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Quantification of Negundoside in Vitex negundo Linn. Leaf by High-Performance Thin-Layer Chromatography

A sensitive, simple, accurate, and economic thin-layer chromatographic method has been established for analysis of negundoside in leaf powder of Vitex negundo Linn. A methanol extract of the leaf powder was used for the experimental work. Separation was performed on aluminum-backed silica gel 60 F254 HPTLC plates with ethyl acetate—methanol—water—glacial acetic acid 7.8:1.2:0.7:0.3 as mobile phase. After development, quantification was performed by densitometry at 267 nm. The limits of detection and quantitation were 0.4 and 1.2 μg mL−1, respectively. The response was linearly dependent on the amount of negundoside in the range 1.2 to 22.5 μg mL−1. The regression coefficient for negundoside was 0.9998. The validated TLC densitometric method can be used for routine quality-control analysis of V. negundo leaf powder and for quantitative analysis of negundoside.