rise at 30 min and dropped slowly at 60, 90 and 120 min and finally reaching baseline at 90 and 120 min in normal subjects. 112

124 Glycaemic index of kheer in obese subjects Data in respect to area under glucose curve and glycaemic index of kheer incorporating sucralose and Stevia powder in comparison to glucose load of 50 g in obese subjects are presented in Table Table 4.75 : Area under blood glucose response curve (mg/dl) of kheer prepared using sugar, sucralose and Stevia powder in obese subjects S.No. Glucose Sugar (Control) GI Sucralose GI Stevia GI Mean Area under blood glucose response curve for control, sucralose and Stevia powder based kheer in obese subjects were found to be 4262, 3532 and 3082, respectively. The mean values of glycaemic index of control, sucralose and Stevia powder based kheer were observed to be 71.27, and 51.53, respectively. The glycaemic index of sucralose and Stevia powder based kheer had lower glycaemic index in comparison to control in obese subjects. Blood glucose curve of obese subjects in response to kheer in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig. 4.22: Blood glucose response curve for kheer in obese subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based kheer. After the feeding of 113

125 glucose load blood glucose raised to 183 mg/dl which was higher than the peak rise in blood glucose with control kheer (154 mg/dl) at 30 min. Whereas, consumption of sucralose and Stevia powder based kheer resulted in the rise of blood glucose level to 128 and 123 mg/dl, respectively at 30 min. Blood glucose level after the consumption of sucralose and Stevia powder based kheer dropped at 60 min which was found to be 107 and 103 mg/dl. Blood glucose level dropped slightly and reached baseline at 90 min in Stevia powder based and sucralose based kheer. Blood glucose level after consumption of sucralose and Stevia powder based kheer at 90 and 120 min was found to be 93 and 94 and 89 and 92 mg/dl, respectively. After consumption of sucralose based kheer the blood glucose was found to be 107, 93 and 89 mg/dl at 60, 90 min and 120 min, respectively. Data depicted states that both sucralose and Stevia based kheer raised blood glucose level increased and decreased sharply and reached baseline quickly Glycaemic index of kheer in diabetic subjects Data in respect to area under glucose curve and glycaemic index of kheer incorporating sucralose and Stevia powder in comparison to glucose load of 50 g in diabetic subjects are presented in Table Table 4.76 : Area under blood glucose response curve (mg/dl) of kheer prepared using sugar, sucralose and Stevia powder in diabetic subjects S.No. Glucose Control GI Sucralose GI Stevia G.I Mean The mean area under blood glucose response curve for glucose was 7112 mg/dl followed by control kheer (5652 mg/dl) sucralose kheer (4152 mg/dl) and Stevia powder based kheer (4043 mg/dl). The mean values of glycaemic index of control, sucralose and Stevia powder based kheer were found to be 79.47, and 56.85, respectively. The glycaemic index of Stevia powder based kheer was found to be lower than the control and sucralose based kheer in diabetic subjects. Whereas sucralose based kheer exhibited higher 114

126 glycaemic index than Stevia powder based kheer but lower than control kheer in diabetic subjects. Blood glucose curve of diabetic subjects in response to sweet porridge in comparison to glucose load of 50 g exhibited in Fig Mg/dl Time (Min.) Fig : Blood glucose response curve for kheer in diabetic subjects The peak rise in blood glucose curve occurred at 30 min after the consumption of glucose as well as control, sucralose and Stevia powder based kheer. After the consumption of glucose load blood glucose raised to 296 mg/dl which was higher than the peak rise in blood glucose with control kheer (261 mg/dl). Whereas, consumption of sucralose and Stevia powder based kheer resulted in the rise of blood glucose level to 213 and 183 mg/dl, respectively at 30 min. After the consumption of sucralose and Stevia powder based kheer the blood glucose level dropped after 60 min (183 and177 mg/dl). After the consumption of control, sucralose and Stevia powder based kheer the blood glucose level was observed to be 203, 167 and 154 mg/dl, respectively, at 90 min in diabetic subjects. After 120 min of consumption of all types of kheer blood glucose level was found to be 173, 137 and 139 mg/dl, respectively. Consumption of glucose load, control, sucralose and Stevia powder based kheer resulted in sharp and continuous dropping of blood glucose level in the diabetic subjects. Stevia powder based kheer after 120 min of consumption reached the baseline value in diabetic subjects. Glycaemic index of kheer in normal, obese and diabetic subjects A comparative data regarding glycaemic index of kheer prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects are presented in Table

127 Table 4.77 : Glycaemic index of kheer in normal, obese and diabetic subjects Normal Obese Diabetics CD(P<0.5) Control 66.71± ± ± Sucralose 47.22± ± ± Stevia powder 44.37± ± ± CD (P<0.05) Values are mean ± S.E of ten independent determinations Glycaemic index of kheer prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 66.71, and 44.37, respectively. Kheer prepared using sugar had significantly higher (66.71) GI value, whereas kheer prepared using sucralose and Stevia powder did not vary significantly in normal, obese and diabetic subjects. However, GI values of kheer (control) in obese (71.27) and in diabetic (79.47) subjects differed significantly. GI values of kheer based on sugar and Stevia powder differed significantly in obese as well as diabetic subjects. Kheer prepared using Stevia powder and sucralose showed a non-significant difference in GI of normal subjects when compared to GI of obese subjects. GI values of all the three types of kheer were found higher in diabetic subjects followed by obese and normal subjects. 4.5 Cost calculation of developed food products The data on the products prepared using sugar, sucralose and the products prepared using Stevia powder are depicted in Table Table 4.78 : Cost (Rs./100g) of the food products prepared using sugar, sucralose and acceptable level of Stevia powder S. No. Products Sugar (Control) Sucralose Stevia powder 1. Tea Coffee Milkshake RTS Biscuits Cake Buns Sweet porridge Carrot Halwa Kheer Sugar based products such as tea, coffee, milk shake, RTS, biscuits, cake, buns, sweet porridge, carrot halwa and kheer had higher cost as compared to the products prepared using Stevia powder and sucralose. 116

128 CHAPTER-V DISCUSSION S. rebaudiana leaves contain a zero-calorie diterpene glycosides (stevioside and the rebaudiosides) 300 times sweeter than sucrose with superior solubility in water and a positive taste profile that are safely metabolized by the body without any effect (Megeji et al., 2005; Geuns et al., 2007). Globally, scientists have concluded that Stevia sweeteners are safe for people of all ages. Stevia leaf or extracted forms like stevioside, rebaudioside A and steviol glycosides was approved by US FDA as a dietary supplement considered (Generally recognized as safe) rating in the US (GRAS Notification 287 for Steviol Glycosides with Rebaudioside A and Stevioside as Principal Components) as appears to have an adequate daily intake (ADI) of 25 mg/ kg (Genus et al. 2003). Stevia is a nutrient rich herb containing substantial amount of nutrients, like 80 to 85 per cent water, protein, fibre, aminoacids, free sugars, iminosugar Steviamine, lipids, essential oils, ascorbic acid, beta carotene, riboflavin, thiamine, austroinulin, chromium, cobalt, magnesium, iron, potassium, phosphorus and trace elements (Komissarenko et al., 1994; Choudhary and Bandyopadhyay, 1999; Konoshima and Takasaki, 2002; Sharma et al., 2006; Jayaraman et al., 2008; Esmat and Ferial, 2009; Hu et al., 2010). Based on nutritional benefits and therapeutic implications of Stevia powder, the present study was undertaken to develop various food products incorporating Stevia powder, sucralose (artificial sweetener) and sugar (control) and analysed for nutrient composition and glycaemic response in normal, obese and healthy adults. The results of present study are discussed under the following heads: 5.1 Nutritional composition of Stevia leaves and Stevia powder 5.2 Organoleptic evaluation of developed food products prepared using sugar, sucralose and Stevia powder 5.3 Nutritional composition of developed food products 5.4 Glycaemic response of developed food products 5.1 Nutritional composition of Stevia leaves and Stevia powder Stevia leaves contained significantly (p<0.05) higher (6.21%) moisture as compared to Stevia white powder (2.92 %). This might be due to the fact that leaves contain higher moisture content as compared to its clarified extract powder. Almost similar moisture content (7 %) in Stevia leaves was reported by Mishra et al. (2010), whereas Goyal et al. (2010) reported lower (4.65 %) moisture content. In contrary, Kaushik et al. (2010) reported higher moisture (7.7 %) content in Stevia leaves. Stevia leaves contained significantly higher (11.16 %) ash content as compared to Stevia white powder (3.24 %). This might be due to the fact that the Stevia white powder (extract of Stevia) being more refined, contains fewer of these phytonutrients. Bawane et al. 117

