Evaluation of Diuretic Potential of Drakshasava Prepared by Traditional and Modern Methods in Experimental Albino Rats

 

Preeti Tiwari1* and Rakesh K. Patel2

1Department of Pharmacognosy, Shri Sarvajanik Pharmacy College, Mehsana-384001, Gujarat, India

2Head of Department of Pharmacognosy, Shri S. K. Patel College of Pharmaceutical Education and Research, Kherva-382711, Gujarat, India

 

ABSTRACT:

The objective of the present study was to evaluate the diuretic potential of Drakshasava-T and Drakshasava-M prepared by traditional and modern methods respectively and its marketed formulation in experimental rats using furosemide (10 mg/kg p.o.) as a standard diuretic drug. Oral administration of Drakshasava-T, Drakshasava-M and its marketed formulation at the dose of 2.0 ml/kg over a period of 5 h showed a significant increase in urine volume as compared to control group. Both types of Drakshasava as Drakshasava-T and Drakshasava-M prepared by traditional and modern methods respectively and its marketed formulation showed significant increase in sodium, potassium and chloride level in urine sample as compared to control group. The maximum diuretic effect was produced by furosemide. Thus, both types of Drakshasava as Drakshasava-T and Drakshasava-M and its marketed formulation showed significant diuretic, natriuretic and kaliuretic effects.

 

KEYWORDS: Diuretic potential, furosemide, Drakshasava, natriuretic effect, kaliuretic effect

 

1.INTRODUCTION:

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 vinifera1. 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 procyanidins2-3. 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 property4-11.

However, no study has been carried out for the diuretic activity of Drakshasava in order to confirm its assumed beneficial property. Therefore, we have undertaken the present study to verify the efficacy of all the test formulations of Drakshasava as Drakshasava-T and Drakshasava-M prepared by traditional and modern methods respectively and its marketed formulation as diuretic agent in experimental albino rats

 

2. MATERIALS AND METHOD:

2.1 Preparation of Drakshasava-T

This was prepared by the method as given in Ayurvedic Formulary of India, Part-II1. 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 33șC± 1șC.  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 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 fermentation13.

 

2.3 Animals

Adult wistar albino rats, weighing between 200-220g of either sex were acclimatized to normal environmental conditions in the animal house for one week. The animals were housed in standard polypropylene cages and maintained under controlled room temperature (22șC±2șC) and humidity (55±5%) with 12:12 hour light and dark cycle. All the animals were given a standard chow diet (Hindustan Lever Limited) and water ad libitum. The guidelines of the Committee for the Purpose of Control and Supervision of Experimentals on Animals (CPCSEA) of the Government of India were followed and prior permission was granted from the Institutional Animals Ethics Committee (CPCSEA No. 07/09).

 

2.4. Experimental Procedure

The method of Lipschitz et al., (1943) was employed for the assessment of diuretic activity. Twenty four hours before testing the animals were transferred to metabolic cages14.Then only water was made accessible ad libitum without food.

All the animals were randomly divided into the five groups with six animals in each group as follows:

Group I  Control group received normal saline as vehicle (25 ml/kg, p.o.)

Group II  Animals received furosemide (10 mg/kg, p.o.)

Group III  Animals received Drakshasava-T (2 ml/kg, p.o.)

Group IV Animals received Drakshasava-M (2 ml/kg, p.o.)

Group V Animals received marketed Drakshasava (2 ml/kg, p.o.)

 

The second group received same volume of normal saline (25 ml) in which furosemide (10 mg/kg bw) was dissolved. The animals of Group III, IV and V received Drakshasava-T, Drakshasava-M and marketed Drakshasava at the dose of 2 ml/kg bw orally, after diluting to all of them up to 25 ml with normal saline to maintain the fluid intake same in all the cases. Immediately after dosing the rats were placed in metabolic cages and kept at room temperature of 25 șC±0.5 șC for 5 h. During this period, no food and water was made available to them. At the end of 5 h the animals were taken out of the cages and the total volume of urine excreted by each group was noted. Urine samples were analysed thereafter for Na+ and K+ concentration by flame photometer while chloride (Cl-) was determined by using standard kit containing chloride reagent from span diagnostics, Surat, India.

 

2.5. Statistical analysis

The results have been expressed as mean ± SEM. Statistical analysis of data among the various groups was performed by using one way analysis of variance (ANOVA) followed by the Tukey’s test using Graph Pad Prism software of Statistics. Significance value (P<0.05) was considered statistically significant.

 

3. RESULTS:

Diuretic effect

Total urine output

Both types of Drakshasava as Drakshasava-T and Drakshasava-M were prepared by traditional and modern methods respectively showed significant (P<0.001) increase in urine volume, as compared to control group. The diuresis was almost equal to that produced by furosemide (Fig.1).

 

Urinary electrolyte concentration

Urinary sodium: All the test formulations of Drakshasava as Drakshasava-T, Drakshasava-M and its marketed formulation were found to produce significant (P<0.001) increase in natriuresis but the maximum natriuresis was produced by furosemide (Fig.2).

