Simultaneous determination of jasmonic acid epimers as phytohormones by chiral liquid chromatography–quadrupole time-of-flight mass spectrometry and their epimerization study

Jasmonic acid (JA) is an essential plant hormone involved in plant development and defense system. There are four stereoisomeric forms of JA and they act quite differently in vivo. In this work, a normal phase liquid chromatography–quadrupole time-of-flight mass spectrometry (NPLC–QTOF-MS) method using cellulose tris (4-methylbenzoate) coated silica gel as the chiral stationary phase was first established for the simultaneous discrimination and direct analysis of all the four JA stereoisomers without need of derivatization. A non-endogenous JA stereoisomer was introduced as the internal standard to ensure the reliability of the developed method. Satisfactory results were obtained in terms of sensitivity (limit of detection, 0.5 ng mL−1 or 2.4 fmol), linearity (R2 = 0.9996) and repeatability (run-to-run RSD of migration time and peak area, 0.37% and 5.9%, respectively, n = 6). Endogenous rise of two natural JA stereoisomers was detected in tobacco leaves and their variations in response to mechanical wounding were monitored. In addition, the configurational stability of JA stereoisomers was investigated using the stereoisomerically pure forms which were not commercially available but easily obtained by our semi-preparative chiral LC method. Experimental evidence indicated that both of the two naturally existing JA stereoisomers were putative signals for wounding response, and the epimerization between them was not a spontaneous process simply promoted by the thermodynamical instability as expected before.

► Direct analysis of four JA stereoisomers by LC–QTOF-MS without derivatization.
► A non-endogenous JA stereoisomer as internal standard to ensure the reliability.
► Endogenous rise of two natural JA stereoisomers was detected in tobacco leaves.
► The variations of JA amount in response to mechanical wounding were monitored.
► The configurational stability of JA stereoisomers was further investigated.