CHAPTER III MATERIALS AND METHODS

CHAPTER III MATERIALS AND METHODS An investigation entitled Standardization and storage of Ready to serve and candy of ber fruit (Zizyphus mauritiana Lamk.) was carried out in the Department of Horticulture, College of Agriculture, Junagadh Agricultural University, Junagadh (Gujarat) during the year 2016. The details of techniques followed and materials used during the course of experimentation are described below. 3.1 MATERIALS Fresh, fully matured and uniform size ber fruits cultivars of umran and apple ber were purchased from the local market and used for experimentation. They were immediately brought to the laboratory and stored out. The unripen, diseased, damaged and off type fruits were discarded. The selected fruits were thoroughly washed with tap water to remove dirt and dust particles adhering to the surface of fruit and were allowed to surface dry and used for both experiments. 3.1.1 EXPERIMENT No.1: Standardization and storage of Ready to serve of ber (Zizyphus mauritiana Lamk.) The experiment entitled Standardization and storage of Ready to serve of ber fruit (Zizyphus mauritiana Lamk.) was carried out during the year 2016. 3.1.2 EXPERIMENTAL DETAILS 1. Experimental site/location Post graduate laboratory, Dept. of Horticulture, College of Agriculture, J.A.U., Junagadh. 2. Year of experiment 2016 3. Experimental design C.R.D. (Factorial) 4. Number of Factors 3 (Three) 1. Juice proportions 2. Total Soluble Solids (TSS) 3. Method of storing

5.No.of treatments 12 (Twelve) combination 6. No. of repetitions 3 (Three) 7. Name of crop and variety Ber Umran 8. Total No. of combinations 36 (Thirty six) 9. Vol. of each bottle 200 ml 10. Time period for storage Up to 6 months 3.1.3 TREATMENT DETAILS FOR RTS The experiment comprised of 12 treatment combinations consisting 3 levels of juice proportions, 2 levels of sugar percentage and 2 levels of method of storage. The details of various treatments and their combinations are presented below: Factor A: Proportion of fruit juice 1) R 1 : 10% of juice 2) R 2 : 15% of juice 3) R 3 : 20% of juice Factor B: Percentage of TSS 1) S 1 : 10% TSS 2) S 2 : 15% TSS Factor C: Method of storage 1) T 1 : Room temperature 2) T 2 : Cold storage 3.1.4 TREATMENT COMBINATIONS FOR RTS 1. R 1 T 1 S 1 : 10% Juice + 10% TSS + Room Temperature 2. R 1 T 1 S 2 : 10% Juice + 10% TSS + Cold Storage 3. R 1 T 2 S 1 : 10% Juice + 15% TSS + Room Temperature 4. R 1 T 2 S 2 : 10% Juice + 15% TSS + Cold Storage 5. R 2 T 1 S 1 : 15% Juice + 10% TSS + Room Temperature 6. R 2 T 1 S 2 : 15% Juice + 10% TSS + Cold Storage 7. R 2 T 2 S 1 : 15% Juice + 15% TSS + Room Temperature 8. R 2 T 2 S 2 : 15% Juice + 15% TSS + Cold Storage 9. R 3 T 1 S 1 : 20% Juice + 10% TSS + Room Temperature 26

10. R 3 T 1 S 2 : 20% Juice + 10% TSS + Cold Storage 11. R 3 T 2 S 1 : 20% Juice + 15% TSS + Room Temperature 12. R 3 T 2 S 2 : 20% Juice + 15% TSS + Cold Storage 3.1.5 EXPERIMENTAL DESIGN The experiment was laid out in a Completely Randomized Design with factorial concept and treatments repeated three times. 3.1.6 TECHNIQUES OF PRODUCT PROCESSING: 3.1.6.1 Method of juice extraction: The peeled fruits were cut into slices and seeds were removed manually. The slices were crushed into pulp with the help of electronic derived mixture grinder. Then, juice was filtered through a fine meshed silk cloth. This juice containing some pulp particles was allowed to keep in refrigerator at low temperature overnight in airtight condition to settle down the coarse particle at bottom. After that, the upper layer of pure juice content separated out. 3.1.6.2 Recipe treatments: After extraction of ber juice, its total soluble solids (TSS) and acidity were measured. Then according to different recipe treatments, the quantity of juice, sugar, citric acid and water was calculated. Then its TSS and acidity were measured. 3.1.6.3 Preparation of Ready-To-Serve (RTS) beverage: For the preparation of RTS of different recipe, sugar syrup was prepared by adding water and citric acid. For this purpose required quantity of sugar was added to measured quantity of water and boiled it to dissolve the sugar. During heating the required quantity of citric acid was added. The prepared syrup was strain through coarse muslin cloth and then according to recipe required quantity of ber juice. For the preparation of different recipe of RTS beverage, the amount of juice required as per FPO specification was calculated. The TSS was measured using refractometer and adjusted with the table. 27