129 (2014) reported ash content in Stevia leaves which ranged from 8.20 to 10.2 per cent. Hence, the results of ash content in Stevia leaves are in agreement with the results reported by Bawane et al. (2014) in Stevia leaves. Similar ash content was also reported by Mishra et al. (2010) and Yadav and Guleria (2012) reported ash content i.e 13.1 per cent in Stevia leaves and 6.3 per cent ash in Stevia white powder. Stevia leaves contained significantly higher contents of crude fibre, fat, crude protein and total carbohydrates as compared to its clarified extract white powder. The results of present study are in agreement with those reported earlier proximate composition of Stevia leaves and powder (Tadhani and Subhash, 2006; Goyal et al., 2010; Atteh et al., 2011; Pande and Khetmalas, 2011). Total, reducing and non reducing sugars were found to be significantly higher in Stevia leaves (on dry weight basis) as compared to Stevia powder. The values of total, reducing and non reducing sugars were reported to be 15.63, 5.81 and 9.82 per cent and 1.78, 0.84 and 0.93 per cent, respectively in Stevia leaves and Stevia powder. Similarly, Abou-Arab et al. (2010) also reported almost similar total sugar, reducing and non reducing sugar contents i.e 15.65, 5.58 and 9.77 per cent, respectively in dried Stevia leaves, whereas 1.96, 0.86 and 1.10 per cent, respectively in Stevia powder. Whereas other workers reported lower sugar contents in Stevia leaves and Stevia powder (Pande and Khetmalas, 2011) as compared to the results obtained in the present study. Stevia leaves contained significantly higher amount of calcium, iron, sodium, magnesium and potassium in comparison to Stevia white powder. Lower amount of mineral content in Stevia powder might be due to the fact that Stevia white powder undergoes electrolytic techniques and ion exchange technique at the time of manufacturing. Slightly higher mineral contents in Stevia leaves powder have been reported by Serio (2010) and lower mineral contents have been reported by Goyal et al. (2010) and Mishra et al. (2010). Other workers also reported higher mineral contents in Stevia leaves as compared to Stevia powder (Serio, 2010; Atteh et al., 2011) 5.2 Organoleptic evaluation of developed food products prepared using sugar, sucralose and Stevia powder Beverages, baked and traditional products were prepared using sugar, sucralose and Stevia powder. In test recipes, Stevia powder was added in two levels in 100 g of developed recipes. Similarly, in another test recipe, sucralose (artificial sweetner) was used, whereas in basic recipe sugar (control) was added for comparison. The organoleptic evaluation was done to select the most acceptable level of Stevia powder in all the recipes and also to compare acceptability with sugar based recipe (control) and sucralose incorporated recipes. 118

130 5.2.1 Beverages The mean scores of sensory evaluation for colour, appearance, aroma, consistency, taste and overall acceptability of beverages such as tea, coffee, milk shake and RTS prepared using Stevia powder and sucralose showed no perceivable variation between the products. All the beverages with Stevia powder (T 1 ) i.e for tea 6 mg, for coffee 8 mg, for milk shake 12 mg and for RTS 10 mg were found acceptable by the panel members which ranked higher in terms of all sensory characteristics as compared to higher concentration of Stevia powder (T 2 ). Mean scores of colour, appearance, aroma, consistency and taste of acceptable level of Stevia based beverages found in the category of liked moderately. On increasing the concentration of Stevia powder caused significant reduction in mean scores of aroma and taste of tea, coffee, milk shake, and RTS and fell in the category of liked slightly by the judges. It might be due to bitter and prominent after taste. Singh et al. (2012) also developed low calorie RTS by replacing half of the sugar with Stevia powder without affecting the quality of beverages with respect to colour, appearance, taste, flavor and overall acceptability. Similarly, other workers (Mogra and Dashora, 2009; Mishra, 2011; Singh et al., 2012; Chaturvedi et al., 2013; Balaswamy et al., 2014) also developed low calorie ready-to-serve fruit beverages using natural sweetener (Stevia) in comparison to artificial sweetener and standard recipe which found equally comparable in terms of sensory characteristics Baked products Baked products like biscuits, cake and buns were prepared using Stevia powder in two concentrations and for comparison using sucralose (artificial sweetener) and sugar as control. Mean scores of all sensory characteristics of biscuits, cakes and buns prepared using Stevia powder (T 1 ) significantly higher when compared with control products and even superior as compared to control and sucralose based baked products. The acceptable level of Stevia powder in biscuits, cakes and buns were 60, 60 and 30 mg, respectively. But as the concentration of Stevia powder was increased in baked products, higher amount was not found acceptable in terms of aroma and taste of the products which fell in the category of liked slightly. However, these products scored lowest scores of all sensory attributes as compared to control and sucralose based baked products. Sucralose incorporated biscuits, cakes and buns obtained poor scores of aroma, texture and taste as compared to acceptable level of Stevia based and sugar based products, which fell in the category of liked slightly to liked moderately by the judges. The results of present study are in conformity with the results of Salem et al. (2009), Savita et al. (2004b) and Serna et al. (2014) who formulated bun, biscuits and cake by 100 per cent replacement of sugar with Stevia powder and found good organoleptic characteristics of the products. 119

131 5.2.3 Traditional products The mean scors of colour, appearance, aroma, texture and taste of sweet porridge, carrot halwa and kheer ranged from 6.10 to 8.70, with the highest for control carrot halwa and lowest for sweet porridge prepared using high concentration of Stevia powder T 2. The mean score of colour, appearance, aroma, taste and overall acceptability of sweet porridge prepared using sucralose and Stevia powder T 1 fell in the category of liked moderately. However, sweet porridge prepared using sugar (control) was liked very much for colour, aroma, taste and overall acceptability. Carrot halwa prepared using sugar (control), sucralose and Stevia powder T 1, the mean scores for colour, appearance, aroma, texture, taste and overall acceptability was liked very much. Kheer prepared using sugar (control), sucralose, Stevia powder T 1 did not differ statistically for colour, appearance, texture and taste attributes. The three types of kheer prepared using sugar, sucralose and Stevia powder (T 1 ) was found in the category of liked moderately in terms of aroma and taste by the judges. Hence, the acceptable levels of Stevia powder in sweet porridge, carrot halwa and kheer were 30, 30 and 20 mg, respectively. The results depicted in the present study are in close agreement with the study reported earlier by Chaturvedi et al. (2013) and Agarwal et al. (2010) who prepared sweet products and found that Stevia powder upto 25 mg is highly acceptable and found that with the increase in the concentration of Stevia, the acceptability level in terms of aroma, texture and taste was significantly affected. Similar results were also reported by Sujith and Chitra (2009) in Stevia powder incorporated kheer, wheat halwa and bread pudding. Agarwal et al. (2009) showed that Stevia powder added to various cereal products were highly acceptable. Yet another study of Kerzicnic et al. (2009) reported higher acceptability of Stevia recipes in comparison to sugar recipes. Additionally Stevia is completely safe for human consumption which had been reported in various studies (Sharma et al., 2009; Mishra, 2011; Alizadeh et al. 2014). 5.3 Nutritional composition of developed food products On the basis of organoleptic acceptability, the acceptable products incorporating Stevia powder (acceptable level), sucralose and sugar (control) were nutritionally analyzed Beverages Four types of beverages such as tea, coffee, milk shake and RTS were analysed for proximate composition, sugars, starch and minerals Proximate composition In case of tea, Stevia powder based tea had highest amount of moisture among its counterpart whereas non-significant differences were observed in crude protein, crude fat, crude fibre and ash content in control, sucralose and Stevia powder tea. The high moisture content in Stevia powder based tea might be due to the fact that Stevia powder is in amorphous form and had high water absorption capacity resulting in higher moisture 120

