Pharmacognostical Investigations on Berries of Solanum xanthocarpum

Transcription

1 Pharmacognostical Investigations on Berries of Solanum xanthocarpum Manita Demla 1 *, Harjinder Verma 2 and Tanveer Naved 1 1 Amity Institute of Pharmacy, Amity University- Noida-UP 2 R.K.S.D College of Pharmacy, Kaithal, (Haryana) Abstract The main objective of this research is based on a physicochemical and phytochemical investigation of S. xanthocarpum (berries). Physicochemical parameters like Total ash, Acid insoluble ash, Water soluble ash, Extractive values, Successive Extraction, Loss on Drying, Foreign Matter determination, Fluorescence Analysis, Toxicological parameters like Pesticide residue and Preliminary Phytochemical Screening were carried out for standardization and authentication of the berries and further preparation of monograph of S. xanthocarpum ( berries). Keywords-Solanum xanthocarpum, Berries, Solanaceae, Phytochemical investigation Introduction Solanum xanthocarpum (Family: Solanaceae) commonly known as Indian night shade or yellow berried night shade plant, is a very prickly diffuse bright green perennial herb found throughout India in dry places as a weed on roadsides and waste lands. The fruit is used as anti-asthmatic [1] hypoglycemic [2], hepatoprotective [3],antitumor, anti-tussive, anti-pyretic, anti- spasmodic, anti anaphylactic and cytotoxic activity [4]. The objective of present study was to establish various Pharmacognostic standards and to evaluate preliminary Phytochemical and Physiochemical analysis of Solanum xanthocarpum berries (Fig.: 1) Taxonomic Classification Kingdom - Plantae Subkingdom - Tracheobionta Division - Class - Subclass - Order - Family - Genus - Magnoliophyta Magnoliopsida Asteridae Solanales Solanaceae Solanum Fig. 1: S. xanthocarpum (Berries) *Corresponding Author Manita Demla - demla.manita@ gmail.com 77 P a g e

2 Materials and Methods Materials Dried whole plant of S. xanthocarpum (Solanaceae) was procured from supplier in New Delhi. The identity of the whole plant was verified by Dr. H. B. Singh, Head, National Institute of Science Communication and Information Resources (Council of Scientific and Industrial Research) 14, Satsang Vihar marg (Near Pusa Gate), New Delhi. A voucher specimen number NISCAIR/RHMD/Consult/ /1602/202 was deposited at the Herbarium of National Institute of Science Communication and Information Resources, New Delhi and also in the Pharmacognosy laboratory, Department of Pharmacy, Amity University, Noida, Uttar Pradesh. All the chemicals and reagents used were obtained from Qualigens Fine Chemicals, Mumbai, Merck Specialties Private Ltd., Mumbai and Changshu Yangyuan Chemical, China and were of analytical reagent (AR) grade. Methods 1. Botanical parameters Morphological Characters [5] In some cases, general appearance of the herb is similar to related species. Detailed study of the morphological characters can be helpful in differentiating them. The macroscopy of a drug includes its visual appearance to the naked eye. Visual inspection provides the quickest and simplest mean by which to establish identity, purity, and possibly the quality. Macroscopic identity of a medicinal plant material is based on shape, size, color, taste, surface characteristic, texture, fracture and appearance of cut surface. For each particular morphological group, a particular systemic examination can be carried out tabulated in Table 1. Table 1: Observations of Morphological Characters S. N Parameters Fruit 1 Color Brownish yellow 2 Odor Characteristic 3 Taste Bitter and acrid 4 Size mm in diameter 5 Shape Berry, globose 6 Touch Surface rough 2. Physio -chemical parameters Foreign matter [5] About 250 gm of the original sample was weighed accurately and spread out in a thin layer. The sample was inspected with the unaided eye or with the use of a magnifying lens (6X or 10X) and the foreign organic matter was separated manually as completely as possible and weighed. The percentage of foreign organic matter was weighed and determined with reference to the weight of the drug taken (Table-2). 78 P a g e

