高效液相色谱法测定生物胺衍生条件的优化研究

Optimization for determining derivative conditions of biogenic amines by HPLC

摘要: 利用荧光检测器（FLD），采用柱前衍生高效液相色谱法（HPLC）对组胺（HIS）、色胺（TRP）、2-苯乙胺（2-PHE）、腐胺（PUT）、尸胺（CAD）、酪胺（TYR）、亚精胺（SPD）和精胺（SPM）8种生物胺（BA）进行测定分析。通过探究丹酰氯（Dns-Cl）质量浓度、反应体系的pH、衍生时间及衍生温度对生物胺衍生反应的影响，确定最优的衍生条件。结果显示。当Dns-Cl的质量浓度为BA质量浓度的10倍、pH 11.0、40 ℃下避光反应45 min，所有BA均能有效分离。BA的相对标准偏差（RSD）在3%以内，能够满足分析要求。

Abstract: Using high performance liquid chromatography with fluorescence detector and pre-column derivatization, wedetermined 8 biogenic amines( including histamine, trptamine, 2-phenylethylamine, putrescine, cadaverine, tyramine, spermidine and spermine). The optimal derivative conditions are determined by investigating the effects of dansyl chloride concentration, pH, reaction time and reaction temperature on the derivatization of biogenic amines. The result shows that all amines are well resolved with dansyl chloride whose concentration is 10 times of that of biogenic amines at pH 11.0 and 40 ℃ in darkness for 45 min. The good repeatability of all biogenic amines with relative standard deviation 3% is considered acceptable.