Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization, Huanggang Normal University , Huangzhou 438000, China.
J Agric Food Chem. 2012 May 23;60(20):5107-11. doi: 10.1021/jf3018047. Epub 2012 May 9.
An ultrasensitive and selective high-performance liquid chromatographic method for the volatile signaling hormone, jasmonic acid, has been developed based on precolumn derivatization with 1,3,5,7-tetramethyl-8-aminozide-difluoroboradiaza-s-indacene (BODIPY-aminozide). The derivatization reaction was carried out at 60 °C for 30 min in the presence of phosphoric acid. The formed jasmonic acid derivative was eluted using a mobile phase of methanol/pH 6.50 ammonium formate buffer/tetrahydrofuran (67:30:3, v/v/v) in 10 min on a C(18) column and detected with fluorescence detection at excitation and emission wavelengths of 495 and 505 nm, respectively. The detection limit (signal-to-noise ratio = 4) reached 1.14 × 10(-10) M or 2.29 fmol per injection (20 μL), which is the lowest of the existing methods. The proposed method has been successfully applied to the direct determination of trace jasmonic acid in the crude extracts of soybean leaves from soybean mosaic virus-infected and normal plants with recoveries of 95-104%.
建立了一种基于柱前衍生化的、用于痕量挥发性信号激素茉莉酸（jasmonic acid）分析的超灵敏和选择性高效液相色谱方法，衍生试剂为 1,3,5,7-四甲基-8-氨基氮杂二氟硼（BODIPY-氨基氮杂）。衍生反应在 60°C 下进行 30 分钟，在磷酸存在下进行。形成的茉莉酸衍生物在 C(18)柱上，以甲醇/ pH 6.50 甲酸铵缓冲液/四氢呋喃（67:30:3，v/v/v）为流动相在 10 分钟内洗脱，并在激发和发射波长分别为 495nm 和 505nm 处进行荧光检测。检测限（信噪比=4）达到 1.14×10(-10) M 或 2.29 fmol 每次进样（20 μL），这是现有方法中的最低值。该方法已成功应用于感染和正常大豆花叶病毒的大豆叶片粗提物中痕量茉莉酸的直接测定，回收率为 95-104%。