3 Loss on Drying [5] This parameter determines the amount of moisture as well as volatile components present in a particular sample (i.e. water drying off from the drug). Prepared air dried material (2 gm) was placed on a tarred evaporating dish and dried at 105ºC for 1 hour and weighed. The drying was continued until two successive reading matches each other or the difference between two successive weighing was not more than 0.25%. Constant weight was reached when two consecutive weighing after drying for 30 minutes in a desiccator, showed not more than 0.01 gm difference (Table-2). Ash Values Ash value is an important characteristic of a drug and with the help of this parameter we can detect the extent of adulteration as well as establish the quality and purity of the drug. The acid insoluble ash consists mainly of silica and thereby indicating the contamination with earthy materials. The water-soluble ash is used to estimate the amount of inorganic elements [6].About 2g of accurately weighed air dried drug was taken in a previously ignited and tared silica crucible, and incorporated at a temperature not exceeding 450 until free from carbon., washed with hot water, and residue was placed on an ash less filter paper and ignited for fifteen minutes, until white ash was formed. The residue was allowed to cool in desiccator for 30 minute, weighed. The further treatment was carried out to get water soluble and acid insoluble ash values [5] (Table-2). Table 2: Observations of Physiochemical Parameters: S. N Particulars Value (%w/w) 1 Foreign matter LOD 15 3 Total ash 15 4 Acid insoluble ash 3 5 Water soluble ash 7.5 Extractive values It determines the amount of active constituents present in a given amount of plant material when extracted with a solvent. Cold Maceration [5] The air-dried coarse drug powder (4g) accurately weighed in a glass stoppered conical flask is macerated with 100 ml of solvent (pet ether, ethanol, methanol and water) quantitatively, and then allowed to stand for 18 hrs. It is filtered rapidly, taking precaution against loss of solvent, the filtrate evaporated to dryness in a tarred flat bottom dish on a water bath and dried at 105 o C for 6 hrs, and cooled in a desiccator for 30 minutes, to constant weight and weighed content of extractable matter in mg per g of air dried material is calculated and given in Table 3 79 P a g e

4 Hot Extraction/ Soxhelation [7] 20 g of the air dried, coarsely powdered berries will be accurately weighed and subjected to soxhletion for approximately six hours and the extracts will be concentrated under vacuum using rota vapor. This procedure was followed for the preparation of all the extracts like pet ether, ethyl acetate, chloroform, methanol, ethanol and water and values recorded were given in Table 4. [8], [9] Successive Extractive values The berries (drug) collected were cleaned and crushed to a coarse powder. The coarse powder was extracted with different solvents of increasing polarity, petroleum ether, chloroform, ethyl acetate and methanol in soxhlet extractor. Finally the marc was allowed for maceration for 24 hours with distilled water to obtain aq. extract and values were given in Table 5. Table 3: Observations of Extractive residue by cold maceration S. N Extractive solvent Extractable matter (%w/w) 1 Pet ether Ethyl acetate Chloroform Methanol Ethanol Aqueous 1.40 Table 4: Observations of Extractive Residue by hot extraction S. N Extractive solvent Extractable matter (% w/w) 1 Pet ether Ethyl acetate Chloroform Methanol Ethanol Aqueous 30.0 Table 5: Observations of Successive Extractive Values %Extractable matter S. N Extractive solvent 1 Pet ether Ethyl acetate Chloroform Methanol Aqueous P a g e

