Design and evaluation of herbal hepatoprotective formulation against paracetamol induced liver toxicity

Abstract

Aim: To isolate and identify the quercetin from polyherbal hepatoprotective formulation. Polyherbal formulations were developed by using five bioactive fractionated extracts of Butea monosperma, Bauhinia variegata and Ocimum gratissimum for treatment of liver disorders by exploiting the knowledge of traditional system of medicine and evaluated for hepatoprotective activity using acute liver toxicity model of paracetamol induced liver damage in rats.

Methods: Major active fractions were isolated by solvent fractionation and quantified by HPTLC method. Two polyherbal tablet formulations were developed by the wet granulation method using microcrystalline cellulose, aerosil and other excipients and subjected for physicochemical evaluation to assess physical stability followed by pharmacological screening. The prepared tablets were finally subjected to stability testing to assess its shelf-life. The rats were monitored for change in liver morphology, biochemical parameters like serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP) and total bilirubin for polyherbal tablet formulation at 50 mg/kg and polyherbal tablet formulation at 100 mg/kg.

Results: Active principle was isolated, quantified by HPTLC and characterized with IR. Both formulations showed significant hepatoprotective activity. The histological studies were also support the biochemical parameters. From the results of biochemical analysis and histopathological studies, it can be accomplished that polyherbal tablet formulation at 100 mg/kg can be effectively formulated into a suitable dosage form with added benefit of no side effects for control and cure of chronic ailments like liver disorders. A comparative histopathological study of liver exhibited almost normal architecture as compared to toxicant group.

Conclusion: Biochemical marker showed improved results for polyherbal tablet formulation at 100 mg/kg. Polyherbal tablet formulation contains a potent hepatoprotective agent suggested to be a flavone concentrated in polyherbal formulation which may find clinical application in amelioration of paracetamol induced liver damage.

Keywords: Bauhinia variegata; Butea monosperma; Hepatoprotective activity; Ocimum gratissimum; Polyherbal formulation.

Fig. 1

3D-chromatogram (A) and densitogram (B) of standard quercetin (0.5–2.5 μg/spot).

Fig. 2

HPTLC densitogram of acetone fraction of Butea monosperma (A), ethyl acetate fraction of Bauhinia variegata (B), n-butanol fraction of Bauhinia variegata (C), dichloromethane fraction of Ocimum gratissimum (D), ethyl acetate fraction of Ocimum gratissimum (E) showing quercetin.

Fig. 3

IR spectrum of isolated compound from the mixture of fractions of Butea monosperma, Bauhinia variegata and Ocimum gratissimum.

Fig. 4

Graphs showing effect of polyherbal formulation PTF-1 and PTF-2 on biochemical parameters in paracetamol induced hepatic injury in rats.

Fig. 5

The photomicrographs of liver section from rats (A) received saline 0.5 mL as normal control group (10 × 10), (B) received paracetamol (2 g/kg) (10 × 10), (C) received silymarin (100 mg/kg bw) + PCM (10 × 10), (D) received PF1 (50 mg/kg bw) + PCM and (E) received PF2 (100 mg/kg bw).