Development and validation of an HPTLC-densitometric method for the estimation of trigonelline in the 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 the leaves of Abrus precatorius L. and its herbal formulation Chatak® using ICH guidelines. Accelerated solvent extraction (ASE) as an alternative to conventional techniques was also explored for 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 a twin chamber using the mobile phase toluene-ethyl acetate-formic acid-methanol (2:3:1.8:3.8, υ/υ/υ/υ). The plates were scanned at 268 nm (λmax of trigonelline) using a Camag TLC scanner 3 with the CATS 4 software. A linear relationship was obtained between the response (peak area) and the amount of trigonelline in the range 40–200 ng per spot; the correlation coefficient was 0.9957. The ASE method gives higher extraction efficiency in less time compared to Soxhlet extraction. The HPTLC-densitometric method showed good linearity, recovery, and high precision. It is useful to analyze trigonelline in A. precatorius leaves and routine quality control of its marketed formulation.