Liu Xin, Ma Li, Lin Ya-Wei, Lu Ying-Tang
Key Laboratory of MOE for Plant Developmental Biology, College of Life Sciences, Wuhan University, Wuhan 430072, PR China.
J Chromatogr A. 2003 Dec 22;1021(1-2):209-13. doi: 10.1016/j.chroma.2003.09.004.
A novel method based on capillary electrophoresis coupled to laser-induced fluorescence detection (CE-LIF) was developed for the determination of abscisic acid (ABA), which is an essential phytohormone during plant growth and development. ABA was labeled with 8-aminopyrene-1,3,6-trisulfonate via reductive amination in presence of acetic acid and sodium cyanoborohydride. The derivatization yield was maximized by optimizing several derivatization parameters including derivatization reagent concentration, reaction temperature and time. The conjugate was separated and quantitated by CE-LIF. The linearity of ABA was determined in the range from 0.1 to 10 micromol l(-1) with a correlation of 0.9979. The derivatization limit of detection for ABA was found to be 56 fmol (corresponding to the concentration of 2.8 x 10(-8) mol l(-1)). The detection limit for ABA was 5.5 amol for an injection volume of 5 nl. As a preliminary application, the proposed method was successfully applied to determining trace amount of ABA in the crude extracts of tobacco without extra purification and enrichment procedure and showed a better selectivity and sensitivity than those conventional methods used in determination of ABA.
开发了一种基于毛细管电泳-激光诱导荧光检测（CE-LIF）的新方法，用于测定脱落酸（ABA），ABA是植物生长发育过程中的一种重要植物激素。在乙酸和氰基硼氢化钠存在下，通过还原胺化反应，用8-氨基芘-1,3,6-三磺酸对ABA进行标记。通过优化衍生化试剂浓度、反应温度和时间等几个衍生化参数，使衍生化产率最大化。通过CE-LIF对缀合物进行分离和定量。ABA在0.1至10 μmol l(-1)范围内呈线性，相关系数为0.9979。发现ABA的衍生化检测限为56 fmol（对应浓度为2.8×10(-8) mol l(-1)）。进样体积为5 nl时，ABA的检测限为5.5 amol。作为初步应用，该方法成功应用于不经额外纯化和富集程序直接测定烟草粗提物中的痕量ABA，并且比用于测定ABA的传统方法具有更好的选择性和灵敏度。
