issn 2229-3566 research article · 1ph.d. scholar, dept.of shalakya tantra, ipgt & ra, guj....

Chaudhari Varsha M et al / IJRAP 2011, 2 (6) 1681-1684

International Journal of Research in Ayurveda & Pharmacy

ISSN 2229-3566 Research Article www.ijrap.net

A PRELIMINARY PHARMACOGNOSTICAL AND PHYSICO CHEMICAL ASSAY OF

HARIDRA KHANDA GRANULES: A PILOT STUDY Chaudhari Varsha M.1*, Manjusha R.2, Dhiman K.S.3, Harish CR4, Shukla VJ5

1Ph.D. Scholar, Dept.of Shalakya Tantra, IPGT & RA, Guj. Ayurved University, Jamnagar, India 2Associate Professor, Dept.of Shalakya Tantra, IPGT & RA, Guj. Ayurved University, Jamnagar, India

3Professor & Head, Dept of Shalakya Tantra, IPGT & RA, Guj. Ayurved University, Jamnagar, India 4Head, Pharmacognosy laboratory, IPGT & RA, Guj. Ayurved University, Jamnagar, India 5Head, Pharmaceutical laboratory, IPGT & RA, Guj. Ayurved University, Jamnagar, India

Received on: 05/10/11 Revised on: 10/11/11 Accepted on: 21/11/11

*Corresponding author Varsha M. Chaudhari, Ph.D. Scholar, Email: [email protected] ABSTRACT Haridra Khanda is mentioned in the management of skin diseases like Urticaria (Udarda Shitpitta Kotha). Most of the ingredients are having properties to control allergic condition and improve Vyadhikshamatva (Immuno-modulatory). Pratishyaya (Rhinitis) is one of the common problem since ancient days; among all type of Rhinitis, Vataja type simulates the Allergic Rhinitis in modern medical literature of ENT. To overcome the problem of fungus formation in Khanda and AvalehaKalpana, Haridra Khanda has been prepared in granules form. In this study, effort has been done to overcome from the controversy regarding ingredients in the finished product and tried to make it standardized as per guidelines of API. Pharmacognostically authenticated raw drugs were used for the preparation of granules and it was analyzed through qualitative and quantitative analysis of Physico – chemical parameters. It is inferred that the formulation meets the minimum qualitative standards as reported in the API and AFI at a preliminary level. KEY WORDS: Pratishyaya, Allergic Rhinitis, Haridra Khanda, Pharmacognosy, Pharmaceutics INTRODUCTION Haridra Khanda is indicated in the management of skin diseases like Urticaria (Udarda Shitpitta Kotha) in one of the authenticated text of Ayurvedic formulations i.e. Bhaishajya Ratnavali1. It contains Haridra, Triphala, Trikatu, Trijata, Vidanga, Goghrita, Sita etc. (table 1). All the drugs of Haridra Khanda are having properties to control the allergic condition and improve Vyadhikshamatva (Immunomodulatory); encouraging results were observed with Haridra Khanda Granules in Pratishyaya – Allergic Rhinitis in the previous research work2. Pratishyaya (Rhinitis) is a common nasal problem recognized since ancient days; Acharya Sushruta has assigned a chapter3 for this clinical entity which denotes its importance. Among all type of Pratishyaya, Vataja type is recurrent in nature and simulates the Allergic Rhinitis in modern medical literature of ENT; which is very difficult to treat, never a fatal illness but may act as forerunner of Asthma. Hence in the present study, Haridra Khanda Granules has been selected to evaluate its efficacy clinically in Vataja Pratishyaya – Allergic Rhinitis. As Jamnagar is the coastal area, there is common problem of fungus formation in Avaleha & Khanda Kalpana; hence, for better palatability and good life span, in this work Haridra Khanda has been prepared in granules form. Proper identification and standardization of the drug is essential and each drug has its own physical and chemical characteristics that help for separating it from other closely related drugs. To validate the standardization of the drug, physicochemical studies by various parameters are needed. Hence, the raw drugs were identified and authenticated by pharmacognosy. Then the granules were prepared with standard operating procedure and an attempt has been made to standardize the finished product by the Pharmacognostical and Physico chemical standards to overcome the controversy regarding the ingredients of the Haridra Khanda as per guidelines of API and AFI.