3.1.6.4 Bottling: The prepared RTS filled in clean, sterilized glass bottles of 200 ml capacity leaving 2.5 cm head space and corked them air tight with the help of crown corking machine. 3.1.6.5 Pasteurization The filled bottles of RTS were pasteurized in hot water at a temperature of 90 0 C for 20 minutes. Bottles kept at room temperature to cool down. 3.1.6.6 Storage Prepared RTS bottles 6 combinations were stored in dried place at room temperature ranging from 12.1 0 C to 35.5 0 C and relative humidity ranging from 19.4% to 92.0%.and remaining 6 combinations stored in cool chamber at 13 0 c temperature 3.1.7 OBSERVATIONS RECORDED: Prepared Ready to serve (RTS) beverage of ber was subjected to following periodical observations. Prepared Ready To Serve (RTS): The observation on following physical and chemical characteristic of RTS were recorded at monthly interval up to 6 months of storage 3.1.7.1 Sensory Rating: (1) Color (2) Taste (3) Flavor (4) Appearance (6) Product setting at bottom (7) Overall acceptance 3.1.7.2 Biochemical Parameters: (1) Total soluble solids ( 0 Brix) (2) Titrable acidity (%) (3) Ascorbic acid (mg/100 ml RTS) (4) Sugars: 28

(i) Reducing sugars (%) (ii) Non-reducing sugars (%) (iii) Total sugars (%) 3.1.7.3 Microbial count of the RTS: Bacterial and fungal colony was carried out by serial dilution methods. 3.1.7.4 Economic feasibility: the treatment. The economics of different treatments was calculated on the basis of cost of 3.1.8 METHODOLOGY USED FOR OBSERVATION 3.1.8.1 SENSORY RATING: Prepared juice and RTS under different treatments were evaluated for sensory characteristics viz., color, taste, flavor, appearance, cloudiness, product setting at bottom and overall acceptability. Each attribute was given a separate score of 10 points. Product scoring less than 5 points out of 10 points was treated as unacceptable from the quality point of view. Sensory panel consisted of 5 trained panelists and the panelists were instructed to evaluate the samples as per hedonic scale procedure as described by Ranganna (1979) in format detailed below: Treatment Color Taste Flavor Appearance Product setting at bottom 1 to 12 Overall acceptance 3.1.8.2 CHEMICAL CHARACTERISTICS: 1) Total soluble solids (TSS) The total soluble was measured with the help of hand refractometer at room temperature and the method suggested by Ranganna, 1979 was followed. The few drops of the RTS were put on prism of the hand refractometer to obtain the total soluble solids. The results were expressed in terms of 0 Brix 29

2) Acidity (%) To estimate Titrable acidity the analytical method given by Ranganna (1979) was followed. One gram of crushed candy sample was taken in a beaker and diluted by adding 10 ml of distilled water and the solution filtered with the help of muslin cloth. In a burette 0.1 N NaOH was filled and the sample was titrated against the standard 0.1 N NaOH solution till the light pink color appears. The estimated value then expressed in terms of percent acid by adopting the following formula. T N Q E Acidity (%) = 100 V W Whereas, T = Titer value (ml) N = Normality of alkali solution V = Volume made up (ml) E = Equivalent weight of acid (g) Ws = Weight of sample (g) Vs = Volume of sample taken for titration (ml) 3) Ascorbic acid (mg / 100g) Ascorbic acid was determined by using 2-6 dichlorophenol indophenol method as described by Sadasivam and Manickam (1996). The crushed candy pulp of 10 g was transferred to volumetric flask and volume was made up to 100 ml by adding 3 % metaphospheric acid solution. After 30 minutes, the suspension was filtered through Whatman No. 1 filter paper. From the filtrate, 5 ml aliquot was taken and titrated against standardized dye solution. Titration was continued till the light pink color persisted for 15 seconds. Initially, the dye solution was standardized by titrating against standard ascorbic acid solution and the dye factor was calculated. Ascorbic acid content was calculated by using following equation. Titrate Dye factor Volume made up Ascorbic acid (mg 100 g -1 ) = Aliquot taken for estimation Weight or volume of sample taken for estimation 100 30

