Development and validation of HPTLC densitometric method for estimation of charantinin Momordica charantia fruits and herbal formulation

Abstract

A simple, sensitive, precise, rapid, and reliable HPTLC method for the estimation of charantin in Momordica charantia fruits as well as in herbal formulation was developed. In this method, Aluminum backed silica gel 60 F254 plates (20x10 cm: 200 µm thickness) were used as stationary phase and toluene-ethyl acetate-methanol-formic acid (68:20:10:02, v/v) as a optimized mobile phase. Developed chromatogram was scanned at 525 nm, the wavelength of maximum absorption for charantin after derivatization with anisaldehyde-sulphuric acid reagent. Regression analysis of the calibration data showed an excellent linear relationship between peak-area vs drug concentration. Linearity was found to be in the range of 100-500 ng/band. The suitability of developed HPTLC method for estimation of charantin was established by validating it as per the ICH guidelines. The limits of detection (LOD) and quantification (LOQ) for charantin were found to be ≈30 and ≈90 ng/band. The developed method was found to be linear (r2=0.9943), precise (RSD < 1.5% and < 2% for intra-day and inter-day precision), accurate (mean recovery of within the range of 98.68-100.20%), specific and robust. The developed method has been successfully applied for the determination of charantin in M. charantia fruits and herbal formulation without any interference.