5 Fluorescence Analysis [10] 1-2 mg of the dried powdered berries of S. xanthocarpum were taken and placed on a microscopic slide and observed in day light as well as in short wave UV light (254nm) and long wave UV light (366 nm).the powdered drugs were then treated with different reagents. The reagents used were 1 N sodium hydroxide (aqueous), 1 N sodium hydroxide (alcoholic), 1 N hydrochloric acid, ammonia, 5% iodine, 5% ferric chloride, acetic acid, 1 N sulphuric acid, 1 N nitric acid Fluorescence of powdered berries of S.xanthocarpum were observed in day light and in UV given in Table 6. Table 6: Observations of Fluorescence Analysis S.N Material/Treatment Daylight UV light 254 nm UV light 366nm 1 Powder as such Brown Brown Green 2 Powder treated with Brown Brown Brown distilled 3 Powder treated with 1N NaOH in water Dark brown Blackish brown Light brown 4 Powder treated with Dark brown Dark brown Green 1NaOH in methanol 5 Powder treated with Dark brown Brown Bluish green 50% HNO % HCl Light brown Dark brown Light green 7 Conc. HCl Light brown Dark brown Yellowish green 8 H 2 SO 4 Dark brown Black Bluish green 9 Acetone Brown Black Dark brown Preliminary phytochemical screening [11, [12] The extracts obtained were subjected to Qualitative test for the identification of various constituents. Detection of Alkaloids To a small portion of each extract, few drops of dil. HCl acid were added separately and filtered. The filtrates were tested with various reagents such as Mayer s, Dragendorff s, Hager s and Wagner s reagents to detect the presence of alkaloids. Detection of Glycosides Small quantity of each extract was separately subjected to Legal s test, Kellerkilliani s test and Baljet s test for cardiac glycosides. Detection of Carbohydrates Small quantity of each extract was subjected to Molisch s test, Fehling s 81 P a g e

6 test, Benedict s test and Saliwnoff s test. Detection of Steroids Small amount of each extract was subjected to Salkowaski s test and Libermann test. Detection of Saponins Small amount of each extract was shaken with water to check foam formation and its time of stability. Detection of Flavonoids Extracts was subjected to Shinoda test Detection of tannins and phenolic compounds Small amount of each extract was tested with few drops of freshly prepared 5% FeCl 3 solution and lead acetate solution to detect the presence of phenolic compounds. Detection of Amino Acids Small amount of each extract was treated with Ninhydrin s test for detection of amino acids. Results of preliminary phytochemical screening were given in Table 7. Table 7: Preliminary phytochemical screening: S. No Phytoconstituents Pet.ether Chloroform Aqueous Ethanolic 1. Alkaloids +ve +ve +ve +ve 2. Carbohydrates -ve +ve +ve +ve 3. Proteins -ve -ve -ve +ve 4. Amino acids +ve +ve +ve +ve 5. Steroids -ve +ve +ve +ve 6. Phenolic -ve -ve +ve +ve compounds 7. GLYCOSIDES +ve +ve +ve +ve 8. Flavanoids -ve +ve +ve +ve 9. Saponins -ve -ve +ve +ve +VE PRESENT, -VE ABSENT Pesticide residue [13] Sample was crushed to coarse powder and homogenized, then 10g of sample was mixed vigorously with 120 ml of acetonitrile water mixture using glass rod in a 250 ml beaker and kept overnight then it was filter via suction using non-absorbent cotton pad pre rinsed with acetonitrile on a Buchner funnel, filtrate was transferred into separating funnel add 120 ml NaCl solution, 82 P a g e

