Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, Liebigstr. 27, 04103 Leipzig, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 May 15;927:209-13. doi: 10.1016/j.jchromb.2013.03.012. Epub 2013 Mar 21.
Profiling of polyunsaturated fatty acids (PUFAs) and their oxidized metabolites, mainly eicosanoids, in human plasma by fast liquid chromatography-mass spectrometry is described. Sample preparation involved protein precipitation of 200μL plasma followed by on-line solid-phase extraction. 7 PUFAs and 94 oxidized metabolites were separated utilizing a C-18 column packed with 2.6μm core-shell particles in 7min. The analytes and deuterium-labeled standards were detected via scheduled multiple reaction monitoring transitions (123 sMRM). Simultaneously, linear ion trap fragment spectra were acquired for confirmation, if necessary. The lower limit of quantitation ranged between 200 and 1000ng/mL for the PUFAs and 10-1000pg/mL for the metabolites. The method was applied to a study on plasma samples from 50 healthy subjects.
本文描述了一种快速液相色谱-质谱法对人血浆中多不饱和脂肪酸(PUFAs)及其氧化代谢产物(主要为类二十烷酸)进行分析的方法。样品制备包括 200μL 血浆的蛋白沉淀,然后进行在线固相萃取。利用填充有 2.6μm 核壳颗粒的 C-18 柱,在 7min 内分离了 7 种 PUFAs 和 94 种氧化代谢产物。通过预定的多重反应监测转换(123 sMRM)检测分析物和氘标记标准品。如有必要,同时获取线性离子阱碎片谱进行确认。PUFAs 的定量下限范围为 200-1000ng/mL,代谢产物的定量下限范围为 10-1000pg/mL。该方法应用于 50 名健康受试者的血浆样品研究。