Korytowski W, Geiger P G, Girotti A W
Department of Biochemistry, Medical College of Wisconsin, Milwaukee 53226, USA.
J Chromatogr B Biomed Appl. 1995 Aug 18;670(2):189-97. doi: 10.1016/0378-4347(95)00182-4.
Lipid hydroperoxide species can be analyzed with high sensitivity and specificity, using reversed-phase high-performance liquid chromatography with reductive mode electrochemical detection on a mercury drop cathode [HPLC-ED(Hg)]. The purpose of this study was to examine different variables in the operation of HPLC-ED(Hg) and to select optimal conditions for the analysis of several biologically relevant peroxides, including species derived from cholesterol, cholesteryl linoleate, oleate, linoleate, and two synthetic phosphatidylcholines. Parameters such as operating potential and mobile-phase solvent proportions, electrolyte composition, and ionic strength were evaluated for each peroxide class. Under optimal conditions, we have achieved baseline separation of four cholesterol hydroperoxide species, not only from one another, but also from phospholipid hydroperoxides; detection limits were < 0.3 pmol and < 30 pmol for the cholesterol and phospholipid hydroperoxides, respectively.
脂质氢过氧化物可以通过反相高效液相色谱法结合汞滴阴极的还原模式电化学检测 [HPLC-ED(Hg)] 进行高灵敏度和高特异性分析。本研究的目的是研究HPLC-ED(Hg) 操作中的不同变量，并选择分析几种具有生物学相关性的过氧化物的最佳条件，这些过氧化物包括源自胆固醇、亚油酸胆固醇酯、油酸酯、亚油酸酯以及两种合成磷脂酰胆碱的物质。对每种过氧化物类别评估了操作电位、流动相溶剂比例、电解质组成和离子强度等参数。在最佳条件下，我们实现了四种胆固醇氢过氧化物的基线分离，不仅彼此分离，还与磷脂氢过氧化物分离；胆固醇和磷脂氢过氧化物的检测限分别 < 0.3 pmol 和 < 30 pmol。
