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Pharmacognostic and Antifungal Investigations of Elaeocarpus Ganitrus (Rudrakasha)

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Pharmacognostic and Antifungal Investigations of Elaeocarpus Ganitrus (Rudrakasha) www.ijpsonline.com the analysis of bulk samples of ketoprofen and salicylic acid. Asian J 2006;43:516-8. Chem 2006;18:393-6. 10. Maheshwari RK, Chaturvedi SC, Jain NK. Application of hydrotropy 2. Maheshwari RK. New application of hydrotropic solubilization in the in spectrophotometric analysis of pharmaceutical dosage form. Indian spectrophotometric estimation of ketoprofen in tablet dosage form. Drugs 2005;42:760-3. Pharm Rev 2005;3:123-5. 11. Jain NK, Agrawal RK, Singhai AK. Hydrotropic solubilization of 3. Maheshwari RK. Application of hydrotropic solubilization in the nifedipine. Pharmazie 1990;45:221-5. analysis of aceclofenac. Asian J Chem 2006;18:1572-4. 12. Da Silva RC, Spitzer M, Da Silva LHM, Loh W. Investigations on the 4. Maheshwari RK. Analysis of frusemide by application of hydrotropic mechanism of aqueous solubility increase caused by some hydrotropes. solubilization phenomenon. Indian Pharma 2005;4:55-8. Thermochim Acta 1999;328:161-7. 5. Maheshwari RK. Spectrophotometric determination of cefixime in 13. Etman MA, Hada AH. Hydrotropic and cosolvent solubilization of tablets by hydrotropic solubilization phenomenon. Indian Pharma indomethacin. Acta Pharm 1999;49:291-8. 2005;4:63-8. 14. Poochikian GD, Cradock JC. Enhanced chartreusin solubility by 6. Maheshwari RK. Novel application of hydrotropic solubilization in hydroxyl benzoate hydrotropy. J Pharm Sci 1979;68:728-32. the spectrophotometric analysis of tinidazole in dosage form. Asian J 15. Shukla RS, Patel A, Soni ML, Modi V, Jaliwala YA. Quantitative Chem 2006;18:640-4. spectrophotometric estimation of cefadroxil using hydrotropic 7. Maheshwari RK. Spectrophotometric analysis of amoxycillin in tablets solubilization technique. Asian J Pharm 2008;2:146-7. using hydrotropic solubilization technique. Asian J Chem 2006;18: 3194-6. 8. Maheshwari RK, Chaturvedi SC, Jain NK. Application of hydrotropic Accepted 21 March 2010 solubilization phenomenon in spectrophotometric analysis of Revised 16 November 2009 hydrochlorothiazide tablets. Indian drugs 2005;42:541-4. Received 30 June 2008 9. Maheshwari RK, Chaturvedi SC, Jain NK. Analysis of aceclofenac in tablets using hydrotropic solubilization technique. Indian Drugs Indian J. Pharm. Sci., 2010, 72 (2): 258-261 Pharmacognostic and Antifungal Investigations of Elaeocarpus ganitrus (Rudrakasha) B. SINGH*, A. CHOPRA, M. P. S. ISHAR, A. SHARMA1 AND T. RAJ Pharmacognosy and Phytochemistry Laboratory, Department of Pharmaceutical Sciences, Guru Nanak Dev University, Amritsar-143 005, India, 1University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh-160 014, India Singh, et al.: Pharmacognostic and Antifungal Activity of Rudrakasha Rudrakasha is the dried bead obtained from the ripe fruit of Elaeocarpus ganitrus Roxb. (Family: Elaeocarpaceae). Microscopic studies revealed the presence of a hard endocarp with lignifi ed isodiametric sclereids, seeds with membranous seed coat, which enclosed a dense cellular endosperm comprising of calcium oxalate druses. Physicochemical parameters showed that total ash was 1.36 times and 1.56 times more than the acid insoluble ash and water-soluble ash, respectively. Further, ethanol had a maximum extractable value of 2.4% and moisture content was found to be 9.7%. Different extracts, petroleum ether, chloroform, ethanol and water were prepared. Chemically the extracts showed the presence of phytosterols, fats, alkaloids, fl avonoids, carbohydrates, proteins and tannins. The extracts were evaluated for antifungal activity on different fungal strains. Chlorofom and ethanol extracts have high antifungal activity against Candida albicans. Whereas, chloroform, ethanol and water extracts showed moderate inhibition against Aspergillus niger. Key words: Rudrakasha, Elaeocarpus ganitrus, antifungal activity, Candida albicans, Aspergillus niger Elaeocarpus ganitrus Roxb. (Syn. E. sphaericus also called blueberry beads[1]. It finds a prominent Gaertn; family Elaeocarpaceae), is popular for its place in Hindu religion and Ayurveda, the ancient attractive fruit stones. Its beads are covered by an Indian system of medicine. In Hindi it is known as outer shell of blue color on fully ripening, hence Rudraksha[2]. Rudrakasha fruits are thermogenic, sedative and are useful in cough, bronchitis, neuralgia, *Address for correspondence cephalagia, anorexia, migraine, manic conditions E-mail: [email protected] and other brain disorders[3]. The flesh or pulp March - April 2010 Indian Journal of Pharmaceutical Sciences 261 www.ijpsonline.com of drupe is given in epilepsy, diseases of head blue as per well documented method[24]. Wherever and in mental illness[4]. Besides it is