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Physico-Chemical and Phyto-Chemical Study of Rhizome of S. Das 1,2 *, S. Sharma 1,2 and S. Lariya 1,3 Research Article 1 Pacific Academy Of Higher Education And Research University, Udaipur, India. 2 Department of Pharmacology, Malhotra College of Pharmacy, Bhopal, Madhya Pradesh, India. 3 Radha raman Institute of Pharmaceutical Science, Bhopal, Madhya Pradesh, India. ABSTRACT The pteridophytes constitute the primitive vascular plant groups which are found spread all over world including India. is a fern belongs to the family Dryopteridaceae is considered to possess great medicinal value. It has been reported to possess wide ethanomedical use. The investigation carried out by us was on the Pharmacognostical, Physiochemical and Phytochemical aspects of the rhizome part of the plant. Morphological studies of rhizome showed the presence of various diagnostic characters. Moisture content, Ash value and extractive value were determined for quality standard of drugs. During the experimental work the rhizome part showed the presence of various phytoconstituents like flavonoids, tannins, protein and amino acid etc. Keywords: Pteridophytes, Dryopteridaceae, Phytochemical, Moisture content, Flavonoids. INTRODUCTION Herbal drugs have gained importance and popularity in recent years because of their safety, efficacy and cost effectiveness. Traditional medicine using herbal drugs exists in every part of the world. The major areas are Chinese, Indian and European traditions. The philosophies of these traditional medicines have some resemblance to each other but differ widely from modern Western medicine. In view of the progress of Western medicine not only new synthetic drugs but also herbal drugs have to fulfill the international requirements on quality, safety and efficacy. Herbal drugs have the advantage of being available for patients in the geographical area of the special traditional medicine. The development procedure of herbal drugs for world-wide use has to be different from that of synthetic drugs 1. Medicinal plants are of great importance to the health of individual and communities. The medicinal value of these plants lies in some chemical active substances that produce a definite physiological action on the human body. The most important of these chemically active (bioactive) constituents of plants are: alkaloids, tannins, flavonoid and phenolic compounds. Many of these indigenous medicinal plants are also used for medicinal purposes 2. Out of 1,200 species of pteridophytes occurring in India, about 170 species have been found to be used as food, flavor, dye, medicine, bio-fertilizers, oil, fiber and bio-gas production 3. Ethnobotany and medicinal properties of Indian fern and their allies have been described by various workers time to time 4-8. Likewise 160 species of useful pteridophytes in India on the basis of phytochemical, pharmacological and ethnobotanical studies have been reported 9. An overview on medicinal uses of 110 Indian pteridophytic has been published 10. that belongs to the family Dryopteridaceae is considered to possess great medicinal value. It has been reported to possess wide ethanomedical use, whose rhizome has antifungal property and is used as an antidote. It also has variety of applications against the disorders like epilepsy, leprosy, cuts, wounds, ulcers, swelling, pains and snake bites. The decoction of the dried rhizome, stem and stripe is used for blood purification 11. The leaves of the plant are used for the treatment of epilepsy 12-13. Pounded rhizomes are used in swellings and pain and have antifungal properties 14. A small portion of the rhizome of the plant is powdered and taken with water twice a day in rheumatism, epilepsy and leprosy 15. is used in mental disorder. Filtered water extract Vol. 3 (2) Apr-Jun 2014 www.ijpcsonline.com 517

