Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche "Togo Rosati", 06126, Perugia, Italy.
Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche "Togo Rosati", 06126, Perugia, Italy.
Free Radic Biol Med. 2019 Nov 20;144:134-143. doi: 10.1016/j.freeradbiomed.2019.04.017. Epub 2019 Apr 19.
The role of vitamin E in both enzymatic and free radical-dependent metabolism of polyunsaturated fatty acids (PUFAs) has been well demonstrated. This study proposed a new LC-MS/MS method to quantify the main vitamin E forms, their metabolites and main PUFA species in human blood, since, at present, there are not procedures able to simultaneously determine these two classes of compounds. After the optimization of sample treatment and reverse-phase separation conditions, tandem mass spectrometry detection was evaluated experimenting both positive and negative electrospray ionisation modes. The procedure was also preliminarily adapted to assess five arachidonic acid-derived eicosanoids that could be under the influence of vitamin E function, such as LTB4 (leukotriene B4), 20-HETE (20-hydroxyeicosatetraenoic acid) and their ω-oxidation metabolites. After the validation study, the performance characteristics were confirmed analysing a certified reference material (SRM 1950 - frozen human plasma by NIST). Finally, an application of the method in the analysis of lipid abnormalities of chronic kidney disease patients was shown.
维生素 E 在多不饱和脂肪酸（PUFAs）的酶促和自由基依赖代谢中的作用已得到充分证实。本研究提出了一种新的 LC-MS/MS 方法，用于定量测定人血液中的主要维生素 E 形式、其代谢物和主要 PUFAs 种类，因为目前尚无能够同时测定这两类化合物的程序。在优化样品处理和反相分离条件后，通过实验比较正、负离子喷雾电离模式评估了串联质谱检测。该程序还初步适用于评估五种可能受维生素 E 功能影响的花生四烯酸衍生的类二十烷酸，如 LTB4（白三烯 B4）、20-HETE（20-羟二十碳四烯酸）及其 ω-氧化代谢物。验证研究后，通过分析 NIST 的认证参考物质（SRM 1950-冷冻人血浆）确认了性能特征。最后，展示了该方法在分析慢性肾脏病患者脂质异常中的应用。
