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  • Title: Determination of endogenous brassinosteroids using sequential magnetic solid phase extraction followed by in situ derivatization/desorption method coupled with liquid chromatography-tandem mass spectrometry.
    Author: Ding J, Mao LJ, Guo N, Yu L, Feng YQ.
    Journal: J Chromatogr A; 2016 May 13; 1446():103-13. PubMed ID: 27072523.
    Abstract:
    In this study, a sequential magnetic solid phase extraction followed by in situ derivatization/desorption method was proposed for the fast, selective and sensitive determination of brassinosteroids (BRs) in plant tissues. Magnetic sorbent for quick, easy, cheap, effective, rugged and safe method (mQuEChERS) and polymer(4-vinylphenylboronic acid-co-ethylene glycol dimethacrylate) coated Fe3O4@SiO2 (p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2) were prepared and characterized. Using them as sorbents, pigments and hydrophilic interferents were firstly removed from plant extract by mQuEChERS, and then endogenous BRs were selectively enriched by p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2 through boronate affinity interaction. After loading BRs on p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2, instead of directly eluting free BRs, the adsorbed BRs were released by adding 4-(N,N-dimethyamino)phenylboronic acid (4-DMAPBA) solution for in situ derivatizaiton/desorption of BRs based on a transesterification reaction between the boronate moieties of p(4-VPBA-co-EGDMA) coated Fe3O4@SiO2 and 4-DMAPBA, finally the resultant solution was submitted to LC-MS/MS for quantification. The whole procedure of the sequential MSPE could be accomplished within 1h, and the matrix effect to MS signal after the sample pretreatment was estimated to be in the range of 93.0-97.4%. The established method provided broad linear dynamics ranges (1.0-100.0pg/mL) with correlation coefficients (R) >0.9978, substantial sensitivity (limits of detection ranged from 0.27 to 1.29pg/mL), high reproducibility (intra-day and inter-day relative standard deviations (RSDs) less than 14.8%) and satisfactory accuracy (recoveries ranged from 74.0%-116.6%). Furthermore, endogenous BRs were successfully detected in one flower of Brassica napus L. (22.5-542.7pg/g fresh weight) and other plant tissues (13.7-289.8pg/g fresh weight).
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