Quantification of jasmonic acid, methyl jasmonate, and salicylic acid in plants by capillary liquid chromatography electrospray tandem mass spectrometry.

Jasmonic acid, methyl jasmonate, and salicylic acid have been reported to occur in plants and are thought to be essential for the regulation of systemic defense responses. This work describes a method for the quantitation in plant tissue of these regulators by reverse-phase capillary liquid chromatography interfaced to an electrospray tandem mass spectrometer. Inclusion during sample preparation of hydrogenated and/or deuterated internal standards corresponding to analogs of the regulators compensated for sample loss and permitted quantitation using the multiple reaction monitoring mode of the mass spectrometer. The free acids were analyzed in a negative-ion mode, whereas methyl jasmonate was analyzed in a positive-ion mode. Using these procedures an extract of fresh hybrid poplar leaves was found to contain per gram of leaf tissue 2.6 micrograms of jasmonic acid, 1.3 micrograms of methyl jasmonate, and 31.0 micrograms of salicylic acid. The techniques used should be applicable to other plant materials.