Separation and quantitation of jasmonic acid using HPTLC.

A rapid, simple, and stringent protocol for the detection and quantitation of jasmonic acid (JA) is designed using high-performance thin-layer chromatography. Acidified culture filtrate of Lasiodiplodia theobromae is extracted with an equal volume of ethyl acetate and spotted on silica gel 60 F(254) foil using Linomat-5 spray-on applicator. Standard JA is also spotted either internally or adjacent to the sample, and the foils are developed with isopropanol-ammonia-water [10:1:1 (v/v)] as the mobile phase. A quantitative estimation of the separated JA is performed by measuring the absorbance at 295 nm in the reflective mode. The sensitivity of the method is improved by adding internal standard to obtain a detection limit of 1 microg. The limit of quantitation is found to be 80 microg with this method. The method is shown to have selectivity, accuracy, precision, and high sample throughput, making it useful for the routine analysis of JA in basic science and perfumery industries.