1. INTRODUCTION:

In India, medicinal plants form the backbone of several indigenous traditional systems of medicine. Pharmacological studies have acknowledged the value of medicinal plants as potential source of bioactive compounds¹. Phytochemicals from medicinal plants serve as lead compounds in drug discovery and design². Medicinal plants are rich source of novel drugs that forms the ingredients in traditional system of medicine, modern medicines, nutraceuticals, food supplements, folk medicines, pharmaceutical intermediates, bioactive principles and lead compounds in synthetic drugs³.

WHO, report depicts that more than 80% of world’s population rely on plant based products to meet health care needs. Nearly, 25 to 45% of modern prescriptions contain plant derived lead molecules as a basic source in drug formulations. The value of plant based prescribed drugs in 1990 was estimated at $ 15.5 billion which has been on the raise since then. Furthermore, about 42% of 25 top selling drugs marketed worldwide are either directly obtained from natural sources or entities derived from plant products⁴.

Furthermore the active components of herbal remedies have the advantages of being combined with many other substances that appear to be inactive. However, these complementary components give the plant as a whole safety and efficiency much superior to that of its isolated and pure active components. Presently, in the developing countries, synthetic drugs are not only expensive and inadequate for the treatment of diseases but are also often with adulteration and side effects⁵. Therefore, there is the need to search for plants and plant derived formulations of medicinal value.

Balarishta is a polyherbal hydroalcoholic preparation and is used to cure gastric problems (vata vyadhi), as restorative and as a prime tool for the treatment of arthritis, rheumatism and all other autoimmune disorders⁶. The chief ingredients of Balarishta are dried roots of Sida cordifolia which have been extensively investigated and it was reported that they contain alkaloids as ephedrine, vasicine, vasicinone^7-8, phytosterols, fatty acids, a rich quantity of phenolic substances and flavonoids as rutin, flavones⁹ as well as saponins as kaempferol and sitoindosides¹⁰. Roots of Withania somnifera have been found to contain withanolides, a group of steroidal lactoneswhich is known for its usefulness in the treatment of hypercholesterolemia, arthritis in combination with other drugs which is also credited to be hypoglycemic and diuretic^11-12. All these compounds have many favourable effects on human health such as lowering of human low density lipoproteins, reduction of heart disease and cancer because of their antioxidant property.

Therefore, we undertook the present investigation to evaluate the antimicrobial activity of Balarishta-T, Balarishta-M prepared by traditional and modern methods respectively and marketed Balarishta against common human pathogens.

2. MATERIALS AND METHODS:

2.1 Preparation of Balarishta-T:

This was prepared by the method as given in The Ayurvedic Formulary of India, Part-I⁶. All the ingredients of Balarishta were procured from local market, Jamnagar while jaggery was procured from local market, Mehsana. Authentication of all the ingredients of Balarishta was done by Dr. G. D. Bagchi, Scientist, Department of Taxonomy and Pharmacognosy, Central Institute of Medicinal and Aromatic Plants, Lucknow. Prepared herbarium has been deposited in the Central Institute of Medicinal and Aromatic Plants, Lucknow for future reference. Identification of all the individual plant material was done as per The Ayurvedic Pharmacopoeia of India.

According to this method, dried roots of Sida cordifolia and Withania somnifera were coarsely powdered and then placed in polished vessel of brass along with prescribed quantity of water (12.288l) and allowed to steep. After 12 h of steeping, this material was warmed at medium flame until the water for decoction reduced to one fourth of the prescribed quantity(3.072 l) , then the heating was stopped and it was filtered in cleaned vessel and after that jaggery was added and mixed properly. Then, dhataki flowers (Woodfordia floribunda) and prescribed quantity of coarsely powdered prakshepa dravyas as Ipomoea digitata (roots), Ricinus communis (roots), Alpinia galangal (roots), Eletteria cardamomum (seeds), Ipomoea tridentate (entire plant ), Eugenia caryophyllus (flower bud), Andropogon muricatus (roots) and Tribulus terrestris (fruits) were added and this sweet filtered fluid was placed for fermentation in incubator for fifteen days at 33±1°C. After 15 days, completion of fermentation was confirmed by standard tests¹³. The fermented preparation was filtered with cotton cloth and kept in clean covered vessel for further next seven days. Then, when the fine suspended particles settled down, it is strained again and poured in amber colored glass bottles previously rinsed with ethyl alcohol, packed and properly labeled.

2.2 Preparation of Balarishta-M:

Method of preparation of Balarishta-M was same as followed with Balarishta-T only dhataki flowers were replaced with yeast for inducing fermentation¹⁴.

2.3 Antimicrobial Activity Test

Antimicrobial activity of Balarishta-T, Balarishta-M and marketed Balarishta was tested using a modified disc diffusion assay (DDA) method originally described by Baurer (1966)¹⁵. Test preparations of Balarishta were dissolved in 20% DMSO treated water. The inoculums for each microorganism were prepared from broth cultures (10⁵ CFU/ml). A loop of culture from the slant stock was cultured in nutrient agar medium overnight and spread with a sterile swab into Petri-plates. Sterile disc (6 mm dia, Hi-media Mumbai, India) impregnated with test preparations (100µl/disc) and Kanamycin (30µg/disc) were placed on the culture plates and incubated for 24h at 37ºC. The solvent (DMSO) loaded disc without test preparations served as control in the study. The results were recorded by measuring the zones of growth inhibition. Clear inhibition zones around discs indicated the presence of antimicrobial activity. All data of antimicrobial activity were taken as average of triplicate.

3. RESULTS:

All types of Balarishta as Balarishta-T, Balarishta-M prepared by traditional and modern methods respectively and marketed Balarishta showed significant antibacterial activity by exhibiting significant zone of inhibition against common human pathogens as Staphylococcus aureus, Bacillus subtilis, Salmonella typhii, Escherichia coli and Pseudomonas aeruginosa as shown in Table 1.

Table1. Diameter of Zone of Inhibition (mm) of Balarishta-T, Balarishta-M and marketed Balarishta

Sample	Zone of Inhibition (mm)
Sample	Staphylococcus aureus	Bacillus subtilis	Salmonella typhii	Escherichia coli	Pseudomonas aeruginosa
Balarishta-T (100µl/disc)	22.46±0.63	27.62±0.91	25.86±1.19	24.91±0.78	25.18±1.56
Balarishta-M (100µl/disc)	21.57±0.82	25.76±0.48	26.94±0.82	23.87±0.93	23.42±1.18
Marketed Balarishta (100µl/disc)	21.14±0.56	26.24±0.78	26.25±0.73	22.45±0.68	24.15±0.96
Kanamycin (30µg/disc)	28±1.24	34±0.98	33.14±0.87	34.91±1.42	32.64±0.59
Negative Control (DMSO)	-ve	-ve	-ve	-ve	-ve

All values are shown as mean± SD of three replicates

4. DISCUSSION:

Plants are known to have beneficial therapeutic effects documented in Traditional Indian System of Medicine. Though bioactive products of Bala and its preparations as Balarishta have been used in treatment of various ailments since time immemorial, role of phytochemicals in inhibition of growth of microorganisms has gained less prominence¹⁶. In the present study, preparations of Balarishta as Balarishta-T, Balarishta-M and marketed Balarishta exhibited significant antibacterial activity against common human pathogens. Further investigations may lead to the development of naturally derived new antibiotics of high potency.

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Received on 22.06.2014 Modified on 25.06.2014

Research J. Pharm. and Tech. 7(7): July 2014 Page 789-791