血清普罗布考与脂溶性抗氧化剂的同步测定

Simultaneous measurement of serum probucol and lipid-soluble antioxidants.

作者信息

Schäfer Elinder L, Walldius G

机构信息

King Gustaf V Research Institute, Karolinska Institutet, Stockholm, Sweden.

出版信息

J Lipid Res. 1992 Jan;33(1):131-7.

PMID:1552229

Abstract

A method is described for the simultaneous measurement of probucol, retinol, tocopherols, lycopene, and carotenes by reverse phase high performance liquid chromatography. A high sensitivity was achieved by use of a microbore column and by monitoring the effluent at the optimum wavelengths of each substance with a diode array detector. The detection limits were lycopene 0.5 ng; alpha-carotene, beta-carotene, and retinol 1 ng; probucol 2 ng; alpha-tocopherol and gamma-tocopherol 15 ng. The eluent was acetonitrile-water-tetrahydrofuran 81.3:5.7:13 (v/v/v) and the flow rate was 0.4 ml/min. Quantitation was performed by use of the four internal standards retinol acetate, 2-pentanone bis(3,5-di-tert)mercaptole, alpha-tocopherol acetate, and retinol palmitate, which resemble the respective analytes in structure and/or polarity. In order to attain a reproducible recovery of particularly the carotenes, the total lipid content of the samples had to be controlled by dilution of the sample before extraction. The coefficients of variation for between-day determinations of a serum pool were 3.8% for retinol, 4.5% for probucol, 11.2% for gamma-tocopherol, 4.5% for alpha-tocopherol, 10.4% for lycopene, 8.0% for alpha-carotene, and 7.0% for beta-carotene.

摘要

本文描述了一种用反相高效液相色谱法同时测定普罗布考、视黄醇、生育酚、番茄红素和类胡萝卜素的方法。通过使用微径柱并利用二极管阵列检测器在每种物质的最佳波长下监测流出物，实现了高灵敏度。检测限为番茄红素0.5 ng；α-胡萝卜素、β-胡萝卜素和视黄醇1 ng；普罗布考2 ng；α-生育酚和γ-生育酚15 ng。洗脱液为乙腈 - 水 - 四氢呋喃81.3:5.7:13（v/v/v），流速为0.4 ml/min。使用视黄醇乙酸酯、2-戊酮双（3,5-二叔丁基）巯基醚、α-生育酚乙酸酯和视黄醇棕榈酸酯这四种内标进行定量，它们在结构和/或极性上与各自的分析物相似。为了获得特别是类胡萝卜素的可重复回收率，必须在提取前通过稀释样品来控制样品的总脂质含量。血清混合液日间测定的变异系数为：视黄醇3.8%，普罗布考4.5%，γ-生育酚11.2%，α-生育酚4.5%，番茄红素10.4%，α-胡萝卜素8.0%，β-胡萝卜素7.0%。