reported to necessary sections were also stained with safranin, exhibit multifarious pharmacological activities that fast-green and iodine solution. Powdered material was include antiinflammatory[5], analgesic[6], sedative[6], cleared with chloral hydrate (S. D. Fine Chemicals antidepressant[7], antiasthmatic[8], hypoglycemic[4], Ltd., Mumbai, India) and mounted in glycerine antihypertensive[9-11], smooth muscle relaxant[12], (Glaxosmithkline Pharmaceuticals Ltd. Mumbai, hydrocholeretic[12], antiulcerogenic[7] and India) and stained where ever necessary. The fruit anticonvulsant[13]. A detailed study on pharmacognostic had a hard, stony endocarp or sclerotesta. The surface standards of Rudraksha is lacking. In ancient Indian of the endocarp was black, deeply folded, and hard medicine, the fruits were employed to ward off evil with ridges and furrows. Mesocarp was not seen spirits and omens[14] which can be considered to be in the fruit. The fruit included five or six carpels, at least partly manifestations of microbial infection. each carpel having a single large seed (fig. 1). In In addition, the Rudrakasha extracts also known to sectional view, the seed was elliptical and consisted exhibit very high antimicrobial activity[15]. Therefore, of a membranous seed coat. The seed coat enclosed it was decided to evaluate the drug (beads) for a dense cellular endosperm (fig. 2). The seed coat, various pharmacognostic parameters and beads though membranous, is differentiated into outer darkly extracts for antifungal activity against different fungal staining cell layer and inner vertically oblong compact strains. dark cell layer. They have dense accumulation of tannins. The middle seed coat has two or three layers Dried fruits (beads) of Elaeocarpus ganitrus were of circular brachysclereids or stone cells. Their walls procured from a cultivated source, Hind Herb Store, are highly lignified. The sclereids have dark cell Saharanpur, Uttar Pradesh, India, in the month of contents. The endosperm cells are in parallel compact September 2007. Identity of dried fruit was confi rmed rows. They extend from periphery to the centre; through Plant Anatomy Research Centre, Medicinal the cells are squarish and thin walled. The cells Plant Research Unit, Chennai. Voucher specimen no. towards the periphery are smaller and they become PARC/2008/164 has been deposited in the herbarium gradually larger towards the centre of the seed. The of the same institute. Coarsely powdered dried fruits endosperm cells have large calcium oxalate druses or were successively extracted with petroleum ether, sphaerocrystals. The crystals are either one or two chloroform and ethanol for 48 h each using Soxhlet per cell. They are random in distribution (fi g. 3). The apparatus and finally boiled with distilled water druses are 10 μm in diameter. The stony endocarp or for 6 h. The extracts were filtered, concentrated in sclerotesta consists of only sclereids. No other cell vacuo and dried in an oven at 40-50º. After removal types or cell inclusions are evident in the powder of solvents under vacuo from various extracts, the (fi g. 4). The sclereids may be short or elongated. The percentage of various extractives obtained was: short sclereids are isodiametric. The sclereid walls petroleum ether extract (PE) 1.655, chloroform are highly lignifi ed; the cells have wide lumen with extract (CE) 0.458, ethanol extract (EE) l.538 and brownish content. water extract (WE) 1.013. Other physicochemical parameters[16,17] showed total ash of 1.5%, acid In vitro antifungal activity of all the extracts was insoluble ash 1.1%, water-soluble ash 0.96%, pet carried out using the disk-diffusion assay[25] and ether soluble extractive 1.3%, chloroform soluble broth dilution test[26,27]. The disk-diffusion assay was extractive 0.5%, ethanol soluble extractive 2.4%, applied to determine the growth inhibition of fungi water soluble extractive 1.8% and loss on drying by extracts to be tested. Overnight fungal cultures 9.7%. Phytochemical screening[18-22] gave positive (100 μl) were spread onto SDA. The extracts were test for phytosterols, fats, alkaloids, flavonoids, applied to 8 mm disks (Whatman paper No.1). After carbohydrates, proteins and tannins. 48 h of incubation at 25o, the diameter of growth inhibition zones was measured. MIC of all extractives For microscopic studies the paraffin embedded was determined by broth dilution test which was specimens were sectioned with the help of rotary performed in test tubes. The conidial suspension, microtome. Thickness of the sections was 10-12 μm. which gave the fi nal concentration of 1×105 CFU/ml, De-waxing of the sections was done by customary was prepared. A growth control tube and a sterility procedure[23]. The sections were stained with toluidine control tube were used in each test. After 24-72 h 262 Indian Journal of Pharmaceutical
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