52 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*** Pharmaceutical Sciences ICV 5.13*** RESEARCH ARTICLE!!! INGREDIENT IDENTIFICATION AND ANALYTICAL EVALUATION OF CHITRAKADI GHRITA, AN AYURVEDIC COMPOUND FORMULATION Dr. Deepa Chugh 1* Dr. Anup. B Thakar 2 Harisha C.R 3 Shukla V.J 4 Dr. Mannat Marwaha 5 M. D. Scholar 1 *, Associate Professor and I/C Head 2, Department of Panchakarma, Institute for Post Graduate Teaching & Research in Ayurveda, Gujarat Ayurved University, Jamnagar- 361008, Gujrat India. Head, Pharmacognosy laboratory 3, Head, Pharmaceutical Chemistry Laboratory 4, Institute for Post Graduate Teaching and Research in Ayurveda, Gujrat Ayurved University, Jamnagar, Gujrat, India M. D. Scholar 5, Department of Swasthwritta, Rajiv Gandhi Govt. Post Graduate Ayurvedic College, Paprola-176063, Dist-Kangra, Himachal Pradesh, India. KEYWORDS: Pharmacognosy, analytical, Chitrakadi ghrita, vata rogas For Correspondence: Dr. Deepa Chugh * Address: M. D. Scholar Department of Panchakarma, Institute For Post Graduate Teaching & Research in Ayurveda, Gujarat Ayurved University, Jamnagar- 361008, Gujrat India. E.mail ID: deeps.1110@gmail.com ABSTRACT Chitrakadi Ghrita, an Ayurvedic formulation is recommended in management of various Vata rogas. As Charaka in Chikitsa Sthana 28 mentioned it as Param Vataharam.The present study deals with pharmacognostical identification of ingredients of Chitrakadi Ghrita and its analytical analysis. In pharmacognostical study both macroscopic and microscopic analysis of raw drugs of Chitrakadi Ghrita has done and revealed the quality of constituents of Chitrakadi Ghrita. Specific gravity was 0.8852 at room temperature, acid value was 0.2089, saponification value 227.78 and HPTLC was carried out after organizing appropriate solvent system in which maximum 10 spots at 254 nm and 9 spots at 366 nm were distinguished and most of the R f values were identical in alcoholic extract which shows presence of certain definite constituents in Chitrakadi Ghrita.
53 P a g e International Standard Serial Number (ISSN): 2319-8141 INTRODUCTION: Ayurveda is an applied science which is practiced from the Vedic era, having an unbroken continuity till today. This immortal science of the life represents the totality of life & gives the total knowledge required to maintain the holistic balance of the functioning of the mind & body. Vata is the main Dosha among the three Doshas also the Vata Prakriti is more prone to disease condition. The Ancient scientist Achaya Charaka knows the prevention and treatment of diseases. They have mentioned various formations in Vatavyadhi adhyaya to treat the various disease caused by Vata. There is need of time to standardise these formations and assess the efficacy of theses formations. As Charaka mentioned Chitrakadi Ghrita as a Param Vataharam 1.The main ingradiants of Chitrakadi Ghrita are Chitraka (Plumbago zeylanica Linn.) Shunthi (Zingiber officinale Rosc.), Rasna (Pluchea lanceolata C.B Clarke), Pushkarmula ( Inula racemosa Hook.f.), Pippali ( Piper longum Linn.), Shati ( Hedychium spicatium Buchham), these drugs are ushna virya mostly having vatahara properties and Cow,s ghee is having yogvahi 2 (catalyst) property which It improves the pharmacokinetic of drug. The main aim of present study is to ascertain the genuinity of all ingredients of Chitrakadi Ghrita and presence of components as recommended in Ayurvedic pharmacopoeia of India Chitraka 3,Shunth 4,Rasna 5, Pippali 6, Pushkarmula 7,Shati 8 through pharmacognostical and physicochemical studies. Materials and Methods: Collection of Raw drugs: All the raw drugs of Chitrakadi Ghrita were procured from Pharmacy of Gujrat Ayurveda University,Jamnagar and were identified and authenticated by pharmacognosy laboratory of I.P.G.T & R.A, Gujrat Ayurveda University,Jamnagar. Ghee was procured from Khadi gramodyoga bhandar,jamnagar and Chitrakadi Ghrita was prepared in Pharmacy of Gujrat Ayurveda University, Jamnagar. Pharmacognostical evaluation: Organoleptic study: Individual powders were subjected for various sensory characters colour, taste, odour, nature of powder and were noted down. 9 Powder microscopy: The powders of respective parts of Chitraka, Shunthi, Pippali, Pushkarmoola, Rasna and Shati were studied with or without staining and microphotographs were taken. 10
54 P a g e International Standard Serial Number (ISSN): 2319-8141 Formulation Preparation: Chitrakadi Ghrita was prepared according to Sharangdhar 11 as he mentioned in Sharangdhar Samhita madhayam khand. Phytochemical evaluation-various phytochemical parameters like refractive index, acid value, iodine value, specific gravity of Chitrakadi Ghrita were done. HPTLC was also carried out. 12 HPTLC: Instrument used was CAMAG make HPTLC with WINCATS 1.4.3 software and Linomat 5 Sample Applicator. The Stationary Phase used was HPTLC plates silica gel 60 F 254 and mobile phase was Toluene: Et. Acetate (8:2). The sample was prepared in Methanol, and 2 l Sample was applied as 8mm band for each Spot. Detection was done @254nm, @366nm in Densitometry TLC Scanner 3. 