Antimicrobial Activity of Ashwagandharishta Prepared by Traditional and Modern Methods

 

Preeti Tiwari*

Head of Department of Pharmacognosy, Dr. K. N. Modi Institute of Pharmaceutical Education and Research, Modinagar (U. P.)

*Corresponding Author E-mail: preetitiwari198311@yahoo.com

 

ABSTRACT:

In the present investigation, different types of test preparations of Ashwagandharishta as Ashwagandharishta-T, Ashwagandharishta-M prepared by traditional and modern methods respectively and marketed Ashwagandharishta were evaluated for antimicrobial activity against common human pathogens. It was observed that all the test preparations of Ashwagandharishta exhibited significant zone of inhibition against selected common human pathogens. The results indicate that all the test preparations of Ashwagandharishta as Ashwagandharishta-T, Ashwagandharishta-M and marketed Ashwagandharishta might be used as natural drug for the treatment of several infectious diseases caused by these organisms.

 

KEYWORDS: Ashwagandharishta-T, Ashwagandharishta-M, Antimicrobial activity

 

 


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 worlds 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 ahs been on the raise since then. Furthermore, about 42% of 25 top selling drugs marketed world wide 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.

 

Ashwagandharishta is a polyherbal hydro alcoholic preparation and is used as rasayana. Rasayanas are used to promote health and longevity by increasing defense against disease, arresting the ageing process and revitalizing the body in debilitated conditions6. The chief ingredient of Ashwagandharishta is roots of Ashwagandha, Withania somnifera, commonly known for its usefulness in the treatment of hypercholesterolemia, arthritis in combination with other drugs, is also credited to be hypoglycemic and diuretic7. The pharmacological effect of the roots of Withania somnifera is attributed to withanolides, a group of steroidal lactones8.

 

Besides Withania roots, the other ingredients of Ashwagandharishta as arjuna (bark of Terminalia arjuna), liquorice (roots of Glycyrrhiza glabra), majith (roots of Rubia cordifolia), rasna (roots of Alpinia chinensis), taj (inner bark of Cinnamomum zeylanicum), nagarmotha (rhizomes of Cyperus rotundus), haritaki (fruits of Terminalia chebula), turmeric (rhizomes of Curcuma longa), nagakesara (stamens of Mesua ferrea) etc. contain a rich quantity of polyphenolic compounds and flavonoids and possess significant antioxidant activity9-10. Therefore, we undertook the present investigation to evaluate the antimicrobial activity of Ashwagandharishta-T, Ashwagandharishta-M prepared by traditional and modern methods respectivelyand marketed Ashwagandharishta against common human pathogens.

 

2. MATERIAL AND METHODS:

2.1 Preparation of Ashwagandharishta-T

This was prepared by the method as given in the Ayurvedic Formulary of India6. The ingredients of Ashwagandharishta were procured from local market, Jamnagar. Identification of all the 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 the CIMAP for future reference.

 

According to this method, coarsely powdered ashwagandha roots (Withania somnifera) with prescribed ingredients were placed in polished vessel of brass along with prescribed quantity of water (24.576 l), and allowed to steep. After 12 h of steeping, this material was warmed at medium flame until the water for decoction reduced to one eighths of the prescribed quantity (3.072 l), then the heating was stopped and it was filtered in cleaned vessel and after that honey was added. Then, dhataki flowers (Woodfordia floribunda) and prakshepa dravyas as sonth, marich, pippali, tvak, tejpatra, priyangu and nagakesara were added and this sweet filtered material was placed for fermentation in incubator for fifteen days at 33oC1oC. After 15 days, completion of fermentation was confirmed by standard tests11. The fermented preparation was filtered with cotton cloth and kept in cleaned covered vessel for further next seven days. Then, the preparation was poured in amber colored glass bottles, packed and properly labeled.

 

2.2 Preparation of Ashwagandharishta-M

Method of preparation was same as followed with Ashwagandharishta-T only dhataki flowers were replaced with yeast for inducing fermentation12.

 

2.3 Antimicrobial Activity Test

Antimicrobial activity of Ashawagandharishta-T, Ashwagandharishta-M and marketed Ashwagandharishta was tested using a modified disc diffusion assay (DDA) method originally described by Baurer (1966)13. Test preparations of Ashwagandharishta 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 (100l/disc) and Kanamycin (30g/disc) were placed on the culture plates and incubated for 24h at 37C. 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 Ashwagandharishta as Ashwagandharishta-T, Ashwagandharishta-M prepared by traditional and modern methods respectively and marketed Ashwagandharishta 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 Ashwagandharishta-T, Ashwagandharishta-M and marketed Ashwagandharishta

Sample

Zone of Inhibition (mm)

Staphylococcus aureus

Bacillus subtilis

Salmonella typhii

Escherichia coli

Pseudomonas

aeruginosa

Ashwagandharishta-T

(100l/disc)

21.420.93

24.780.49

22.421.19

25.620.73

24.640.49

Ashwagandharishta-M

(100l/disc)

20.681.14

22.960.57

21.350.61

24.480.85

22.850.74

Marketed Ashwagandharishta (100l/disc)

20.521.26

23.460.69

21.420.59

23.971.16

23.531.12

Kanamycin (30g/disc)

281.24

340.98

33.140.87

34.911.42

32.640.59

Negative Control (DMSO)

-ve

-ve

-ve

-ve

-ve

All values are shown as meanSD of three replicates


 

4. DISCUSSION:

Plants are known to have beneficial therapeutic effects documented in Traditional Indian System of Medicine. Though bioactive products of Ashwagandha and its preparations as Ashwagandharishta have been used in treatment of various aliments since time immemorial, role of phytochemicals in inhibition of growth of microorganisms has gained less prominence14. In the present study, preparations of Ashwagandharishta as Ashwagandharishta-T, Ashwagandharishta-M and marketed Ashwagandharishta 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:

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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 I. Controller of Publications, Delhi, 2000;8-9.

7. Andallu B, Radhika B. Hypoglycaemic, Diuretic and Hypocholesterolemic effect of Winter cherry (Withania somnifera, Dunal) root. Indian J Exp Biol 2000; 38:607-9.

8. Budhiraja RD, Sudhir S. Review of biological activity of Withanolides. J Sci Ind Research 1987;46:488.

9. Jadhav PD, Laddha KS. Estimation of gallic and ellagic acid from Terminalia chebula Retz. Indian Drugs 2004; 41(4):200-06.

10. Tuba AK, Ilhami G. Antioxidant and free radical scavenging properties of curcumin. Chem Biol Interact 2008;174:27-37.

11. Mishra S. Bhaisazya Kalpana Vigyan, Chaukambha Surbharati Prakashan. Varanasi. 2005;253-54.

12. Alam M, Radhamani S, Ali U, Purushottam KK. Microbiological screening of dhataki flowers. J Res Ayurveda Siddha 1984;2(4):371-5.

13. 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.

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

 

 

Received on 19.06.2014 Accepted on 29.06.2014

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Asian J. Res. Pharm. Sci. 4(3): July-Sept. 2014; Page 115-117