咔唑-9-乙基氯甲酸酯柱前衍生胺类化合物高效液相色谱分离

Determination of Amines by High Performance Liquid Chromatography with a Pre-column Derivatization and Fluorescence Detection

采用一种新的氯甲酸酯类荧光衍生试剂咔唑 9 乙基氯甲酸酯进行柱前衍生胺类化合物并通过荧光检测的方法进行高效液相色谱 (HPLC)分析。衍生物的荧光激发和发射波长为λex λem =2 93 36 5nm。色谱柱 :HypersilBDSC1 8柱 ,柱温 :35℃ ;流速 :1mL min ;流动相A为水 乙腈 (2 0 80 ,V/V) ,流动相B为乙腈 水 (95 5 ,V/V)。衍生试剂用量 (mol)是胺类化合物总量的 3～ 4倍时 ,衍生化产率最大且恒定。该方法具有检测灵敏度高 ,衍生化反应简单、快速、不受样品基质盐分的干扰 ,尤其适合天然生物样品 ,食品及饲料中的胺类化合物的分析.

2-(9-Carbazole) ethyl chloroformate (CEOC), a novel fluorescence derivatization reagent, has been firstly developed for the determiantion of amines. In a one-step procedure, CEOC reacts with the amines to form stable derivatives. Maximum yield is observed with a molar reagent excess of approximately 3 to 4 times for each of the amines. The method in conjunction with a gradient elution offers a baseline separation on a reversed-phase Hypersil BDS C-18 column. Mobile phase: A = acetonitrile/water (20/80, V/V); B = acetonitrile/water (95/5, V/V) flow rate = 1.0 mL/min; column temperature was set at 35 degreesC. The fluorescent derivatives are detected at lambda(ex)/lambda(em) = 293/365 nm. It was proved that CEOC-high performance liquid chromatography (HPLC) not only had excellent precision and accuracy but also high tolerance of the matrix of samples (including salt), therefore the method is suitable for the determination of biological materials, foods and feeds.