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PHARMACOLOGICAL STUDY
Year : 2013  |  Volume : 34  |  Issue : 3  |  Page : 327-330  

Acute toxicity study of Vasaguduchyadi Kwatha: A compound Ayurvedic formulation


1 Lecturer and Head, Department of Pharmacology, Institute of Ayurvedic Pharmaceutical Sciences, Gujarat Ayurved University, Jamnagar, Gujarat, India
2 Drug Discovery Team, Research and Development, The Himalaya Drug Company, Makali, Bangalore, India
3 Head, Pharmaceutical Chemistry Laboratory, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, India
4 Professor and Head, Department of Rasa Shastra and Bhaishjya Kalpana, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, Gujarat, India
5 Director, Reseach and Development, SDM Centre for Research in Ayurveda and Allied Sciences, Kuthpady, Udupi, Karnataka, India

Date of Web Publication17-Dec-2013

Correspondence Address:
Kalpu N Kotecha
Lecturer and Head, Department of Pharmacology, Institute of Ayurvedic Pharmaceutical Sciences, A. K. Jamal Bldg, Gujarat Ayurved University, Jamnagar, Gujarat
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0974-8520.123138

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   Abstract 

Vasaguduchyadi Kwatha is a compound Ayurvedic formulation, indicated for the treatment of liver diseases, especially for Kamala (jaundice) and Panduroga (anemia). However, till date, no safety profile of this formulation has been reported; hence, in the present study, freshly prepared and market samples of Vasaguduchyadi Kwatha were evaluated for acute toxicity. Acute toxicity test was evaluated as per OECD 425 guidelines with 5 000 mg/kg as limit test in Wistar strain albino rats. Test formulations were administered to overnight fasted animals and parameters like body weight, behavioral changes, and mortality were assessed for 14 days. Hematological and biochemical parameters were assessed on 14 th day. Results showed no significant changes in terms of behavioral changes, mortality, and body weight. Both the samples did not affect any of the hematological parameters. However, increase in blood urea level was observed. This study shows that both the samples of Vasaguduchyadi Kwatha are relatively safe up to the dose of 5 g/kg. However, further chronic toxicity evaluation is necessary to establish its safety profile on chronic administration.

Keywords: Acute toxicity, hepatoprotective, Kamala, Vasaguduchyadi Kwatha


How to cite this article:
Kotecha KN, Kotecha B K, Shukla VJ, Prajapati P, Ravishankar B. Acute toxicity study of Vasaguduchyadi Kwatha: A compound Ayurvedic formulation. AYU 2013;34:327-30

How to cite this URL:
Kotecha KN, Kotecha B K, Shukla VJ, Prajapati P, Ravishankar B. Acute toxicity study of Vasaguduchyadi Kwatha: A compound Ayurvedic formulation. AYU [serial online] 2013 [cited 2020 Jan 19];34:327-30. Available from: http://www.ayujournal.org/text.asp?2013/34/3/327/123138


   Introduction Top


Liver disease is still a worldwide health problem. Unfortunately, conventional or synthetic drugs used in the treatment of liver diseases are inadequate and sometimes can have serious side effects. [1] In absence of a reliable liver protective drug in modern medicine, there are number of medicinal preparations in Ayurveda recommended for the treatment of liver disorders. [2] However, only a small proportion of hepatoprotective plants as well as formulations used in traditional medicine are pharmacologically evaluated for their safety and efficacy. [3] Vasaguduchyadi Kwatha is a compound Ayurvedic formulation explained in Astangahridaya [4] for the treatment of liver diseases, especially for Kamala (jaundice) and Panduroga (anemia). This formulation is extensively used by physicians to treat various liver diseases, especially in alcoholic liver disease. However, till date, no safety profile of this formulation is available. Many physicians use market samples, while some opt for freshly prepared formulation. Hence, in the present study, both fresh and market samples of Vasaguduchyadi Kwatha were selected for acute toxicity study.


   Materials and Methods Top


Test formulation

The plant materials [Table 1] of the test formulation were collected from pharmacy department and adjacent area of Jamnagar city after careful botanical identifications by referring to various botanical floras and with the help of pharmacognosist of the institute. These samples were converted to coarse powder (sieve no. 44) form and from the powder samples Kwatha (decoction) was prepared freshly according to the classical method [5] just prior to administration to the animals. In brief, 16 parts deionized water and one part drug which were boiled on low flame till 1/4 th part was remaining. This was filtered and allowed to cool before administration. The prepared Kwatha contained 25 g of solid material in 100 ml. The rat weighing 200 g received 4 ml of Kwatha (1 g/200 g or 5 g/kg). The market sample of Vasaguduchyadi Kwatha was procured from market.
Table 1: Formulation composition of Vasaguduchyadi Kwatha per liter

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Animals

Female Wistar strain albino rats weighing 150 to 220 g were obtained from animal house attached to Pharmacology laboratory. Five animals in each group were housed in separate cage made up of poly-propylene with stainless steel top grill. The dry wheat (post hulled) waste was used as bedding material and was changed every morning. The animals were acclimatized for seven days before commencement of the experiment in standard laboratory conditions, 12 ± 01 hour day and night rhythm, maintained at 25 ± 3°C and 40 to 60% humidity. Animals were fed with Amrut brand rat pellet feed supplied by Pranav Agro Mills Pvt. Limited. For their drinking purpose, tap water ad libitum was used. Protocol used in this study for the use of animals was approved by the institutional animal ethics committee -Approval number; (IAEC - 04/08-10/PhD/02).

