PHARMACOGNOSTICAL AND PHYSICO-CHEMICAL EVALUATION ON MAYAPHALA GRANULES AN AYURVEDIC FORMULATION

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1 62 P a g e International Standard Serial Number (ISSN): International Journal of Universal Pharmacy and Bio Sciences 3(1): January-February 2014 INTERNATIONAL JOURNAL OF UNIVERSAL PHARMACY AND BIO SCIENCES IMPACT FACTOR 1.89*** ICV 5.13*** Pharmaceutical Sciences RESEARCH ARTICLE!!! PHARMACOGNOSTICAL AND PHYSICO-CHEMICAL EVALUATION ON MAYAPHALA GRANULES AN AYURVEDIC FORMULATION 1 Bhavbhushan, 2 Mandip Goyal, 3 Harisha C.R, 4 V.J.Shukla, 5 Chandola H.M. 1 P G Scholar, Department of Kayachikitsha, 2 Assistant professor, Department of Kayachikitsha, 3 Head, Pharmacognosy Laboratory, 4 Head, Pharmaceutical chemistry Laboratory, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India, 5 Director, Chaudhary Brahm Prakash Ayurved Charak Sansthan, Khera Dabar, Najafgarh, New Delhi. ABSTRACT KEYWORDS: Quercus infectoria Olivier. Commonly known as Mayaphala in Mayaphala, Quercus infectoria Olivier.Pharmacognosy, Phytochemistry For Correspondence: Bhavbhushan * Address: PG Scholar, Department of Kayachikitsha, I.P.G.T. and R.A., Gujarat Ayurved University, Jamnagar Gujarat, India Mobile no drbhavbhushan2003@gm ail.com Sanskrit, is most untouched and unexplored drug till today by most of the researchers. Since the plant is reported for many biological activities, it has been selected for detailed Pharmacognostical study of its Gall granules including Physico-chemical analysis following standard guidelines of Ayurvedic Pharmacopoeia of India. The microscopic characteristics of granules shows prismatic crystals of calcium oxalate, large quantity of tannin materials, oil globules, simple starch grains, stone cells, pitted vessels. Qualitative analysis of granules shows that, loss on drying was % w/w, ash value % w/w, P H 3.5.Preliminary phyto-chemical screening shows the presence of Tannin,Flavonoid,Steroids, etc. Thin layer chromatography result show 5 and 4 spots respectively, at 254 nm and 366 nm. These observed parameters can be used as the identifying tool for the drug Mayaphala.

2 63 P a g e International Standard Serial Number (ISSN): INTRODUCTION: Quercus infectoria Olivier (Fagaceae), commonly known as Magic Nut, is a small tree or shrub mainly found in Greece, Asia Minor, Syria, Iran and in Himalayan region of India. In Sanskrit it is known as Mayaphala while Majuphala in Hindi and Gujarati. Tree possesses rough brown gall or fruit that emerge on its shoots as a consequence of assault of Gall Wasp Cypnisgallae tincotoriae. The Gall is collected in August and October. A small hole is often seen in the Gall, Which is the escape route of Gall wasp. These galls are spherical or pear shaped and measure 6-50 mm. in diameter.the surface of the mature dry gall may be smooth and shining as though varnished and chestnut brown color, when the galls are gathered at the correct stage i.e. before the insect emerges, the inner tissue is soft, of a deep greenish yellow colour, with a very astringent taste and slightly sweet after taste. The galls vary generally in size, colour, and general appearance depending upon the producing Region or Country etc i.the galls are collected before the escape of the insect and well dried. According to Ayurveda, it is good drug of choice in Grahani and Pravahika and also used in healing ulcer in the mouth as well as on the skin. The external application of Mayaphala gall powder or paste heals the ulcer in the mouth as well as on the skin, and the paste applied as gum paint which relieves bleeding from gums due to the presence of high amount of tannin in gall ii. It is also extensively used in textile industry. Galls of Quercus infectoria possess many therapeutic activities such as Anti-diabetic iii, Anti-parkinsonism, Anti-tremorine, Anti-inflammatory iv, Anti-viral, Antibacterial v, Anti-fungal, and Larvicidal activity vi.though the plant is reported for many such activities, the plant has not been studied yet for Pharmacognostical characters. So present study was carried out to set some Pharmacognostical and physicochemical standards on gall granules and powder for authentification of plant. MATERIAL AND METHODS Collection and authentification of the sample The Gall of Mayaphala were used as material and collected from Pharmacy of Gujarat Ayurved University, Jamnagar. The sample was authentified by the Pharmacognosy Lab. of the University. Preparation of granules Granules were prepared by wet granulation method in which powdered drug was moistened with water, lumped and then passed through sieves vii. Pharmacognostical Evaluation: Organoleptic characters i.e. taste, odour and colour of granules were recorded. Microscopic study carried out by 10g of granules dissolved in small quantity of distilled water, filtered and filtrate was dried then