132 absorption. These results are in close agreement with those reported by various workers (Byanna and Gowda, 2010; Mishra, 2011; Balaswamy et al., 2014) With regard to proximate composition of coffee prepared using Stevia powder had significantly higher (85.88 %) moisture and lower (7.31%) total carbohydrate content in comparison to control. The crude fat, crude protein, crude fibre and ash content did not differ significantly among all three types of coffee prepared using sugar (control), sucralose and Stevia powder. High moisture and low total carbohydrate content of Stevia powder coffee might be due to the fact that Stevia powder is added in very minute amount so the solid mass content is more in the developed Stevia coffee resulting in high moisture and low total carbohydrates. Similar trend was also observed by Mogra and Dashora (2009) and Agarwal et al. (2009) they reported higher moisture and low carbohydrate contents in modified recipes prepared using Stevia powder in comparison to basic recipes prepared using sugar. Milk shake prepared using sugar, sucralose and Stevia powder did not differ significantly for crude fat, crude protein, crude fibre and ash content. However milk shake prepared using sucralose and Stevia powder had significantly high amount of moisture (82.17 and %). Whereas Stevia powder milk shake had significantly lower total carbohydrate (5.73 %) content in comparison to control (8.42 %). The results are in close agreement to the study reported by Agarwal et al. (2010) and Mishra (2011). They reported that when sugar was replaced with Stevia powder completely, then the carbohydrate content also decreased significantly. In case of proximate composition of RTS, significant difference was found in moisture content of control and sucralose and Stevia powder incorporated RTS. Moisture content of both RTS incorporated sucralose and Stevia powder was higher (90.97 %) than that of control RTS (87.46 %). Sugar, sucralose and Stevia powder based RTS did not differ significantly for protein, fat, fibre and ash content. Maximum amount of total carbohydrates was exhibited by control RTS (10.85 %) followed by sucralose based RTS (7.67 %) and Stevia based RTS (7.18 %). The results are in close agreement with the results of previous studies conducted by various workers in ready-to-serve beverages prepared using Stevia powder (Gisleine et al., 2006 and Singh et al., 2012) Sugars and starch Among three types of tea, sugar content of Stevia powder based tea was significantly lower as compared to control. This might be due to the fact that Stevia powder contained diterpene sweet glycosides which are times sweeter than sugar and Stevia powder is required in very minute amount to impart appropriate sweetness (IDF, 2013; Gupta et al, 2014). Total, reducing and non reducing content was observed to be significantly higher in control (sugar based) coffee, followed by sucralose and Stevia powder based coffee. 121

133 However, sucralose and Stevia powder coffee did not differ significantly for sugars content. Starch content of all the three types of coffee prepared using sugar, sucralose and Stevia powder ranged from 1.92 to 1.96 per cent, respectively which did not differ significantly. Similar trend was also observed in case of milk shake and RTS. Control milk shake and RTS had significantly higher content of total, reducing and non reducing sugars as compared to Stevia based and sucralose based milk shake and RTS. However, non-significant difference was observed in starch content. It might be due to that Stevia powder and sucralose powder are times sweeter than sucrose apart from being a calorie free sugar (Chaturvedi et al., 2013) Minerals Among three types of tea non-significant differences were observed in total calcium, iron and zinc content. Whereas total potassium and magnesium content were found higher in Stevia powder incorporated tea followed by sucralose and control tea. This might be due to the fact that higher potassium and magnesium content of Stevia powder although Stevia is incorporated in minute amount still can increase the mineral content in product significantly. Similar findings were also observed by Chaturvedi et al. (2013) and Kaushik et al. (2010) in papaya shake and aonla shake. In case of coffee, coffee prepared using sugar (control), sucralose and Stevia powder did not differ statistically for their calcium, zinc, iron and sodium content. Highest iron content was observed in Stevia based coffee and lowest was observed in sugar based coffee. Stevia based coffee was found to have higher magnesium content followed by sucralose based coffee and sugar based coffee. Similarly, the potassium content of Stevia powder based coffee was observed to be significantly higher than the other two types of coffees. Whereas control coffee and sucralose based coffee did not differ significantly among each other. These findings are in agreement with those reported earlier by Sharma et al. (2006). Similar trend was also observed in milk shake and RTS prepared using Stevia powder which had significantly higher content of magnesium and potassium content. However, calcium, iron, zinc and sodium content of beverages did not differ significantly. It might be due to the fact that stevioside as white fine powder form had high magnesium and potassium content (Sharma et al., 2006). Similar results were also reported by other workers in beverages based on Stevia powder (Singh et al., 2012; Chaturvedi et al., 2013) Baked products Three types of baked products such as biscuits, cake and bun prepared using sugar (control), sucralose and Stevia powder were analyzed for their nutritional composition Proximate composition Stevia powder based biscuits had significantly higher moisture content followed by sucralose and sugar based biscuits. Biscuits prepared using sugar, sucralose and Stevia 122

134 powder did not vary significantly for their protein and fat content. The control biscuit exhibited 2.85 per cent for crude fibre which increased with the incorporation of sucralose and Stevia powder but non-significantly. Total carbohydrates content in sugar based biscuits was found to be significantly (P<0.01) highest and Stevia powder based biscuits being the lowest. Stevia based biscuits showed significantly (P<0.01) higher ash content followed by control and sucralose based biscuits. This might be due to the fact that Stevia is a mineral loaded plant. So incorporation of Stevia powder in the developed recipes significantly increase the mineral contents. Similar trend was observed in the ash content (Shafi et al., 2011). Incorporation of Stevia powder increased ash content when replaced with sucrose in the biscuits. Similarly, in case of cake and buns prepared using Stevia had lower sugar content in comparison to sugar based cake and buns. However, there was non-significant difference was found in protein, fat and fibre content of cake and buns prepared using Stevia powder, sucralose and sugar. But significant difference was found in ash and total carbohydrate contents. As control cake and buns had significantly higher content of total carbohydrates but Stevia based cake and bun had significantly higher content of ash might be due to higher mineral contents in Stevia extract powder (Atteh et al., 2011). The results of present study are in close agreement with those reported earlier in baked products based on Stevia powder, artificial sweetener and control (sugar) (Mishra et al., 2010; Kaushik et al., 2010). A similar trend was also observed by Savita et al. (2004) and Mogra and Dashora (2009) in cereal based products Sugars and starch With regard to sugar content Stevia based biscuits, cake and buns had significantly lower content of total, reducing and non reducing sugar contents followed by sucralose and sugar based biscuits, cake and buns. Starch content differed non-significantly. Hence, addition of Stevia and sucralose in recipes significantly reduced caloric content. Similar results were also reposted earlier by Agarwal et al. (2009) in Stevia based sweet products. Savita et al. (2004) also reported low sugar content in besan ladoo incorporating 50 mg Stevia powder Minerals Maximum calcium content was observed in Stevia based followed by sucralose based and control biscuits. Non-significant differences were found in iron and zinc content among all types of biscuits. Sucralose based biscuits were observed to have highest amount of total sodium content and control the lowest. This might be due to the fact that minerals which are beneficial to human health like potassium, calcium, magnesium are present in abundance in Stevia (Barathi et al., 2003; Serio, 2010; Atteh et al., 2011). Similar trend in mineral content was also observed in cake and buns prepared using acceptable level of Stevia powder, sucralose and sugar. 123