of rhizome is given to the unconscious persons suffering from epilepsy in India 16. MATERIALS AND METHODS Plant Selection The rhizome part of was collected from Amarkantak forest district Bilaspur, Chattisgarh, in the month of July 2013.It was further identified and authenticated by the Botanical Department, Saifia Science College, Bhopal,Madhya Pradesh, Ref no. 476/Bot/Saf/13. The rhizomes were washed with tap water, prior to distilled water, shade dried and powdered. The powdered plant materials were subjected to successive extraction with methanol using Soxhlet extractor. The extracts were dried and stored in freezer at 0 0 C for future use. Macroscopic study The rhizome was taken for various macroscopic organoleptic evaluation like colour, odour, shape, taste, appearance etc. ash, water soluble ash, water soluble extractive value, alcohol soluble extractive value, moisture content, ph were calculated as per Indian Pharmacopoeia. Preliminary phytochemical screening of rhizome of For preliminary phytochemical screening, 100 g of powder drug was extracted with Methanol. The mother extract obtained from successive solvent extraction were then subjected to various qualitative chemical tests to determine the presence of various phytoconstituents like glycosides, tannins, phytosterols, fixed oils and fats, proteins and amino acids, flavonoids, saponins, etc. The above phytoconstituents were tested as per the standard methods 17-18. RESULT AND DISCUSSION Microscopy Study (Wallis T E, 2002): Qualitative microscopic evaluation was carried out by taking transverse sections of fresh rhizome of. Free hand transverse sections of rhizome were first observed with distilled water and then with phloroglucinol and concentrated HCl. Physicochemical parameters The determination of various physicochemical parameters such as total ash, acid insoluble rhizome Vol. 3 (2) Apr-Jun 2014 www.ijpcsonline.com 518

Macroscopy of rhizome of The macroscopic character were always served as useful keys in faster and early identification of plant material and also serves as an important standardization parameter. The macroscopic features of rhizome of are described here. The organoleptic evaluation is discussed in Table no.1. Microscopy of T.S. of rhizome of The rhizome show the outermost surface, a single-layered barrel shaped epidermal cells embedded with tannin content. The outer cortex consisting of a few layers of cells with lignified thick walls and inner cortex made up of several layers of parenchyma cells with thin cellulose walls. Vascular bundles are loosely distributed around the perimeter of a central pith. The xylem vessels possess ligneous secondary wall thickenings. The remainder of the rhizome vascular system is scattered in small bundles throughout the cortex. Pith is composed of parenchymatous cells containing starch grains and few filled with oleoresinous contents. Physicochemical parameters To evaluation of physicochemical parameters gives a clear idea about the specific characteristics of crude drugs under examination, besides it s macro-morphological or cyto morphological characters, microscopical natures in both its entire and it s powder form. While these diagnostic features enable the analyst to know the nature and characteristics crude drugs, further evaluation of different parameters indicate their acceptability by criteria other than the morphological characteristics. The procedures normally adopted to get qualitative information about the purity and standard of a crude drug include the determination of various parameters. The determination of physicochemical parameter is important in determination of adulterantsand improper handling of drugs. Table- 2 shows the result of various physico- chemical parameter of powdered drug carried out using standard methods. Phytochemical Analysis of rhizome of The results of preliminary phytochemical analysis are tabulated in Table 3. The phytochemical study revealed the presence of various phytocompounds in methanolic solvent extract. Table 1: Organoleptic Evaluation of rhizome of S. No. Oraganoleptic Parameters Result 1 Colour Brown 2 Odour Aromatic 3 Taste Astringent 4 Surface Rough with striations Table 2: Physical Evaluation Parameters S. No Parameter Values(%)w/w 1 Loss on Drying 8.45 2 Ash Values Total Ash 9.10 Acid insoluble ash 4.87 Water soluble ash 6.18 3 Extractive Values Water soluble extractive 11.6 Alcohol soluble extractive 20.4 Vol. 3 (2) Apr-Jun 2014 www.ijpcsonline.com 519