4) Reducing sugar (%) Reducing sugar was estimated by using Nalson Smogyi methodas reported by Sadasivam and Manickam (1996). In this method, DNSA reagent was used and absorbance was noted at 565 nm and accordingly graph was plotted against standard glucose solution and this process repeated 3 to 4 times and calculated the % reducing sugar by using standard formula given formula. Reducing sugar (%) = Glucose equivalent 0.05 x Total volume made up Titer x Weight of sample taken 100 5) Total sugar (%) Total sugars estimation was performed by phenol sulphuric acid method with one gram of pulp was taken in a 100 ml volumetric flask in which 2 ml of lead acetate solution was added and kept for 10 minutes. The solution then deluded by adding 1 ml potassium oxalate solution and volume was made up to 100 ml by adding distilled water. The solution was filtered through Whatman No. 1 filter paper. The filtrate was diluted in ratio of 1:20 with distilled water and 1 ml aliquot was taken for the estimation of sugar. Then, 1 ml of 5% phenol solution and 5 ml of 96 % H 2 SO 4 was added quickly and allowed to stand for 10 minutes after mixing. The colour of solution was read at 490 nm on spectrophotometer. The estimation of total sugar was carried out using standard graph prepared with glucose (0-150 µg) using following expression (Sadasivam and Manickam, 1992).Percentage of total sugar was calculated according to following formula. Total sugar (% ) = Sample reading Glucose equivalent Volume made up 100 10 6 Weight of pulp Aliquot taken 31

6) Non reducing sugar (%) The amount of non-reducing sugar was obtained by subtracting reducing sugars from the amount of total sugars and non-reducing sugar content was expressed in terms of percentage Non reducing sugar (%) = Total sugar (%) Reducing sugar (%) 3.1.8.3 METHOD OF MICROBIAL COUNTS The stored candy was subjected to microbial analysis for total viable count using Nutrient agar for bacterial analysis and Potato Dextrose Agar for fungus analysis as per the standard procedure given by Ranganna, (1979). Preparation of dilutions Sample of one gram was aseptically taken into a sterilized glass test tube and transferred to 9 ml sterile phosphate buffer dilution blank to obtain 1:10 dilution. Subsequently, 0.1 ml of above prepared dilution was used for making further dilutions in 9.9 ml phosphate buffer tubes. Suitable dilutions were prepared and poured in a set of sterile petri plates in duplicates. Total viable count One ml of suitable dilution from each sample prepared as described in previous section was used for plating in duplicates and thereafter 15 ml of molten Nutrient Agar was poured aseptically to plates. The contents were mixed and plates were cooled. The plates then were inverted and incubated in an incubator maintained at 37 ± 0.5 0C for 24 hour and number of colony forming units (CFU/g) was recorded. Similarly, for fungus analysis, 1 ml of suitable dilution from each sample prepared as described in previous section was used for plating in duplicates and thereafter 15 ml of molten Potato Dextrose Agar was poured aseptically to plates. The contents were mixed and cooled. The plates then were inverted and incubated in an incubator maintained at 37 ± 0.5 0C for 96 hour and number of colony forming units (CFU/g) was recorded. Number of colonies Total CFU/ml = Volume plated x Dilution 32

3.1.8.4 ECONOMICS OF THE TREATMENTS The economics of different treatments was calculated on the basis of the treatment. The cost benefit ratio (CBR) was calculated on the basic of the formula given below. Price of the product Rs. / Kg CBR = Total cost Rs. / Kg 3.14 STATISTICAL ANALYSIS The data were analyzed statistically according to the analysis of variance techniques as suggested by Cochran and Cox (1950). The analysis of variance for different parameters is presented in Appendices. The critical difference (CD) was calculated to access the significance, non- significance of difference between treatments mean, wherever it was found significant through F test at 5% level of significance. 33

Flow Chart showing the procedure to prepare ber RTS Ber pulp Straining Preparation of syrup (Boiling sugar and citric acid in water) Straining Ber juice + syrup Bottling Homogenization Corking Pasteurization Storage 34

