高效液相色谱-化学发光检测法同时测定人血浆中的磷脂氢过氧化物和胆固醇酯氢过氧化物

Simultaneous determination of phospholipid hydroperoxides and cholesteryl ester hydroperoxides in human plasma by high-performance liquid chromatography with chemiluminescence detection.

作者信息

Yasuda M, Narita S

机构信息

Chemical Analysis Laboratories, Shionogi and Co., Fukushima-ku, Osaka, Japan.

出版信息

J Chromatogr B Biomed Sci Appl. 1997 May 23;693(1):211-7. doi: 10.1016/s0378-4347(97)00034-0.

DOI:10.1016/s0378-4347(97)00034-0

PMID:9200537

Abstract

A method was developed for the simultaneous determination of phosphatidylcholine hydroperoxides (PCOOH) and cholesteryl ester hydroperoxides (CEOOH). Lipid hydroperoxides (LOOH) were quantitatively extracted from human plasma with a mixture of n-hexane and ethyl acetate, and separated by column-switching high-performance liquid chromatography using one aminopropyl column and two octyl columns followed by chemiluminescence detection. LOOHs could be completely separated from each other and detected at picomole levels. The results of method validation tests were satisfactory. This method was then applied to determine LOOH in normal human plasma; the levels of PCOOH and CEOOH found were 36.0+/-4.0 nM (mean+/-S.D., n=6) and 12.3+/-3.1 nM (mean+/-S.D., n=6), respectively.

摘要

开发了一种同时测定磷脂酰胆碱氢过氧化物（PCOOH）和胆固醇酯氢过氧化物（CEOOH）的方法。脂质氢过氧化物（LOOH）用正己烷和乙酸乙酯的混合物从人血浆中定量提取，并通过柱切换高效液相色谱法分离，使用一根氨丙基柱和两根辛基柱，随后进行化学发光检测。LOOHs可以彼此完全分离，并在皮摩尔水平进行检测。方法验证试验的结果令人满意。然后将该方法应用于测定正常人血浆中的LOOH；所发现的PCOOH和CEOOH水平分别为36.0±4.0 nM（平均值±标准差，n = 6）和12.3±3.1 nM（平均值±标准差，n = 6）。