Comparison of sample preparation methods for the quantitative analysis of eicosanoids and other oxylipins in plasma by means of LC-MS/MS
Comparison of sample preparation methods for the quantitative analysis of eicosanoids and other oxylipins in plasma by means of LC-MS/MS
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DOI：
10.1007/s00216-014-8377-4
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发表时间：
2015-02-01
影响因子：
4.3
通讯作者：
Schebb, Nils Helge
中科院分区：
文献类型：
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作者：
Ostermann, Annika I.;Willenberg, Ina;Schebb, Nils Helge
氧脂素是有效的脂质介质。为了评价其生物学作用，已经开发了几种基于LC-MS的方法。虽然这些方法相似，但所描述的用于提取氧脂素的样品制备程序差异很大。为了推导出分析人血浆中非酯化氧脂素的最合适方法，我们评价了7种已建立的样品制备方法的性能。比较了6种常用的固相萃取（SPE）和1种液-液萃取（LLE）方法对血浆中13种内标物的回收率、氧脂素的萃取效果和离子抑制基质的还原情况。在所有三个参数的性能显着的差异被发现。液液萃取与乙酸乙酯是整体上不是一个足够的样品制备策略。使用Oasis和StrataX材料的方案不能充分去除干扰基质化合物。氧脂素在阴离子交换BondElut柱上的提取效率较低，而基质的去除几乎完美。在去除所有干扰基质组分的同时，没有一种方案导致分析物的高提取效率。然而，SPE上的C18-材料与去除基质的水和正己烷之前用甲酸甲酯洗脱显示出最好的性能，用于分析血浆中的氧化脂质的广谱。
Oxylipins are potent lipid mediators. For the evaluation of their biological roles, several LC-MS based methods have been developed. While these methods are similar, the described sample preparation procedures for the extraction of oxylipins differ considerably. In order to deduce the most appropriate method for the analysis of non-esterified oxylipins in human plasma, we evaluated the performance of seven established sample preparation procedures. Six commonly used solid phase extraction (SPE) and one liquid-liquid extraction (LLE) protocol were compared based on the recovery of 13 added internal standards, extraction efficacy of oxylipins from plasma and reduction of ion-suppressing matrix. Dramatic differences in the performance in all three parameters were found. LLE with ethyl acetate was overall not a sufficient sample preparation strategy. The protocols using Oasis-and StrataX-material insufficiently removed interfering matrix compounds. Extraction efficacy of oxylipins on anion-exchanging BondElut cartridges was low, while removal of matrix was nearly perfect. None of the protocols led to a high extraction efficacy of analytes while removing all interfering matrix components. However, SPE on a C18-material with removal of matrix by water and n-hexane prior elution with methyl formate showed the best performance for the analysis of a broad spectrum of oxylipins in plasma.