98 P a g e International Standard Serial Number (ISSN): 2319-8141 International Journal of Universal Pharmacy and Bio Sciences 3(5): September-October 2014 INTERNATIONAL JOURNAL OF UNIVERSAL PHARMACY AND BIO SCIENCES IMPACT FACTOR 2.093*** Pharmaceutical Sciences ICV 5.13*** RESEARCH ARTICLE!!! PHARMACOGNOSTICAL AND PHARMACEUTICAL STUDY OF NAVAYASA CHURNA IN THE MANAGEMENT OF PANDU W.S.R. TO IRON DEFICIENCY ANAEMIA (IDA) Hetal D. Vyas 1*, Alankruta R. Dave 2, Harisha C R 3, V J Shukla 4 1* Ph.D. Scholar, Dept of Kayachikitsa, 2 Associate Professor and I/C Head of Kayachikitsa Department, 3 Head, Pharmacognosy Laboratory 4 Head, Pharmaceutical Chemistry Laboratory, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar- India. ABSTRACT KEYWORDS: Pandu is a disease characterized by pallor of body which we can correlate with Anaemia of modern science. The commonest type of Navayasa churna, Anaemia is met in practice is iron Deficiency Anaemia is treated by Pharmacognostical, oral iron formulations those are also available in Ayurvedic classical books. Navayasa Churna is mentioned in Bhaishajyaratnavali in Pharmaceutical, HPTLC. Pandu roga chikitsa 1. ingredients of this formulation having Deepan For Correspondence:,Pachan,Krimighna,Anuloman, rasayana properties and Lauha bhasma Hetal D. Vyas * improves rakta dhatu work on Pandu. In the present study a systematic approach has been evolved and effort has been made to develop well Address: Ph.D. scholar, designed methodology for the standardization of this Formulation. The Dept of Kayachikitsa finished product was subjected to organoleptic, microscopic characterization, physico-chemical analysis and HPTLC studies. The I.P.G.T. & R.A. G.A.U., pharmacognostical evaluation shows fragments of mesocarp cells, Jamnagar, Gujarat, India sclereids, tannin contents of Haritaki; Silica deposition of Amalaki: 361008 Unicellular covering Trichomas, stone cells, Rosset cristal of bibhitaki. In physico-chemical evaluation hardness, weight variation, moisture E mail: content, ash values viz., acid insoluble ash, alcohol soluble extractive vyashetal1979@gmail.com value, water soluble extractive value as well as ph value etc. were determined. Spots obtained in HPTLC were found 5 spots of maximum drugs at Rf values also reported in previous study.
99 P a g e International Standard Serial Number (ISSN): 2319-8141 INTRODUCTION: Accurate identification and quarantee of purity through pharmacognosy and pharmaceutical chemistry measure is inescapable ladder needed for the quality assurance and standardization of any of the herbal medicine whether it is single drug or formulation. Navayasa churna is a herbomineral compound preparations mentioned in Bhaishjya Ratnavali in the management of Pandu. Nine out of ten anaemia suffers live in developing countries, about 2 billion people suffer iron deficiency anaemia. 2 The subject of herbal drug standaedization is massively wide and deep. India can emerge as the major country and play the lead role in production of standardized, therapeutically effective Ayurvedic formulations. The report on the standardization of Navayasa churna in present study is based on organoleptic, microscopic, Analytical and HPTLC study. Materials and Methods Collection and preparation of drug Dried specimens of mature fruits of Haritaki, Bibhitaki, Amalaki, Marich, Pippali, Vidanga, Rhyzome of Sunthi, Musta, root of chitraka were procured from the Pharmacy of I.P.G.T. & R.A. G.A.U. Jamnagar. Navayasa Lauha was obtained from the pharmacy and tablet was prepared in the Dept. of RS & BK, I.P.G.T. & R.A. Jamnagar, India..The obtained fruits, Roots and Rhyzomes were shade dried and made in to fine powder separately with help of mechanical grinder and mix Lauha bhasma binding agent and make the tablet form as per description in ayurvedic classics. (Table-1) Pharmacognostical study Pharmacognostic study gives the scientific information regarding the purity and quality of the plant drugs. The detailed pharmacognostical evalution gives valuable information regarding the morphology, microscopical and physical characteristics of crude drugs. Orgrnoleptic evaluation Various parameters such as colour, odour, taste, touch and texture of the finished product were observed and recorded. (Table-2). Microscopic Evaluation Sample drug was powdered, dissolved in a small amount of distilled water for a while and then mountedin glycerin. Microscopical examination was carried out with and without staining.vi By powder microscopy, to observe the characters, determine the chemical nature of the cell wall along with the determination of the form and chemical nature of the cell contents. Microphotographs were taken by using Carl Zeiss binocular microscope attached with camera.3