 

Urinary potassium:

Both types of Drakshasava as Drakshasava-T and Drakshasava-M have been shown to produce significant (P<0.001) increase in the excretion of potassium in urine as compared to the control group. Furosemide also significantly increased the excretion of potassium. Thus, all the test formulations of Drakshasava showed significant kaliuretic effect (Fig.2).

 

Urinary chloride:

All the test formulations of Drakshasava as Drakshasava-T, Drakshasava-M and its marketed formulation showed significant (P<0.001) increase in the excretion of chloride in urine as compared to control. Furosemide also showed significant increase in the excretion of chloride in urine (Fig.2).

 

 

Fig. 1. Effect of Drakshasava-T, M and marketed Drakshasava on urine volume

All values are expressed as mean ± SEM (n = 6) b  P<0.001 as compared to control

 

 

Fig. 2. Effect of Drakshasava-T, M and marketed Drakshasava on urinary electrolyte concentration

All values are expressed as mean ± SEM (n = 6)  b  P<0.001 as compared to control

 

4. DISCUSSION:

This study shows that both types of Drakshasava as Drakshasava-T and Drakshasava-M prepared by traditional and modern methods respectively and its marketed formulation produced striking increase in total urine output over a period of 5 h. All these test formulations of Drakshasava also showed significant (P<0.001) increase in the excretion of sodium, potassium and chloride in urine as compared to control group. Therefore, both types of Drakshasava as Drakshasava-T and Drakshasava-M have been shown to possess significant diuretic, natriuretic and kaliuretic effects which may be one of the basis of their therapeutic application in various ailments, such as nephritis, burning micturation etc. and different oedematous diseases. Their diuretic effects have been shown to be more or less equal to that produced by furosemide.

 

Preliminary phytochemical studies have confirmed the presence of phenolics, particularly hydrolysable tannins and flavonoids and other nonphenolic constituents as steroidal saponins in all the test formulations of Drakshasava as Drakshasava-T, Drakshasava-M and its marketed formulation, promoting the hypothesis that these type of polar compounds may also be responsible for the diuretic effects. It is known that this type of compounds increase renal circulation, and thus the rate of glomerular filtration which promotes increased urine formation15-17. Thus, presence of self generated alcohol helps in the faster absorption of biologically active compounds as tannins, flavonoids and steroidal saponins which by their chemical nature are antioxidants, might contribute to the prevention of cardiac diseases as hypertension by acting as diuretics18.

 

5. ACKNOWLEDGEMENT:

The authors are immensely thankful to the department of Pharmacology, Shri Sarvajanik Pharmacy College, Mehsana for providing the requisite facilities.

 

6. REFERENCES:

1)       The Ayurvedic Formulary of India, Part-II, 2000, 1st edition, The Controller of Publications, Delhi, p.35.

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5)       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). Journal of Agricultural and Food Chemistry 1997; 45:1638-1643.

6)       Teissedre PL, Frankel EN, Waterhouse AL, Peleg H, German GB. Inhibition of in vitro human LDL oxidation by phenolic antioxidants from grapes and wines. Journal of the Science of Food and Agriculture 1996; 70:55-61.

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

8)       Renaud S, Lorgeril MD. Wine, alcohol, platelets and the French paradox for coronary heart disease. The Lancet 1992; 339:1523-1526.

9)       Davalos A, Bortolome B, Gomez-cordoves C. Antioxidant properties of commercial grape juices and vinegars. Food Chemistry 2005; 93(2):325-330.

10)     Orhan DD, Orhan N, Ergun E, Ergun F. Hepatoprotective effect of Vitis vinifera L. leaves on carbon tetrachloride-induced acute liver damage in rats. Jornal of Ethnopharmacology 2007; 112:145-151.

11)     Corder R, Mullen W, Khan NQ, Marks SC, Wood EG, Carrier MJ, Crozier A.  Red wine procyanidins and vascular health. Nature 2006;444:566.

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

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

14)      Lipschitz WL, Hadidian Z, Kerpcsar A. Bioassay of Diuretics. Journal of Pharmacology and Experimental Therapeutics 1943; 79:97-110.

15)     Afzal M, Khan NA, Ghufran A, Iqbal A, Inamuddin M. Diuretic and nephroprotective effect of Jawarish Zarooni Sada- a polyherbal Unani formulation. Journal of Ethnopharmacology 2004;91:219-223.

16)    Loew D, Heimsoth V, Erwin K, Schilcher H. 1991. Diureticos: Quimica, Farmacologiay Therapeutica incluida Fitoterapia, Barcelona, Salvat Editores S.A.:270.

17)      Das PK, Goswami S, Chinniah A. Woodfordia fruticosa: Traditional uses and recent findings. Journal of Ethnopharmacology 2007; 110:189-199.

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Received on 04.08.2012

Modified on 18.08.2012

Accepted on 25.08.2012

© A&V Publication all right reserved

Research J. Pharmacology and Pharmacodynamics. 4(5): September –October, 2012, 281-284