Air-assisted liquid-liquid microextraction and enantioselective gas chromatography-tandem mass spectrometry quantification of methyl jasmonate stereoisomers in tea (Camellia sinensis L.)

Different stereoisomers of phytohormones may have different roles in plants while the quantification for stereoisomers still remains a challenge. The chiral separation method for methyl jasmonate (MeJA) in tea (Camellia sinensis L.) was developed by air-assisted liquid-liquid microextraction (AALLME) with enantioselective gas chromatography-mass spectrometry (Es-GC-MS/MS). The Plackett-Burman factorial design was introduced to optimize the multiple parameters of AALLME. Sensitive Es-GC-MS/MS enabled the quantification of the MeJA stereoisomers, which were separated from each other with resolutions of 3.5, 2.7, and 2.5. The linearities were R2 > 0.9971, the limits of detection were 0.08–0.12 ng/mL, the limits of quantification (LOD) were 0.28–0.39 ng/mL, and the recovery range was 80.60–107.03%. The method was applied to analyze the MeJA stereoisomer profiles in tea plants after geometrid feeding and mechanical wounding. (+)-MeJA appeared to be the main bio-active monomer that responded to injury stress. (+)-MeJA similarly increased in tea shoots at only 30 s after geometrid attack or mechanical wounding. The proposed method is superior as air-assisted liquid-liquid microextraction could induce the loss of volatile MeJA in a much shorter time, compared to the conventional enrichment procedure. A novel assay protocol for relative quantification of MeJA stereoisomers in plants responding to different injuries was applied.