100 P a g e International Standard Serial Number (ISSN): 2319-8141 Analytical Study: Physico- chemical Analysis: In physical evaluation hardness, weight variation, ash values viz., total ash, acid insoluble ash and extractive values viz., alcohol soluble extractive values, water soluble extractive value as well as ph value etc. were determined. 4 (Table 3) High performance thin layer Chromatography (HPTLC) HPTLC was performed as per the guidelines provided by API. Methanolic extract of drug sample was used for spotting.hptlc was performed using toluene + Ethyl acetate + acetic acid (8 : 1: 1) solvent system and observed under visible light after Spray of vanillin sulfuric acid followed by heating the plate at 110 C. The colour and Rf values of the resolved spots were noted. 5 (Table 4) Results Orgenoleptic characters Tablet characterized as hard in texture, hard in touch brown in colour characteristic in odour and astringent in taste. Microscopical characters Diagnostic characters of microscopic analysis of test drug shows the present of Simple and circular starch grains of sunthi, Fibers of Chitraka, Trichome of Bibhitaki, Scleroids of Haritaki, Rosette crystal of Bibhitaki, starch grains of Musta, colouring matter of Vidanga, stone cells of Vidanga, starch grains of Maricha, pitted stone cells of Chitraka, stone cells of Haritaki, Silica crystals of Musta, starch grains of Chitraka, oil globule of Musta, Prismatic crystals of Chitraka, oilresin contain of Shunthi. Scleroids of Amalaki, Bottle neck shaped stone cells of Pippali. Physico chemical parameters Physicochemical test of Navayasa churna revealed loss on drying 9.56% w/w, total ash content 49.80% w/w, acid insoluble ash 1.84%w/w, water soluble extract 12.7% w/w, methanol soluble extract 9.7% w/w, and ph 4.56. High performance thin (HPTLC) layer chromatography The HPTLC profiles of methanolic extract of the formulation are super - imposable indicating the presence of all the constituents as per the ingredients. Some of the spots at Rf 0.02,0.14,0.24,0.61,0.73,0.92 were observed in[254]nm the short UV light and some spots at Rf.0.02,0.14,0.23,0.61,0.73 were observed in [366]nm long UV light spectrum. HPTLC of the test drug after spray of with vanniline sulphuric acid reagent showed three major spots. After analysis of Navayas churna by different parameters such as foreign matter, moisture content, total ash, acid insoluble ash, water and alcohol soluble extractives and HPTLC densitograms shows good corelation between them and are similar as per the previous reported works. The study of microscopic characters of present formulation shows the presence of diagnostic identifying characters of
101 P a g e International Standard Serial Number (ISSN): 2319-8141 ingredients which are used. Purity and potency of the materials and formulations following the procedures given could be performed in laboratory of pharmaceutical house. Discussion Pharmacognocy study helps in aunthentication of the commonly used drugs through morphological, histological and physic-chemical parameters. This can prevent the accidental misuse of drugs and adulteration to a greater extent. 6 Total ash is designed to measure the total amount of material remaining after ignition. In present study total ash value of Navayas churna is very high (49.80%w/w) because Lauha bhasma is a mineral & not ignited. ph value is measure the acidic & basic solution of the sample.ph value of Navayas churna is 4.56 is determine the acidic nature of the solution. Loss on drying method determines the amount of water, the part of water for crystalization or in the sample. Loss on drying of test drug is 9.56%w/w. HPTLC is the most common form of chromatography method used by ayurvedic research works to detect the number of compounds present in a product. After careful analysis & discussion with experts the mobile phase was fixed to be Toluene+Ethylacetate in the proportion of 8:1:1 respectively. The spots produced by HPTLC were observed in day light short