Pharmacognostic studies of selected medicinal plants

Transcription

1 CHAPTER-III Pharmacognostic studies of selected medicinal plants 3.1 Introduction Standardization of natural products is a complex task due to their heterogeneous composition, which is in the form of whole plant, plant part or s obtained thereo f. To ensure reproducible quality of herbal products, proper control of starting material is utmost essential. The first step towards ensuring quality of starting material is authentication. Thus, in recent years there has been a rapid increase in the standardization of selected medicinal plants of potential therapeutic significance (Reddy and Venkatesh, 2004). Despite the modern techniques, identification of plant drugs by pharmacognostic studies is more reliable. According to the World Health Organization, (WHO, 1998), the macroscopic and microscopic description of a medicinal plant is the first step towards establishing the identity and degree of purity of such materials and should be carried out before any tests are undertaken. The term Pharmacognosy was coined by Seydler, a German scientist, which has root from the green word Pharmakon a drug and gignoso, to acquire knowledge is actually deals with the structural, physical, chemical and biological properties of crude drugs along with their history, method of cultivation and its preparation (Evans,1989). During the Vedic age any such person who collected drugs was much reverend and was known as "parameshti prajapati". In Greece, the same people were known as "Rhizotomi". Charaka stressed the importance of correct identification of a plant as vital for treatment. He states that simply confirming the name of the plant does not help in any ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 243

2 way for treatment and that one should be well versed with morphological characters. To acquire knowledge of plants especially through its name, morphological characters, uses, etc., one should associate/interact with the hill tribes and cowherds of the forest. The forest dwellers have a clear idea about the measurements (pramana), colour (varna), physical characters (akriti) and the specific reproductive characteristic (jatilinga) of each plant. Dhanvantri nighantu, the ancient master-mind had the unique intuitive and innovative powers in fixing up the identity of various plants with reference to the latent properties of the plant on one side and prescribing them for particular ailments all in scientific objectivity and precision. In addition, the raw material standardization can be done by correct identification and evaluation of organoleptic characters like colour, odour, taste, ash and material content, and the fluorescent studies of powdered drug are of utmost importance to distinguish the different varieties of the plant. Selected 07 medicinal plants i.e., Annona reticulata L., Annona squamosa L., Corchorus olitorius L., Euphorbia tirucalli L., Ficus racemosa L., Pongamia pinnata L., Vitex negundo L. parts were having many minor variation creating difficulties in their proper identification. Hence, the present attempt was made to distinguish the plant parts through pharmacognostic studies. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 244

3 3.2 Review of literature The selected seven ethno medicinal plants for the present investigation is used not only in this region for curative purposes but also used all over the world. Goel and Aswual (1990) have made an effort to document the less known medicinal plants which are used in indigenous folklore from Northern India. Murthi and Sheshadri (1941) have studied the root characters of Decalepis hamiltoniii and K. indicus and revealed that these two belong to taxonomically different genera. Chase and Pratt (1949) have investigated the fluorescence of 151 identified powdered drugs. They also presented a key for the identification of powdered vegetable drugs based on fluoresence studies. Prasad et al. (1959) made a comparative study of roots of Withanla somnifera and Withania coagulans. Dhalla et al. (1961) have studied the pharmcognosy of the roots of both wild Withania somnifera and cultivated Withania ashwagandha. Prasad and Wahi (1965) have studied pharmacognosy characteristics of the root of Hemidesmus indicus. Krishnamurthy and Sundaram (1967) have showed the Morphology of the foliar epidermis of Hemidesmus indicus in which they have described the type of stomata, cell type and trichomes. Wahi et al. (1971) and Nayar (1979) made comparative studies of macroscopic and microscopic characters of the root of four plants. Yoganarasimhan et al. (1981) have given the key characters of some medicinally important plants like, Tagara, Bharangi, Ishwari, which will help to identify these plants scientifically to avoid adulterations. Surunge and Deokule (1986) and Mulla and Datwani (1994) have reported pharmacognstic characters and evaluated the ash values, soluble and insoluble ash, ive value of rhizome of Trichodesma indicum R.Br. and Curculigo recurvata DR. respectively. Fadeyi et al. (1989) have studied four species of the genus Boerhavia occurring in Nigeria with respect to anatomical and phytochemical aspects. Dave and Kurjachen (1991) have made an extensive comparative anatomical work on the fruits of Cryptolepis buchanani and Hemidesmus indicus. Deokule (1991 and 1995) ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 245

4 have also emphasized on morphological, histological and phytochemical studies in his work on some members of Leguminaceae like Vigna trilobata, Crotalaria return and Wagatea spicata. Wang et al. (1994) have made thorough investigation of original plants and commercial products of the traditional Chinese drug Jiuyandu huo. Wang et al. (1995) have published a review article on recent advances in pharmacognosy research in China. Lohar et al. (1995) have done standardization of Ptelia trifoliaia Linn by using some phytochemical and pharmacognostical methods. Fillippini et al. (1995) have done comparative investigations of finely powdered sample of commercially available capsules of Herba equiseti with that of authenticated plant material Equisetum arvense L. They concluded that with the help of scanning electron microscopic technique it is possible to identify the drug even at the species level. Hamano et al. (1996) have evaluated 48 samples of Cinnamon barks sold in the markets in Japan. Lim et al. (1996) have done comparative morphological and histological studies on the Chinese crude drug Huang qi and its adulterants. Dutta and De (1995) have studied pharmacognostic characters of pods of Cassia fistula and Cassia javanica as the later is often found as adulterant. Krishnan Nambiar et al. (1998) have published a paper on pharmacognostic studies on Saraca asoca in comparison with Polyalthia longifolia a common adulterant of asoca. Madhavan (1999) made an attempt to suggest a few appropriate standardization techniques to strengthen Ayurvedic system. Krishnan Nambiar (2000) have studied pharmacognostical characters of Asparagus racemoses collected from different parts of Kerala. Satakopan (2000) has focused on a few modern techniques which can be implied to control the quality of Ayurvedic, Siddha and Unani crude drugs. There are many more reports on standardization of crude drugs, and to mention a few, the works of Jaffer et al. (1998), Joshi and Jayshree (1988), Pattanshetty et al. (1989) Barthwal and Srivastava (1990), Toker et al. (1995) are noteworthy. They found that the ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 246

