Misregulation of oxidative and antioxidative processes in the organism - oxidative stress - contributes to the pathogenesis of different diseases, e.g. inflammatory or neurodegenerative diseases. Oxidative stress leads to autoxidation of polyunsaturated fatty acids giving rise to prostaglandin-like isoprostanes (IsoP) and isofurans (IsoF). On the one hand they could serve as biomarker of oxidative stress and on the other hand may act as lipid mediators, similarly as the enzymatically formed oxylipins. In the present paper we describe the development of an LC-ESI(-)-MS/MS method allowing the parallel quantification of 27 IsoP and 8 IsoF derived from 6 different PUFA (ALA, ARA, EPA, AdA, n6-DPA, DHA) within 12 min. The chromatographic separation was carried out on an RP-C18 column (2.1 × 150 mm, 1.8 μm) yielding narrow peaks with an average width at half maximum of 3.3-4.2 s. Detection was carried out on a triple quadrupole mass spectrometer operating in selected reaction monitoring mode allowing the selective detection of regioisomers. The limit of detection ranged between 0.1 and 1 nM allowing in combination with solid phase extraction the detection of IsoP and IsoF at subnanomolar concentrations in biological samples. The method was validated for human plasma showing high accuracy and precision. Application of the approach on the investigation of oxidative stress in cultured cells indicated a distinct pattern of IsoP and IsoF in response to reactive oxygen species which warrants further investigation. The described method is not only the most comprehensive approach for the simultaneous quantification of IsoP and IsoF, but it was also integrated in a targeted metabolomics method (Ostermann et al. (2015) Anal Bioanal Chem) allowing the quantification of in total 164 oxylipins formed enzymatically and non-enzymatically within 30.5 min.

中文翻译：

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开发一种 LC-ESI(-)-MS/MS 方法，用于同时定量来自主要 n3- 和 n6-PUFA 的 35 种异前列腺素和异呋喃

生物体中氧化和抗氧化过程的错误调节——氧化应激——促成了不同疾病的发病机制，例如炎症或神经退行性疾病。氧化应激导致多不饱和脂肪酸的自氧化，产生前列腺素样异前列腺素 (IsoP) 和异呋喃 (IsoF)。一方面，它们可以作为氧化应激的生物标志物，另一方面可以作为脂质介质，类似于酶促形成的氧脂。在本文中，我们描述了 LC-ESI(-)-MS/MS 方法的开发，该方法允许对来自 6 种不同 PUFA（ALA、ARA、EPA、AdA、n6-DPA、DHA）的 27 IsoP 和 8 IsoF 进行平行定量) 12 分钟内。色谱分离在 RP-C18 色谱柱 (2.1 × 150 mm, 1. 8 μm) 产生窄峰，平均半峰宽为 3.3-4.2 s。在选择反应监测模式下操作的三重四极杆质谱仪上进行检测，从而允许选择性检测区域异构体。检测限范围在 0.1 到 1 nM 之间，允许结合固相萃取检测生物样品中亚纳摩尔浓度的 IsoP 和 IsoF。该方法在人血浆中得到验证，显示出高精度和高精度。该方法在培养细胞中的氧化应激研究中的应用表明，IsoP 和 IsoF 响应活性氧的不同模式，值得进一步研究。所描述的方法不仅是同时量化 IsoP 和 IsoF 的最全面的方法，