Acute oral toxicity study of polyherbal formulation AV/KPC/10

The current study was designed to study acute oral toxicity study of polyherbal formulation AV/KPC/10 (M/S Ayurvet Limited, Baddi, India) according to OECD guidelines. For new substances it is the recommended stepwise testing approach for developing scientifically sound data on the safety of the substance. Three wistar rats (2 females and 1 male) were used for the study. AV/KPC/10 in single oral dose (2000mg/kg) supplemented to all rats. The parameters like general appearance, behavior, body weight, mortality and necropsy were studied. No changes in general appearance and mortality was observed. AV/KPC/10 was found to be safe at dose of 2000mg/kg.


Introduction
Traditional and alternative medicine is extensively practiced in the prevention, diagnosis, and treatment of various diseases [1] [2]. Despite the widespread use of plants for treatment of several ailments there is a little known about their toxicity and safety. The evaluation of the toxic action of the plant extracts or herbal formulations is important in order to consider them safe before used as medicines [3]. A key stage in ensuring the safety of drugs is to conduct toxicity tests in appropriate animal models [4]. The acute oral toxicity test aims at establishing the therapeutic index, i.e. defined as the ratio LD 50 : ED 50 . In general, the narrower this margin, the more likely it is that the drug will produce unwanted effects, the greater the index the safer the compound. However, the term acute oral toxicity is most often used in connection to lethality and lethal dose determinations [5]. The objective of the current experiment was to study the Acute Toxicity of AV/KPC/10 (M/S Ayurvet Limited, India), an herbal formulation containing herbs viz. Phyllanthus niruri, Tephrosia Purpurea, Glycyrrhiza glabra etc. recommended for the treatment of ketosis in cattle.

Materials & methods
Acute Toxicity study of "AV/KPC/10" was performed following OECD Guideline 423 in the Department of Pharmacology and Toxicology, College of Veterinary & Animal Sciences, Bombay Veterinary College, Mumbai, Maharashtra, India

Experimental animals
The animals for the current study were approved by Institutional Animal Ethical Committee of Bombay Veterinary College, approval no: BVC/IAEC/50/2012. Wistar rats (2 females and 1 male) of age 9 weeks and average weight of 250 gms were used for the study.

Experimental design
The rats were acclimatized to their surroundings 5 days prior to the test by separating them from the rest of the animals and allotting different cages to them. The rats were identified by color marking. Rats had free access to standard commercial rat feed, except on the day prior to dosing. Water was provided ad libitum. The rat feeding needle was used to dose the animals. Weights were recorded prior to dosing (2ml/animal/single oral dose) (2000 mg/kg) on day 7 and prior to sacrifice on IJBAR (2015) 6 (03) www.ssjournals.com day 14. All the procedures were carried out at ambient temperature (27°C).

General appearance and behavioral observations
The appearance and behavioral parameters of animals after drug administration is indicator of the toxicity of the test drug [6] [7]. The behavioral patterns of animals were observed first 6 h and followed by 14 h after the administration. No significant changes were observed in wellness parameters used for evaluation of toxicity. Skin, fur, eyes, mucous membrane, behavioral pattern, salivation and sleep pattern parameters of the treated animals were found to be normal (table 1). No toxic symptom or mortality was observed in any animal. All treated animals lived up to 14 days after the administration of AV/KPC/10.

Body Weights
An increase in body weight of the animal after test drug administration is indicator of its toxic effect [8]. Table 2 showed the change observed before and after the administration of the AV/KPC/10. Although, the body weights of all the rats were increased after the oral administration of AV/KPC/10. But, the changes of the body weights were found to be statistically insignificant. Insignificant increase in body weight of test animals indicates that the administration of the AV/KPC/10 had no toxic effect on animals.

LD 50 Value
An LD 50 value is the dose at which 50 percent of the test animals can be expected to die [9][10] [11]. As per calculations from Acute Oral Toxicity (Category 5 as per OECD guidelines 420, 423 & 425 for acute Toxicity Studies) the LD 50 value of AV/KPC/10 was found to be more than 2000 mg/kg body weight.

Conclusion
All the three animals survived by the end of the study; Clinical signs symptoms and gross necropsy did not reveal any major findings. The LD 50 of the AV/KPC/10 was greater than 2000mg/kg (Category 5 as per OECD guidelines 420, 423 & 425 for acute Toxicity Studies) and hence it is practically nontoxic.