7 extract was shaken with 50 ml n-hexane thrice. Organic phase was dried over anhydrous Sodium sulphate and it was concentrated to 5 ml using water bath at 55 o C. Extract was cleaned with gm preactivated florosil and 5 gm anhydrous sodium sulphate. Column was pre rinsed with petroleum ether. It was eluted using 150 ml mixture comprising of n-hexane (141 ml) and diethyl ether (9 ml) at a flow rate of 1 drop per second. The extract was concentrated close to dryness on water bath and volume was made up to 1 ml. with n- hexane. Table 8: Observations of Pesticide Residue S.N Types of pesticides Max. residual Conc. (ppb) limits (ppb) 1 Alpha - NAD HCH 2 Beta HCH - NAD 3 Alpha + 10 NAD Beta HCH 4 Gamma HCH 5 P.P DDE P.P DDT P.P DDE P.P DDT Discussion The whole plant is used traditionally for curing various ailments as antimicrobial [14], anti-urolithiatic and natriuretic [15] -, hypoglycemic [16], anti-asthmatic [17], larvicidal [18] [19], anthelmentic and antispasmodic, antitumor, cardiotonic, hypotensive, anti-anaphylactic, cytotoxic activities are also reported [4]. For this purpose mainly leaves, root and stem bark are used. The literature survey revealed that the berries has not been fully investigated pharmacognostically/ pharmacologically. Hence this study was conceived to evaluate and standardized the berries of S.xanthocarpum, so that we have the standardize material for the pharmacognostical investigations. Conclusion Berries of S. xanthocarpum was collected and analyzed for the various standardization parameters which will be useful for the authentification and identification or the quality control of the drug. Macroscopy, Determination of foreign matter, Physiochemical parameters, phytochemical screening of different extracts revealed the presence of different category of phytochemical like alkaloids, carbohydrates, glycosides, phenols, tannins, triterpenoids, flavonoids. Ethanolic extract and aqueous extract showed the presence of maximum group of compounds, which may provide a guide line for further phytochemical investigation. Pesticide residue as alpha HCH and beta HCH was found to be absent. Through the extensive literature survey on S. xanthcarpum we came to know that not much work done on this plant and especially berries, hence there is a need to explore the drug for pharmacological actions and if 83 P a g e

8 found so; can be used to cure various ailments in the same way as we used the other parts as this easily available medicinal plant. References 1. Mohan L, Sharma P and Srivastava CN: Southeast Asian J Trop Med Public Health 2007; 2: Kar DM, Maharana L, Pattnaik S and Dash GK :Studies on hypoglycemic activity of Solanum xanthocarpum Schrad. & Wendl. fruit extract in rats. Journals of Ethno pharmacology 2006; 2: Najmi AK, Pillai KK, Pal SN, Aqil M. Journal of Ethnopharmacology 2005; 97: Gupta SS, Rai M, Gupta NK. Curr Sci 1967; 36: Anonymous (1998). Guidelines for the Assessment of Herbal Medicines. World Health Organization, Geneva, Document No. WHO/TRM/ Anonymous (1996). Sectoral study an Indian Medicinal plants-status, perspective and strategy for growth. Biotech Consortium India Ltd., New Delhi. 7. Mukherjee PK: Quality Control Of Herbal Drugs-An Approach to Evaluation of Botanicals, Business Horizons, New Delhi, India Edition 1,2005: Harborne JB:Phytochemical methods A guide to modern techniques of plant analysis, Third edition, Evans WC: Trease and Evans Pharmacognosy, WB Saunders Company Ltd, London,, Edition14, Kokoski J., Kokoski R. and Slama FJ: Fluorescence of powdered vegetable drugs under ultraviolet radiation. J Am Pharmacol Assoc. 1958; 47: Khandelwal KR: Practical Pharmacognosy technique and experiments, Edition 23, 2005,: Kokate C.K: Practical Pharmacognosy, Vallabh Prakashan,1994: Association of official analytical chemist.fourteen Edition 1984: Salar Raj K and Suchitra: Evaluation of antimicrobial potential of different extracts of S. xanthocarpum Schrad. and Wendl. African Journal of Microbiology 2009, 3(3): Patel Vina B, X. Rathod Vina B, Patela Jaymin M, Brahmbhatta V:Anti-urolithiatic and natriuretic activity of steroidal constituents of S.xanthocarpum Der Pharma Chemica, 2010, 2(1): Gupta Soumya, Mal Mainak, Bhattacharya Plaban: Evaluation of hypoglycemic potential of S.xanthocarpum (Solanaceae) fruits 84 P a g e

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