135 5.3.3 Traditional Products Three types of traditional products such as sweet porridge, carrot halwa and kheer were prepared using sugar (control), sucralose and Stevia powder. The acceptable level of Stevia powder in sweet porridge, carrot halwa and kheer were 30, 30 and 20 mg, respectively Proximate composition With regard to proximate composition of sweet porridge prepared using sugar (control), sucralose and Stevia powder, moisture was found to be per cent which increased to per cent in sucralose and per cent in Stevia powder sweet porridge. Protein, fat and fibre content of sweet porridge prepared using sugar (control), sucralose and Stevia powder exhibited a non-significant difference in the present study. However, a significant increase in ash content of sweet porridge prepared using Stevia powder is reported in the present study. Total carbohydrates content of Stevia powder based sweet porridge was significantly (P<0.05) lower than control sweet porridge. This might be due to the fact that sucralose and Stevia powder are a low calorie sweetner so incorporation of sucralose and Stevia powder instead of sugar in sweet porridge did not increase the carbohydrate content of the developed sweet porridge. Similar trend was also observed in carrot halwa and kheer. Similarly, other workers also observed the similar results in traditional products prepared using Stevia powder (Savita et al., 2004; Agarwal et al., 2009; Parimalavalli and Radhaisri, 2011) Sugars and starch Stevia powder sweet porridge showed significantly (P<0.05) lower amounts of total (15.09 g/100 g), reducing sugars (4.30 g/100 g) and non-reducing sugar (10.79 g/100 g) content, respectively. Significantly (P<0.05) higher amount of total, reducing and nonreducing sugars was found to be in control sweet porridge followed by porridge prepared using sucralose and Stevia powder sweet porridge. Starch content of all three types of sweet porridge did not differ significantly. Similar trend was also observed in carrot halwa and kheer Minerals Stevia powder based sweet porridge contained significantly higher amount of calcium content followed by sucralose based sweet porridge and control sweet porridge. Highest iron content was found to be in control sweet porridge (2.21 mg/100 g) and similar value for iron content was found to be in sucralose and Stevia powder sweet porridge (2.18 mg/100 g). A non-significant difference was found in iron content of all types of developed sweet porridge. Similar zinc content was found in control and sucralose sweet porridge (1.23 mg/100 g) followed by Stevia powder sweet porridge (1.21 mg/100 g). A non-significant difference was found to be among all sweet porridge for total sodium content. Maximum magnesium content 124

136 was observed in Stevia powder sweet porridge and minimum was found in control sweet porridge. Significant difference in total potassium content was found to be in Stevia powder sweet porridge in comparison to sucralose sweet porridge and control. Similar trend was also observed in mineral contents of carrot halwa and kheer. Mogra and Dashora (2009) also reported higher mineral contents in sweet preparations. 5.4 Glycaemic response of developed food products Glycaemic respose of beverages, baked and traditional products was assesed in normal, obese and diabetic subjects. As beverages were low in available carbohydrates and in order to provide 50 g of equicarbohydrate portion the amount was exceeding the portion size so, amount given to subjects was adjusted in order to provide 25 g of equicarbohydrates/ Serving for beverages and 50 g of equicarbohydrates/ Serving for baked and traditional products of carbohydrates, as in the form of test foods and glucose was used as the reference food Beverages The mean glycaemic response of milkshake prepared using Stevia powder (12 mg) was found to be 111, 101, 89 and 93 mg/dl in normal subjects, 110, 97, 95 and 97 mg/dl in obese and 148, 138, 132 and 130 mg/dl in diabetic subjects at 30, 60, 90 and 120 min, respectively. A continuous reducing trend was observed for milkshake prepared using sugar (control), sucralose and Stevia powder in all three types of subjects with the peak at 30 min. A comparative data exhibited glycaemic index of milk shake prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 48.35, and 33.06, respectively. According to glycaemic index, the foods can be categorized in to low (0-55), medium (56-70), and high (>70) glycaemic index (Premakumari et al., 2013). Milk shake in normal and obese subjects were observed to be in low GI food. Whereas, milk shake prepared using sugar had high GI in diabetics. As sugar was replaced with sucralose and Stevia powder the milk shake was found to be in low GI category. This might be due to the fact that as mik protein has the strong insulinotropic effect (Nilsson et al., 2004). So when milk is ingested milk protein stimulates insulin production, which inturn facilitated glucose utilization, leading to lower GI (Van loon et al., 2000). The results reported in the present study are in close agreement to the studies reported by earlier workers (Ostman et al., 2001; Lenner et al., 2004). The mean glycaemic response of Stevia powder based RTS (10 mg) was found to be 102, 97, 93 and 93 mg/dl in normal, 100, 97, 92 and 93 mg/dl in obese and 154, 151, 145 and 139 mg/dl in diabetic subjects at 30, 60, 90 and 120 min, respectively. A continuous dropping in blood glucose level was observed for RTS prepared using sugar (control), sucralose and Stevia powder in all three types of subjects with the peak at 30 min. RTS prepared using sugar had significantly higher (37.38) GI value, whereas GI value of RTS prepared using sucralose 125

137 (27.91) and Stevia powder (25.68) were found almost similar in normal subjects. In case of obese and diabetic subjects, GI values of RTS based on Stevia powder and sucralose were ranged from to in obese and to in diabetic subjects. However, GI values of RTS (control) in obese and in diabetic subjects differed significantly. The GI values were 37.38, ans in normal, obese and diabetic subjects. GI values of all the three types of RTS were found higher in diabetic subjects followed by obese and normal subjects but still found in the category of low GI food. Gregerson et al. (2004) reported that stevioside suppresses the post pradial blood glucose level in type-2 diabetic subjects by an average of 18 per cent. Similarly, Atteh et al. (2011) and Ali et al. (2013) also reported that stevioside and steviol stimulate insulin secretion by steviol stimulate insulin secreation by direct action on beta cells and these compounds have potential role as antihyperglycaemic effect in the treatment of type-2 diabetes Baked Products Glycaemic response of baked products like biscuits, cake and buns were assessed in normal, obese and diabetic subjects. The mean glycaemic response of biscuits prepared using Stevia powder (60 mg) was found to be 120, 109, 97 and 91 mg/dl in normal, 126, 110, 99 and 93 mg/dl in obese and 185, 162, 151 and 149 mg/dl in diabetic subjects at 30, 60, 90 and 120 min, respectively. The peak in blood glucose level was observed at 30 min after the consumption of biscuits prepared using sugar (control), sucralose and Stevia powder in all the three types of subjects. Glycaemic index of biscuits prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 64.99, and 49.19, respectively. Biscuits prepared using sugar and sucralose was found to have a medium GI in normal and obese subjects. However, biscuits prepared using sugar had high GI in diabetic subjects. However, biscuits prepared using sucralose exhibited a medium GI whereas biscuits prepared using Stevia powder exhibited low GI. This might be due to the fact that Stevia powder contains stevioside and is not absorbed in body by the intestinal tracts. The steviol is excreted in the faeces by the billary way. It was also demonstrated that the steviol remains unabsorbed in the body results in no increase in the blood glucose level (Jeppesen et al., 2000; Atteh et al., 2011; Ali et al., 2013). The mean glycaemic response of cake prepared using Stevia powder (60 mg) were found to be 113, 115, 95 and 92 mg/dl in normal, 126, 127, 103 and 92 mg/dl in obese and 178, 192, 169 and 158 mg/dl in diabetic subjects at 30, 60, 90 and 120 min, respectively. However, cake prepared using sugar (control) showed peak at 30 min whereas sucralose and Stevia based cake showed peak at 60 min after ingestion. A comparative data regarding glycaemic index of cake prepared using sugar (control), sucralose and Stevia powder in normal, obese and diabetic subjects revealed that the cake prepared using sugar had high GI 126

