Evaluation of Ayurvedic Compound Formulation – Krimimudgara Rasa
Saluja AK1, Shah UD1, Mona R Kukkar*1, Rajiv R Kukkar2 and Shukla SH2
1A. R. College of Pharmacy, Vallabh Vidyanagar, Gujarat
2Indukaka Ipcowala College of Pharmacy, New Vallabh Vidyanagar, Gujarat
*Corresponding Author E-mail: rajivkukkar@rediffmail.com
ABSTRACT
Quality assurance is an integral part of all systems of medicine to ensure quality medicament. Thus, there is an urgent need to evaluate such parameters which can be adopted by the pharmaceutical industries. In the communication, attempts have been made to evaluate Krimimudgara Rasa, an Ayurvedic compound formulation. one sample procured from market (MF) and one sample prepared in laboratory (LF) were subjected to physicochemical analysis, HPTLC fingerprinting, and botanical characterization, and compared using authentic ingredients as reference. It was observed that the microscopic and chromatographic analyses compliment each other in their findings, and can be used effectively for the identification of raw materials in the compound formulation (s).
KEYWORDS: Krimimudgara Rasa, Ayurvedic formulation, Quality control parameters, Drug standardisation, Pharmacognosy
INTRODUCTION:
Herbal medicine has been enjoying renaissance among the customers throughout the world. However, one of the impediments in the acceptance of the Ayurvedic or Siddha formulations is the lack of standard quality control profiles.1
Due to lack of proper quality control methods, there are batch to batch variations amongst the same product obtained from different sources. The main problem encountered while working with compound formulations is that most of them consist of several ingredients, and the presence of each ingredient has to he confirmed in the final product2. The ongoing research has led to the development of methods for the evaluation of Krimimudgara Rasa. (KMR)
Krimimudgara Rasa as prescribed in the Ayurvedic formulary claimed to be used as Kriminaaashaka i.e. anthelmintic and which consists of a moderately fine powder of one part of Shudh Parad (pure mercury), two parts of Shudh Gandhak (pure sulphur), three parts of Ajmod (Apium leptophyllum fruits), five parts of Vidang (Embelia ribes Burm.f. fruits), six parts of Kuchla seeds (Strychnous nux-vomica seeds), seven parts of Palashbij ( Butea monosperma Taub.seeds)3,4
MATERIAL AND METHOD:
Samples of Krimimudgara Rasa of one batch was procured from one Ayurvedic pharmacy (Market Formulation,MF) and one sample of Krimimudgara Rasa was prepared in laboratory (Lab Formulation, LF) based on traditional methods in accordance with procedure given 5,6
Authentic samples of Ajmod (Apium leptophyllum fruits), Palashbij (Butea monosperma seeds), Vidang (Embelia ribes Burm.f. fruits) and Kuchla seeds ( Strychnous nux-vomica seeds) were used as controls.
Organoleptic characters and particle size of all the samples were recorded. Quantitative analysis for total ash, acid insoluble ash, volatile matter were carried out according to the prescribed standard methods in Indian Pharmacopoeia and elemental analysis was done by energy dispersive X-ray analysis (EDAX) instrument in both samples of KMR.
For microscopic analysis a small quantity representative of the KMR, along with the genuine samples i.e. Ajmod, Palasbij,Vidang and Jaharkuchla were mixed with water, stained with iodine and mounted in glycerine were used to examine the starch grains and its type.
Another small quantity of samples cleared by heating with chloral hydrate and mounted in glycerin was used to identify diagnostic microscopical characters of the ingredients.
Further small quantity of the KMR cleared with dilute KOH 5% was mounted in glycerine.
