Department of Analytical Chemistry, Chemical Faculty, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, Poland.
Int J Mol Sci. 2013 Mar 18;14(3):6157-69. doi: 10.3390/ijms14036157.
Isoprostanes are stable products of arachidonic acid peroxidation and are regarded as the most reliable markers of oxidative stress in vivo. Here we describe the LC-MS/MS procedure enabling simultaneous determination of four regioisomers (8-iso prostaglandin F2α, 8-iso-15(R)-prostaglandin F2α, 11β-prostaglandin F2α, 15(R)-prostaglandin F2α) in plasma samples collected from mice. The four plasma isoprostanes are determined by LC-ESI-MS/MS with deuterated 8-iso-PGF2α-d4 as an internal standard (I.S.). For plasma samples spiked with the isoprostanes at a level of 200 pg/mL each, the method imprecision has been below 7.1% and mean inaccuracy equaled 8.7%. The applicability of the proposed approach has been verified by the assessment of changes in isoprostane levels in plasma samples derived from mice exposed to tert-butyl hydroperoxide (TBHP), a model inducer of oxidative stress, or to antitumor drug doxorubicin (DOX) known for potent stimulation of redox cycling. Compared to the control group of mice, both oxidative stress inducers tested increased the levels of three out of four isoprostanes in exposed animals; 11β-prostaglandin F2α being the exception. The greatest rise was observed in the case of 15(R)-prostaglandin F2α, by about 50% and 70% in plasma samples derived from mice exposed to DOX and TBHP, respectively.
异前列烷是花生四烯酸过氧化的稳定产物，被认为是体内氧化应激的最可靠标志物。本文描述了一种 LC-MS/MS 方法，能够同时测定血浆样品中的四种区域异构体（8-异前列烷 F2α、8-异-15(R)-前列腺素 F2α、11β-前列腺素 F2α、15(R)-前列腺素 F2α）。用 LC-ESI-MS/MS 测定 4 种血浆异前列烷，以氘代 8-异-PGF2α-d4 为内标（I.S.）。对于每个浓度为 200 pg/mL 的异前列烷加标血浆样品，该方法的精密度低于 7.1%，平均准确度等于 8.7%。通过评估暴露于叔丁基过氧化物（TBHP）（一种氧化应激诱导剂）或抗肿瘤药物阿霉素（DOX）（一种已知的氧化还原循环强烈刺激物）的小鼠血浆样品中异前列烷水平的变化，验证了所提出方法的适用性。与对照组小鼠相比，两种氧化应激诱导剂均增加了暴露动物中四种异前列烷中的三种水平；11β-前列腺素 F2α 是例外。在 DOX 和 TBHP 暴露的小鼠血浆样品中，15(R)-前列腺素 F2α 的增加最为显著，分别约为 50%和 70%。
