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.

Drakshasava is a polyherbal hydroalcoholic preparation and is used to improve digestion, as blood purifier, in the treatment of anaemia and advised as a choice of remedy in respiratory problems. The chief ingredient of Drakshasava is draksha, dried fruits of Vitis vinifera⁶. The composition and properties of fruits of Vitis vinifera have been extensively investigated and it was reported that they contain large amount of phenolic compounds as catechins, epicatechin, quercetin and gallic acid, dimeric, trimeric and tetrameric procyanidins^7-8. 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^9-16.

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

2. MATERIALS AND METHOD:

2.1 Preparation of Drakshasava-T:

This was prepared by the method as given in Ayurvedic Formulary of India, Part-II⁶. The ingredients were procured from local market, Jamnagar. Identification of individual plant material was done as per Ayurvedic Pharmacopoeia of India. Authentication of all these ingredients 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 CIMAP for future reference.

According to this method, dried fruits of Vitis vinifera were crushed and then placed in polished vessel of brass along with prescribed quantity of water (16.384 L), and allowed to steep overnight. After overnight steeping, this material was warmed at medium flame until the water for decoction reduced to one fourth of the prescribed quantity (4.096 L), then the heating was stopped and it was filtered through unstarched muslin cloth in cleaned and fumigated vessel and after that jaggery and honey were added and mixed properly. Then dhataki flowers (Woodfordia floribunda) and prescribed quantity of coarsely powdered prakshepa dravyas as Myristica fragrans (flowers), Eugenia caryophyllus (flower bud), Cubeba officinalis (fruits), Santalum album (heart wood), Piper nigrum (fruits), Cinnamomum zeylanicum (stem bark), Eletteria cardamomum (seeds) and Cinnamomum tamala (leaves) were added and this sweet filtered fluid was placed for fermentation in incubator for fifteen days at 33ºC± 1ºC. After fifteen days, completion of fermentation was confirmed by standard tests¹⁷. The fermented preparation was filtered with unstarched muslin cloth and kept in cleaned covered vessel for further next seven days. Then, it was poured in clean amber colored glass bottles previously rinsed with ethyl alcohol, packed and labelled properly.

2.2 Preparation of Drakshasava-M:

Method of preparation was same as followed with Drakshasava-T, only dhataki flowers were replaced with yeast for inducing fermentation¹⁸.

2.3 Antimicrobial Activity Test:

Antimicrobial activity of Drakshasava-T, Drakshasava-M and marketed Drakshasava was tested using a modified disc diffusion assay (DDA) method originally described by Baurer (1966)¹⁹. Test preparations of Drakshasava 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.

2. RESULTS:

All types of Drakshasava as Drakshasava-T, Drakshasava-M prepared by traditional and modern methods respectively and marketed Drakshasava 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.

4. DISCUSSION:

Plants are known to have beneficial therapeutic effects documented in Traditional Indian System of Medicine. Though bioactive products of Draksha and its preparations as Drakshasava have been used in treatment of various ailments since time immemorial, role of phytochemicals in inhibition of growth of microorganisms has gained less prominence²⁰. In the present study, preparations of Drakshasava as Drakshasava-T, Drakshasava-M and marketed Drakshasava exhibited significant antibacterial activity against common human pathogens. Further investigations may lead to the development of naturally derived new antibiotics of high potency.

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

Sample	Zone of Inhibition (mm)
Sample	Staphylococcus aureus	Bacillus subtilis	Salmonella typhii	Escherichia coli	Pseudomonas aeruginosa
Drakshasava-T (100µl/disc)	25.63±0.75	28.76±1.41	29.58±0.93	27.49±0.88	25.72±0.84
Drakshasava-M (100µl/disc)	24.46±0.86	27.59±0.41	28.47±0.59	26.64±0.96	24.66±0.72
Marketed Drakshasava (100µl/disc)	23.62±1.21	25.82±0.97	26.68±0.79	25.72±0.61	23.59±1.43
Kanamycin (30µg/disc)	28±1.24	34±0.98	33.14±0.87	34.91±1.42	32.64±0.59
Negative Control (DMSO)	Negative	Negative	Negative	Negative	Negative

All values are shown as mean± SD of three replicates

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Received on 22.07.2014 Modified on 25.07.2014

Res. J. Pharmacognosy & Phytochem. 6(3): July-Sept.2014; Page 126-128