Aims & Objectives 1. Pharmacognostical Study of finished product – Haridra Khanda Granules 2. Physico-chemical analysis of Haridra Khanda Granules MATERIALS & METHODS Collection Of Raw Drugs Raw drugs except Shunthi, Nagakesara and Tamalapatra for the study were procured from the Pharmacy, Gujarat Ayurved University, Jamnagar. Shunthi and Tamalapatra were purchased from local market of Jamnagar whereas Nagakesara Pushpa was procured from Alva Pharmacy, Karnataka. Ingredients Of Haridra Khanda Granules The formulation composition of Haridra Khanda along with the proportion and part used of individual components’ is depicted at Table 1. Identification And Authentification Of Raw Drugs Raw drugs were identified and authenticated by the department of Dravyaguna and Pharmacognosy laboratory IPGT &RA, Gujarat Ayurved University, Jamnagar. The identification was carried out based on the morphological features, organoleptic features and powder microscopy of the individual drugs. The API4standards were referred for the authentification. Method Of Preparation Of The Haridra Khanda Granules Haridra Khanda Granules were prepared at Rasashastra and Bhaishajya Kalpana Department of I.P.G.T. & R.A., Gujarat Ayurved University, Jamnagar. Dried raw ingredients (Kashtha Aushadhi) of Haridra Khanda Granules were powdered separately and passed through sieve 85# to obtain fine powder and mixed thoroughly in specified quantity (Except Haridra) to obtain a homogeneous mixture along with Lauha Bhasma. Then Haridra Powder was fried in Go – Ghrita maintaining the temperature between 80 to 90°c till Haridra turned brown and its typical smell emanated; after that Go – Dugdha was added and stirred. Sugar syrup was prepared and filtered while hot through muslin cloth. Fried Haridra was added to the syrup and heated with constant stirring maintaining the temperature at about 90°c. After that mixture of fine powder was added and mixed thoroughly to



prepare a homogeneous blend. Then the blend was passed through 40# sieve to obtain granules and dried at room temperature, packed in airtight containers. Material For Pharmacognosy Of Finished Product Haridra Khanda Granules were dissolved in small quantity of distilled water, filtered through filter paper, filtrate air dried and observed under microscope with and without stain; microscopic

characters were recorded and took micro photographs under corl zeiss binocular microscope attached with camera. Material For Physico - Chemical Study Of Finished Product Haridra Khanda Granules were analyzed by using, qualitative and quantitative parameters as per the guideline of API5 at Pharmaceutical Chemistry Laboratory of I. P. G.T & R. A., Gujarat Ayurved University, Jamnagar.

Preparation Of Methanol Extract Granules weighing 5 gm were taken with 100 ml of alcohol kept for twenty-four hours. Filtrate was prepared and evaporated till it gets dried in a flat-bottomed shallow dish and concentrated on water bath to volume of requirement.

RESULTS & DISCUSSION Pharmacognostical Study Of Haridra Khanda Granules Organoleptic and Microscopic study of the powdered drug was done as per the guidelines of Ayurvedic pharmacopoeia of India6 at Department of Pharmacognosy, I.P.G.T & R.A, Jamnagar.

