C_(30)-HPLC-PDA分离与鉴定β,β-胡萝卜素几何异构体

Separation and Identification of β,β-carotene Geometrical Isomers by C_(30)-HPLC-PDA

在装备了二极管阵列(PDA)检测器的HPLC上,应用C30固定相,β,β-胡萝卜素的几何异构体获得了良好的分离。分离条件为:固定相=YMCCarotenoidS-5(4.6×250mm)柱;流动相A=乙腈:甲醇:三乙胺(75:25:0.05,V/V);流动相B=甲基叔丁基醚(MTBE)-三乙胺(100:0.05,V/V);线性梯度:B在20min内由0%增至80%;流速=1.0ml/min;PDA波长范围=260～700nm;进样量=20μl。根据各组分的色谱行为、光谱特征和在碘催化下发生几何异构的产物分析,13,13’-、11,11’-、9,9’-顺式和全反式异构体被鉴定。本项研究的结果奠定了应用C30-HPLC-PDA定量检测β,β-胡萝卜素的几何异构体的基础。

The geometrical isomers of β, β-carotene were separated by HPLC equipped with a photodiode array （PDA） detector on a C30 column under conditions as below： stationary phase=YMCTM Carotenoid S-5 column （250 × 4.6mm）; mobile phase -TCA（75：25：0.05, V/V）; mobile phase B=MTBE-TCA（ 100： 0.05, V/V）; linear gradient： B increased from 0 to 80%（V/V） in 20 minutes; flow rate： 1.0ml/min. PDA wavelength range： 260-700nm; sample injection volume=20μ1. According to their chromatographic behavior, spectral characters and geometrical transition catalyzed by iodine, 4 isomers of 13,13＇ Z-, 11,11＇ Z-, 9,9＇ Z- and all E-isomers were identified. Result from this study is going to be the base to quantify β,β -carotene geometrical isomers by C30-HPLC-PDA.