使用高效液相色谱-电子自旋共振联用仪（HPLC-ESR）和高效液相色谱-电子自旋共振-质谱联用仪（HPLC-ESR-MS）鉴定氧化磷脂酰胆碱与亚铁离子反应混合物中形成的自由基。

Identification of a radical formed in the reaction mixtures of oxidized phosphatidylcholine with ferrous ions using HPLC-ESR and HPLC-ESR-MS.

作者信息

Kumamoto Kazumasa, Hirai Tomihiro, Kishioka Shiroh, Iwahashi Hideo

机构信息

Department of Pharmacology, Wakayama Medical University, 811-1 Kimiidera, Wakayama, 641-8509, Japan.

出版信息

Free Radic Res. 2005 Sep;39(9):987-93. doi: 10.1080/10715760500232073.

DOI:10.1080/10715760500232073

PMID:16087480

Abstract

Identification of free radicals was performed for the reaction mixtures of autoxidized 1,2-dilinoleoylphosphatidylcholine (DLPC) with ferrous ions (or DLPC hydroperoxide with ferrous ions) and of DLPC with soybean lipoxygenase using electron spin resonance (ESR), high performance liquid chromatography (HPLC)--ESR and HPLC--ESR-mass spectrometry (MS) combined use of spin trapping technique. ESR measurements of the reaction mixtures showed prominent signals with hyperfine coupling constants (a(N)=1.58 mT and a(H)beta=0.26 mT). Outstanding peaks with almost same retention times (autoxidized DLPC, 36.9 min; DLPC hydroperoxide, 35.0 min; DLPC with soybean lipoxygenase, 37.1 min) were observed on the elution profile of the HPLC--ESR analyses of the reaction mixtures. HPLC--ESR--MS analyses of the reaction mixtures gave two ions at m/z 266 and 179, suggesting that 4-POBN/pentyl radical adduct forms in these reaction mixtures.

摘要

利用电子自旋共振（ESR）、高效液相色谱（HPLC）-ESR联用以及HPLC-ESR-质谱（MS）联用的自旋捕集技术，对自氧化1,2-二亚油酰磷脂酰胆碱（DLPC）与亚铁离子的反应混合物（或DLPC氢过氧化物与亚铁离子的反应混合物）以及DLPC与大豆脂氧合酶的反应混合物进行自由基鉴定。反应混合物的ESR测量显示出具有超精细偶合常数（a(N)=1.58 mT和a(H)β=0.26 mT）的显著信号。在反应混合物的HPLC-ESR分析的洗脱图谱上观察到具有几乎相同保留时间的突出峰（自氧化DLPC，36.9分钟；DLPC氢过氧化物，35.0分钟；DLPC与大豆脂氧合酶，37.1分钟）。反应混合物的HPLC-ESR-MS分析给出了m/z 266和179处的两个离子，表明在这些反应混合物中形成了4-POBN/戊基自由基加合物。