Organoleptic Characters Organoleptic characters of the sample are tabulated in table 2. Powder Microscopy Diagnostic microscopic characters of Haridra Khanda Granules are - starch grains and olioresin from Haridra; stone cells of Pippali; compound starch grain, pollen grain and brownish coloring matter of Nagakesar; prismatic and acicular crystals of calcium oxalate of Tamalpatra, brownish red colouring matter of Vidanga, stone cells and stone cells in group from Haritaki, oil globules and vessels along with oil globules, lignified fibres of Twaka, scleroids of Amalaki and trichomes from Bibhitaki were observed (Plate No – 1,2) All these microscopic characteristics identified were equivalent to the standard individual profile as per API. Physico - Chemical Study Organoleptic Characters Organoleptic characters of the sample are tabulated in table 2 as mentioned in Pharmacognostical study. Physico-Chemical Parameters Granules were evaluated for physico-chemical parameters like Total Ash Value, Loss on drying, pH value, Sugar estimation (Total sugar, Reducing Sugar, Non - Reducing Sugar), Alcohol soluble and water-soluble extractive values. Presence of more moisture content in a sample can create preservation problem. Hence loss on drying was also selected as one of parameters. Since, the sample was in the form of granules, which contains significant quantity of sugar; sugar estimation was considered as another parameter. The results are placed at table 3. Qualitative Test Of Haridra Khanda Granules Methanol extract of the sample was analyzed qualitatively for different functional groups. Details are placed in table 4. Thin Layer Chromatography TLC is mentioned as a primary tool for identification as part of monographs on all medicinal plants. Alkaloid fraction was used for the spotting of the TLC plate (Silica gel G Precoated plates). Then the spotted TLC was run with the solvent systems (Toluene (7 ml), Ethyl acetate (2 ml), Glacial acetic acid (0.5 ml), Formic Acid (0.5 ml)) separately; the resulting TLC pattern was viewed under long wave ultra violet light at 366 nm or Short wave ultra violet light at 254 nm. Repeated TLC was done to develop proper solvent system and above mentioned solvent system was finalized. High Performance Thin Layer Chromatography Methanol extract of Haridra Khanda Granules were spotted on pre coated silica gel GF 60254 aluminium plate as 5mm bands, 5mm apart and 1cm from the edge of the plates, by means of a Camag Linomate V sample applicator fitted with a 100 μL Hamilton syringe. Toluene (7 ml), Ethyl acetate (2 ml), Glacial acetic acid (0.5

ml), Formic Acid (0.5 ml) (v/v) (20ml) was used as a mobile phase. After development, Densitometric scanning was performed with a Camag T.L.C. scanner III in reflectance absorbance mode at 254 nm and 366 nm under control of win CATS software (V 1.2.1 Camag). The slit dimensions were 6 mm x 0.45 mm and the scanning speed was 20 mm s-1. Then the plate was sprayed with Anisaldehyde Sulphuric acid followed by heating and then visualized in day light. The findings of High – performance Thin Layer Chromatography of Methanolic extract of Haridra Khanda Granules in 254 nm, 366 nm and in visible day light after spraying Anisaldehyde Sulphuric acid are depicted in table 5. CONCLUSION The formulation Haridra Khanda Granules was subjected to pharmacognostical, physico-chemical studies. It is inferred that the formulation meets the minimum qualitative standards as reported in the API at a preliminary level. AFI advocates preparing Haridra Khanda Granules in presence of water. In this attempt, Go – Dugdha was used instead of water considering the classical reference. Hence, slight differences have been observed in physico– chemical parameters of the granules. The observations may be considered in further studies as reference. ACKNOWLEDGMENT Authors wish to thank Prof. Prajapati PK, Director of Pharmacy, Head Dept. of Rasashastra and Bhaishajya kalpana including drug research; Dr. Galib, Assistant Prof. Dept. of Rasashastra and Bhaishajya kalpana including drug research IPGT & RA, Gujarat Ayurved University, Jamnagar for their sincere guidance in the preparation of granules. REFERENCES 1. Bhaishajya Ratnavali of Shri Govind das (2005), 8th revised edited and enlaged

by Bhishagratna Shri Brahmashanka Mishra, ‘Vidyotini’ Hindi Commetary Analysis with Appendixes by Shri Kaviraja Ambikadatta Shastri Ayurvedacharya, ed. Shri Rajeshwadatta Shastri, Chaukhambha Sanskrit Sansthan, Varanasi, 55/13-17.

2. Bhakti Chhaya et.al.(2008): A Clinical Evaluation of Haridra Khanda & Pippalyadi Taila Nasya on Pratishyaya – Allergic Rhinitis, in Dept. of Shalakya, I.P.G.T. & R.A., Jamnagar.

