Riso P, Porrini M
Dipartimento di Scienze e Technologic Alimentari e Microbiologiche, sez., Nutrizione, Università degli Studi di Milano, Italy.
Int J Vitam Nutr Res. 1997;67(1):47-54.
In this paper a HPLC method for the determination of lutein, zeaxanthin, beta-cryptoxanthin, alpha-carotene, beta-carotene and lycopene in mixed vegetables and fruit and in human plasma is described. The carotenoids were well separated and the separation was achieved within fifteen minutes using a HPLC system consisting of a 5 microns Vydac 201TP54C18 column, an UV detector, methanol-tetrahydrofuran (95:5 v/v) as mobile phase and a flow rate of 1.0 ml/min. The validity of the separation method was determined by evaluating the linearity of the calibration graphs of each carotenoid (between 0.1 and 1.0 microgram/ml for all compounds except lycopene between 0.1 and 0.8 microgram/ml, r = 0.999) and the accuracy of the chromatographic response (CV < 10%). The reproducibility of the retention times was also good. In the foods samples the extraction procedure was very effective whereas, the saponification step significantly damaged some of the carotenoids. In the plasma the extraction and separation of these compounds were also effective and the qualitative data obtained comparable with those reported in literature. The use of echinenone as internal standard helped to improve quality control.
本文描述了一种用于测定混合蔬菜和水果以及人体血浆中叶黄素、玉米黄质、β-隐黄质、α-胡萝卜素、β-胡萝卜素和番茄红素的高效液相色谱法。使用由5微米的Vydac 201TP54C18柱、紫外检测器、甲醇 - 四氢呋喃（95:5 v/v）作为流动相且流速为1.0毫升/分钟的高效液相色谱系统，类胡萝卜素得到了很好的分离，并且在15分钟内完成了分离。通过评估每种类胡萝卜素校准曲线的线性（除番茄红素在0.1至0.8微克/毫升之间外，所有化合物在0.1至1.0微克/毫升之间，r = 0.999）以及色谱响应的准确性（CV < 10%）来确定分离方法的有效性。保留时间的重现性也很好。在食品样品中，提取过程非常有效，而皂化步骤显著破坏了一些类胡萝卜素。在血浆中，这些化合物的提取和分离也很有效，获得的定性数据与文献报道的数据相当。使用海胆酮作为内标有助于提高质量控制。