TLC of the methanolic extracts of all the samples and the reference ingredients was carried out on silica gel 60 F254. Precoated plates. (0.2 mm thickness, Merck)
Table-1-Physicochemical analysis of samples of Krimimudgara Rasa
|
Standard Parameter |
Formulation |
|
|
LF (% w/w) |
MF(% w/w) |
|
|
Total ash value |
7.25 |
25.4 |
|
Acid insoluble ash |
0.74 |
11.4 |
|
Volatile matter |
9.20 |
8.60 |
|
Elemental analysis 1.silicon 2.calcium 3.mercury 4.sulphur 5.pottasium 6.sodium 7.magnesium |
0.20 0.37 6.44 8.41 1.0 - - |
5.01 4.64 2.12 1.64 0.53 0.31 0.59 |
|
Percentage of embelin |
0.254 |
0.135 |
|
Percentage of strychnine |
0.080 |
0.046 |
Fig. 1 Microscopical Characters of Krimimudgara Rasa
(a) Characters of Ajmod
Endosperm cells containing aleurone grain
Vittae along with mesocarp, Lignified fibrovascular tissue
Characters of Palashbij
(c) Characters of vidang
Stone cells, Xylem vessels with spiral thickening
Mesocarp with reddish brown cells and pseudo fibres ,
Testa consisting rectangular cells with brown pigment
Characters of Kuchla
Endosperm, lignified fragment of trichomes, pigment layer ,
Endosperm with red brown content
Rod shaped fragment of lignified trichomes ,
Fig.2: HPTLC fingerprint profile of embelin and strychnine
(a) HPTLC chromatogram for embelin (scanned at 333 nm)
(b) HPTLC chromatogram for strychnine (scanned at 254 nm)
Table-2-TLC fingerprint characteristics (scanned under visible light)
|
Ingredients |
Identifying RFS Colour |
LF |
MF |
|
Ajmod |
0.86(green),0.74(pink),0.69(magenta),0.64(brown),0.40(violet) |
+ |
+ |
|
Palash |
0.26(light blue) (yellowish violet)(palmitic acid),0.84(pinkish orange) (stearic acid) |
+ |
+ |
|
Vidang |
0.44, Pink |
+ |
+ |
|
Kuchla |
0.35,(Orangebrown),0.42(Orange brown) |
+ |
+ |
The mobile phase used was toluene: ethyl acetate (7:3) for Apium leptophyllum, pet ether: diethyl ether (4.2:2) for Butea monosperma, n-propanol: n-butanol: 4N ammonia for Embelia ribes, toluene: ethyl acetate: diethylamine (7:2:1) for Strychnous nux-vomica
The plates was developed over a distance of 8 cm and visualized under visible light after derivatization.
High Performance Thin Layer Chromatography studied was done for quantification of strychnine and embelin in both the sample of Krimimudgara Rasa LF and MF.
Standard embelin and strychnine were procured from Regional Research Laboratory (RRL) Jammu, India.
RESULTS AND DISCUSSION:
Krimimudgara Rasa samples of laboratory formulation (LF) and Market Formulation (MF) were subjected to analysis as above. Both the samples were black in colour, smooth with very fine granules.
Results of ash values, volatile matter, and elemental analysis were calculated. (Table-1)Variations were observed in some of the parameters studied.Total ash, acid insoluble ash values for Market Formulation were found to be higher when compared with the corresponding values for the LF.
Volatile matter values for LF was found to be comply with standard values while for MF it was found to be lower than LF.Elemental analysis revealed the presence of higher concentration of silicon and calcium in MF, while comparison with LF and mercury and sulphur content were found to be higher in LF than MF. These variations may be due to change in the quality of the raw materials used in terms of place and period of collection and storage period and conditions.
Microscopic examinations were also carried out to see the presence of the different ingredients in both samples of Krimimudgara Rasa. (Fig.1)
Presence of yellowish brown vittae, associated with parenchyma of Mesocarp, lignified groups of fibro vascular tissues, confirm the presence of Ajmod.
Fragments of pigment layers, endosperm cell filled with oil globules and vessel elements indicate presence of Palashbij.
Likewise testa containing rectangular cells with brown pigment, Mesocarp with reddish brown cells indicates presence of Vidang.
In case of kuchla, rod shaped fragment of lignified trichomes, bunch of lignified trichomes, endosperm with red brown content were observed.
The TLC profile of the authentic samples of the ingredients i.e. Ajmod, Palashbij, Vidang and Jaharkuchla were compared with the profiles of both the samples of Krimimudgara Rasa. (Table-2)
The HPTLC finger print profiles are depicted in (Fig.2)
The LF was found to contain 0.0254% of embelin and 0.080% of strychnine while the MF showed 0.135% of embelin and 0.046% of strychnine.
CONCLUSION:
Thus from ongoing observations it can be concluded that the characteristic microscopical features and the quantification of active principle by HPTLC profiles may be utilized as marker parameter for monitoring the quality of formulation. Hence, the physicochemical parameters, quantitative analysis, HPTLC fingerprinting profiles and the microscopic characteristics together may be used for quality evaluation and the standardization of compound formulations.
REFERENCES:
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2. Rastogi S, Khatoon S, Pandey M.M, Rathi Anshu, Rawat AKS and Mehrotra S. Evaluation of Ayurvedic compound formulation 2-Palasabijadi curna, Indian Journal of Traditional Knowledge. 2008; 7; 384-388.
3. Gopal K.Rastantarsar and siddhprayog sangrah, Kaleda bhavan, Ajmer, India.1991.
4. Gupta N.The Ayurvedic system of medicine.logas press.1906
5. Shastri Ambika Dutt.Rasratansamuchhaya,Chaukhambha amarbharti prakashan.1988.
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Received on 18.03.2009 Modified on 24.05.2009
Accepted on 12.06.2009 © RJPT All right reserved
Research J. Pharm. and Tech.2 (3): July-Sept. 2009,;Page 534-536