3.2. EXPERIMENT No.: Standardization and storage of candy of ber fruit The experiment entitled Standardization and storage of candy of ber fruit (Zizyphus mauritiana Lamk.) was carried out during the year 2016. 3.2.1. EXPERIMENTAL DETAILS 1. Experimental site/location Post graduate laboratory, Dept.of Horticulture, College of Agriculture, J.A.U., Junagadh. 2. Year of experiment 2016 3. Experimental design C.R.D. (Factorial) 4. Number of Factors 3 (Three) 1. Salt concentration 2. Varieties 3. Method of sugar addition 5. No. of treatments combination 12 (Twelve) 6. No. of repetitions 3 (Three) 7. Name of crop and variety Ber Umran and Apple Ber 8. Total Number of combinations 36 (Thirty six) 9. Wt. of each packet per combination 500 g 10. Time period for storage Up to 6 months 3.2.2 TREATMENT DETAILS FOR CANDY The experiment comprised of 12 treatment combinations consisting 3 level of salt concentration, 2 levels of varieties and 2 levels of method of sugar addition. The details of various treatments and their combinations are presented below: Factor A: Salt concentration 1. S 1 : Without salt (NaCl) 2. S 2 : 2% salt (NaCl) solution 3. S 3 : 4% salt (NaCl) solution 35

Factor B: Varieties 1. V1: Umran 2. V2: Apple Ber Factor C: Method of sugar addition 1. P 1 : Direct addition 2. P 2 : Sugar syrup (50 0 Brix) 3.2.3 TREATMENT COMBINATIONS FOR CANDY 1. S 1 V 1 P 1 : Without NaCl + Umran + Direct sugar addition 2. S 1 V 1 P 2 : Without NaCl + Umran + Sugar syrup 50 0 Brix 3. S 1 V 2 P 1 : Without NaCl + Apple ber + Direct sugar addition 4. S 1 V 2 P 2 : Without NaCl + Apple ber +Sugar syrup 50 0 Brix 5. S 2 V 1 P 1 : 2% NaCl + Umran + Direct sugar addition 6. S 2 V 1 P 2 : 2% NaCl + Umran + Sugar syrup 50 0 Brix 7. S 2 V 2 P 1 : 2% NaCl +Apple ber + Direct sugar addition 8. S 2 V 2 P 2 : 2% NaCl + Apple ber + Sugar syrup 50 0 Brix 9. S 3 V 1 P 1 : 4% NaCl + Umran + Direct sugar addition 10. S 3 V 1 P 2 : 4% NaCl + Umran + Sugar syrup 50 0 Brix 11. S 3 V 2 P 1 : 4% NaCl + Apple ber + Direct sugar addition 12. S 3 V 2 P 2 : 4% NaCl + Apple ber + Sugar syrup 50 0 Brix 3.2.4 EXPERIMENTAL DESIGN: The experiment was laid out in a Completely Randomized Design with factorial concept and treatments repeated three times. 3.2.5 SAMPLE PREPARATION The ber fruits reached to full maturity stage with green, pale yellow, golden yellow colors respectively were selected. These fruits were dipped in water for discarding light and infested fruits. These fruits were then cleaned under running tap water to remove impurities and dust from the surface of the ber fruits and they were graded manually. 36

3.2.5.1 Peeling and cutting in to slices for ber candy Ber fruits were peeled with the use of hand peeler. After peeling the whole fruit, pieces were cut in to slices of uniform size. 3.2.5.2 Soaking in salt solution for one day The ber fruit slices are after cutting soaking in salt solution of 2 percent, 4 percent and without salt for one day 3.2.5.3 Blanching The ber slices were blanched in boiling water at 98 ± 2 0 C for 2-3 min as per the method suggested by Kadam et al., 1991. 3.2.5.4 Preparation of sugar syrup The experiment was carried out with two methods of sugar addition concentrations of i.e., direct addition of sugar and sugar syrup at 50 0 Brix. The sugar powder direct added in slices and osmotic solution of 50 0 Brix sucrose syrup was prepared by dissolving 1 kg sugar in one liter of water respectively whereas, The hand refractometer having range from 50 to 80 0 Brix (Erma, Japan) were used for the measurement of sucrose syrup concentration. 3.2.5.5 Slices are drained with sugar syrup Ber fruit slices are after blanching removal of total salt from the slices are addition of sugar two methods, first one with direct addition of sugar powder for 6 combinations and remaining 6 combination are dipped in sugar syrup at 50 0 Brix all the combinations are for increasing 10 0 Brix by heating up to 50 0 Brix for 3-4 days 3.2.5.6 Oven drying The laboratory scale oven dryer available in the Department of Horticulture was used for the drying of candies of different fruits. Drying was carried out at 60 0 C and the drying was done up to 24-36 hours. 3.2.5.7 Packaging and storage of candy The packaging of candy of each treatment under study was done in polypropylene (pp) pouches of 50 μ thickness of 100 g packets and was stored during 37

