Milne Ginger L, Gao Benlian, Terry Erin S, Zackert William E, Sanchez Stephanie C
Eicosanoid Core Laboratory, Vanderbilt University School of Medicine, Nashville, TN 37232-6602, USA.
Free Radic Biol Med. 2013 Jun;59:36-44. doi: 10.1016/j.freeradbiomed.2012.09.030. Epub 2012 Oct 5.
F2-Isoprostanes (IsoPs) are isomers of prostaglandin F2α formed from the nonenzymatic free radical-catalyzed peroxidation of arachidonic acid. Since discovery of these molecules by Morrow and Roberts in 1990, F2-IsoPs have been shown to be excellent biomarkers as well as potent mediators of oxidative stress in vivo in humans. Isofurans (IsoFs) are also oxidation products generated from the nonenzymatic oxidation of arachidonic acid. IsoFs are preferentially formed instead of F2-IsoPs in settings of increased oxygen tension. The protocol presented herein is the current methodology that our laboratory uses to quantify F2-IsoPs and IsoFs in biological tissues and fluids using gas chromatography/mass spectrometry (GC/MS). A variety of analytical procedures to measure F2-IsoPs, including other GC/MS methods and liquid chromatography/MS and immunological approaches, are reported in the literature. This method provides a very low limit of quantitation and is suitable for analysis of both F2-IsoPs and IsoFs from a variety of biological sources including urine, plasma, tissues, cerebral spinal fluid, exhaled breath condensate, and amniotic fluid, among others.
F2-异前列腺素（IsoPs）是由花生四烯酸的非酶自由基催化过氧化反应形成的前列腺素F2α的异构体。自1990年Morrow和Roberts发现这些分子以来，F2-异前列腺素已被证明是人体体内氧化应激的优秀生物标志物和强效介质。异呋喃（IsoFs）也是花生四烯酸非酶氧化产生的氧化产物。在氧张力增加的情况下，异呋喃优先于F2-异前列腺素形成。本文介绍的方案是我们实验室目前使用气相色谱/质谱联用（GC/MS）技术对生物组织和液体中的F2-异前列腺素和异呋喃进行定量的方法。文献中报道了多种测量F2-异前列腺素的分析方法，包括其他GC/MS方法、液相色谱/质谱联用方法和免疫方法。该方法的定量下限非常低，适用于分析来自多种生物来源的F2-异前列腺素和异呋喃，包括尿液、血浆、组织、脑脊液、呼出气冷凝物和羊水等。
