TY - JOUR T1 - Determination of endogenous brassinosteroids in plant tissues using solid-phase extraction with double layered cartridge followed by high-performance liquid chromatography-tandem mass spectrometry. AU - Ding,Jun, AU - Mao,Li-Jing, AU - Wang,Shao-Ting, AU - Yuan,Bi-Feng, AU - Feng,Yu-Qi, Y1 - 2013/02/25/ PY - 2012/09/25/received PY - 2012/12/17/revised PY - 2012/12/26/accepted PY - 2013/2/26/entrez PY - 2013/2/26/pubmed PY - 2014/2/5/medline SP - 386 EP - 94 JF - Phytochemical analysis : PCA JO - Phytochem Anal VL - 24 IS - 4 N2 - INTRODUCTION: Brassinosteroids (BRs) are a group of important phytohormones that play vital roles in plant growth, development and a series of physiological phenomena. In order to understand biosynthesis, degradation and metabolic pathways of BRs, a reliable analytical method of BRs with effective sample pre-treatment process is favourable. OBJECTIVE: The development of a quick and effective method for the quantification of endogenous BRs in plant tissue with the aid of double layered solid-phase extraction (SPE) cartridges (graphite carbon black and primary secondary amine silica sorbent: GCB/PSA). METHOD: The method involved an initial extraction of BRs with acetonitrile, a dehydration process with anhydrous MgSO4 and NaCl, a SPE purification process with a double layered cartridge, and a further clean-up step utilising liquid-liquid extraction (LLE). The purification process was mainly realised on the GCB/PSA cartridge. GCB could eliminate hydrophobic compounds, especially those containing a π system, and PSA was introduced to remove the polar interferences. Endogenous BRs were quantified by HPLC-ESI-MS/MS. RESULTS: Good linearities were obtained in the range of 0.4-500 ng/mL (0.0124-15 ng), with the correlation coefficients above 0.9957. The relative recoveries of BRs of this method were in the range of 71.1-113.1%, with intra- and interday relative standard deviations (RSDs) less than 16.3%. With the proposed method, the requirement of plant tissue amount was minimised to 1 g fresh weight, which is the smallest amount reported so far, to our knowledge. SN - 1099-1565 UR - https://www.unboundmedicine.com/medline/citation/23436553/Determination_of_endogenous_brassinosteroids_in_plant_tissues_using_solid_phase_extraction_with_double_layered_cartridge_followed_by_high_performance_liquid_chromatography_tandem_mass_spectrometry_ L2 - https://doi.org/10.1002/pca.2421 DB - PRIME DP - Unbound Medicine ER -