Development and validation of HPTLC-densitometric method for estimation of trigonelline in leaves of Abrus precatorius L. and its formulation

A new, simple and reproducible HPTLC-densitometric method has been established and validated for estimation of trigonelline in leaves of Abrus precatorius L. and its herbal formulation Chatak® using ICH guidelines. Accelerated solvent extraction (ASE) as an alternative to convention techniques was also explored for the rapid extraction. The methanol extracts of leaves, its formulation, and standard solution of trigonelline were applied on silica gel F254 HPTLC plates and developed in twin chamber using mobile phase toluene-ethyl acetate-formic acid-methanol (2:3:1.8:3.8, v/v/v/v). The plates were scanned at 268 nm (λmax of trigonelline) using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area) and amount of trigonelline in the range 40–200 ng spot−1; the correlation coefficient was 0.9957. ASE method has higher extraction efficiency in less time as compared to Soxhlet extraction. The HPTLC-densitometric method showed good linearity, recovery and high precision of compound. It is useful to analyze trigonelline in A. precatorius leaves and routine quality control of its marketed formulation.