Gansu Provincial Key Laboratory of Aridland Crop Science, Gansu Agricultural University, Lanzhou 730070, China.
Molecules. 2022 Jan 27;27(3):831. doi: 10.3390/molecules27030831.
A determination method for trace 24-epibrassinolide (EBL) in plant tissues was developed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The plant tissue samples were extracted using a methanol-formic acid solution, and the corresponding supernatant was purified with ODS C18 solid-phase extraction column. The extracts were separated using a Zorbax Eclipse Plus C18 (2.1 mm × 50 mm, 1.8 μm) column with methanol and 0.1% formic acid as the mobile phase. The ion source for the mass spectrometry was an electrospray ionization source with positive ion mode detection. The linear range of the target compound was 0.7104 μg/kg, the limit of detection (LOD) was 0.110.37 μg/kg, the limit of quantification (LOQ) was 0.361.22 μg/kg, the recovery rate was 84.0116.3%, and the relative standard deviation (RSD%) was 0.8~10.5. The samples of maize plumule, brassica rapeseed flower, and marigold leaf were detected using the external standard method. The optimization of the extraction method and detection method of EBL improved the detection sensitivity, laid a foundation for the artificial synthesis of EBL, improved the extraction rate of EBL, and provided a theoretical basis for the study of EBL in many plants.
建立了一种利用超高效液相色谱-串联质谱法（UPLC-MS/MS）测定植物组织中痕量 24-表油菜素内酯（EBL）的方法。采用甲醇-甲酸溶液提取植物组织样品，相应的上清液用 ODS C18 固相萃取柱纯化。采用甲醇和 0.1%甲酸作为流动相，在 Zorbax Eclipse Plus C18（2.1mm×50mm，1.8μm）柱上进行分离。质谱的离子源为正离子模式电喷雾电离源。目标化合物的线性范围为 0.7104μg/kg，检测限（LOD）为 0.110.37μg/kg，定量限（LOQ）为 0.361.22μg/kg，回收率为 84.0116.3%，相对标准偏差（RSD%）为 0.8~10.5。采用外标法对玉米幼芽、油菜花花和万寿菊花叶进行了检测。EBL 的提取方法和检测方法的优化提高了检测灵敏度，为 EBL 的人工合成奠定了基础，提高了 EBL 的提取率，为研究多种植物中的 EBL 提供了理论依据。
