HPTLC Method Development and Validation of Antidiabetic Marker Compound from Polyherbal Formulation

Abstract:

The objective of this research work was to develop a simple, rapid and reliable HPTLC method for standardization of anti-diabetic polyherbal formulation and to carry out validation of Trigonelline in formulation. Development of method was carried out by using Quercetin, Gallic Acid, Curcumin and Trigonelline as bioactive markers reported to have an anti-diabetic activity. Chromatographic analysis was performed using silica gel 60 F254 TLC plate, CAMAG Linomat 5 applicator and solvent system consisting of Isopropyl Alcohol: Ammonia: Acetone in the ratio 1:1:1. Densitometry scanning was performed under reflectance absorbance mode at 254 nm and 366 nm to identify the spots. Rf value of the marker compounds Quercetin, Gallic Acid, Curcumin and Trigonelline was found to be 0.66, 0.42, 0.81 and 0.34 respectively. Validation of Trigonelline was carried out in formulation as per ICH guidelines in terms of Linearity, Precision, Repeatability, Specificity, Robustness, LOD, LOQ and Accuracy. No analytical method has been reported so far associated with a polyherbal formulation containing Quercetin, Gallic acid, Curcumin and Trigonelline focusing on anti diabetic activity. Thus this method can be used for routine quality control of raw material as well as formulation containing Trigonelline as one of its component.

Key words: Polyherbal formulation, Marker Compounds, HPTLC Method development, Validation.