UV and long UV and Rf value was calculated. Track showed 6 spots under 254nm with Rf values and 5 spots were seen under 366nm with Rf values. After completion of chromatographic procedure spraying of the plate was done with vanillin sulfuric acid and the spots obtained were observed in day light. Conclusion The groundwork requisites for the standardization of Navayasa churna tab is covered in the current study additional cavernous analysis and investigation are required for the identification of all the active chemical constituents of the test drug to substantiate the clinical efficacy Pharmacognostical findings confirm the ingredients present in the Navayasa churna and raw drugs cross verified with API no major change in the microscopic structure during the pharmaceutical processes of preparation of Tablet 7. TABLE 1 INGREDIENTS OF NAVAYASA CHURNA No. Name of Drug Botanical name Family Part used Parts 1 Haritaki Terminelia chebula Retz. Combrataceae Fruit 1 Part 2 Bibhitaki Terminelia bellirica Roxb. Combrataceae Fruit 1 Part 3 Amalaki Emblica officinalis Gaertn. Euphorbiaceae Fruit 1 Part 4 Sunthi Zingiber Officinale Rosc. Zingiberaceae Rhizome 1Part 5 Maricha Piper nigrum.l Piperaceae Fruit 1Part 6 Pippali Piper longum L Piperaceae Fruit 1 Part 7 Musta Cyperus rotundus L Cyperaceae Rhizome 1 Part 8 Vidanga Embelia ribs Burm f. Myrsinaceae Fruit 1 Part 9 Chitrak Plumbago Zeylanica L. Plumbaginaceae Root 1 Part 10 Navayasa Lauha Incenereted Iron 9 Part Total 18 Parts
102 P a g e International Standard Serial Number (ISSN): 2319-8141 TABLE -2: ORGANOLEPTIC CHARACTERS OF NAVAYASA CHURNA No Parameters Observation 1 Colour Brown 2 Odour Characteristic 3 Taste Kashaya and Tikta 4 Consistency Solid TABLE 3 PHYSICO CHEMICAL ANALYSES No. Test Navayasa churna - Tablet 1 Uniformi tyof Tablet (a) Average Weight (mg) 345 (b) Highest Weight (mg) 385 (c) Lowest Weight (mg) 296 2 Tablet Hardness (Kg/cm²) 0.99 3 Tablet Disintegration Time (min.) 4 4 Loss on drying % w/w 9.56 5 Ash value % w/w 49.80 6 Acid insoluble Ash %w/w 1.84 7 Water soluble extract% w/w 12.7 8 Methenol soluble extract % w/w 9.7 9 ph (5% v/w Aqua solution) 4.56 TABLE 4 HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHY (HPTLC) Drugs Name (Tablet) Navayasa churna Under U. V. Radiation Post chromatographic 254 nm 366 nm visualization ( After spray of Vanillin sulfuric acid) No. of Rf nm No. of Spot Rf nm No. of Spot Rf nm Spot 1 0.02 1 0.02 1 0.32 2 0.14 2 0.14 2 0.34 3 0.24 3 0.23 3 0.44 4 0.61 4 0.61 5 0.73 5 0.73 6 0.92
103 P a g e International Standard Serial Number (ISSN): 2319-8141 PHOTO PLATE - 1 Simple Starch grains of Shunti Fibres of Chitraka Trichome of Bibhitaki Scleroids of Haritaki Rosette crystal of Bibhitaki Starch grains of Musta Colouring matter of Vidanga Stone cells of Vidanga Starch grains of Maricha
104 P a g e International Standard Serial Number (ISSN): 2319-8141 Pitted stone cells of Chitrak Stone cells of Haritaki Silica crystals of Musta Starch grains of Chitraka Oil globule of Musta Prismatic crystals of Chitraka Olioresine contain of Shunti Scleroids of Amalaki Bottle neck shaped stone cells of Pippali
105 P a g e International Standard Serial Number (ISSN): 2319-8141 PHOTO PLATE NO 2 DENSITOGRAM Densitogram curve of Methanol extract Navayasa churna at 254 nm Densitogram curve of Methanol extrct of Navayasa churna at 366 nm PHOTO PLATE NO. 3 HPTLC PLATE OF METHANOLIC EXTRACT OF NAVAYASA CHURNA 254nm 366nm
106 P a g e International Standard Serial Number (ISSN): 2319-8141 REFERENCES: 1. WHO Turning the tide of malnutrition ; responding to the challenge of the 21 st century. Geneva; WHO, 2000 (WHO/NHD,007). 2. Bhaishjyaratnavali Pandu Rogadhikar 22. 3. Trease and Evans, Pharmacognosy, 15 th Ed., W.B. Sunders Company Ltd. 1996 p.569,570. 4. Anonymous, The Ayurvedic Pharmacopoeia of India, 1st Edition, Ministry of Health and Family Welfare, Govt. of India, Part I, Vol. I1999,Appendix - 3, Pg -230 (3.3). 5. Wagener H and Bladt S ; Plant drug analysis A thin layer Chromatography atlas, Berlin: Sprnger 1996, 230-231. 6. Brain K.R and Turner T.D, in the practical evaluation of Phytopharmaceuticals, Wright Scientenchnica, Bristol, 1975, 1 st edition. Page 142. 7. Anonymous. The Ayurvedic Pharmacopoeia of India. Part 1, Vol. 1, 2, 3, 4; 1st ed. New Delhi: Ministry of Health and Family welfare, Department of AYUSH Government of India; 2008.