Determination of ABA Content in Wheat Leaves with the Method of Improved SPE-LC-ESI-MS/MS

Su Shi;Li Ruihang;Lang Danying;Wang Junwei;Guo Aiguang;Xu Hong;College of Life Sciences,Northwest A&F University;College of Agronomy,Northwest A&F University;State Key Laboratory of Crop Stress Biology for Arid Area;

The quantity determination of abscisic acid,Liquid chromatography-tandem mass,Wheat albino line,Leaves of seedling stage

Here we present a optimize liquid phase chromatography with electro spray tandem mass spectrometry(LC-ESI-MS/MS) method for fast and highly sensitive determination of endogenous ABA content in wheat leaves.The ultrapure water was used as solvent,and the leaf tissues were extracted in dark at 4℃ for 12 h. Then,ABA fraction was enriched by C18 solid phase extraction column and was quantified by LC-ESI-MS/MS system. The methanol/0.2% formic acid(4:1,v/v) were used as mobile phase for isocratic elution of C18 reversed-phase column.The electro spray ionization(ESI) operated in negative ion mode,and the monitor mode was multiple reaction monitoring(MRM),themonitoredion pairswas 262.8 152.7. ABA standard test indicated good linearityin7.5~60 ng/m L and 4.7 ng/m L detection limits on a signal-to-noise ratio of three. We got the average recovery rate of this system98.61% by the method of 20 ng ABA standards addition. Using this system,we determined the endogenous ABA content in the leaves of wheat albino line and parent variety Aibian1 seedlings at room and low temperature,and with exogenous ABA treatment. The results showed the differences between two cultivars. So,we can conclude that the optimized SPE-LC-ESI-MS/MS system can be used for the analysis of trace amounts of ABA in wheat leaves,and has the advantages of simple sample pretreatment,higher recovery and sensitive detection.