5 crude drugs were contaminated by Altemaria, Aspergillus, Curvularia, Fusarium, Pennicillium and Rhizopus. Out of 37 samples screened 30 were found to be aflatoxin positive. The same authors in the year 1990 found that the active principles have been deteriorated due to fungal association and stored under different relative humidities. Kulkarni and Lipnis (1992) have made an attempt to decontaminate some crude drugs by gama radiations and revealed that the irradiation at 10 kg and 15 kg dose of gama radiation could completely decontaminate the samples without altering their physicochemical parameters and active phytochemical constituents. Fischer et al. (1993) have analyzed phototherapeutic products of oral dosage forms marketed in Brazil regarding microbiological quality. Huang and Blume (1994) have tested a few medicinal plant materials for the microbial contamination. Kivman et al. (1998) have studied the problems in the standardization of the microbial purity of raw medicinal plant material. Limyati and Juniar (1998) have assessed Jamu Gendong, a kind of traditional medicine in Indonesia for microbial contamination of its raw materials and end products. Krishnaveni and Saaanth Rani have studied pharmacognostic parameters for the leaves of Sapindus trifollliatus Vahl have studied with the aim of drawing the pharmacopial standard for the species identification. Sugumaran and Vetrichelvan have pharmacognostic profile of Bauhinia purpurea. According them ash value and ive values are helpful in the differentiation from other related species. Divyakanth and Vimal Kumar, 2008 have focused on pharmacognostic characteristic of Neolamarkia cadamba (roxb) in that they have studied microscopic, macroscopic characteristics of the plant viz., morphological studies, such as shape, size, apex, surface, base, margin, venation, taste and odour of leaves, as a part of quantitative microscopy, stomatal number, stomatal index, vein islet number and vein termination number were determined by using fresh leaves of the plant. The total ash, water-soluble ash, acid-insoluble ash, sulphated ash, alcohol- and water-soluble ive ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 247

6 value and foaming index were determined. Satkopan (2000) have focused on a few modern technique which can be implied to control the quality of Ayurvedic, Siddha and Unani crude drugs. Nirmaladevi and Periyanayagam (2008) have studied the micro morphological studies of the plant Plectranthus amboinicus for identification of medicinal plants and also have studied SEM for better understanding of structural details and to assist in the solution of taxonomic problem. Nawagish et al., 2007 have studied the preliminary and pharmacological standardization of Lawsonia inermis Lin. Venkatesh et al. (2008) have studied the pharmacognostic studies of Dodonaea viscose to overcome the taxonomic confusion among Dodonaea viscosae varieties. In endemic plant of Jatropa species pharmacognostic studies were performed by Uthayakumari and Sumathy (2011). Prasad, Kadam et al., (2012) have studied the pharmacognostic studies of Mimusops elengi to overcome drug standardisation among Mimusops elengi varieties. Sumitra Chanda (2014) has given detailed review on importance of pharmacognosy in medicinal plants. Kamaruz Zaman and Kalyani Pathak (2013) have studied the pharmacognostic studies of A. reticulta to overcome drug standardisation among Annona species, Gorwadiya (2010) drug standardised in F. racemosa, Kumar et al., (2013) have studied the pharmacognostic studies of P. pinneta, Ahirrao et al., (2011) have pharmacognostic profile of V. negundo. Seed fractions of Corchorus olitorious pharmacognostic studies were recorded by Maxwell Osaronowen Egua (2014). ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 248

7 3.3 Materials and methods Organoleptic evaluation In organoleptic evaluation, various sensory parameters of the plant material, such as weight, colour, odour, and taste of the leaves were recorded. It includes conclusions drawn from studies resulted due to impressions on organs of senses. Florescent studies of powdered drug (Chase, Pratt, 1948) The dried powder of selected plants parts was sieved through the sieve plate No.120 and was used for fluorescent studies. A pinch of this powder was taken in a clean test tube with about 10ml of solvent. Likewise, several tubes were made by adding various solvents like, alcohol, methanol, ethyalacetate, chloroform, benzene, hexane, acetone, water, hydrochloric acid and conc. sulphuric acid. All the tubes were shaken well and incubated for about 30min. The colour of the drug solution thus obtained were observed for their characteristic colour reaction under the visible light(fluorescent tube) and the ultraviolet light(uv 356 ) and were recorded by comparing with standard colour charts(chase and Pratt, 1949). Determination of ive value: (Raghunathan, 1982) Accurately weight 100g of powdered plant materials were ed successively in a Sox-let or with solvent Petroleum ether, Chloroform, Methanol, Alcohol and distilled water separately. The obtained s were allowed to dry at room temperature. After complete evaporation of the solvent, weight, colour and nature of the s were noted. Weight of the residue obtained Extractive value (%) = x 100 Weight of the plant material taken ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 249