Table 3: Phytochemical analysis S. No. Name of the Phytochemical Qualitative Test Result 1 Carbohydrates Molisch s test - Fehling s test - 2 Alkaloids Mayers test - Wagners test - 3 Proteins & Amino acids Ninhydrin Test + Millon s Test + 4 Tannin and Phenolic compound FeCl 3 Test + Lead acetate Test + 5 Flavonoids Shinoda Test + Alkaline reagent Test + 6 Fixed Oils & Fats Spot Test - Saponification Test - 7 Phytosterol Salkowski Test - Libermann s Test - 8 Glycoside Keller-Killiani Test + 9 Saponin Foam Test - CONCLUSION All studied standardization parameters such as macroscopy, microscopy, physicochemical parameters, and phytochemical screening was carried out and it could provide the knowledge in authentication of rhizome. Phytochemical screening results will be useful to find out the genuine drug. Physicochemical parameters such as Loss on drying, Total ash, Acid insoluble ash, Water soluble ash, and Successive extractive values were observed with solvents of Alcohol, Aqueous. These values can be useful to detect adulteration. This report would help in the identification of the crude drug in future. ACKNOWLEDGEMENTS The author is grateful to Dr. Zia ul Hasan,H.O.D.,Department of Botany,Saifia Science College, Bhopal, for identifying plant specimen. REFERENCES 1. Vogel HG. Similarities between various systems of traditional medicine considerations for the future of ethnopharmacology. J Ethnopharmacol 1991; 35:179-90. 2. Aiyelaagbe OO and Paul M. Osamudiamen :Phytochemical screening for active compounds in Mangifera indica leaves from Ibadan, Oyo State. Plant science Research 2009;2(1):11-13. 3. Manickam VS and Irudayaraj V: Pteridophytic flora of the Western Ghats, South India. BI Publications Private Limited New Delhi, 1992. 4. Kirtikar KR and Basu BD. Indian Medicinal Plants I-IV. International Book Distributors, Dehra Dun, India, 1935. 5. Nayar BK. Medicinal ferns of India. Bulletin of National Botanical Garden, Lucknow. 1959;29:1-36. 6. Nadkarni BK. Indian Materia Medica with Ayurvedic, Unantibii, Siddha, Allopathic, Homeopathic, Naturopathic and home remedies. Popular Book Depot, Bombay, 1954. 7. Chopra RN, Nayar SL and Chopra LC. Glossary of Indian Medicinal Plants. CSIR, New Delhi, 1956. 8. Puri HS. Indian Pteridophytes used in folk remedies. American Fern Journal 1970;56:79-81. 9. Singh HB. Potential medicinal Pteridophytes of India and their chemical constituents. Journal of Economic and Taxonomic Botany. 1999;23(1):63-78. 10. Dixit RD and Singh S. Medicinal Pteridophytes-An Overview. Medicinal Plants Utilization and Conservation, Aavishkar Publication, Jaipur. 2004: 268-297. 11. Singh S. Ethnomedicinal uses of pteridophytes of Amarkantak, Madhya Pradesh. Indian Journal of Traditional Knowledge. 2005;4(4):392-395. 12. Kaushik P and Dhiman A. Common medicinal pteridophytes. Indian Fern J.1995;12(1):139-45. 13. Vasudeva SM. Economic importance of pteridophytes.indian Fern J. 1999; 16(1):130-52. 14. Asolkar LV, Kakkar KK and Chakre OJ. Glossary of Indian Medicinal Plants CSIR, New Delhi. 1992;65 15. Shah NC and Singh SC. Hitherto Unreported phytotherapeutical uses from Tribal pockets of Madhya Pradesh (India). Ethnobotany. 1990; 2(1):91-95. Vol. 3 (2) Apr-Jun 2014 www.ijpcsonline.com 520

16. Shanker R and Khare PK. Ethnobotnical studies of some ferns from Panchmarhi hills(m.p).higher plants of Indian sub-continant. 1994;111:289-294. 17. Kokate CK, Khandelwal KR, Pawar AP and Gokhale SB. Practical Pharmacognosy, Nirali Prakashan,Pune, 3rd ed., 1995:137. 18. Trease and Evans. Text Book of Pharmacognosy,ELBS Publications,12th ed., 1989: 49, 126, 132-137, 205, 248. Vol. 3 (2) Apr-Jun 2014 www.ijpcsonline.com 521