138 value and fell in the category of medium GI food in normal and obese subjects. Whereas, in diabetic subjects the GI value of cake prepared using sugar was high it can be categorized as high GI food. On the other hand, sucralose based cake as medium GI food for diabetic subjects. Stevia powder incorporated cake had 47.25, and GI values in normal, obese and diabetic subjects. As Stevia powder cake was found in low GI category in all the subjects in comparison to sucralose based and sugar based cake. Similar trend as in biscuits was also observed in glycaemic response of buns prepared using Stevia powder, sucralose and sugar. The mean glycaemic response of buns prepared using Stevia powder (30 mg) were found to be 134, 117, 97 and 94 mg/dl in normal, 137, 198, 111 and 99 mg/dl in obese and 200, 165, 155 and 147 mg/dl in diabetic subjects at 30, 60, 90 and 120 min, respectively with the peak at 30 min in all the three types of subjects. Sugar incorporated buns had high GI value (84.41) in diabetic subjects and medium GI values in obese subjects (69.24) and in normal (64.05) subjects. Whereas, Stevia powder sweet buns had 49.08, and GI values in normal, obese and diabetic subjects. Stevia powder sweet bun was found in medium GI food category for diabetics. These results are in agreement with those reported by Savita et al. (2004). They also reported low glycaemic response of sweet buns incorporated Stevia powder. Glycaemic index of Stevia based buns found to be lower in diabetic as well as in normal subjects. Other workers also reported hypoglycaemic response of Stevia based cakes, chapatti, mathi and chikki in diabetic subjects (Ferri et al., 2006; Anton et al., 2010; Parimalavalli and Radhaisri, 2011; Alizadeh et al., 2014) Traditional Products A comparative data regarding glycaemic response and glycaemic index of traditional products (sweet porridge, carrot halwa and kheer) prepared using sugar (control), sucralose and acceptable level of Stevia powder in normal, obese and diabetic subjects are reported in the present study. The mean glycaemic response of sweet porrdige prepared using Stevia powder (30 mg) was found to be 117, 107, 99 and 96 mg/dl in normal, 126, 109, 93 and 99 mg/dl in obese and 184, 163, 159 and 148 mg/dl in diabetic subjects at 30, 60, 90 and 120 min, respectively. Thereafter a continuous decrease was observed in the blood glucose level after the consumption of Stevia based sweet porridge whereas fluctuation i.e. a rise in blood glucose at 90 min after a decrease at 60 min was observed after the consumption of control and sucralose based sweet porridge. The highest GI was observed by the sweet porridge prepared using sugar (62.52) followed by the sweet porridge prepared using sucralose (49.81) and Stevia powder (47.12) based porridge in normal subjects. Sweet porridge prepared using Stevia powder was observed to have a lowest GI in normal (47.12), obese (49.07) and diabetic (55.85) subjects. For normal and obese subjects porridge incorporated sugar was observed as medium GI food but in diabetic subjects it was found to be a high GI food. The results 127

139 reported in the present study are in close agreement to the results reported by Parimalavalli and Radhaisri (2011) and Alizadeh et al. (2014). The mean glycaemic response of carrot halwa prepared using Stevia powder (30 mg) were found to be 112, 117, 103 and 98 mg/dl in normal, 109, 120, 103 and 99 mg/dl in obese and 164, 172, 154 and 148 mg/dl in diabetic subjects at 30, 60, 90 and 120 min, respectively. However, a continuous dropping of glycaemic response for control carrot halwa 106 and 101 mg/dl in normal and 98 and 91 mg/dl in obese subjects were observed whereas diabetic subjects showed a drop of 185 mg/dl at in glycaemic resonse 60 min and rise of 190 mg/dl at 90 min after the consumption of control halwa. Similar trend as in sweet porridge was also observed in case of glycaemic response of kheer prepared using sugar (control), sucralose based (10 mg) and Stevia powder based (20 mg) in normal, obese and diabetic subjects. The glycaemic index of kheer for sugar (control) was found to be 66.71, and in normal, obese and diabetic subjects. Wheras glycaemic index for sucralose and Stevia powder was found to be 47.22, and and 44.37, and in normal, obese and diabetic subjects, respectively. However, in case of diabetic subjects a fluctuation in mean glycaemic response was found after the consumption of kheer prepared using sugar. A continuous reducing trend in glycaemic response was found after consumption of kheer prepared using sucralose and Stevia powder in diabetic patients. Similar trends were also exhibited by Sujith and Chithra (2009) and reported that Stevia incorporated rice kheer was found to be low GI food in comparison to control rice kheer. In the present study also the results indicated that Stevia incorporated food products has hypoglycaemic response. Among developed beverages (milkshake and RTS), RTS prepared using Stevia powder (10 mg) was found to have the lowest GI i.e , and in normal, obese and diabetic subjects, respectively in comparison to milkshake. Among developed baked products (biscuit, cake and bun), cake prepared using Stevia powder (60 mg) were observed to have lowest GI i.e and in normal and obese subjects, respectively. Though, also found to have slight high glycaemic response in diabetic subject with peak at 60 min after consumption. Among traditional products (sweet porridge, carrot halwa and kheer) the GI of carrot halwa prepared using Stevia powder (30 mg) were found to have lower GI i.e , and in normal, obese and diabetic subjects, respectively. 128

140 CHAPTER-VI SUMMARY AND CONCLUSION Stevia rebaudiana is a natural herb native of North-Eastern Paraguay, is fast becoming a major source of high potency sweetener which produces sweet taste but has no calorific value. Considering its sweetness potentialities and possible health implications, the present study was undertaken to develop various types of beverages, baked and traditional products by incorporating Stevia powder, artificial sweetener (sucralose) and sugar (control). Stevia based products were organoleptically evaluated in comparison to sucralose and sugar based products. The acceptable products were nutritionally analysed and studied for their glycaemic response in normal (n=10), obese (n=10) and diabetic (n=10) subjects. The results of the present study are summarized as below: With regard to proximate composition of Stevia leaves and Stevia powder, higher amount of moisture and ash was found in Stevia leaves as compared to Stevia powder. Stevia leaves contained significantly higher contents of crude fibre, fat, crude protein and total carbohydrates as compared to its clarified extract white powder. In case of sugar content, maximum amount of total sugars, reducing and non-reducing sugar content was observed in Stevia powder. Stevia leaves contained significantly higher amount of calcium, iron, sodium, magnesium and potassium to comparison to Stevia white powder. The organoleptic acceptability of beverages, tea, coffee, milk shake and RTS prepared using two levels of Stevia powder, sucralose and sugar (control) results indicated that mean score of colour, appearance, aroma, consistency and taste of tea prepared using Stevia powder T 1 (6 mg) were 7.20, 7.10, 7.40, 7.00 and 7.70, respectively, which fell in the category of liked moderately. Similarly, mean scores of all sensory attributes of tea prepared using sugar and sucralose also fell in the category of liked moderately. Mean score of colour, appearance, aroma, consistency and taste of tea prepared using 8 mg Stevia powder (T 1 ) were 7.20, 7.10, 7.10, 7.10 and 7.30 which fell in the category of liked moderately. Mean scores of all sensory characteristics of Stevia powder (T 1 ) based coffee were found comparable with the coffee prepared using sucralose and sugar. Whereas coffee prepared using higher concentration of Stevia powder 16 mg (T 2 ) had lowest mean scores of all sensory attributes. As on increasing the concentration of Stevia powder in coffee significantly affected the aroma and taste which was neither liked nor disliked by the judges. Milk shake prepared using Stevia powder (12 mg) had 7.70, 7.80, 7.40 and 7.30 mean scores of colour, appearance, aroma and consistency which found in the category of liked moderately whereas mean score of taste was 8.20 which fell in the category of liked very much. The means cores of all sensory attributes of milk shake based on the Stevia powder (T 1 ) was comparable and even superior than control and sucralose based milk shakes. Higher 129

