A novel HPTLC method for quantitative estimation of biomarkers in polyherbal formulation

Objective: To explore the quantitative estimation of biomarkers gallic acid and berberine in polyherbal formulation Entoban syrup.Methods: High performance thin layer chromatography was performed to evaluate the presence of gallic acid and berberine employing toluene: ethyl acetate: formic acid:methanol 12:9:4:0.5(v/v/v/v) and ethanol: water: formic acid 90:9:1(v/v/v), as a mobile phase respectively.Results: The R f values(0.58) for gallic acid and(0.76) for berberine in both sample and reference standard were found comparable under UV light at 273 nm and 366 nm respectively. The high performance thin layer chromatography method developed for quantization was simple, accurate and specific.Conclusions: The present standardization provides specific and accurate tool to develop qualifications for identity, transparency and reproducibility of biomarkers in Entoban syrup.

Evaluation of Acute and Repeated Dose Toxicity of the Polyherbal Formulation Linkus Syrup in Experimental Animals

The objective of the present study was to evaluate the pre-clinical efficacy and toxicity of polyherbal cough syrup Linkus. Method: Animals (healthy Wistar albino rats;(150 - 250 g) of either sex) were housed under standard environmental conditions;i.e. 25°C ± 1°C and 12 h dark/light cycle. Food and water were available at libitum. The rats were treated orally with the recommended doses of the test drug (Linkus). After 15 minutes, they were individually placed in a closed Plexiglas chamber (20 × 10 × 10 cm) and exposed to citric acid (0.1 g/ml) inhalation for 7 minutes. The cough reflexes were produced and counted for the last 5 minutes and compared with those of the control animals. The following studies were conducted to evaluate the toxicity of the test drug in healthy Wistar albino rats: lethal dose50 (LD50);rats of either sex (n = 10/sex) were treated orally with doses (1 or 5 g/kg) of the test drug. Mortality and behavioral changes were observed for 1 week. Repeated dose toxicity on the healthy Wistar albino rats of both sexes (n = 5/dose/sex) was treated orally with doses of 20 mg/kg (adult human dose = ~1400 mg), 500 mg/kg (adult human dose = ~35,000 mg) and 1000 mg/kg (adult human dose = ~70,000 mg) of test drug (Linkus) for 14 days. Additionally, the control animals were treated orally with water for 14 days. Results: In female rats, the test drug (Linkus) at the dose of 300 mg/kg caused significant (p < 0.01) reduction in the cough reflexes as compared to the control. However, in male rats, a significant reduction was observed at the tested dose of 200 mg/kg (p < 0.05) and 300 mg/kg (p < 0.01). The test product did not cause mortality in rats at the given doses of 1 or 5 g/kg. Other signs of toxicity like hair loss and weight reduction were not observed. In female and male rats, the test drug (Linkus) at different doses did not show any abnormal effects on complete blood count profile of rats. Serum enzyme markers, i.e. alanine aminotransferase (ALT), alakaline phosphatase, gamma glutamyle transferase (GGT), direct bilirubin, creatinine, and proteins were also observed and found that the test drug at a higher dose did not cause any of the abnormality and had shown significant p value as compared to the control. Conclusion: The test drug (Linkus) could be an effective and safe cough syrup because it did not show any of the side effects or toxicity on experimental animals.

Design, Development and Phytochemical Evaluation of a Poly Herbal Formulation Linkus Syrup

The present work has been carried out on polyherbal formulation named as Linkus Syrup. The herbal formulation consists of Glycyrrhiza glabra, Hyssopus officinalis, Piper longum and Alpinia galangal. The Linkus syrup physico-chemical evaluation such as pH, density, identification of polysaccharide, tanning agents, ascorbic acid and shelf life was complied. The TLC and quantitative determination of alkaloid were quantified. Determination of biomarker has been validated for the analysis of vasicine. The study result revealed that Linkus syrup formulation was well standardized at different levels such as physic-chemical consistency and assay of bio marker compound.

Quantitative Estimation of Gallic Acid as Biomarker in Lipitame Tablets by HPTLC Densitometry for Diabetic Dyslipidemia

Lipitame is a poly herbal formulation comprised of Terminalia arjuna, Terminalia belerica, Commiphora mukul and Phyllanthus emblica. The formulation is investigated for its analysis evaluation. Biomarkers of three herbs such as Terminalia arjuna, Terminalia belerica, and Phyllanthus emblica have been already cited to contain gallic acid, which was qualitatively and quantitatively estimated. In the present study rapid and inexpensive qualification methods for the quality control of Terminalia arjuna, Terminalia belerica, Commiphora mukul and Phyllanthus emblica on thin layer chromatography (TLC) were developed and validated. The solvent system used was toluene:ethyl acetate:formic acid:methanol (12:9:4:0.5). The scanning of plate was performed linearly at 273 nm (absorption) by use of a TLC Scanner III CAMAG with a deuterium source, and the area of spots corresponding to Gallic acid standard was integrated. It was found that gallic acid has been found in the Lipitame tablets on HPTLC densitometry assessment compared with authentic gallic acid reference standard.

Determination of the heavy metal content of Entoban herbal medicinal product

The levels of As, Cd, Pb and Hg were investigated in the medicinal herbal syrup Entoban, which is widely used in the management of non-specific diarrhea and acute intestinal infections of various etiologies. The ingredients in Entoban syrup include 6 potent herbal drugs: Aegle marmelose, Berberis aristata, Butea frondosa, Holarrhena antidysenterica, Myrtus communis, and Quercus infectoria. Entoban medicinal plant syrup was analyzed for As, Cd, Pb and Hg by flame atomic absorption spectroscopy(FAAS). Contents of heavy metals in the examined samples were in the range: As(0.074–10.0 ppm); Cd(0.020–0.3 ppm); Pb(0.00–10.0 ppm) and Hg(0.00–1.0 ppm). Results were compared with permissible limit acceptability intake(AHPA). According to determined amounts of heavy metals, the investigated Entoban syrup samples were validated and considered safe for human consumption.