8 Determination of Ash value: (W.H.O. 2002) Take a known weight of the dried plant material in a pre weighed silica crucible. Heat it on a Bunsen burner at a low flame, till the plant material gets charred. Then it is transferred to a muffle furnace and heated strongly at a dull red heat ( C) till a white ash is obtained. It is cooled in a desiccators for about 15 to 30min, and weighed using electronic balance and the readings are noted down (Raghunathan, 1982). Z-X Total ash value of the sample = x 100 Y Where, Weight of the empty crucible = Z, Weight of crucible with Ash = X, Weight of the plant material (g) = Y Z-X Weight of the dish + ash = x 100 Determination of acid insoluble ash: Y The above obtained total ash was boiled for 15 ml with 25ml of 25% hydrochloric acid. This was filtered and the insoluble matter was collected on ash less Whatman filter paper, ignited in sintered crucible, cooled and then kept in a desiccators for 15minute. The residue was weighed in electronic balance and the acid insoluble ash was calculated. 100 Acid insoluble ash value of the sample = x A Where, A = Weight of the residue obtained, Y = Weight of the plant material. Y ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 250

9 3.4 Experimental results Annona reticulata L. Organoleptic studies The detailed study of organoleptic characters of s, ash values and the florescent behaviour of Annona reticulata L. leaf can be easily identified and further distinguished from their possible adulterants and substituent s by these parameters. Organoleptic studies of s The successive Soxhlet s viz. petroleum ether. Chloroform ethyalacetate and 98% methanol s of A. reticulata L. yields were determined and their differential values were recorded in the Table Among all the s obtained, the 98% methanol was having higher percentage of yield of compounds then other s. The weight of leaf s are 1.50 (pet. ether), 1.80 (chloroform), 1.50 (ethyl acetate) and 2.80 (98% methanol) percent respectively. It is evident from these results, that the 98% methanol solvent has higher ive values followed by chloroform, pet-ether and ethyl acetate. Table-3.1: Extractive values and organoleptic characters of Annona reticulata L. s. Sl no Plant part used Solvents Weight of the s Colour of the Taste of the Nature of the Odour of the 1 LEAVES PE 1.50 Yellow Bitter waxy Sweet 2 CHCL Dark Green Bitter sticky Aromatic 3 EtOH 1.50 Light green Bitter sticky Aromatic 4 98% Methanol 2.80 Dark brown Salty Bitter waxy Aromatic The crude s of A. reticulata leaf have shown a wide range of colour. The petroleum ether shows yellow. While, the chloroform exhibited dark green colour. Whereas, the ethyl acetate shows light green colour and 98% methanol shows dark brown colour (Table: 3.1). A. reticulata leaf s taste varies from to such as tasteless, sweet. Bitter, light bitter and salty. Of these the petroleum ether, chloroform and ethyl acetate exhibited bitter and 98% methanol show salty bitter in taste. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 251

10 Further the nature of these s varies, pet. ether is waxy in nature whereas chloroform and ethyl acetate s shows sticky. Whereas the 98% methanol is waxy. Similarly, the s were exhibiting variety of odour. The petroleum ether exhibiting sweet and chloroform, ethyl acetate and 98% methanol s exhibited the aromatic odour. Determination of total ash and acid insoluble ash contents In the present investigation, the total ash and acid insoluble and water insoluble ash contents of A. reticulata leaf was studied (Table: 3.2). Table-3.2: Total ash and acid insoluble ash content in Annona reticulata L. (mg/100g) Sl no parts Total ash Acid insoluble ash contents Water insoluble ash contents 1. Leaf The total ash value recorded as (07%), acid insoluble ash value determined as 3.61% and water insoluble ash value recorded 2.42%. This variation in the content of ash is probably due to the nature of metabolites they possess. Fluorescent studies of powered drugs The characteristic colour behaviour of dried and powdered drugs and powder dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone, ethyl acetate, Conc. HC1 and H 2 So 4 (5%) i.e., leaf of A. reticulata was observed both under the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions thus emitted fluorescence light are tabulated in Table: 3.3. Table-3.3: Fluorescent studies of powdered drugs Annona reticulata L. Sl no Treatment visible UV 365 nm 1 Powder Dark green Onion pink 2 Pet. Ether Blackish green Ash 3 Chloroform Yellow green Opaline green 4 Ethanol Light green Light pink 5 Eather Dark green Light pink 6 Acetone Light green Light pink 7 Ethyl acetate Brownish green Light onion ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 252

11 8 Benzene Light green Black 9 Conc. HCl (5%) Light brown Black 10 Conc. H2So4 (5%) Light brown Pale white The powdered drug solutions have exhibited a wide range of fluorescence colours under the UV light and visible light. The petroleum ether drug solutions have shown the fluorescence colours such as blackish green in visible light and ash in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence colours as yellow green in visible in Opaline green UV light. While ethanol (95%v/v) treated drug solutions exhibited wide range of fluorescence colours light green in visible in UV light pink. However, the ether drug solutions exhibited dark green in visible light and light pink in UV light. In acetone drug solution shows the colours light green in visible light and light pink in UV light, In ethyl acetate drug solution shows the colours chassis red in visible light and light pink in UV light, similarly benzene drug solution exhibited light green and black. Whereas, dilute HC1 (5%) acid drug solution indicating the light brown and black and in dilute H 2 So 4 (5%) acid drug solution shows light brown and pale white respectively. The colour reaction varies from non-polar to high polar solvents (Plate-3.1) Annona squamosa L. Organoleptic studies The detailed study of organoleptic characters of s, ash values and the fluorescent behaviour of A. squamosa L. leaf can be easily identified and further distinguished from their possible adulterants and substituent s by these parameters. Organoleptic studies of s The successive Soxhlet s viz. petroleum ether. Chloroform ethyalacetate and 98% methanol s of A. squamosa L. yields were determined and their differential values were recorded in the Table Among all the s obtained, the 98% methanol was having higher percentage of yield of compounds then other s. The weight of leaf s are 0.85 (pet. ether), 1.6 (chloroform), 0.45 (ethyl acetate) and 3.23 (98% methanol) percent respectively. It is evident from these results, that the 98% ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 253