3 64 P a g e International Standard Serial Number (ISSN): slides prepared with stain and without stain studied under the Carl-zeiss binocular microscope attached with camera 8. The microphotographs were also taken. The characters of Mayaphala were cross verified with API for further confirmation 9. Physico- chemical Study: The physico-chemical parameters like, LOD, Ash value, P H etc. and phytochemical analysis for the separation of constituents 10. HPTLC profile: HPTLC of the sample was carried out in Pharmaceutical chemistry Lab. following standard procedure used for the standardization of samples mentioned in API. High performance thin layer chromatography was performed for the normal phase separation of components of methanol extract of sample. Solvent system was prepared by taking Toluene and Ethyl acetate in a proportion of 8:2 respectively. The spots obtained from both the extracts were examined under ultra violet light of wavelength 254nm and 366nm with the stationary phase of Silica gel G. chromatographic derivation was done with Methanol sulphuric-acid 11 RESULTS AND OBSERVATION: Pharmacognostical study Detailed Pharmacognostical evaluation of Granules was carried out for the sample of Mayaphala granules; Organoleptic characters of powder shows the presence of light brown color with astringent odor and taste. Table 1: Organoleptic characters of granules Sr. No Characters Result 1 Color Light Brown 2 Odor Astringent 3 Taste Astringent 4 Nature of Drug Granular The powder microscopy of Mayaphala granules (fig,2 ) revealed presence of prismatic crystals of calcium oxalate (fig,5),oil globules(fig,10),annular vessel(fig,9), Tannin content with parenchyma cells (Fig,7),starch grains (Fig,6),Thick-walled Stones cells (fig,8), group of lignified fibers (fig,3) and pitted stone cells (Fig,4) etc. Physicochemical parameters Qualitative analysis Results on Mayaphala granules such as, Loss on drying, Ash value, P H, Water soluble and Alcohol soluble extractive values are shown in Table 2

4 65 P a g e International Standard Serial Number (ISSN): Table 2. Physicochemical parameters Sr. No. Parameters Result 1 Loss on drying % w/w 2 Total Ash % w/w 3 P H Value Water soluble extract 71.0% w/w 5 Alcohol soluble extract 74.7% w/w 6 Acid insoluble Ash Not required Preliminary Qualitative Analysis Preliminary Qualitative Analysis showed presence of Tannin, carbohydrate, steroid and flavonoid 12. Details are tabulated in the table no. 3 Table 3. Preliminary Qualitative Analysis Material Test/ Reagent Functional gp Observation Result Dragendorff s Test Alkaloids Orange Brown ppt. Wagner s Test Alkaloids Reddish brown ppt. 5% FeCl 3 solution Tannin & Phenolic compound Deep blue-black color. Biuret Test Protein No color change -ve Molisch s test Carbohydrate Violet ring was observed at the junction Alcoholic extract of Mayaphala Granules Fehling s test Salkowski reaction Liebermann- Burchard reaction Carbohydrate Steroids Steroids First yellow, then brick red ppt. observed Greenish yellow fluorescence First red, then blue and finally green color appears Lead Acetate Flavonoid Yellow ppt. Shaking in test-tube Saponin-Glycosides Frothing with honeycomb appearance -ve Baljet s test Glycosides Yellow to Orange color appears Ninhydrin test Amino acids Bluish or Purple color appears -ve Alcoholic extract when made alkaline Coumarin Glycosides Blue or Green fluorescence -ve Presence

