Study On The Efficient Analysis Method Based On Mass Spectrometry For Plant Hormones Detection

Plant hormones are important small molecules that act at lower concentrations to regulate numerous plant physiological and developmental processes. For the study of biological functions of plant hormones, it is significant to develop a high-throughput and highly selective and sensitive method for determination of plant hormones. Based on SPE-HPLC-MS/MS method, the method for simultaneous determination of six plant hormones in leaf tissue of oilseed rape was developed in our study.1.The quantivative method was developed for analysis of six plant hormones in the leaf tissue of oilseed rape stimulated by oxalic acid, based on the SPE-HPLC-MS/MS. 1-propanol /H2O/ concentrated HCl (2:1:0.002, V/V/V) and dichloromethane were used to extract the plant hormones. The extraction was performed with C18 solid-phase extraction cartridge and the sample was subsequently analyzed by HPLC-ESI-MS/MS. Methanol and H2O with 0.05% formic acid were selected as mobile phase for HPLC, using a gradient of increasing methanol content. The plant hormones were quantified in MRM mode and identified in IDA mode by the hybrid triple quadruple/linear ion trap mass spectrometry. The external standard method was employed for quantitative analysis. Good linearities were obtained for six plant hormones with the correlation coefficients above 0.9924. The detection limits of the target compounds were in the range of 0.005-0.2ng/mL. The extraction recovery yields of plant hormones under SPE conditions ranged from 67.03%-119.83%. Compared with previous methods, sample preparation time and amount of sample required for analysis of plant hormones were reduced, and more classes of hormones were quantitatively measured.2. A novel method for the determination of plant hormones in the fertile oilseed rape samples and the stertile oilseed rape samples has been developed using microwave– assisted– soild phase extraction coupled with HPLC-MS/MS. The influences parameters of extraction, microwave extraction temperature, power and time were all investigated. And the optimal pretreatment condition was microwave temperature 70℃, microwave power 500 W and microwave pretreatment time 8min. The extraction was also performed with C18 solid-phase extraction cartridge. Compared with the conventional extraction, this novel method was simple, rapid and highly selective. Under the optimal conditions, solutions containing varying amounts of each standard compound and deuterium internal standard of the target plant hormones were used to create calibration curves. Good linearities were obtained and in the range of 0.025-200ng/mL for the six plant hormones. The correlation coefficients were in the range of 0.9961-0.9994 and the limit detection was 0.025ng/mL. The recoveries of target analyte spiked in leaf samples were all above 66.49%. This method was feasible. This method was subsequently applied to the determination of phytohormones in the real sample and our results were consistent with related research.