12 methanol solvent has higher ive values followed by chloroform, pet-ether and ethyl acetate. Table-3.4: Extractive values and organoleptic characters of Annona squamosa L. s Sl. no Plant part used Solvents Weight of the s(% dry weight in gm) Colour of the Taste of the Nature of the Odour of the 1 Leaf PE 0.85 yellow Slight Bitter Light sticky Sweet 2 CHCL3 1.6 Light green 3 EtOH 0.45 Brownish green 4 98% Methanol 3.23 Dark Brown Sweet bitter sticky Sweet Bitter Light sticky Aromatic Bitter waxy Aromatic The crude s of A. squamosa L. leaf have shown a wide range of colour. The petroleum ether shows yellow. While the chloroform exhibited light green colour. Whereas, the ethyl acetate shows brownish green colour and 98% methanol shows dark brown colour (Table: 3.4). A. squamosa L. leaf s taste varies from to such as tasteless, sweet. Bitter, light bitter and salty. Of these the petroleum ether and chloroform s exhibited slight bitter, sweet bitter respectively. The ethyl acetate and 98% methanol s bitter in taste. Further the nature of these s varies, pet. ether and ethyl acetate s were light waxy in nature. Whereas chloroform in nature sticky. While the 98% methanol was in waxy. Determination of total ash and acid insoluble ash contents In the present investigation, the total ash and acid insoluble and water insoluble ash contents of A. squamosa L. leaf was studied (Table: 3.5.). Table-3.5: Total ash and acid insoluble ash content in Annona squamosa L. (mg/100g) Sl no parts Total ash Acid insoluble ash contents Water insoluble ash contents 1. Leaf ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 254

13 The total ash value recorded as 6.84%, acid insoluble ash value determined as 0.94% and water insoluble ash value recorded 3.54%. This variation in the content of ash is probably due to the nature of metabolites they possess. Fluorescent studies of powered drugs The characteristic colour behaviour of dried and powdered drugs and powder dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone, ethyl acetate, Conc. HC1 and H 2 So 4 (5%) i.e., leaf of A.squamosa L. was observed both under the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions thus emitted fluorescence light are tabulated in Table: 3.6. Table-3.6: Fluorescent studies of powdered drugs Annona squamosa L. Sl no Treatment visible UV 365 nm 1 Powder Green Dark brown 2 Pet. Ether Pastel green Light ash 3 Chloroform Mint green Sky blue 4 Ethanol Light green Slight ash 5 Ether Bus Green Opaline green 6 Acetone Pista green Dark blue 7 Ethyl acetate Slight green Onion 8 Benzene Pastel green Pink 9 Conc. HCl (5%) Pale cream Blackish 10 Conc. H2So4 (5%) Gold brown Light blackish The powdered drug solutions have exhibited a wide range of fluorescence colo urs under the UV light and visible light. The petroleum ether drug solutions have shown the fluorescence colours such as pastel green in visible light and light ash in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence co lours as mint green in visible in sky blue UV light. While ethanol (95%v/v) treated drug solutions exhibited wide range of fluorescence colours light green in visible in UV light ash. However, the ether drug solutions exhibited bus green in visible light and opaline green in UV light. In acetone drug solution shows the colour pista green in visible light and dark blue in UV light, In ethyl acetate drug solution shows the colours Slight green in visible light and Onion in UV light, similarly benzene drug solution exhibited Pastel green and Pink. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 255

14 Whereas, dilute HC1 (5%) acid drug solution indicating the light Pale cream and Blackish and in dilute H 2 So 4 (5%) acid drug solution shows Gold brown and Light blackish respectively. The colour reaction varies from non-polar to high polar solvents (Plate-3.1) Corchorus olitorius L. Organoleptic studies The detailed study of organoleptic characters of s, ash values and the florescent behaviour of Corchorus olitorius L. seed can be easily identified and further distinguished from their possible adulterants and substituent s by these parameters. Organoleptic studies of s The successive Soxhlet s viz. petroleum ether. Chloroform ethyalacetate and 98% methanol s of C. olitorius L. yields were determined and their differential values were recorded in the Table Among all the s obtained, the 98% methanol was having higher percentage of yield of compounds then other s. The weight of seed s are 0.92 (pet. ether), 1.83 (chloroform), 2.51 (ethyl acetate) and 4.74 (98% methanol) percent respectively. It is evident from these results, that the 98% methanol solvent has higher ive values followed by ethyl acetate chloroform and pet-ether. Table-3.7: Extractive values and organoleptic characters of Corchorus olitorius L. seed s. Sl. no Plant part used Solvents Weight of the s Colour of the Taste of the Nature of the Odour of the 1 Seed PE 0.92 Creamy Bitter Creamy Sweet yellow 2 CHCL Slight Bitter Creamy Sweet greenish 3 EtOH 2.51 Light Bitter waxy Aromatic Brownish 4 98% Methanol 4.74 Brown Bitter waxy Aromatic The crude s of C. olitorius L. seed have shown a wide range of colour. The petroleum ether shows creamy yellow. While, the chloroform expose the slight greenish colour. Whereas, the ethyl acetate shows light brownish colour and ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 256

