Kuksis Arnis, Suomela Jukka-Pekka, Tarvainen Marko, Kallio Heikki
Banting and Best Department of Medical Research, University of Toronto, ON, Canada.
Mol Biotechnol. 2009 Jun;42(2):224-68. doi: 10.1007/s12033-009-9146-z. Epub 2009 Mar 3.
Thin-layer chromatography (TLC), gas chromatography (GC), and liquid chromatography (LC) in combination with mass spectrometry (MS) have been adopted for the isolation and identification of oxolipids and for determining their functionality. TLC provides a rapid separation and access to most oxolipids as intact molecules and has recently been effectively interfaced with time-of-flight (TOF) MS (TOF-MS). GC with flame ionization (FI) (GC/FI) and electron impact (EI) MS (GC/EI-MS) has been extensively utilized in the analysis of isoprostanes and other low-molecular-weight oxolipids, although these methods require derivatization of the analytes. In contrast, LC with ultraviolet (UV) absorption (LC/UV) or evaporate light scattering detection (ELSD) (LC/ELSD) as well as electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) MS (LC/ESI-MS) or LC/APCI-MS has proven to be well suited for the analysis of intact oxolipids and their conjugates without or with minimal derivatization. Nevertheless, kit-based colorimetric and fluorescent procedures continue to serve as sensitive indicators of the presence of hydroperoxides and aldehydes.
薄层色谱法(TLC)、气相色谱法(GC)以及液相色谱法(LC)与质谱法(MS)联用,已被用于氧脂类的分离与鉴定及其功能的测定。TLC能快速分离并获取大多数完整分子形式的氧脂类,且最近已有效地与飞行时间质谱仪(TOF-MS)联用。配备火焰离子化检测器(FI)的气相色谱仪(GC/FI)和配备电子轰击电离源(EI)的质谱仪(GC/EI-MS)已广泛用于异前列腺素和其他低分子量氧脂类的分析,不过这些方法需要对分析物进行衍生化处理。相比之下,配备紫外吸收检测器(UV)或蒸发光散射检测器(ELSD)的液相色谱仪(LC/UV或LC/ELSD)以及配备电喷雾电离源(ESI)或大气压化学电离源(APCI)的质谱仪(LC/ESI-MS或LC/APCI-MS),已证明非常适合分析完整的氧脂类及其共轭物,无需衍生化或只需进行最少的衍生化处理。尽管如此,基于试剂盒的比色法和荧光法仍是氢过氧化物和醛类存在的灵敏指标。