5 66 P a g e International Standard Serial Number (ISSN): Table 4. HPTLC Profile Results of HPTLC study shows that 5 and 4 spots were observed under 254 nm and 366 nm respectively. The reported R f values of HPTLC are shown in the table no 4 and fig no.13, and 14. Solvent system 254nm(Short UV) 366nm(Long UV) Toluene:Ethylacetate (8:2) No of Spots R f Value No of spots R f Value ,0.08,0.49,0.54, ,0.02,0.08,0.49 DISCUSSION The present drug was proved to be genuine by assessing the Organoleptic characters and powder microscopy features. Tannin content found in microscopy while in quality assessment is supporting the already proven clinical data also Physico-chemical and phyto-chemical parameters were also applied for assessing the drug. The Total Ash 1.5% w/w indicates higher inorganic matter while extractive values with water and methanol as a solvents shows good amount of solubility. The P H conventionally represents the acidity and alkalinity; the ph of sample was detected by using P H meter and it was 3.5 showing the acidic nature of the drug. Thin layer chromatography results shows that 5 and 4 spots respectively were observed when plate was scanned at 254 nm and 366 nm. The results were same for both detection wavelengths which show that the components are sensitive for both wavelengths. CONCLUSION Preliminary Organoleptic features and results of powder microscopy revealed the presence of important characters like, oil globules, prismatic crystals of calcium oxalate, tannin content, starch grains, fibers and annular vessel in the tested sample. While Tannin, Flavonoid, steroids are the qualitatively found compounds for the same. Thin layer chromatography results shows that 5 and 4 spots were observed when plate was scanned at 254 nm and 366 nm respectively. All the above discussed parameters can be used as the identifying tools for Pharmacognostical and preliminary phytochemically for Mayaphala. REFERENCES: 1. Pandey Gyanendra Dravyaguna vigyana Varanasi ChaukhambaKrishnadas Academy Reprint- 2004; Nishteshwar K & Hemadri Koppula Dravyaguna vijnana,1 st ed.delhi Chaukhamba Sanskrit prathishthan 2010;

6 67 P a g e International Standard Serial Number (ISSN): Hwang JK, Kong TW, Baek NI, Pyun YR. Alpha-glycosidaseinhibitory activity of hexagalloyl glucose from the galls of Quercus infectoria. Planta Med 2000; 66: Kaur G, Hamid H, Ali A, Alam MS, Athar M. Anti-inflammatory evaluation of alcoholic extract of galls of Quercus infectoria. J Ethnopharmacol 2004; 90: Basri DF, Fan SH. The potential of aqueous and acetone extracts of galls of Quercusinfectoria as antibacterial agents. Indian J Pharmacol 2005; 37: Redwane A, Lazrek HB, Bouallam S, Markouk M, Amarouch H, Jana M. Larvicidal activity of extracts from Quercus lusitanica var.infectoria galls (Oliv.). J Ethnopharmacol 2002; 79: Mithal BM, Textbook of pharmaceutical formulation, Vallabhprakashan, Delhi; Kokate CK, Purohit AP, Gokhle SB, Pharmacognosy. 42 nd edition. Pune : Nirali Prakashan;2008: Anonymous, 2007,The Ayurvedic Pharmacopoeia of India Part-1, vol.-4, Govt. of India publication New Delhi, 1 st edition,vol.-4: Anonymous, 1999, The Ayurvedic Pharmacopoeia of India Part-1, vol.-4, Govt. of India publication New Delhi, 1 st edition,vol.-1,appendix Stahl E, thin layer chromatography a laboratory hand book.berlin:springer-verlag;1969: Khandel KR. Preliminary phytochemical screening. Practical Pharmacognosy Techniques and Experiments.19 th edition Pune: Nirali Prakashan Publishers; 2008: PLATE 1 Mayaphala gall (Fig. 1) Mayaphala granules (Fig. 2 ) Fibre(Fig. 3)

7 68 P a g e International Standard Serial Number (ISSN): Pitted stone cell (Fig. 4) Prismatic crystal (Fig. 5 ) Starch grains (Fig. 6 ) Tannin with parenchyma cells (Fig. 7 ) Thick walled stone cells (Fig. 8 ) Annular vessels (Fig. 9 ) Oil globule (Fig. 10 ) Pitted stone with wide lumen (Fig. 11 ) Piitted vessel (Fig. 12 )

8 69 P a g e International Standard Serial Number (ISSN): HPTLC Peak display at 254 nm (Fig. 13 ) Peak display at 366 nm (Fig. 14 )