3. Sushruta, Sushruta Samhita Dalhana Comm. - Nibandhasangraha, Gayadasacharya comm.-Nyayachandrika Panjika on Nidanasthana, Ed. By Vd. Jadavaji Trikamji Acharya & Narayana Ram Acharya, Chaukhamba Surbharati Prakashana, Varanasi, 2008.

4. Anonymous, The Ayurvedic Pharmacopoeia of India, Part- I, Vol I (2001), Vol. II (1999), Vo. III (20001), Vol. IV (2004), 1st Edition, Ministry of Health And Family Welfare, Department of AYUSH (Government of India).

5. Anonymous, The Ayurvedic Formulary of India, Part I, Vol. I, 2nd Edition, 2001, Ministry of Health And Family Welfare, Department of AYUSH (Government of India).

6. Anonymous, The Ayurvedic Pharmacopoeia of India, Part- I, Vol I, 40, 1st Edition, 2001, Ministry of Health And Family Welfare, Department of AYUSH (Government of India) .

7.



Table 1: Ingredients Of Haridra Khanda Granules Ingredient Part used Proportion Goghrita - 6 part

Godugdha - 64 part Sita - 50 part

Haridra (Curcuma longa Linn.) Rhizome 8 part Shunthi (Zingiber officinale Rosc.) Rhizome 1 part

Maricha (Piper nigrum Linn.) Fruit 1 part Pippali (Piper longum Linn.) Fruit 1 part

Twak(Cinnamomum zeylanica Blume.) Bark 1 part Ela (Elettaria cardamomum Maton) Seed 1 part

Tamala patra (Cinnamomum tamala Nees & Eberm) Leaf 1 part Vidanga (Embelia ribes Burm.f.) Seed 1 part

Trivrita (Operculina turpethum Linn. (Silva Manso)) Root 1 part Haritaki (Terminalis chebula Retz.) Pericarp 1 part

Bibhitaki (Terminalia bellirica Roxb.) Pericarp 1 part Amalaki (Emblica officinalis Gaertn.) Pericarp 1 part

Nagkesara (Mesua ferrea Linn.) Anthers 1 part Musta (Cyperus rotundus Linn.) Rhizome 1 part

Lauha bhasma - 1 part

Table 2: Organoleptic Characters Of Haridra Khanda Granules S.No Parameters Sample - Haridrakhanda Granules

1 Texture Rough 2 Color Yellowish 3 Taste Sweet, Bitter 4 Odor Non Irritant

Table 3: Physico – Chemical Parameters Of Haridra Khanda Granules

S. No. Parameters Sample - Haridrakhanda Granules 1. Loss on drying 3 % w/w 2. Water soluble Extract 71.20 % w/w 3. Alcohol soluble Extract 32 % w/w 4. Total Ash 8.50 % w/w 5. pH Value 4.96 (Acidic)

6. Sugar Estimation Total Sugar – 59.18%w/w Reducing Sugar – 7.20%w/w

Non - Reducing Sugar – 51.98% w/w

7. Iron (Fe2O3)content 0.49%

Table 4: Qualitative Test Of Haridra Khanda Granules Sr.No Test Name of Reagents Results

1 Carbohydrate Molisch’s test Positive 2 Steroid / Sterol Libermann – Burchard test Positive 3 Saponin Glycosides Foam Test Abscent 4 Flavonoids Shinoda test Positive 5 Alkaloids Dragendroff’s test Abscent 6 Starch Iodine test Abscent 7. Tannin& Phenolic Compound Ferric chloride test Positive

Table 5: HPTLC Findings Of Haridra Khanda Granules

Sr. No. Visualization No. of the spots Rf value

1. 254 nm 8

0.02 0.26 0.31 0.41 0.49 0.57 0.66 0.76

2. 366 nm 7

0.02 0.27 0.40 0.49 0.66 0.88 0.93

3. After spraying

Anisaldehyde sulphuric acid

5

0.22 0.26 0.40 0.70 0.82

issn 2229-3566 research article · 1ph.d. scholar, dept.of shalakya tantra, ipgt & ra, guj....

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