Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, China.
J Agric Food Chem. 2010 Apr 28;58(8):4560-4. doi: 10.1021/jf100581u.
In phytohormone analysis, mass spectrometry (MS)-based methods are primary and powerful tools. However, complex sample preparation and high cost are problems for their application. As a complement for MS-based methods, a new fluorescent labeling reagent for carboxylic acids, 6-oxy(acetylpiperazine) fluorescein (APF), has been used for the determination of endogenous phytohormones, including indolebutyric acid, 1-naphthaleneacetic acid and 2,4-dichlorophenoxyacetic acid. The derivatization yield was maximized by optimizing derivatization conditions in detail, and the derivatives of three phytohormones could be separated completely in 15 min on a C18 column with fluorescence detection at lambda(ex)/lambda(em) = 467/512 nm. The derivatization limits could reach 0.1 microM, and the detection limits (signal-to-noise ratio = 3) were 4.43-14.2 nM. The proposed method has been applied to the determination of the exogenous phytohormones in the crude extracts of vegetable samples without extra purification and enrichment with recoveries of 94.2-102.4%.
在植物激素分析中，基于质谱（MS）的方法是主要且强大的工具。然而，复杂的样品制备和高成本是其应用的问题。作为基于 MS 方法的补充，一种新的羧酸荧光标记试剂，6-氧（乙酰哌嗪）荧光素（APF），已被用于测定内源性植物激素，包括吲哚丁酸、1-萘乙酸和 2,4-二氯苯氧乙酸。通过详细优化衍生化条件，使衍生化产率最大化，三种植物激素的衍生物可以在 15 分钟内在 C18 柱上完全分离，荧光检测波长为 lambda(ex)/lambda(em) = 467/512nm。衍生化限度可达到 0.1 microM，检测限度（信噪比=3）为 4.43-14.2 nM。该方法已应用于蔬菜样品粗提取物中外源植物激素的测定，无需额外的纯化和富集，回收率为 94.2-102.4%。