15 98% methanol shows brownish colour. (Table: 3.7). C. olitorius L. seed all the four solvent s shows bitter in taste. Further the nature of these s varies, pet.ether and chloroform s were creamy in nature. Whereas the ethyl acetate and 98% methanol s shows waxy in nature. Determination of total ash and acid insoluble ash contents In the present investigation, the total ash, acid insoluble and water insoluble ash contents of C. olitorius L. seed was studied (Table: 3.8). Table-3.8: Total ash and acid insoluble ash content in Corchorus olitorius L. seed (mg/100g) Sl. no part Total ash Acid insoluble ash contents Water insoluble ash contents 1. Seed The total ash value recorded as (6.18%), acid insoluble ash value determined as 2.43% and water insoluble ash value recorded 1.14%. This variation in the content of ash is probably due to the nature of metabolites they possess. Fluorescent studies of powered drugs The characteristic colour behaviour of dried and powdered drugs and powder dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone, ethyl acetate, Conc. HC1 and H 2 So 4 (5%) i.e., seeds of Corchorus olitorius L. was observed both under the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions thus emitted fluorescence light are tabulated in Table: 3.9. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 257

16 Table-3.9: Fluorescent studies of powdered drugs in Corchorus olitorius seed Sl. no Treatment visible UV 365 nm 1 Powder Pale cream with Blackish Light yellowish 2 Pet. Ether Transparent Green Pale Yellow precipitate 3 Chloroform Light green Light greenish 4 Ethanol Transparent Transparent 5 Ether Light brownish Light yellowish 6 Acetone Transparent Transparent 7 Ethyl acetate Blackish Blue 8 Benzene Transparent Transparent 9 Conc. HCl (5%) Highly Transparent White creamy 10 Conc. H2So4 (5%) Slight Pinkish Transparent Creamy The powdered drug solutions have exhibited a wide range of fluorescence colours under the UV light and visible light. The petroleum ether drug solutions have shown the fluorescence colours such as transparent green in visible light and pale yellow precipitate in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence colours as light green in visible in light greenish UV light. While ethanol (95%v/v) treated drug solutions exhibited wide range of fluorescence colours transparent in visible in UV light Transparent. However, the eather drug solutions exhibited light brownish in visible light and light yellowish in UV light. In acetone drug solution shows the colours transparent in visible light and transparent in UV light, In ethyl acetate drug solution shows the colour blackish in visible light and blue in UV light, similarly benzene drug solution exhibited transparent and transparent. Whereas, dilute HCl (5%) acid drug solution indicating the light brown and black and in dilute H 2 So 4 (5%) acid drug solution shows highly transparent and white creamy respectively. The colour reaction varies from non-polar to high polar solvents (Plate-3.1) Euphorbia tirucalli L. Organoleptic studies The successive Soxhlet s viz. petroleum ether. Chloroform ethyalacetate and 98% methanol s of E. tirucalli L. yields were determined and their differential values were recorded in the Table Among all the s obtained, the ethyl acetate was having higher percentage of yield of compounds then other s. The ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 258

17 weight of leaf s are 0.97 (pet. ether), 1.21 (chloroform), 2.94 (ethyl acetate) and 2.31 (98% methanol) percent respectively. It is evident from these results, that the ethyl acetate solvent has higher ive values followed by 98% methanol, chloroform and pet-ether. Table-3.10: Extractive values and organoleptic characters of E. tirucalli L. s. Sl no Plant part used Solvents Weight of the s Colour of the Taste of the Nature of the Odour of the 1 Leaf PE 0.97 Creamy Bitter waxy Sweet yellow 2 CHCL Greenish Bitter Creamy Sweet 3 EtOH 2.94 Light Bitter waxy Aromatic Brownish 4 98% Methanol 2.31 Brown Bitter waxy Aromatic The crude s of E. tirucalli L. leaf have shown a wide range of colour. The petroleum ether shows creamy yellow. While, the chloroform exhibited greenish colour. Whereas, the ethyl acetate shows light brownish colour and 98% methanolic shows brown colour (Table: 3.10). The four solvent s of E. tirucalli L. leaf shows bitter in taste. Further the nature of these s varies, pet. ether, ethyl acetate and 98% methanol s were waxy in nature whereas chloroform shows creamy in nature. Similarly, the s were exhibiting variety of odour. The petroleum ether and chloroform s exhibiting sweet odour. Whereas the ethyl acetate and 98% methanolic s exhibited the aromatic odour. Determination of total ash and acid insoluble ash contents In the present investigation, the total ash and acid insoluble and water insoluble ash contents of E. tirucalli L. leaf was studied (Table: 3.11). Table-3.11: Total ash and acid insoluble ash content in E. tirucalli L. (mg/100g) Sl no parts Total ash Acid insoluble ash contents Water insoluble ash contents 1. Leaf ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 259

