Physicochemical, Preliminary Phytochemical and Powder Microscopical studies of “M-Swadishta Virechana Churna” – An Ayurvedic Polyherbal Formulation

Ayurvedic formulation M-Swadista virechana churna is known to be an effective laxative in diseases like Arsha (heamorrhoids), Amasangraha (accumulation of Ama-undigested toxic substance) and Vibandha (constipation). Literature reviews revealed that there are no reports available on the physicochemical standardization & Powder microscopy of “M-Swadishta virechana churna. Hence, analytical studies have been taken up to derive the Pharmacopoeial standards for quality control. The ayurvedic compound formulation, M-Swadishta virechana churna contains Swarnapatri (Cassia angustifolia), Madhuka (Glycerrhiza glabra) and Ikshu Sarkara (cane sugar- Saccharam officinarum). The present communication deals with the physico chemical, preliminary phyto chemical and chromatographic with powder microscopical studies gave valuable parameters for laying down Pharmacopoeial standards of the compound formulation.


Introduction
Ayurveda is an ancient medical science for the restoration of health and prevention of diseases. Swadista virechana churna is one among the popularly used Ayurvedic compound formulation administered as a laxative in diseases like malavaroda (constipation) , amavruddhi(increase of undigested toxic substance) , shiroruja(headache), arsha (haemorrhoids), raktavikara(blood disorders) , charmaroga (skin diseases), pama(scabies), udara shodanartha action and frequently used in Indian systems of medicine (2).
The active component of senna leaves Anthraquinone glycoside molecules are converted by the normal bacteria in the colon into rhein-anthrone, which in turn has two effects. It first stimulates colon activity and thus speeds bowel movements. Second, it increases fluid secretion by the colon. Together, these actions work to get a sluggish colon functional again. Several controlled studies have confirmed the benefit of senna in treating constipation (3).
The other ingredient of formulation, Madhuka (Glycyrrhiza glabra Linn) is commonly known as liquorice and sweet wood belonging to leguminosae family. The liquorice roots are sweet, refrigerant, tonic, mild laxative, aphrodisiac, haemostatic and an active anti-allergic, anti-inflammatory, spasmolytic, antiulcer and liver protective herb is widely used Ayurveda. Glycerrhiza glabra Linn contains tri terpene, saponins, flavonoides, polysaccharides, pectines, simple sugars, amino acids, mineral salts and various other substances(4). Glycyrrhizin, a triterpenoid compound accounts for the sweet taste of Glycyrrhiza glabra Linn. roots (Madhuka). The isoflavones, glabridin and hispaglabridines A and B have significant antioxidant activity (5).In vitro studies have demonstrated liquorice isoflavones, hispaglabridines A and B inhibit Fe 3+ induced mitochondrial lipid peroxidation in rat liver cells (6).
Another adjuvant ingredient of the compound formulation, Ikshu sarkara (cane sugar-Saccharam officinarum) have the properties of sweet, unctuous, coolant, nourishing, aphrodisiac, mild laxative, antiemetic (7). It offers a host of essential nutrients and minerals, which contains magnesium, phosphorus, calcium, potassium and iron. Cane sugar also have mild laxative action, thus helps in prevent cancers of colon, prostate and breast cancer and also helps revitalize, energize and hydrate the body (8).
Though, the leaves of Swarnapatri or Senna (Cassia angustifolia) have an excellent laxative quality, but afterwards have a binding effect. It can also cause nausea and griping pains when used alone due to inhibition of water and electrolyte absorption from the large intestine, which increases volume and pressure of the intestinal contents and thus stimulate the colon motility resulting in propulsive contractions (9). However, the adverse effects of the senna can be counter balanced by using Madhuka (Glycerrhiza glabra Linn.) and Ikshu sarkara (Cane sugar-Saccharam officinarum).
Organoleptic evaluation was used for identification of sensory characteristics powder like colour, odour (smell), touch and taste.
For powder microscopic analysis, about 1 to 3 pinch of the sample was warmed with 2 to 4 drops of chloral hydrate solution, water and little safranin stain and observed under the microscope to identify the diagnostic features of the compound formulation for the presence of different fragments of tissues in curna. It was also observed by putting a pinch of phloroglucinol, 2 drops of concentrated hydrochloric acid and few drops of alcohol for the presence of lignin containing tissues. (10) Fluorescence analysis of the powder was observed under UV light according to Chase and Pratt et al. (11) Physico chemical analysis like determination of percentage of moisture content (loss on drying at 105 o C ), Ash value, water soluble ash, total insoluble ash in various solvent, sulphated ash, percentage of soluble extractives in methanaol, ethanol & ether. pH of filtrate of 10% w/v aqueous solution was carried out as per methods described in Quality control methods for medicinal plants (WHO, 1998) (12) TLC studies have been carried out according to Igon Stahlet al. (13) and preliminary phytochemical studies were carried out according to Kokate.C.K et al. (14) Phyto chemical constituents of the ingredients of M-Swadista virechana curna has been reviewed and preliminary phytochemicals of the compound formulation have been screened.

