摘要
对氨基酸的反相高效液相色谱的双荧光检测作了介绍。采用邻苯二甲醛 /巯基乙醇(OPA/MCE)与 9-芴甲基氯甲酸酯 (FMOC)柱前衍生 ,同时采用两台串联的荧光检测器对伯氨基酸和仲氨基酸进行分析 ;利用高半胱氨酸 (HCYS) ,高丝氨酸 (HSER) ,正缬氨酸 (NVAL) ,硫代脯氨酸(TPRO)四种内标消除了系统误差。同时 ,由样品各组分保留时间重现性的相对误差及标准氨基酸衍生物的回收率实验来确定该方法的准确性。结果表明 :此检测氨基酸方法灵敏度高、检出限高且重现性好 ,克服了单一衍生剂及单检测分析法的不足 ;消除了NH3 峰对色谱柱影响 ;样品保留时间重现性相对误差 <5 % ,标样回收率 >90 % 。
This paper introduces analysis of RP HPLC for amino acids,which uses OPA/MCE and FMOC derivatives with two fluorescence detectors.The system's error is removed through Internal Standard,i.e.HCYS,HSER,NVAL,TPRO.Meanwhile,the method' accuracy is determined by relative error of recoverity with all kinds of amino acid's ramification.The result shows:it is an ideal method for amino acids'analysis with better sensitivity,detection limits and recoverity,overcoming the limitation by single ramification & only one detector.In addition,it gets over influent of the peak of NH 3 to column.And the relative error of retentive time is <5% & experiment of recycling is >90%,which explains credibility of the method.
出处
《重庆大学学报（自然科学版）》
CAS
CSCD
2000年第4期83-85,共3页
Journal of Chongqing University
关键词
氨基酸
衍生化
荧光分析
色谱分析
amino acid
derivative
analysis of fluorescence
internal standard
HPLC