Acute oral toxicity study

Acute oral toxicity study for both freshly prepared sample as well as market samples were carried out following OECD guideline 425 (modified, adopted March 23, 2006) with 5 000 mg/kg as limit test. Food, but not water, was withheld for overnight before the experiment and further 2 hours after administration of test drug. The animals were observed continuously for 6 hours after the dosing for behavioral changes if found any. The careful cage side observation was done without disturbing the animal attention and at the end of every hour the animals were individually exposed to open arena for recording the behavioral changes like increased or decreased motor activity, convulsions, Straub's reactions, muscle spasm, muscle relaxation, salivation, diarrhea, hypoactivity, passivity, relaxation, ataxia, narcosis, etc., Furthermore, all the animals were observed for mortality during the entire period of the study. The body weight of each animal was recorded just prior to dosing on day one and 14 th day. On 14 th day, blood was collected by puncturing supra-orbital plexus by capillary tubes under ether anesthesia for estimation of hematological and biochemical parameters. To estimate hematological parameters, 0.08 ml blood was mixed with 0.02 ml of Ethylene Diamine Tetraacetic Acid-EDTA (33.33 mg/ml) and fed to the auto analyzer (Sismes KX-21, Trans Asia). The parameters measured were as follows: Total WBC, neutrophils percentage, lymphocyte percentage, eosinophils percentage, monocyte percentage, hemoglobin content, packed cell volume (PCV), total RBC, platelet count, mean corpuscular volume, mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC).

For estimation of biochemical parameters, serum was separated from collected blood and requisite quantity of serum was fed to the auto analyzer (Fully automated Biochemical Random Access Analyzer, BS-200; Lilac Medicare Pvt. Ltd., Mumbai) which was automatically drawn into the instrument. Biochemical parameters measured were blood sugar, [6] serum cholesterol, [7] serum triglyceride, [8] blood urea, [9] serum creatinine, [10] serum glutamic pyruvic transaminase, [11] serum glutamic oxaloacetic transaminase, [12] serum total protein, [13] serum albumin and serum globulin [14] , serum alkaline phosphatase, [15] total bilirubin, [16] direct bilirubin, [17] uric acid, [18] and serum calcium. [19]

Statistical analysis

The results were presented as Mean ± SEM for five rats in each group. Statistical comparisons were performed by unpaired student's t test and One-Way ANOVA for all the groups with the level of significance set at P < 0.05.


   Results Top


In the acute toxicity study no mortality was observed in treated rats and no toxic effect was observed throughout the 14 days study period at a dose of 5 000 mg/kg in both the samples of Vasaguduchyadi Kwatha. There were no changes observed in normal gross behavior of animals in both the treated groups.

The changes in body weight of the treated and control rats have been shown in [Table 2]. A normal progressive weight gain was observed in control group. In fresh sample of Vasaguduchyadi Kwatha administered group, marginal gain was observed, whereas in market sample administered group, significant increase in body weight was observed.
Table 2: Effect on body weight

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None of the hematological parameters was affected to significant extent in both the samples of Vasaguduchyadi Kwatha in comparison to control group [Table 3]. Among the 15 serum biochemical parameters, only one parameter was affected to significant extent [Table 4] and there was significant increase in blood urea level in both of the sample administered groups.
Table 3: Effect on hematological parameters

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Table 4: Effect on serum biochemical parameters

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   Discussion Top


The drugs intended to be used therapeutically should be subjected to toxicity evaluation before they are considered safe for use in the human beings. This is important because incomplete knowledge about the toxicity profile of a putative drug will entail certain amount of risk to the recipient. [20] As explained in introductory part, the classical formulation Vasaguduchyadi Kwatha is extensively used clinically to treat various liver diseases; however, its acute toxicity profile is not reported till date. Hence, in the present study, acute toxicity of Vasaguduchyadi Kwatha has been evaluated.

Change in body weight is an important factor to monitor the health of an animal. Frequent Loss of body weight is the first indicator of the onset of an adverse effect and the dose, at which body weight loss is by 10% or more is considered to be a toxic dose, irrespective of whether or not it is accompanied by any other changes. In the present study, in test drug administered groups like control group, gain in body weight was observed and the magnitude of body weight gain was comparatively high in market sample of Vasaguduchyadi Kwatha administered group. Furthermore, at 5 000 mg/kg, sample did not produce any observable toxic effects during entire duration of study and all animals survived 14 days of observation. Both the samples did not affect any of the hematological parameters studied, while only the matter of concern in this study which can be taken as toxic effect is significant increase of blood urea; however, reason behind this change is a matter of further research.