141 concentration of Stevia powder i.e. 24 mg (T 2 ) was not found acceptable in the milk shake which was fell in the category of liked slightly. Mean scores of all sensory characteristics of RTS prepared by using Stevia powder T 1 (10 mg), sucralose, and sugar (control) were in the category of liked moderately. Whereas RTS prepared using 20 mg Stevia powder (T 2 ) had lowest mean scores of aroma (4.70) and taste (4.50) which found in the category disliked slightly. Overall acceptability of mean scores of tea, coffee, milk shake and RTS prepared using Stevia powder (T 1 ) were 7.28, 7.16, 7.78 and 7.50, respectively and found in the category of liked moderately. However, Stevia powder (T 1 ) based beverages found equally acceptable in terms of all sensory characteristics in comparison to standard beverages (sugar based) and sucralose based beverages. With regard to sensory characteristics of baked products, mean scores of colour (7.50), appearance (7.60), aroma (7.40), texture (7.40), taste (7.50) and overall acceptability (7.48) of biscuits prepared using Stevia powder T 1 (60 mg) followed by biscuits prepared using sugar and sucralose. Mean scores of sensory parameters of all three types of biscuits were in the category of liked moderately. Whereas, on increasing the amount of Stevia powder T 2 (120 mg) caused significant reduction in mean scores of aroma, texture and taste which fell in the category of liked slightly. Similar tends was also observed in cake prepared using Stevia powder T 1 (60 mg) and T 2 (120 mg), sucralose and sugar. All the three types of cakes were found in the category of liked moderately except cake prepared using high amount of Stevia powder (120 mg) which was not found acceptable in terms of sensory characteristics. In case of buns, mean scores of all sensory parameters like colour, appearance, aroma, texture, taste and overall acceptability was in the category of liked moderately by the judges. Whereas, cake prepared using 60 mg of Stevia powder (T 2 ) was in the category of liked slightly. Similar trend was also observed in sensory attributes of traditional products such as sweet porridge, carrot halwa and kheer. With regard to nutritional composition of beverages, it was observed that tea, coffee, milk shake and RTS prepared using acceptable level of Stevia powder (T 1 ) has significantly higher content of moisture and ash as compared to control and sucralose based beverages. Whereas, total carbohydrate i.e , 10.81, and per cent, respectively were found significantly higher in sugar based recipes. Non-significant difference was observed in crude protein, fat and fibre contents of all the beverages. Similarly, total, reducing and nonreducing sugar contents of control beverages were also found significantly higher as compared to other test beverages. With regard to mineral contents like potassium and 130

142 magnesium were found significantly higher in acceptable level of Stevia powder based beverages followed by sucralose and sugar based beverages. Data regarding proximate composition of baked products like biscuits, cake and buns prepared using Stevia powder (T 1 ), sucralose and sugar revealed that moisture and ash content of Stevia based biscuits, cake and bun was significantly higher followed by sucralose and sugar based baked products. Whereas, total carbohydrate i.e , and per cent, respectively were found significantly higher in control biscuits, cake and buns. Crude protein, fat and fibre contents differed non-significantly in all the three types of baked products based on Stevia powder, sucralose and sugar. Similarly, total, reducing and non-reducing sugar ranged from to 35.45, 3.76 to 5.76 and to per cent, respectively in biscuits, cake and buns prepared using sugar. The sugar contents of control baked products were found significantly higher as compared to Stevia powder based and sucralose based baked products. In case of mineral contents, Stevia powder based biscuits, cake and buns had higher amount of calcium, magnesium and potassium as compared to control and sucralose based baked products. Similar trend was also observed in proximate composition, sugar and mineral contents of traditional products (sweet porridge, carrot halwa and kheer) based on acceptable level of Stevia powder, sucralose and sugar. It may be concluded that Stevia powder being calorie-free, nutritious and acceptable in various food products, can readily be used as an ideal sweetener. Glycaemic response of most acceptable products milk shake, RTS, biscuits, cake, buns, sweet porridge, carrot halwa and kheer was assessed in normal, obese and diabetic subjects. The glycaemic index of food products prepared using most acceptable level of Stevia powder in comparison to sugar based (control) and sucralose based products was assessed in normal (n=10), obese (n=10) and diabetic (n=10) subjects. The average height, weight and BMI of normal, obese and diabetic subjects were cm, kg and kg/m 2, , and kg/m 2 and cm, 69.3 kg and kg/ m 2, respectively. The amount of food given to subjects was adjusted in order to provide 25 g for beverages and 50 g for baked and traditional products of carbohydrates, as in the form of test foods and glucose was used as the reference food. The mean glycemic response of sucralose and Stevia powder based milkshake showed 84 and 86 at fasting, 118 and 111 at 30, 107 and 101 at 60 and 91 and 89 at 90 min, respectively in normal subjects. The mean glycemic response of control milkshake, sucralose and Stevia powder based milkshake was found to be 158, 128 and 110 mg/dl at 30 min, 128, 100 and 97 mg/dl at 60 min and 104, 91 and 95 mg/dl at 90 min, respectively in obese subjects. The mean glycemic response of Stevia powder (12 mg) based milkshake was found 131

143 to be 148, 138, 132 and 130 mg/dl at 30, 60, 90 and 120 min, respectively. The peak value was 210, 170, 154 and 148 mg/dl for reference, control, sucralose and Stevia based milkshakes in diabetic subjects. Thereafter the levels showed a reducing trend at 120 min. Although Stevia and sucralose were found to have almost similar mean post parandial blood glucose level i.e. 130 and 128 mg/dl at 120 min, respectively. All three types of milkshakes were found to have the highest peak at 30 min. Glycaemic index of milk shake prepared using sugar (control), sucralose and Stevia powder in normal subjects were found to be 48.35, and 33.06, respectively. Milk shake prepared using sugar had significantly higher (48.35) GI value, whereas milk shake prepared using sucralose and Stevia powder were comparable in normal subjects. Similarly, in case of obese and diabetic subjects, GI values of milk shakes based on sucralose and Stevia powder were comparable. However, GI values of milk shake (control) in obese (52.80) and in diabetic (59.15) subjects significantly, higher as compared to the GI values of milk shake based on sucralose and Stevia powder in obese and diabetic subjects. GI values of all the three types of milk shakes were found higher in diabetic subjects followed by obese and normal subjects. The mean glycemic response of sucralose (5 mg) and Stevia powder RTS (10 mg) was found to be 106 and 102 at 30 min, 100 and 97 at 60 min and 96 and 93 mg/dl at 90 min, respectively after concumption in normal subjects. The mean glycemic response in obese subjects after ingestion of control, sucralose and Stevia based RTS was found to be 118, 104 and 100 mg/dl at 30 min, 110, 100 and 97 mg/dl at 60 min and 104, 96 and 92 mg/dl at 90 min, respectively. However, mean post parandial blood glucose level was found to be 95, 87 and 93 mg/dl, respectively for control, sucralose and Stevia powder based RTS in obese subjects. The mean glycemic response of reference food was found to be 210, 184 and 178 mg/dl at 30, 60 and 90 min, respectively in diabetic subjects. Whereas, the mean glycaemic response of RTS prepared using sucralose was found to be 161, 156 and 142 mg/dl and Stevia powder was found to be 154, 151 and 145 mg/dl at 30, 60 and 90 min, respectively in diabetic subjects. Therefore, a reducing trend was observed in all three types of RTS in diabetic subject with the peak at 30 min after the consumption. Stevia powder based RTS recorded the lowest post parandial blood glucose level i.e. 139 mg/dl at 120 min. Glycaemic index of RTS prepared using sugar (control), sucralose and Stevia powder in normal subjects were 37.38, and 25.68, respectively. GI of RTS in normal subjects was in the low category. Whereas, GI value of control RTS in obese (38.43) and in diabetes (47.38) was significantly higher as compared to GI value in normal subjects but still found in the low category of glycaemic index. On the other hand, GI values of Stevia powder based RTS in normal, obese and diabetic subjects were 25.68, and as these values were significantly lower as compared to sucralose and sugar based RTS in all the three types of subjects. Hence, Stevia based RTS can be categorized as low GI food for obese and diabetic subjects. 132