February- September, 2016 at room temperature (18.2-38.2 0 C and 17.2-79.3 % RH) for a period of 6 months & observations were recorded at an interval of 30 days. 3.2.6 OBSERVATIONS RECORDED: Prepared Candy of ber was subjected to following periodical observations. Prepared Candy: The observation on following physical and chemical characteristic of Candy were recorded at monthly interval up to 6 months of storage 3.2.6.1 Sensory Rating: (1) Color (2) Flavor (3) Taste (4) Texture (5) Overall acceptance 3.2.6.2 Biochemical Parameters: (1) Total soluble solids ( 0 Brix) (2) Titrable acidity (%) (3) Ascorbic acid (mg/100 g) (4) Sugars: (i) Reducing sugar (%) (ii) Non-reducing sugar (%) (iii) Total sugar (%) 3.2.6.3 Microbial count of the Candy: Bacterial and fungal colony was carried out by serial dilution methods. 3.2.6.5 Economic feasibility: Economics of treatments was worked out. 38

3.2.7 METHODOLOGY USED FOR OBSERVATION: 3.2.7.1 Physical characteristics: Recovery of slices: To determine the pulp recovery percentage known weight of fruits in each treatment were taken and juice was extracted by different method as per treatment combination. The yield of pulp in each treatment weighted. Then the recovery percentage of pulp was calculated by the formula: Weight of slices (g) Recovery (%) = ------------------------------- X 100 Weight of fresh fruit (g) 3.2.7.2 SENSORY RATING: Prepared juice and RTS under different treatments were evaluated for sensory characteristics viz., color, taste, flavor, appearance, cloudiness, product setting at bottom and overall acceptability. Each attribute was given a separate score of 10 points. Product scoring less than 5 points out of 10 points was treated as unacceptable from the quality point of view. Sensory panel consisted of 5 trained panelists and the panelists were instructed to evaluate the samples as per hedonic scale procedure as described by Ranganna (1979) in format detailed below: Treatment color Taste Flavor Texture Overall acceptance 1 to 12 3.2.7.3 CHEMICAL CHARACTERISTICS: 1) Total soluble solids (TSS) The total soluble was measured with the help of hand refractometer at room temperature and the method suggested by Ranganna, 1979 was followed. The few drops of the crushed candy were put on prism of the hand refractometer to obtain the total soluble solids. The results were expressed in terms of 0 Brix 39

2) Acidity (%) To estimate Titrable acidity the analytical method given by Ranganna (1979) was followed. One gram of crushed candy sample was taken in a beaker and diluted by adding 10 ml of distilled water and the solution filtered with the help of muslin cloth. In a burette 0.1 N NaOH was filled and the sample was titrated against the standard 0.1 N NaOH solution till the light pink color appears. The estimated value then expressed in terms of percent acid by adopting the following formula. T N Q E Acidity (%) = 100 V W Whereas, T = Titer value (ml) N = Normality of alkali solution V = Volume made up (ml) E = Equivalent weight of acid (g) Ws = Weight of sample (g) Vs = Volume of sample taken for titration (ml) 3) Ascorbic acid (mg/100g) Ascorbic acid was determined by using 2-6 dichlorophenol indophenol method as described by Sadasivam and Manickam (1996). The crushed candy pulp of 10 g was transferred to volumetric flask and volume was made up to 100 ml by adding 3 % metaphospheric acid solution. After 30 minutes, the suspension was filtered through What man No. 1 filter paper. From the filtrate, 5 ml aliquot was taken and titrated against standardized dye solution. Titration was continued till the light pink color persisted for 15 seconds. Initially, the dye solution was standardized by titrating against standard ascorbic acid solution and the dye factor was calculated. Ascorbic acid content was calculated by using following equation. Titrate Dye factor Volume made up Ascorbic acid (mg 100 g -1 ) = Aliquot taken for estimation Weight or volume of sample taken for estimation 100 40

