Comprehensive ultra-performance liquid chromatographic separation and mass spectrometric analysis of eicosanoid metabolites in human samples.
Comprehensive ultra-performance liquid chromatographic separation and mass spectrometric analysis of eicosanoid metabolites in human samples.
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DOI：
10.1016/j.chroma.2014.07.006
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发表时间：
2014-09-12
期刊：
影响因子：
--
通讯作者：
Dennis EA
中科院分区：
文献类型：
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作者：
Wang Y;Armando AM;Quehenberger O;Yan C;Dennis EA
在过去的十年中，已知的类二十烷酸的数量大幅增加，我们现在开发了一种超高效液相色谱 - 电喷雾电离三重四极杆质谱 (UPLC-QTRAP/MS/MS) 方法来监测和定量多种类二十烷酸。 UPLC-QTRAP/MS/MS 方法利用预定的多反应监测 (MRM) 来优化灵敏度、可分析的代谢物数量以及分析的时间要求。在一次 5 分钟的 UPLC 运行中，可以分离和监测总共 184 种类二十烷酸，包括 26 种氘代内标。为了演示实际应用，在固相萃取 (SPE) 后对人血浆样品进行了分析，并确定了添加到血浆中的 26 种氘代内标的回收率和基质效应。该方法经过验证，结果显示，对于大多数化合物而言，其定量限在 pg 水平下非常灵敏，回收率高达 70-120%，并且所有化合物的 CV <30 都很精确。此外，该方法在跨越几个数量级的范围内显示出线性响应。在 QC 人血浆样本中，我们鉴定并严格定量了 120 多种类二十烷酸。
Over the past decade, the number of known eicosanoids has expanded immensely and we have now developed an ultra-performance liquid chromatography - electrospray ionization triple quadrupole mass spectrometric (UPLC-QTRAP/MS/MS) method to monitor and quantify numerous eicosanoids. The UPLC-QTRAP/MS/MS approach utilizes scheduled multiple reaction monitoring (MRM) to optimize sensitivity, number of metabolites that can be analyzed and the time requirement of the analysis. A total of 184 eicosanoids including 26 deuterated internal standards can be separated and monitored in a single 5 min UPLC run. To demonstrate a practical application, human plasma samples were analyzed following solid-phase extraction (SPE) and the recovery rate and matrix effects were determined for the 26 deuterated internal standards added to the plasma. The method was validated and shown to be sensitive with the limit of quantitation at pg levels for most compounds, accurate with recovery rates of 70-120%, and precise with a CV<30 for all compounds. Also, the method showed a linear response over a range spanning several orders of magnitude. In a QC human plasma sample, we identified and rigorously quantified over 120 eicosanoids.