13 RESULTS AND DISCUSSIONS: Organoleptic characters: Individual powders were subjected for various sensory characters colour, taste, odour, nature of powder and were depicted in the table No 1. Powder microscopy: The diagnostic characters under the microscope were noted are Chitraka: Cork in surface view, Sceleriform vessels, Stone cells, fibres and starch grains. ( Fig 1-a,b,c). Pippali: Starch grains, oil globules, asepted fibre, stone cells and mesocarp cells. (Fig 1-d,e,f). Pushkarmula: Cork in surface, crystal fibre, pitted stone cells, prismatic crystals, sceleriform vessels, simple starch with hilum and tannin material. (Fig 2-a,b,c). Shunthi: Cork in tangential view, oleoresins, sceleriform vessels, simple fibres, and simple starch grains (Fig 2-d,e,f). Rasna: Rhomboidal crystal of calcium oxalate, cork in surface view, Border pitted vessels, lignified fibres, simple and compound starch grains with hilum, Stone cells and tannin content. (Fig 3-a,b,c). Shati: Annular vessels, cork in surface view, multicellular simple trichome, prismatic crystals, simple fibre, simple starch grains and tannin.(fig 3-d,e,f) The ingredients after mixing properly and once again the compound powder subjected to microscopy to confirm the ingredients and cross verified. Preparation of Chitrakadi Ghrita- Chitrakadi Ghrita was prepared according to Sharangdhar. Ghee was poured in a large stainless steel container and when it melts under moderate flame kalka of Chitraka, Shunthi,Rasna, Pushkarmula,
55 P a g e International Standard Serial Number (ISSN): 2319-8141 Pippali, Shati was added and then four times water that of ghee was added and heated until sneha sidhi lakshana were observed. Table No 2. Analytical results of Chitrakadighrita Results of Specific gravity, refractive index, iodine value and saphonification values are tabulated in the table No.3. HPTLC of Chitrakadighrita 10 of spots at 254nm and 9 spots 366nm were recorded and results were tabulated in the table No 4 represented in the diagram (Fig 4 a&b). DISCUSSION: Chitrakadi Ghrita one of the important polyherbal formulation widely using in Vata rogas. The ingredients which are mentioned were identified, authentified. The microscopic characters commonly observed were Calcium oxalate crystals, tannin content cells, cork, border pitted vessels, sceleriform vessels and starch grains are very commom in all the ingredients. Stone cells observed only in Chitraka and Pushkarmula. The microscopic characters were specific and also observed after proper mixing. The iodine value, refractive index, acid value, saponification value of the Ghrita were within the normal reference range. 14. HPTLC shows that nearly 3 spots were merging in both the long and short UV. CONCLUSION: The microscopic characters that the purity of the finished product first time evaluate the raw drugs individually as well as in mixing condition. HPTLC results show specific range of spots in both long and short UV. Thus it is concluded that the values may be considered for further standardisation. Table No. 1. Organoleptic study-individual powders Name Taste Colour Odour Nature of powder Chitraka Astringent Light Brown Characteristic Coarse Shunthi Pungent Creamish yellow Aromatic Fine Pippali Pungent Greenish black Characteristic Coarse Pushkarmula Kashaya Irritant Yellowish brown Aromatic Coarse Rasna Bitter Blackish brown Aromatic Coarse Shati Sweetish Whitish cream Aromatic Fine
56 P a g e International Standard Serial Number (ISSN): 2319-8141 Table. No. 2. Contents and proportion of Chitrakadi Ghrita Sr. No. Botanical Name Part Used Quantity 01 Chitraka (Plumbago zeylanica Linn.) Moola 1/6 part 02 Shunthi (Zingiber officinale Rosc.) Moola 1/6 part 03 Rasna (Pluchea lanceolata C.B Clarke) Moola 1/6 part 04 Pushkarmula ( Inula racemosa Hook.f.) Moola 1/6 part 05 Pippali ( Piper longum Linn.) Moola 1/6 part 06 Shati ( Hedychium spicatium Buchham) Moola 1/6 part 07 Cow,s ghee 4 part Table. No. 3. Analytical results of Chitrakadighrita TEST RESULT Specific gravity 0.8852 Refractive index 1.468 Iodine value 34.77 Acid value 0.2089 saponification value 227.78 Table No. 4. HPTLC Results of Chitrakadighrita Sr. No. No. of Spots R f Values UV SHORT 10 spots 0.02, 0.16, 0.26, 0.30, Track 2-254 nm 0.40,0.47,0.51,0.64,0.81,0.91 UV LONG- 366 09 Spot 0.02,0.26,0.30,0.39,0.44,0.54 0.64,0.85,0.98
57 P a g e International Standard Serial Number (ISSN): 2319-8141 Legends: Fig 1a- Cork-Chitraka, Fig 1b- Sceleriform vessels- Chitraka, Fig 1c- Stone cells- Chitraka, Fig 1d- Stone cells in group-pippali, Fig 1e- Asepted fibre-pippali, Fig 1f- Mesocarp cell- Pippali.
58 P a g e International Standard Serial Number (ISSN): 2319-8141 Fig 2a- Cork-Pushkarmula, Fig 2b- Crystal fibre- Pushkarmula, Fig 2c- Pitted stone cells- Pushkarmula, Fig 2d- Oleoresins-Shunthi, Fig 2e- Simple starch grains-shunthi, Fig 2 f- Cork-Shunthi
59 P a g e International Standard Serial Number (ISSN): 2319-8141 Fig 3a- Rhomboidal crystal- Rasna, Fig 3b- Cork in surface- Rasna, Fig 3c- Border pitted vessel- Rasna, Fig 3d- Prismatic crystal- Shati, Fig 3e- Annular vessel- Shati, Fig3f- Tannin- Shati
60 P a g e International Standard Serial Number (ISSN): 2319-8141 Fig 4a- Spectral comparison at 254 nm, Fig 4b-. Spectral comparison at 366 nm.
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