62 | P a g e International Standard Serial Number (ISSN): 2319-8141 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

1Bhavbhushan, 2Mandip Goyal, 3Harisha C.R, 4V.J.Shukla, 5Chandola H.M. 1P G Scholar, Department of Kayachikitsha, 2Assistant professor, Department of Kayachikitsha, 3 Head, Pharmacognosy Laboratory, 4Head, Pharmaceutical chemistry Laboratory, IPGT & RA, Gujarat Ayurved University, Jamnagar, Gujarat, India, 5Director, Chaudhary Brahm Prakash Ayurved Charak Sansthan, Khera Dabar, Najafgarh, New Delhi. ABSTRACT KEYWORDS: Olivier. Commonly known as “Mayaphala” in

Sanskrit, is most untouched and unexplored drug till today by most Mayaphala, Quercus infectoria Olivier of the researchers. Since the is reported for many biological .Pharmacognosy, activities, it has been selected for detailed Pharmacognostical study Phytochemistry of its granules including Physico-chemical analysis following For Correspondence: standard guidelines of Ayurvedic Pharmacopoeia of India. The Bhavbhushan * Address: PG Scholar, microscopic characteristics of granules shows prismatic crystals of Department of calcium oxalate, large quantity of materials, oil globules, Kayachikitsha, I.P.G.T. simple starch grains, stone cells, pitted vessels. Qualitative analysis and R.A., Gujarat of granules shows that, loss on drying was 5.803 % w/w , ash value H Ayurved University, 1.5248% w/w, P 3.5.Preliminary phyto-chemical screening shows Jamnagar -361008 the presence of Tannin ,Flavonoid ,Steroids, etc. Thin layer Gujarat, India chromatography result show 5 and 4 spots respectively, at 254 nm Mobile no- 9601575811 and 366 nm. These observed parameters can be used as the E-mail- identifying tool for the drug Mayaphala. drbhavbhushan2003@gm ail.com

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INTRODUCTION: Quercus infectoria Olivier (), 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 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 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 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 etci.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 gallii. It is also extensively used in textile industry. Galls of Quercus infectoria possess many therapeutic activities such as Anti-diabeticiii, Anti-parkinsonism, Anti-tremorine, Anti-inflammatoryiv, Anti-viral, Anti- bacterialv, Anti-fungal, and Larvicidal activityvi.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 sievesvii. 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 Full Text Available On www.ijupbs.com

64 | P a g e International Standard Serial Number (ISSN): 2319-8141 slides prepared with stain and without stain studied under the Carl-zeiss binocular microscope attached with camera8. The microphotographs were also taken. The characters of Mayaphala were cross verified with API for further confirmation9. Physico- chemical Study: The physico-chemical parameters like, LOD, Ash value, PH etc. and phytochemical analysis for the separation of constituents10. 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-acid11 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, PH, Water soluble and Alcohol soluble extractive values are shown in Table 2

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Table 2. Physicochemical parameters Sr. No. Parameters Result 1 Loss on drying 5.803 % w/w 2 Total Ash 1.5248% w/w 3 PH Value 3.5 4 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 flavonoid12. 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. +ve Wagner’s Test Alkaloids Reddish brown ppt. +ve Tannin & Phenolic 5% FeCl solution Deep blue-black color. +ve 3 compound Biuret Test Protein No color change -ve Violet ring was observed Molisch’s test Carbohydrate +ve at the junction First yellow, then brick Fehling’s test Carbohydrate +ve Alcoholic extract red ppt. observed of Greenish yellow Mayaphala Salkowski reaction Steroids +ve fluorescence Granules First red, then blue and Liebermann- Steroids finally green color +ve Burchard reaction appears Lead Acetate Flavonoid Yellow ppt. +ve Frothing with honeycomb Shaking in test-tube Saponin-Glycosides -ve appearance Yellow to Orange color +ve Baljet’s test Glycosides appears Bluish or Purple color Ninhydrin test Amino acids -ve appears Alcoholic extract Coumarin Blue or Green -ve when made alkaline Glycosides fluorescence +ve –Presence Full Text Available On www.ijupbs.com

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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 Rf 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 No of Spots Rf Value No of spots Rf Value (8:2) 5 0.03 ,0.08 ,0.49 4 0.01 ,0.02 ,0.08 ,0.54 ,0.96 ,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 PH conventionally represents the acidity and alkalinity; the pH of sample was detected by using PH 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;556-562 2. Nishteshwar K & Hemadri Koppula Dravyaguna vijnana,1st ed .Delhi Chaukhamba Sanskrit prathishthan 2010; 305-306

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3. Hwang JK, Kong TW, Baek NI, Pyun YR. Alpha-glycosidaseinhibitory activity of hexagalloyl glucose from the galls of Quercus infectoria. Planta Med 2000; 66: 273-274. 4. 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: 285-292. 5. Basri DF, Fan SH. The potential of aqueous and acetone extracts of galls of Quercusinfectoria as antibacterial agents. Indian J Pharmacol 2005; 37: 26-29. 6. 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: 261-263. 7. Mithal BM, Textbook of pharmaceutical formulation, Vallabhprakashan, Delhi; 158 8. Kokate CK, Purohit AP, Gokhle SB, Pharmacognosy. 42nd edition. Pune : Nirali Prakashan;2008:6.3 9. Anonymous, 2007 ,The Ayurvedic Pharmacopoeia of India Part-1, vol.-4, Govt. of India publication New Delhi, 1st edition,Vol.-4:72-74. 10. Anonymous, 1999, The Ayurvedic Pharmacopoeia of India Part-1, vol.-4, Govt. of India publication New Delhi, 1st edition,Vol.-1,Appendix 2. 11. Stahl E, thin layer chromatography a laboratory hand book.Berlin:Springer-Verlag;1969:124-241. 12. Khandel KR. Preliminary phytochemical screening. Practical Pharmacognosy Techniques and Experiments.19th edition Pune: Nirali Prakashan Publishers; 2008:149-156.

PLATE 1

Mayaphala gall (Fig. 1) Mayaphala granules (Fig. 2 ) Fibre(Fig. 3)

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Prismatic crystal (Fig. 5 ) Pitted stone cell (Fig. 4) 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. Piitted vessel (Fig. 12 ) 11 )

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HPTLC

Peak display at 254 nm (Fig. 13 )

Peak display at 366 nm (Fig. 14 )

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