Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Busan, Republic of Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Dec 12;911:113-21. doi: 10.1016/j.jchromb.2012.11.004. Epub 2012 Nov 10.
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 0.5mM 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 400pg/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.
二十烷类在各种生物反应中发挥着重要作用，并且可以作为特定疾病的生物标志物。因此，我们开发了一种高选择性、高灵敏度和高稳定性的液相色谱-串联质谱法，用于测量人血浆中的花生四烯酸及其 32 种代谢物。样品制备包括固相萃取，该方法可有效去除人血浆中存在的干扰源。采用 Luna C(8)-柱，以 0.5mM 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。采用电喷雾电离接口的串联质谱法在负离子模式下进行检测。基质不影响测定的重现性和可靠性。所有分析物在研究浓度范围内均表现出良好的线性（r>0.997）。验证的分析物定量下限范围为 10-400pg/mL。所有二十烷类物质在所有浓度范围内的日内和日间精密度（RDS%）均在 16.8%以内，准确度在 88.1%至 108.2%之间。该方法适用于测定基础血浆中二十烷类物质的水平，以及评估阿司匹林对健康受试者中二十烷类物质血浆水平的影响。
