LC-MS/MS for the simultaneous analysis of arachidonic acid and 32 related metabolites in human plasma: Basal plasma concentrations and aspirin-induced changes of eicosanoids
LC-MS/MS for the simultaneous analysis of arachidonic acid and 32 related metabolites in human plasma: Basal plasma concentrations and aspirin-induced changes of eicosanoids
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DOI：
10.1016/j.jchromb.2012.11.004
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发表时间：
2012-12-12
影响因子：
3
通讯作者：
Shin, Jae Gook
中科院分区：
文献类型：
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作者：
Shinde, Dhananjay D.;Kim, Kwon-Bok;Shin, Jae Gook
类二十烷酸在各种生物反应中发挥着重要作用，可用作特定疾病的生物标志物。因此，我们开发了一种高选择性、灵敏且稳定的液相色谱-串联质谱方法来测量人血浆中的花生四烯酸及其 32 种代谢物。样品制备涉及固相萃取，可有效去除人血浆中存在的干扰源。使用 Luna C-8 柱，以 0.5 mM 甲酸铵缓冲液和乙腈作为流动相，在梯度条件下进行色谱分离。使用配备负离子模式电喷雾电离接口的串联质谱法进行检测。基质不影响测定的再现性和可靠性。所有分析物在研究的浓度范围内均表现出良好的线性 (r > 0.997)。经验证的分析物定量下限范围为 10 至 400 pg/mL。所有类二十烷酸浓度范围内的日内和日间精密度 (RDS%) 均在 16.8% 以内，准确度范围在 88.1% 至 108.2% 之间。该测定适用于健康受试者基础血浆类二十烷酸水平的测定以及阿司匹林对类二十烷酸血浆水平的影响评估。 (C) 2012 Elsevier B.V. 保留所有权利。
Eicosanoids play an important role in various biological responses and can be used as biomarkers for specific diseases. Therefore, we developed a highly selective, sensitive, and robust liquid chromatography-tandem mass spectrometric method to measure arachidonic acid and its 32 metabolites in human plasma. Sample preparation involved solid phase extraction, which efficiently removed sources of interference present in human plasma. Chromatographic separation was performed using a Luna C-8-column with 0.5 mM ammonium formate buffer and acetonitrile as the mobile phase under gradient conditions. Detection was performed using tandem mass spectrometry equipped with an electrospray ionization interface in negative ion mode. The matrix did not affect the reproducibility and reliability of the assay. All analytes showed good linearity over the investigated concentration range (r > 0.997). The validated lower limit of quantitation for the analytes ranged from 10 to 400 pg/mL. Intra- and inter-day precision (RDS%) over the concentration ranges for all eicosanoids were within 16.8%, and accuracy ranged between 88.1 and 108.2%. This assay was suitable for the determination of basal plasma levels of eicosanoids and the evaluation of effect of aspirin on eicosanoid plasma levels in healthy subjects. (C) 2012 Elsevier B.V. All rights reserved.