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Development and validation of a rapid, specific and sensitive LC-MS/MS bioanalytical method for eicosanoid quantification - assessment of arachidonic acid metabolic pathway activity in hypertensive rats
Biochimie ( IF 3.9 ) Pub Date : 2020-03-13 , DOI: 10.1016/j.biochi.2020.03.010
Agnieszka Kij , Kamil Kus , Izabela Czyzynska-Cichon , Stefan Chlopicki , Maria Walczak

Lipid mediators such as eicosanoids maintain various physiological processes, and their alterations are involved in the development of numerous cardiovascular diseases. Therefore, the reliable assessment of their profile could be helpful in diagnosis as well as in eicosanoid biomarker-based treatment. Hence, the presented study aimed to develop and validate a new rapid, specific and sensitive LC-MS/MS method for quantification of arachidonic acid-derived eicosanoids in plasma, including lipid mediators generated via COX-, LOX- and CYP450-dependent pathways. The developed method features high sensitivity because the lower limit of quantification ranged from 0.05–0.5 ng mL-1 as well as an accuracy and precision estimated within 88.88–111.25% and 1.03–11.82%, respectively. An application of a simple and fast liquid-liquid extraction procedure for sample cleaning resulted in a highly satisfactory recovery of the analytes (>88.30%). Additionally, the method was validated using artificial plasma, an approach that enabled elimination of the matrix effect caused by endogenous concentration of studied lipid mediators. Importantly, the presented LC-MS/MS method allowed for simultaneous quantitative and qualitative [quan/qual] analysis of the selected eicosanoids, leading to an additional improvement of the method specificity. Moreover, the validated method was successfully applied for eicosanoid profiling in rat, mouse and human plasma samples, clearly demonstrating the heterogeneity of studied lipid mediators between those species.



中文翻译:

快速,特异和灵敏的LC-MS / MS生物分析方法用于类花生酸定量的开发和验证-高血压大鼠中花生四烯酸代谢途径活性的评估

脂质介质例如类二十烷酸维持各种生理过程,其改变参与多种心血管疾病的发展。因此,对其轮廓的可靠评估可能有助于诊断以及基于类花生酸生物标志物的治疗。因此,本研究旨在开发和验证一种新的快速,特异性和灵敏的LC-MS / MS方法,用于定量测定血浆中花生四烯酸衍生的类花生酸,包括通过COX,LOX和CYP450依赖性途径产生的脂质介体。该方法具有较高的灵敏度,因为其定量下限范围为0.05–0.5 ng mL -1以及准确度和精密度分别在88.88–111.25%和1.03–11.82%之间。采用简单,快速的液-液萃取程序进行样品清洁,可得到令人满意的分析物回收率(> 88.30%)。此外,该方法已使用人工血浆进行了验证,该方法能够消除因研究脂质内源性内源性浓度引起的基质效应。重要的是,提出的LC-MS / MS方法允许同时对所选类花生酸类物质进行定量和定性[quan / qual]分析,从而进一步提高了方法的特异性。此外,已验证的方法已成功应用于大鼠,小鼠和人类血浆样品中类花生酸的分析,

更新日期:2020-03-16
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