144 The mean glycemic response after ingestion of glucose (50 g of equicarbohydrate) was 87, 178, 135 and 124 mg/dl at fasting 30, 60, 90 and 120 min, respectively in normal subjects. Whereas mean glycaemic response of sucralose (30 mg) and Stevia powder based (60 mg) biscuits was found to be 128 and 120 mg/dl at 30 min, 113 and 109 mg/dl at 60 min and 105 and 97 mg/dl at 90 min, respectively in normal subjects. However, mean glycaemic response of biscuits prepared using sucralose and Stevia powder was found to be 130 and 126 mg/dl at 30 min, 121 and 110 mg/dl at 60 min and 96 and 99 mg/dl at 90 min, respectively in obese subjects. Control biscuits and Stevia powder biscuits showed similar i.e. 93 mg/dl post parandial glucose level at 120 min in obese subjects. Biscuits prepared using sugar (control), sucralose and Stevia powder showed peak i.e. 219, 197 and 185 mg/dl after 30 min. Thereafter levels showed reducing trends in all three types of biscuits at 60, 90 and 120 min. The mean glycaemic response of biscuits prepared using sucralose and Stevia powder were found to be 156 and 162 mg/dl at 60 min and 155 and 151 mg/dl at 90 min respectively in diabetic subjects. With regard to glycaemic index of biscuits prepared using Stevia powder, sucralose and sugar indicated that GI value of Stevia based biscuits in normal, obese and diabetic subjects were 49.19, and 53.41, respectively followed by sucralose based biscuits and sugar based biscuits. GI value of Stevia based biscuits fell in the category of low glycaemic index. On the other hand, GI values of sucralose based biscuits and sugar based biscuits in normal, obese and diabetic subjects were ranged from to 64.99, to and to 78.80, respectively. Sucralose based biscuits fell in the category of medium GI food in all the three types of subjects. Whereas sugar based biscuits fell in the category of medium GI food in normal and obese and in the category of high GI food in diabetic subjects. Similar trend in glycaemic index and glycaemic response as in biscuits was also observed in case of control, sucralose (15 mg) based and Stevia powder (30 mg) based buns in normal, obese and diabetic subjects. However, in case of cake prepared using sugar (control) showed peak value 178 and 141 mg/dl in normal, 183 and 158 mg/dl in obese and 296 and 246 mg/dl in diabetic subjects, respectively at 30 min. Whereas sucralose and Stevia based cake showed peak value 123 and 115 mg/dl in normal, 137 and 127 mg/dl in obese and 212 and 192 mg/dl in diabetic subjects respectively, at 60 min after ingestion. Stevia powder and sucralose based cakes were found to be medium GI foods for normal, obese and diabetic subjects, whereas sugar based cake was found to be medium GI foods for obese and high GI foods for diabetics. The peak value were found to be highest for control porridge (145 mg/dl) followed by for sucralose based (15 mg) porridge (124 mg/dl) and Stevia powder based (30 mg) porridge (117 mg/dl) at 30 min in normal subjects. Decrease in glycaemic response were observed in control, sucralose and Stevia powder based sweet porridge and was found to be 113, 110 and 133

145 107 mg/dl at 60 min and 101, 95 and 99 mg/dl at 90 min at 99, 93 and 96 mg/dl at 120 min, respectively in normal subjects. The mean glycaemic response for control sweet porridge was found to be 150, 116, 98 and 91 mg/dl at 30, 60, 90 and 120 min, respectively in obese subjects. However peak value for sucralose and Stevia powder based porridge were found to be 138 and 126 mg/dl at 30 min, respectively in obese subjects followed by reducing trend with glycaemic response 96 and 93 mg/dl at 90 min, respectively for sucralose and Stevia powder based porridge. The mean glycaemic response of sweet porridge prepared using sugar (control) was 110, 238, 198, 209 and 189 mg/dl at fasting, 30, 60 and 90 and 120 min and the mean glycaemic response of porridge prepared using Stevia powder was 113, 184, 163, 159 and 148 mg/dl at fasting, 30, 60, 90 and 120 min, respectively in diabetic subjects. Thereafter a continuous decrease was observed in the blood glucose level after the consumption of Stevia based sweet porridge whereas fluctuation i.e. decrease at 60 min and a rise in blood glucose at 90 min was observed after the consumption of control and sucralose based sweet porridge. GI value of sweet porridge based on Stevia powder ranged from to in normal, obese and diabetic subjects which was significantly lower as compared to sweet porridge based on sucralose and sugar. GI value of sugar used sweet porridge had significantly higher GI value in normal (62.52), in obese (65.24 and in diabetic (73.52) subjects. Stevia powder based sweet porridge was considered in the low GI food category in all the three types of subjects. Whereas, sugar based sweet porridge was considered as medium GI food in normal and obese but high GI food in diabetics. A similar trend as in sweet porridge was also observed in case of glycaemic response of kheer prepared using sugar (control), sucralose based (10 mg) and Stevia powder based (20 mg) in normal and obese subjects and diabetic subjects i.e a fluctuation in mean glycaemic response was found after the consumption of kheer prepared using sugar whereas a sharp decrease in blood glucose level after the consumption of sucralose based kheer. A continuous reducing trend in glycaemic response was found after consumption of kheer prepared using Stevia powder in diabetic patients. However, the mean glycaemic response for control carrot halwa was found to be 134 and 142 mg/ dl at 30 min, 126 and 121 mg/dl at 60 min and 106 and 98mg/dl at 90 min in normal and obese subjects, respectively. The mean glycaemic response of carrot halwa prepared using sucralose (15 mg) and Stevia powder (30 mg) showed peak value 120 and 117 mg/dl in normal, 124 and 120 mg/dl in obese and 173 and 172 mg/dl in diabetic subjects at 60 min, respectively after consumption. However, after the consumption of control halwa diabetic subjects showed a drop in glycaemic response of 185 mg/dl at 60 min and again rise in blood glucose level of 190 mg/dl at 90 min. A continuous reducing trend was observed for carrot halwa prepared using sucralose and Stevia powder in all three types of subjects. Carrot 134

146 halwa prepared using Stevia powder (30 mg) were found to have lowest GI i.e , and in normal, obese and diabetic subjects, respectively. Among developed beverages (milkshake and RTS) RTS with 10 mg Stevia powder, among developed baked products (biscuit, cake and bun) cake with 60 mg Stevia powder and among traditional products (sweet porridge, carrot halwa and kheer), carrot halwa with 30 mg Stevia powder scored at par for organoleptically acceptability and were found to have lower GI in comparison to their respective controls and comparative food products in normal, obese and diabetic subjects, respectively. All three products i.e. RTS (beverage), cake (baked product) and carrot halwa (traditional products) on the basis of study can be considered as best foods for normal, obese and diabetic subjects among the developed recipes. It may be recommended from the results that Stevia powder based food products as it would be ideal for inclusion in the diets of obese and especially for diabetics due to their low glycaemic response. Plant based Stevia herb is a low calorie nutritious component has an immense potential in the main stream of food processing industries as a health and dietetic benefactor. Hence, there is need for creating awareness among the people about the nutritional and therapeutic values of natural low calorie sweetener (Stevia) incorporated food products for those who have to restrict carbohydrate/ sugar in diet. 135

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154 ANNEXURE - I 9 Point Hedonic Rating Scale Name Dated Products Test products and check how much you like or dislike each one. Use appropriate scale to show your attitude by assigning points that best describe your feelings about the sample. An honest expression of your feelings will help us. Sr. No. Colour Appearance Aroma Texture Taste Overall Total acceptability Remarks Rate Organoleptic score Like extremely 9 Like very much 8 Like moderately 7 Like slightly 6 Neither like nor dislike 5 Dislike slightly 4 Dislike moderately 3 Dislike very much 2 Dislike extremely 1 I