18 The total ash value recorded as (5.39%), acid insoluble ash value determined as 2.28% and water insoluble ash value recorded 1.08 %. This variation in the content of ash is probably due to the nature of metabolites they possess. Fluorescent studies of powered drugs The characteristic colour behaviour of dried and powdered drugs and powder dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone, ethyl acetate, Conc. HC1 and H 2 So 4 (5%) i.e., leaf of E. tirucalli L. was observed both under the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions thus emitted fluorescence light are tabulated in Table: Table-3.12: Fluorescent studies of powdered drugs in E. tirucalli L. Sl. no Treatment visible UV 365 nm 1 Powder Light green Light pink 2 Pet. Ether Pista green Ash 3 Chloroform yellow Opaline green 4 Ethanol Brownish Light pink 5 Eather Yellow Light ash 6 Acetone Greenish Sky blue 7 Ethyl acetate Light yellowish Light pink 8 Benzene Pastal green Onion pink 9 Conc. HCl (5%) Brownish Dark brownish 10 Conc. H2So4 (5%) Brownish Light black The powdered drug solutions have exhibited a wide range of fluorescence colours under the UV light and visible light. The petroleum ether drug solutions have shown the fluorescence colours such as pista green in visible light and ash in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence colours as yellow in visible in opaline green UV light. While ethanol (95%v/v) treated drug solutions exhibited wide range of fluorescence colours brownish in visible in UV light pink. However, the ether drug solutions exhibited yellow in visible light and light ash in UV light. In acetone drug solution shows the colours greenish in visible light and sky blue in UV light, In ethyl acetate drug solution shows the colours light yellowish in visible light and light pink in UV light, similarly benzene drug solution exhibited pastal green and ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 260

19 onion pink. Whereas, dilute HC1 (5%) acid drug solution indicating the dark brownish and black and in dilute H 2 So 4 (5%) acid drug solution shows brownish and light black respectively. The colour reaction varies from non-polar to high polar solvents (Plate-3.1) Ficus racemosa L. Organoleptic studies The detailed study of organoleptic characters of s, ash values and the fluorescent behaviour of F. racemosa L. leaf can be easily identified and further distinguished from their possible adulterants and substituent s by these parameters. Organoleptic studies of s The successive Soxhlet s viz. petroleum ether. Chloroform ethyalacetate and 98% methanol s of F. racemosa L. yields were determined and their differential values were recorded in the Table Among all the s obtained, the 98% methanolic was having higher percentage of yield of compounds then other s. The weight of leaf s are 1.76 (pet. ether), 1.60 (chloroform), 0.67 (ethyl acetate) and 2.58 (98% methanol) percent respectively. It is evident from these results, that the 98% methanolic solvent has higher ive values followed by, pet-ether, chloroform and ethyl acetate. Table-3.13: Extractive values and organoleptic characters of Ficus racemosa L. s Sl. no Plant part used Solvents Weight of the s (% dry weight in gm) Colour of the Taste of the Nature of the Odour of the 1 Leaf PE 1.76 Pale yellow Tasteless waxy Sweet 2 CHCL3 1.6 Dark Tasteless sticky Sweet brown 3 EtOH 0.67 Dark green Bitter sticky Aromatic 4 98% Methanol 2.58 Dark brown Salty waxy Aromatic The crude s of F. racemosa L. leaf have shown a wide range of colour. The petroleum ether shows pale yellow. While, the chloroform exhibited dark brown colour. Whereas, the ethyl acetate shows dark green colour and 98% methanolic shows dark brown colour. (Table: 3.13). F. racemosa L. leaf s taste varies from to such as tasteless, Bitter and salty. Of these the petroleum ether and chloroform exhibited tasteless. The ethyl acetate having bitter ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 261

20 in taste. Whereas the 98% methanolic shown salty in taste. Further the nature of these s varies, pet. ether and 98% methanolic s were waxy in nature whereas chloroform and ethyl acetate s shown sticky in nature. Similarly, the s were exhibiting variety of odour. The petroleum ether and chloroform s exhibiting sweet odour. Whereas the ethyl acetate and 98% methanolic s exhibited the aromatic odour. Determination of total ash and acid insoluble ash contents In the present investigation, the total ash and acid insoluble and water insoluble ash contents of F. racemosa L. leaf was studied (Table: 3.14). Table-3.14: Total ash and acid insoluble ash content in Ficus racemosa L. (mg/100g) Sl no parts Total ash Acid insoluble ash contents Water insoluble ash contents 1. Leaf The total ash value recorded as (8.43%), acid insoluble ash value determined as 3.33% and water insoluble ash value recorded 7.8%. This variation in the content of ash is probably due to the nature of metabolites they possess. Fluorescent studies of powered drugs The characteristic colour behaviour of dried and powdered drugs and powder dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone, ethyl acetate, Conc. HCl and H 2 So 4 (5%) i.e., leaf of F. racemosa L. was observed both under the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions thus emitted fluorescence light are tabulated in Table-3.15: Fluorescent studies of powdered drugs Ficus racemosa L. Sl. no Treatment visible UV 365 nm 1 Powder Light green Light pink 2 Pet. Ether Pista green Ash 3 Chloroform Bus green Opaline green 4 Ethanol Light green Light pink 5 Eather Mint green Light ash 6 Acetone Pista green Sky blue 7 Ethyl acetate Light green Light pink 8 Benzene Pastal green Onion pink 9 Conc. HCl (5%) Sandal wood Dark brownish 10 Conc. H2So4 (5%) Pale cream Light black ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 262

