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.

Draksharishta is a polyherbal hydroalcoholic preparation and is used as blood purifier , in the treatment of anaemia and advised as a choice of remedy in respiratory problems. The chief ingredient of Draksharishta 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 procyanidins⁷. These compounds have many favourable effects on human health such as lowering of human low density lipoproteins, reduction of heart disease and cancer^8-11.

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

2. MATERIALS AND METHODS:

2.1 Preparation of Draksharishta-T

This was prepared by the method as given in Ayurvedic Formulary of India ⁶.The ingredients of Draksharishta were procured from Local market, Jamnagar. Identification of all the individual plant material was done as per Ayurvedic Pharmacopoeia of India. Authentification of all these ingredients was done in the Botany Department of Central Institute of Medicinal and Aromatic Plants (CIMAP), Lucknow. Prepared herbarium has been deposited in the CIMAP for future reference.

According to this method, dried fruits of Vitis vinifera after proper crushing were placed in polished vessel of brass along with prescribed quantity of water (13L), 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 (3.25L), then the heating was stopped and it was filtered through unstarched muslin cloth in cleaned and fumigated vessel and after that jaggery was added and mixed properly. Then Dhataki flowers (Woodfordia floribunda) and prescribed quantity of coarsely powdered prakshepa dravyas as Cinnamomum zeyleynicum (stem bark), Eletteria cardamomum (seeds), Cinnamomum tamala (leaves) , Mesua ferrea (stamens), Callicarpa macrophylla (flowers), Piper nigrum (fruits), Piper longum (fruits), Embelia ribes (fruits) were added and this sweet filtered fluid was placed for fermentation in incubator for fifteen days at 33^oC±1^oC.After fifteen days completion of fermentation was confirmed by standard tests¹².

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 coloured glass bottles previously rinsed with ethyl alcohol, packed and labeled properly.

2.2 Preparation of Draksharishta-M

Method of preparation was same as followed with Draksharishta -T, only dhataki flowers were replaced with Yeast for inducing fermentation¹³.

2.3 Antimicrobial Activity Test

Antimicrobial activity of Draksharishta-T, Draksharishta-M and marketed Draksharishta was tested using a modified disc diffusion assay (DDA) method originally described by Baurer (1966)¹⁴. Test preparations of Draksharishta 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 Draksharishta as Draksharishta-T, Draksharishta-M prepared by traditional and modern methods respectively and marketed Draksharishta 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 Draksharishta-T, Draksharishta-M and marketed Draksharishta

Sample	Zone of Inhibition (mm)
Sample	Staphylococcus aureus	Bacillus subtilis	Salmonella typhii	Escherichia coli	Pseudomonas aeruginosa
Draksharishta-T (100µl/disc)	23.48±0.92	26.52±1.14	26.74±0.89	25.45±0.96	26.54±0.76
Draksharishta-M (100µl/disc)	22.96±0.84	25.47±0.69	26.98±0.71	24.68±1.12	25.62±0.68
Marketed Draksharishta (100µl/disc)	21.54±1.19	25.64±0.87	25.69±0.56	23.56±0.81	24.43±0.48
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 Draksha and its preparations as Draksharishta have been used in treatment of various ailments since time immemorial, role of phytochemicals in inhibition of growth of microorganisms has gained less prominence¹⁵. In the present study, preparations of Draksharishta as Draksharishta-T, Draksharishta-M and marketed Draksharishta exhibited significant antibacterial activity against common human pathogens. Further investigations may lead to the development of naturally derived new antibiotics of high potency.

5. REFERENCES:

1. Prusti A, Mishra SR, sahoo S and Mishra SK. Antibacterial activity of Some Indian Medicinal Plants. Ethnobotanical Leaflets 2008; 12:227-230.

2. Ebi GC and Ofoefule SI (2000) Antimicrobial Activity of Pterocarpus osun stems. Fitoterapia 71:433-435.

3. Ncube NS, Afolayan AJ, Okoh A. Assessment Techniques of antimicrobial properties of natural compounds of plant origin:current methods and future trends. Africal Journal of Biotechnology 2008; 7(12):1797-1806.

4. Ramya S, Govindaraji V, Kannan NK and Jayakumararaj R. In vitro evaluation of antibacterial activity using crude extracts of Catharanthus roseus L. Ethnobatanical Leaflets 2008; 12:1013-1018.

5. Shariff Z U . Modern Herbal Therapy for Common Ailments. Nature Pharmacy Series, Spectrum Books Limited. Ibadan, Nigeria in Association with Safari Books (Export) Limited, United Kingdom, 2001; vol. 1, 94.

6. The Ayurvedic Formulary of India Part-II. Delhi, India: Controller of Publications; 2000. p. 15-16.

7. Baydar NG, Ozkan G, Sagdic O. Total phenolic contents and antibacterial activities of grape (Vitis vinifera L.) extracts. Food Control 2004; 15:335-339.

8. Frankel EN, Kanner J, German JB, Parks E, Kinsella JE. Inhibition of oxidation of human low-density lipoprotein by phenolic substances in red wine. The Lancet 1993; 341(20): 454-457.

9. Mayer AS, Yi OS, Person DA, Waterhouse DL, Frankel EN. Inhibition of human low-density lipoprotein oxidation in relation to composition of phenolic antioxidants in grapes (Vitis vinifera). J Agri Food Chem 1997; 45:1638-1643.

10. Teissedre PL, Frankel EN, Waterhouse AL, Peleg H, German GB. Inhibition of in vitro human LDL oxidation by phenolic antioxidants from grapes and wines. J Sci Food Agri 1996; 70: 55-61.

11. Waterhouse AL. Wine antioxidants may reduce heart disease and cancer. Presentation of American Chemical Society, Washington; 1994

12. Mishra S. Bhaisazya Kalpana Vigyan. Varanasi, India: Chaukambha Bharati Prakashan; 2005. p. 253-254.

13. Alam M, Radhamani S, Ali U, Purushottam KK. Microbiological Screening of Dhataki Flowers. J Res Ayurveda Siddha. 1984; 2(4):371-375.

14. Bauer RW, Kirby MDK, Sherris JC and Turck M . Antibiotic susceptibility Testingby standard single disc diffusion method. American Journal of Clinical Pathology1966; 45:493-96.

15. Sasidharan VK, Krishnakumar T and Manjula CB. Antimicrobial activity of Nine Common plants in Kerala, India. PJS 1998, 127 (1):59-67.

Received on 23.07.2014 Accepted on 04.08.2014

Asian J. Pharm. Tech. 2014; Vol. 4: Issue 3, Pg 131-133