Development and application of an LC-MS/MS method for the combined quantification of oxysterols and bile acids.

IF 5 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Journal of Lipid Research Pub Date : 2024-11-16 DOI:10.1016/j.jlr.2024.100697
Martin Roumain, Giulio G Muccioli
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Abstract

Oxysterols and bile acids are interconnected bioactive lipids playing pivotal roles in diverse physiological and pathological processes. For this reason, they are increasingly studied together for their implications in various diseases. However, due to analytical challenges inherent to the nature of these analytes, very few methods have been developed for the simultaneous analysis of these lipids. We here report the development of a sensitive LC-MS/MS method for the combined quantification of 18 oxysterols, 11 unconjugated, 15 conjugated bile acids, and 1 bile acid precursor, using 8 isotope-labeled internal standards, addressing the need for a more comprehensive analysis of these interesting lipid families. During the method development, we investigated different extraction protocols, set up a purification step and achieved chromatographic separation for these lipids, overcoming challenges such as the large number of analytes, isomers, and wide range of polarity across the analytes. Finally, the method was successfully applied to the analysis of preclinical and clinical samples, quantifying 12 oxysterols and 14 bile acids in human plasma, 10 oxysterols and 18 bile acids in mouse plasma from the vena cava, and 10 oxysterols and 20 bile acids in mouse plasma from the portal vein within a single chromatographic run.

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开发和应用一种 LC-MS/MS 方法,用于氧甾醇和胆汁酸的联合定量。
羟基固醇和胆汁酸是相互关联的生物活性脂质,在各种生理和病理过程中发挥着关键作用。因此,人们越来越多地同时研究它们对各种疾病的影响。然而,由于这些分析物的性质所固有的分析挑战,很少有方法能同时分析这些脂质。我们在此报告了一种灵敏的 LC-MS/MS 方法的开发情况,该方法使用 8 种同位素标记的内标物对 18 种氧基甾醇、11 种非结合胆汁酸、15 种结合胆汁酸和 1 种胆汁酸前体进行了联合定量分析,从而满足了对这些有趣的脂质家族进行更全面分析的需求。在方法开发过程中,我们研究了不同的提取方案,建立了纯化步骤,并实现了这些脂质的色谱分离,克服了分析物数量多、异构体多、分析物极性范围广等难题。最后,我们成功地将该方法应用于临床前和临床样本的分析,在一次色谱运行中定量分析了人血浆中的 12 种氧固醇和 14 种胆汁酸、小鼠腔静脉血浆中的 10 种氧固醇和 18 种胆汁酸以及小鼠门静脉血浆中的 10 种氧固醇和 20 种胆汁酸。
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来源期刊
Journal of Lipid Research
Journal of Lipid Research 生物-生化与分子生物学
CiteScore
11.10
自引率
4.60%
发文量
146
审稿时长
41 days
期刊介绍: The Journal of Lipid Research (JLR) publishes original articles and reviews in the broadly defined area of biological lipids. We encourage the submission of manuscripts relating to lipids, including those addressing problems in biochemistry, molecular biology, structural biology, cell biology, genetics, molecular medicine, clinical medicine and metabolism. Major criteria for acceptance of articles are new insights into mechanisms of lipid function and metabolism and/or genes regulating lipid metabolism along with sound primary experimental data. Interpretation of the data is the authors’ responsibility, and speculation should be labeled as such. Manuscripts that provide new ways of purifying, identifying and quantifying lipids are invited for the Methods section of the Journal. JLR encourages contributions from investigators in all countries, but articles must be submitted in clear and concise English.
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