   Conclusion Top


From the observations recorded in acute toxicity studies for behavioral changes, hematological and biochemical parameters, body weight changes, and mortality, it is clear that both the samples of Vasaguduchyadi Kwatha were relatively safe. However, further chronic toxicity profiles are necessary to establish its safety profile on chronic administration.

 
   References Top

1.Guntupalli M. Hepatoprotective effects of rubiadin, a major constituent of Rubiacordifolia Linn. J Ethnopharmacol 2006;103:484-90.  Back to cited text no. 1
    
2.Chatterjee TK. Medicinal plants with hepatoprotective properties. In: Herbal Options. 3 rd ed. Calcutta: Books and Allied (P) Ltd; 2000. p. 135.  Back to cited text no. 2
    
3.Subramonium A, Pushpangadan P. Development of phytomedicines for liver diseases. Indian J Pharmacol 1999;31:166-75.  Back to cited text no. 3
    
4.Vagbhata's. Astangahrdaya, translated by Prof. Srikantha Murthy. Chikitsa Sthana, Panduroga Chikitsa, 16/13 Krinshandas Ayurveda Series: 27. Varanasi: Krinshadas Academy; 2000. p. 449.  Back to cited text no. 4
    
5.Sharangadhara Samhita, Jiwanprada hindi commentary, by Dr. Smt. Shailja Srivastava, Madhyam Khanda 9/3-5. 3 rd ed. Varanasi: Chaukhambha Orientalia; 2003. p. 215.  Back to cited text no. 5
    
6.Pennock CA, Murphy D, Sellers J, Longdon KJ. A comparison auto analyzer methods for the estimation of glucose in blood. Clin Chim Acta 1973; 48:193-201.  Back to cited text no. 6
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7.Roeschlau P, Bernt E, Gruber WA. Enzymatic determination of total cholesterol in serum. J Clin Chem Clin Biochem 1974; 12:226.  Back to cited text no. 7
    
8.Fossati P, Prencipe L. Serum triglycerides determined colorimetrically with an enzyme that produces hydrogen peroxide. Clin Chem 1982; 2:2077-80.  Back to cited text no. 8
    
9.Talke H., Shubert G E.: Enzymatic urea determinationin the blood and serum in the Warburg optical test. Klin Wschr 1965;42:174.  Back to cited text no. 9
    
10.Slot C. Plasma creatinine determination. A new and specific Jaffe reaction method. Scand J Clin Lab Invest 1965;17:381-7.  Back to cited text no. 10
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11.Burtis CA, Ashwood ER, editors. Tietz textbook of Clinical Chemistry. 3 rd ed. Philadelphia, P.A.: Moss D.W., Henderson A.R.; 1999. p. 652.  Back to cited text no. 11
    
12.Tietz NW, editor. Clinical guide to laboratory tests. 3 rd ed. Philadelphia, PA: WB Saunders; 1995. p. 76.  Back to cited text no. 12
    
13.Tietz NW, editor. Text book of Clinical Chemistry. Philadelphia, PA: W.B. Saunders; 1986. p. 579.  Back to cited text no. 13
    
14.Doumas BT, Arends RL, Pinto PC. In standard methods of clinical chemistry. Vol. 7. Chicago: Academic Press; 1972. p. 175-89.  Back to cited text no. 14
    
15.Wilkinson JH, Boutwell JH, Winsten S. Evaluation of a new system for kinetic measurement of serum alkaline phosphatase. Clin Chem 1969;15:487-95.  Back to cited text no. 15
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16.Pearlman PC, Lee RT. Detection and measurement of total bilirubin in serum with use of surfactants as solubilising agents. Clin Chem 1974;20:447-53.  Back to cited text no. 16
    
17.In: Burtis CA, Ashwood ER, editors. Tietz Textbook of clinical chemistry. 3 rd ed. Philadelphia, PA: WB Saunders; 1999. p. 1136.  Back to cited text no. 17
    
18.Kabasakalian P, Kalliney S, Wescott A. Determination of uric acid in serum, with use of uricase and tribromophenol-amino anti pyrine chromogen. Clin Chem 1973;19:522-4.  Back to cited text no. 18
    
19.Tietz NW, editor. Textbook of clinical chemistry. Philadelphia, PA: W. B. Saunders; 1986.p. 1350.  Back to cited text no. 19
    
20.Lipnick RL, Cotruvo JA, Hill RN, Bruce RD, Stitzel KA, Walker AP, et al. Comparison of the up-and-down, conventional LD50 and fixed dose acute toxicity procedures. Food Chem Toxicol 1995;33:223-31.  Back to cited text no. 20
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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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