Observations and Result Macroscopic (organoleptic) characters of the powder [Fig.II(1)]
Powder is light green in colour, slightly pleasant odour, smooth and fine in touch, sweet, & bitter in taste.

Microscopic characters [Fig.II(1a-20)]
The polyherbal powder was treated with chloral hydrate, water & glycerin and microscopical examination was carried out for the presence of following different fragments of tissues. 

Physicochemical and Preliminary phytochemical studies
The physicochemical studies were carried out and the observations are given in table3. The test for percentage of moisture content (loss on drying) determines both water and volatile matter. Total ash measures the amount of materials remaining after ignition. Acid insoluble ash measures the amount of silica present especially sand and siliceous matter. Extractive values were examined which are useful for evaluation of nature of chemical constituents present in drug (Table 3).
Active phytochemical constituents in each of the ingredient of the formulation has been reviewed ( Table 2) and Preliminary phytochemical screening of compound formulation was identified through qualitative chemical analysis indicated the presence of alkaloids, carbohydrates, flavonoids, terpenoids, resins, saponins, steroid and tannins etc. (Table 4).

Fluorescence analysis
The fluorescence behavior of the powdered drug in different solutions towards ordinary light and Ultra Violet light (both long 365nm & short 254nm wavelengths) were observed (Table 5).

Thin Layer Chromatographic studies (TLC)
ISSN: 0976-5921 International Journal of Ayurvedic Medicine, 2010, 1(3), 134-142 137 TLC studies of the chloroform and methanol extract (8:2) was carried out in various solvent systems at 30 0 C using Silica gel GF 254 precoated sheets of thickness 2mm. The chloroform and methanol extract (8:2) was found to be the best suitable mobile phase for the extract.

Discussion and Conclusion
The sample of M.Swadista virechana churna was found to be light green in colour moderately fine powder with pleasant smell and sweet & bitter in taste. The churna did not show any fungus or objectionable matter when spread out on the petridish. It was observed that the sample passed through 60-mesh sieve and not less than 50% passed through 80-mesh sieve.
Microscopic characterization of powder revealed abundant different kinds of tissues like Sclerenchymatous fibers, parenchymatous cells containing prism shaped and clusters of calcium oxalate crystals, unicellular warty trichomes. It was also observed polygonal epidermal cells in surface view with paracytic stomata, crystal fibers and simple starch grains and group of lignified fibers, pitted xylem vessels [ Fig.II (1a to 20)].
The physicochemical analysis was carried out and observations are presented in table 3. TLC profile of the sample was depicted in table 6 and the chromatogram was observed ( fig.1). TLC of methanol extract curve using chloroform: methanol The results obtained could be immense useful in laying down the pharmacopoeial standards of M.Swadista virechana churna.