21 The powdered drug solutions have exhibited a wide range of fluorescence colours under the UV light and visible light. The petroleum ether drug solutions have shown the fluorescence colours such as pista green in visible light and ash in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence colours as bus green in visible in Opaline green UV light. While ethanol (95%v/v) treated drug solutions exhibited wide range of fluorescence colours light green in visible in UV light ash. However, the ether drug solutions exhibited mint green in visible light and light ash in UV light. In acetone drug solution shows the colours pista green in visible light and sky blue in UV light, in ethyl acetate drug solution shows the colours light green in visible light and light pink in UV light, similarly benzene drug solution exhibited pastal green and onion pink. Whereas, dilute HCl (5%) acid drug solution indicating the dark sandal wood and dark brownish and in dilute H 2 So 4 (5%) acid drug solution shows pale cream and light black respectively. The colour reaction varies from non-polar to high polar solvents (Plate 3.1) Pongamia pinnata L. Organoleptic studies The detailed study of organoleptic characters of s, ash values a nd the fluorescent behaviour of Pongamia pinnata L. seed can be easily identified and further distinguished from their possible adulterants and substituent s by these parameters. Organoleptic studies of s The successive Soxhlet s viz. petroleum ether. Chloroform ethyalacetate and 98% methanol s of P. pinnata L. yields were determined and their differential values were recorded in the Table Among all the s obtained, the pet-ether s are having higher percentage of yield of compounds then other s. The weight of seed s are 5.21 (pet. ether), 1.42 (chloroform), 1.00 (ethyl acetate) and 0.73(98% methanol) percent respectively. It is evident from these results, that the pet-ether solvent has higher ive values followed by chloroform, ethyl acetate and 98% methanol. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 263

22 Table-3.16: Extractive values and organoleptic characters of Pongamia pinnata L. s. Sl. no Plant part used Solvents Weight of the s Colour of the Taste of the Nature of the Odour of the 1 Seed PE 5.21 yellow Tasteless Oily Sweet 2 CHCL Slight Bitter waxy Aromatic white 3 EtOH 1.00 Ash Bitter Creamy Aromatic 4 98% Methanol 0.73 White Bitter Amorphous Aromatic The crude s of P. pinnata L. seed have shown a wide range of colour. The petroleum ether shows yellow. While the chloroform exhibited slight white colour. Whereas, the ethyl acetate shows ash colour and 98% methanol s hows white colour (Table: 3.16). P. pinnata L. s taste varies from to such as tasteless, sweet. Bitter, light bitter and salty. Of these the petroleum ether exhibited tasteless and chloroform, ethyl acetate, 98% methanol show bitter in taste. Further the nature of these s varies, pet. ether is oily in nature and chloroform shows waxy. Similarly, the ethyl acetate s exhibiting creamy in nature. Whereas the 98% methanol is amorphous. Determination of total ash and acid insoluble ash contents In the present investigation, the total ash and acid insoluble and water insoluble ash contents of seed of P. pinnata L. was studied (Table: 3.17). Table-3.17: Total ash and acid insoluble ash content in Pongamia pinnata L. seed (mg/100g) Sl no parts Total ash Acid insoluble ash contents Water insoluble ash contents 1 Seeds The total ash value recorded as (07%), acid insoluble ash value determined as (2.41%) and water insoluble ash value recorded 1.93%. This variation in the content of ash is probably due to the nature of metabolites they possess. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 264

23 Fluorescent studies of powered drugs The characteristic colour behaviour of dried and powdered drugs and powder dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone, ethyl acetate, Conc. HC1 and H 2 So 4 (5%) i.e., seed of P. pinnata L. was observed both under the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions thus emitted fluorescence light are tabulated in Table: Table-3.18: Fluorescent studies of powdered drugs in Pongamia pinnata L. Seed. Sl. no Treatment visible UV 365 nm 1 Powder White Ash 2 Pet. Ether Pale rose Light pink 3 Chloroform White turbidity Ash turbidity 4 Ethanol Turbidity Blue 5 Eather Transparent Transparent 6 Acetone Pale rose Light pink 7 Ethyl acetate Chassis red Onion 8 Benzene Light sard Transparent 9 Conc. HCl (5%) Pace rose Slight blue 10 Conc. H2So4 (5%) Turbidity White The powdered drug solutions have exhibited a wide range of fluorescence colours under the UV light and visible light. The petroleum ether drug solutions have shown the fluorescence colours such as pale yellow in visible light and ash in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence colours as white turbidity in visible ash turbidity in UV light. While ethanol (95%v/v) treated drug solutions exhibited wide range of fluorescence colours turbidity in visible in UV light blue. However, the eather drug solutions exhibited transparent in visible light and transparent in UV light. In acetone drug solution shows the colours pale rose in visible light and light pink in UV light, In ethyl acetate drug solution shows the colours chassis red in visible light and onion in UV light, similarly benzene drug solution exhibited light sard and transparent. Whereas, dilute Hc1 (5%) acid drug solution indicating the pace rose and slight blue and in dilute H 2 So 4 (5%) acid drug solution shows turbidity and white respectively. The colour reaction varies from non-polar to high polar solvents (Plate 3.1). ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 265

24 3.4.7 Vitex negundo L. Organoleptic studies The detailed study of organoleptic characters of s, ash values and the florescent behaviour of V. negundo L. leaf can be easily identified and further distinguished from their possible adulterants and substituent s by these parameters. Organoleptic studies of V. negundo L. leaf s The successive Soxhlet s viz. petroleum ether. Chloroform ethyalacetate and 98% methanol s of V. negundo L. yields were determined and their differential values were recorded in the Table Among all the s obtained, the 98% methanolic was having higher percentage of yield of compounds then other s. The weight of leaf s are 1.10 (pet. ether), 1.50 (chloroform), 0.94 (ethyl acetate) and 2.05 (98% methanol) percent respectively. It is evident from these results, that the 98% methanolic solvent has higher ive values followed by, chloroform, pet-ether and ethyl acetate. Table-3.19: Extractive values and organoleptic characters of Vitex negundo L. s Sl. no Plant part used Solvents Weight of the s Colour of the Taste of the Nature of the Odour of the 1 Leaf PE 1.10 Green Bitter waxy Sweet 2 CHCL Dark Bitter waxy Sweet Green 3 EtOH 0.94 Light green Bitter sticky Sweet 4 98% Methanol 2.05 Dark green Salty Bitter waxy Aromatic The crude s of V. negundo L. leaf have shown a wide range of colour. The petroleum ether shows green. While, the chloroform exhibited dark green colour. Whereas, the ethyl acetate shows light green colour and 98% methanolic shows dark green colour. (Table: 3.19). V. negundo L. leaf s taste varies from to such as tasteless, Bitter and salty bitter. Of these the petroleum ether, chloroform and ethyl acetate s were exhibited bitter taste. Whereas the 98% methanolic was shown salty bitter in taste. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 266

