氯甲酸芴甲酯柱前衍生高效液相色谱法测定植物中多胺

High Performance Liquid Chromatographic Determination of Plant Polyamines by Pre-column Derivatization With 9-Fluorenylmethyl Chloroformate

建立了一种采用氯甲酸芴甲酯进行柱前衍生，高效液相色谱分离和测定植物中游离 态和结合态多胺的新方法。腐胺、精脒和精胺的分离在 １６ｍｉｎ内完成；检出限分别达到 ０．０３０、０．０１８和０．０２３ｐｍｏｌ；线性动态范围至少是３个数量级。研究了反应温度和时间对荧光 衍生物产率的影响，指出在２５℃下反应２０ｍｉｎ可获得较高产率。多胺总量与衍生试剂的摩 尔比超过１：８时，荧光衍生物产率恒定。本方法用于苹果和番茄样品的测定，并进行了回收 率试验。腐胺、精脒和精胺的５次测定ＲＳＤ％分别为１．１１％～３．４１％，１．４９％～２．９１％， １．５９％～３．１４％；回收率分别为９７．５％～１０３．２％，９８．６％～１０２．２％，９８．４％～１０２．４％。

A new method by which free and bound plant polyamines were separated and determined with high performance liquid chromatography after derivatization with 9-fluorenylmethyl chloroformate was de- veloped. Putrescine, spermidine and spermine might be separated in 16 minutes, the detection limits were 0.030, 0. 018 and 0. 023 pmol, respectively, and the linear dynamic ranges were 103 at least. It was studied for the temperature and time of the reaction how to influence the productivity of fluorescent deriva- nts. The experimental result pointed that higher productivity might be obtained if the reaction time was 20 minutes at 25℃. The productivity of fluorescent derivants would be constant when the molar ratio of poly- amines and 9-fluorenylmethyl chloroformate was beyond 1 : 8. The method has been applied to the deter- minations of apple and tomato samples. The relative standard deviation of 5-run-determinations to putrescine, spermidine and spermine were 1.1 % - 3 .4%, 1.5% - 2 .9%, 1. 6% - 3. 1 %, and the recov- eries of them were 97. 5% - 103. 2%,98. 6% - 102. 2%, 98. 4% - 102.4%, respectively.