4) Reducing sugar (%) Reducing sugar was estimated by using Nalson Smogyi methodas reported by Sadasivam and Manickam (1996). In this method, DNSA reagent was used and absorbance was noted at 565 nm and accordingly graph was plotted against standard glucose solution and the reducing sugar was calculated by using the following formula. The equipment used for the estimation of total sugar is shown in Plate 3.9. Glucose equivalent 0.05 x Total volume made up Reducing sugar (%) = 100 Titer x Weight of sample taken 5) Total sugar (%) For the estimation of total sugars, the phenol sulphuric acid method to estimate total sugar was described by Krishnaveni et al. (1984). In hot acidic medium glucose is dehydrated to hydroxyl methyl furfural. This forms a green colored product with phenol and has absorption maximum at 420 nm. For estimation of total sugars 100 ml of the sample was taken into a boiling tube. Hydrolyze by keeping it in a boiling water bath for three hours with 5 ml of 2.5N-HCl and cool to room temperature. Neutralized it with solid sodium carbonate until the effervescence ceases. Make up the volume to 100 ml and centrifuge. Pipette out 0.2, 0.4, 0.6, 0.8 and 1ml of the working standard into a series of test tubes. Pipette out 0.1 and 0.2ml of the sample solution in two separate test tubes. The volume in each tube made up to 1ml with water. 1 ml of phenol solution added in each tube. 5 ml of 96 % sulphuric acid added in each tube and shakes well. After 10 min shake the contents in the tubes and place in water bath at 25-30 C for 20min. Read the color at 490 nm. The amount of total carbohydrate present in the sample solution calculated by using the standard graph. Percentage of total sugar was calculated according to following formula. Total sugar (% ) = Sample reading Glucose equivalent Volume made up 100 10 6 Weight of pulp Aliquot taken 41

6) Non reducing sugar (%) The amount of non-reducing sugar was obtained by subtracting reducing sugars from the amount of total sugars and non-reducing sugar content was expressed in terms of percentage Non reducing sugar (%) = Total sugar (%) Reducing sugar (%) 3.2.7.4 METHOD OF MICROBIAL COUNTS The stored candy was subjected to microbial analysis for total viable count using Nutrient agar for bacterial analysis and Potato Dextrose Agar for fungus analysis as per the standard procedure given by Ranganna, (1979). a. Preparation of dilutions Sample of one gram was aseptically taken into a sterilized glass test tube and transferred to 9 ml sterile phosphate buffer dilution blank to obtain 1:10 dilution. Subsequently, 0.1 ml of above prepared dilution was used for making further dilutions in 9.9 ml phosphate buffer tubes. Suitable dilutions were prepared and poured in a set of sterile petri plates in duplicates. b. Total viable count One ml of suitable dilution from each sample prepared as described in previous section was used for plating in duplicates and thereafter 15 ml of molten Nutrient Agar was poured aseptically to plates. The contents were mixed and plates were cooled. The plates then were inverted and incubated in an incubator maintained at 37 ± 0.5 0C for 24 hour and number of colony forming units (CFU/g) was recorded. Similarly, for fungus analysis, 1 ml of suitable dilution from each sample prepared as described in previous section was used for plating in duplicates and thereafter 15 ml of molten Potato Dextrose Agar was poured aseptically to plates. The contents were mixed and cooled. The plates then were inverted and incubated in an incubator maintained at 37 ± 0.5 0C for 96 hour and number of colony forming units (CFU/g) was recorded. Total CFU/ml = Number of colonies Volume plated x Dilution 42

3.13 ECONOMICS OF THE TREATMENTS The economics of different treatments was calculated on the basis of the treatment. The cost benefit ratio (CBR) was calculated on the basic of the formula given below. CBR = Price of the product Rs. / Kg Total cost Rs. / Kg 3.14 STATISTICAL ANALYSIS The data were analyzed statistically according to the analysis of variance techniques as suggested by Cochran and Cox (1950). The analysis of variance for different parameters is presented in Appendices. The critical difference (CD) was calculated to access the significance, non- significance of difference between treatments mean, wherever it was found significant through F test at 5% level of significance. 43

Flow chart showing the procedure to prepare ber candy Unripe and cleaned ber fruits Washing Peeling and pricking Cutting into rectangular pieces (3 cm and 2 cm thick) Steeping into 2% salt solution for 24 hrs. Washing thoroughly with fresh water Blanching at 90⁰C for 10 min Cooling into fresh water Steeping pieces in sugar syrup (40⁰ Brix TSS) for a day with different levels of citric acid Removal of pieces Increasing strength of syrup to 60⁰ Brix TSS by evaporation Dipping of removed pieces in syrup 44

Repeating the process and raising strength of syrup by 5⁰ Brix TSS at alternate day to a final strength at 75⁰ Brix TSS Steeping in 75⁰ Brix for 3 days Removal of adhering syrup by dipping in hot water Tray/cabinet drying at 60⁰ for 6-6.5 hrs. Packaging of ber candies in glass jar, PET jars and polypropylene pouches Storing at ambient temperature Draining 45