155 ANNEXURE II Table: Total dietary fibre content of developed food products Products Control (sugar) Sucralose Stevia powder CD(P<0.05) Tea 0.10± ± ±0.01 NS Coffee 1.70± ± ±0.05 NS Milkshake 3.30± ± ±0.02 NS RTS 1.95± ± ±0.05 NS Biscuits 9.22± ± ±0.02 NS Cake 3.09± ± ±0.07 NS Bun 3.31± ± ±0.02 NS Sweet porridge 11.42± ± ±0.01 NS Carrot Halwa 3.21± ± ±0.01 NS Kheer 1.13± ± ±0.06 NS II

156 ANNEXURE- III PARTICIPANT INFORMATION AND CONSENT FORM STUDY TITLE: NUTRIENT EVALUATION AND GLYCEMIC RESPONSE OF FOOD PRODUCTS PREPARED USING STEVIA REBAUDIANA NAME OF RESEARCHER: Suhani Arora, Admission no. 2009HS69D Department of Foods and Nutrition, College of Home Science, CCS HAU, Hisar. WHAT IS THE PURPOSE OF THIS STUDY? 1. To determine nutrient composition of Stevia powder 2. To standardize and develop food products using Stevia powder and to study their organoleptic acceptability 3. To study nutrient composition and glycemic response of the most acceptable developed food products For the glycemic response the participant will be asked ingest the given value added products. Then they will be asked to give blood samples using finger prick method at fasting level and after 30, 60, 90 and 120 min after the load (50g of equicarbohydrate portion). By using this, a glycemic response area under blood glucose response curve will be calculated. WHAT WILL HAPPEN IF I VOLUNTEER? If you agree to participate, you will be requested to complete the enclosed consent form. Name of Researcher Researcher Signature Date Researcher Address: Contact no: III

157 ANNEXURE IV PARTICIPANT CONSENT FORM (Form of consent by a participant volunteering to take part in medical research not associated with clinical treatment) STUDY TITLE: NUTRIENT EVALUATION AND GLYCEMIC RESPONSE OF FOOD PRODUCTS PREPARED USING STEVIA REBAUDIANA 1. I confirm that I have received a copy of the Information Sheet for the above study. I have read it and I understand it. The nature, purpose and what my involvement will be in the study has been described to me in the Information Sheet and has been explained to me by: Suhani Arora, Admission no. 2009HS69D, Department of Foods and Nutrition, College of Home Science, CCS HAU, Hisar. 2. I have had time to consider whether to take part in this study and I have had the opportunity to ask questions. 3. I understand that my participation is voluntary and that I am free to withdraw at any time, without giving any reason, without my medical care or legal rights being affected. 4. By signing this consent form, I agree to take part in the above study and I have discussed these matters with the researcher to my satisfaction. Name of Participants Participants Signature Date Participant s Address: Contact no: IV

158 ABSTRACT 1. Title of thesis : Nutritional Evaluation and Glycaemic Response of Food products Prepared Using Stevia rebaudiana 2. Full name of degree holder : SUHANI ARORA 3. Admission No. : 2009HS69D 4. Title of degree : Doctor of Philosophy in Foods and Nutrition 5. Name and address of Major Advisor : Dr. (Mrs.) Sudesh Jood Professor Department of Foods and Nutrition I.C. College of Home Science CCS Haryana Agricultural University, Hisar (India) 6. Degree awarding University/ Institute : CCS Haryana Agricultural University, Hisar Year of award of degree : Major subject : Foods and Nutrition 9. Total No. of pages in thesis : vii + IV 10. No. of words in the abstract : Approx. 311 Key words: Stevia powder, sucralose, sugar, product development, organoleptic acceptability, nutritional analysis, glycemic response An investigation was conducted to study the nutritional evaluation and glycaemic response of food products prepared using Stevia powder in comparison to artificial sweetener (sucralose) and sugar which served as control. Stevia leaves had significantly higher proximate, sugars and mineral content in comparison to Stevia powder. Results indicated that Stevia leaves had 11.16, 15.63, 5.81 and 9.82 per cent ash, total sugars, reducing sugars and non reducing sugars, respectively. Stevia leaves contained , 32.34, 3.57, , and mg calcium, iron, zinc, sodium, magnesium and potassium, respectively. Whereas, Stevia powder had 3.42 (%) ash, 1.78(%) total sugars, 0.84(%) reducing sugars and 0.93 (%) non reducing sugars, respectively and contained , 6.31, 0.12, 34.92, and mg calcium, iron, zinc, sodium, magnesium and potassium, respectively. Various food products included beverages (tea, coffee, milk shake and RTS), baked products (biscuits, cake and buns) and traditional products (sweet porridge, carrot halwa and Kheer) were standardized and developed using Stevia powder (two levels), sucralose and sugar. Nutrient content of Stevia based products had high mineral and low sugar profile. Products were assessed for glycaemic response. The results showed, recipes prepared using sucralose and Stevia powder had significantly low GI in normal, obese and diabetic subjects. However, among beverages RTS (10 mg Stevia powder) had 25.68, and GI, cake (60 mg Stevia powder) had 47.25, and GI and carrot halwa (30 mg Stevia powder) had 35.99, and GI, respectively in normal, obese and diabetics subjects. Fluctuation in blood glucose level were noticed after ingestion of sugar and sucralose based products whereas, on replacement with Stevia powder in products uniformity was noticed in blood glucose level resulting in low glycaemic index. It can be inferred that incorporation of Stevia powder instead of sucralose and sugar is a best alternative and is a boon for deprived obese and diabetic patients. MAJOR ADVISOR DEGREE HOLDER HEAD OF THE DEPARTMENT

159 CURRICULUM VITAE a) Name of the Student : Suhani Arora b) Date of Birth : 9 th Jan c) Place of Birth : Hisar (Haryana) d) Mother s Name : Mrs. Veena Arora e) Father s Name : Mr. Anil Arora f) Permanent Address : M/S Gajjan Mal Shiv Dayal, g) Telephone : h) Mobile : Commission Agent, Uklana Mandi, Hisar i) suhaniarora966@gmail.com j) Academic qualifications : Degree University/Board Year of Percentage of Subjects passing marks Matric C.B.S.E.Board Science, Maths, English, Social Studies, Hindi 10+2 CBSE Board Psychology, English, Home Science, Fine Arts, Music B.Sc. (Home Sci.) CCSHAU, Hisar FN, HDFS, FRM, CT, HECM M.Sc. ((Foods CCSHAU, Hisar Foods and Nutrition and Nutrition) Ph.D. (Foods and Nutrition) CCSHAU, Hisar Foods and Nutrition k) Co-curricular activities : Community Work- Worked as an active member of NSS Camp organised by ICCOHS, CCSHAU, Hissar i) List of publications : Suhani Arora and Darshan Punia (2012) Glycaemic response of traditional Indian recipes in International conference on food and nutrition technology for public health care at IISMAAS. JNU, New Delhi.held on 4-5 May, 2012, PHN-O-11. Suhani Arora and Darshan Punia (2012) Nutritional evaluation of traditional Indian chapaties in International conference on food and nutrition technology for public health care at IISMAAS. JNU, New Delhi held on 4-5 May, 2012, FPP-O-52. Booklets Suhani Arora and Sudesh Jood (2013) Herbal Sugar Free Baked Products, Department of Foods and Nutrition,CCSHAU, Hisar Medals/honours received Received merit certificate in Ist year of BSc. Home Science Received scholarship throughout Graduation and Post Graduation. Research Experience Research Project on Nutritional evaluation of products developed using Stevia Rebudiana for normal, obese and diabetic patients. Completed a Research Project on Nutritional evaluation and Glycaemic Index of traditional Recipes in M.Sc. Completed Training Project on Bakery Products conducted by foods and nutrition Deptt. in collaboration with NMCP, Netherland in HAU, Hissar Suhani Arora