25 Further the nature of these s varies, pet. ether, chloroform and 98% methanolic s were waxy in nature, whereas ethyl acetate shown salt bitter in nature. Similarly, the s were exhibiting variety of odour. The petroleum ether, chloroform and ethyl acetate s exhibiting sweet odour. Whereas the 98% methanolic s exhibited the aromatic odour. Determination of total ash and acid insoluble ash contents In the present investigation, the total ash and acid insoluble and water insoluble ash contents of V. negundo L. leaf was studied (Table: 3.20). The total ash value recorded as (3.10%), acid insoluble ash value determined as 0.93% and water insoluble ash value recorded 1.24%. This variation in the content of ash is probably due to the nature of metabolites they possess. Table-3.20: Total ash and acid insoluble ash content in Vitex negundo L. (mg/100g) Sl. no parts Total ash Acid insoluble ash contents Water insoluble ash contents 01. Leaf Fluorescent studies of powered drugs The characteristic colour behaviour of dried and powdered drugs and powder dissolved in organic solvents like petroleum ether, chloroform, alcohol, water, acetone, ethyl acetate, Conc. HCl and H 2 So 4 (5%) i.e., leaf of V. negundo L. was observed both under the visible and ultra violet (UV nm) light. The colour reactions of these drugs solutions thus emitted fluorescence light are tabulated in Table: Table-3.21: Fluorescent studies of powdered drugs in Vitex negundo L. Sl. no Treatment visible UV 365 nm 1 Powder Green Light Yellow 2 Pet. Ether Fine Green Pale Yellow 3 Chloroform Blackish green Opaline green 4 Ethanol Yellow green Light pink 5 Eather Light green Light pink 6 Acetone Dark green Light pink 7 Ethyl acetate Blackish Blue 8 Benzene Pastal green Onion pink 9 Conc. HCl (5%) Sandal wood Dark brownish 10 Conc. H2So4 (5%) Pale cream Light black ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 267

26 The powdered drug solutions have exhibited a wide range of fluorescence colours under the UV light and visible light. The petroleum ether drug solutions have shown the fluorescence colours such as fine green in visible light and pale yellow in UV light, the chloroform drugs solutions have exhibited a wide range of fluorescence colours as blackish green in visible in Opaline green UV light. While ethanol (95%v/v) treated drug solutions exhibited wide range of fluorescence colours yellow green in visible in UV light slight pink. However, the eather drug solutions exhibited light green in visible light and light pink in UV light. 1A al 1B al 2A 2B 3B 4A 4B 3A 5A 5B 6A 6B 7A 7B Plate 3.1: Fluorescent studies of powdered drugs of 07 selected plant parts. 1. Annona reticulata L., 2. Annona squamosa L., 3. Corchorus olitorius L., 4. Euphorbia tirucalli L., 5.Ficus racemosa L., 6. Pongamia pinnata L., 7. Vitex negundo L., 2. A: Visible, B: UV 365 nm. ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 268

27 In acetone drug solution shows the colours dark green in visible light and light pink in UV light, In ethyl acetate drug solution shows the colours blackish in visible light and blue in UV light, similarly benzene drug solution exhibited pastal green and onion pink. Whereas, dilute HCl (5%) acid drug solution indicating the sandal wood and dark brownish and in dilute H 2 So 4 (5%) acid drug solution shows pale cream and light black respectively. The colour reaction varies from non-polar to high polar solvents (Plate- 3.1). 3.5 Discussion Pharmacognostic studies of selected 07 medicinal plants have been undertaken to overcome the standardization of drug. Powder analysis Powder analysis of 07 selected medicinal plant parts in the present study reveals the different colours in organoleptic and florescent studies. Yoganarasinhma et al. (1981) have given the key characters of some medicinally important plants like, Tagara, Bharangi, Ishwari, which will help to identify these plants scientifically. Divyakanth and Vimal Kumar (2008) have studied pharmacognostic characters of the two species and according to them these characters will helps identify to the species. Nirmaladevi and Periyanayagam (2008) have also has the opinion that pharmacognostic study will be helpful for the identification of medicinal plants. Fillippini et al. (1995) have done comparative investigation of finally powdered sample of Herba equiseti with the help of Scanning Electron Microscopic technique. Satakopan (2000) has focused on few modern techniques like, SEM which can be implied to control the quality of Ayurvedic, Siddha and Unani crude drugs. Extractive and Ash values Variation in the content of ash and ive values is probably due to the nature of metabolites they posses. The ive values in different solvents viz., petroleum ether, chloroform, ethyalacetate and 98% methanol s were recorded. Among the s obtained, the 98% methanolic showed higher yield. According to the literature the more polar compound will have high ive value and less polar compounds will have less ive value. But, in the present study, it was ETHNOPHARMACOLOGICAL VALIDATION OF MEDICINAL PLANTS TREATING SKIN DISEASES IN HYDERABAD KARNATAKA REGION 269