Abstract Several amino acids, nucleotide metabolites, organic acids, and choline-related metabolites have been identified as possible risk factors for cardiovascular disease (CVD). However, a method that allows rapid processing of a large number of samples while simultaneously quantifying metabolite concentrations is currently unavailable. Here, we developed a simple and reliable method for quantifying metabolomic risk factors of CVD using liquid chromatography–tandem mass spectrometry (LC–MS/MS), and assessed its efficacy using serum samples from patients with angiographically defined coronary artery disease (CAD). Serum samples were mixed with isotopically labeled internal standards, extracted with methanol, and separated by hydrophilic interaction LC. MS detection was simultaneously performed in both positive and negative ionization modes coupled with real-time polarity switching, resulting in markedly improved monitoring capacity and reduced overall run time. In total, 20 metabolites were analyzed in 4 min. The mean linear correlation coefficients were higher than 0.999, with average analytical recoveries of 101.9% (range: 90.1–110.9%). The intra-run and total coefficients of variation (CV) were 1.02-6.11% and 1.25-9.41%, respectively. Several metabolites were significantly associated with traditional CVD risk factors. Multivariable logistic regression analysis revealed that tryptophan and hypoxanthine were negatively correlated with CAD, while trimethylamine-N-oxide was positively correlated with CAD. The method developed in this study is simple, precise, accurate, and provides a potentially convenient tool for CVD risk assessment and future research.

中文翻译：

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使用液相色谱-串联质谱法同时定量心血管疾病相关代谢物的单次运行、快速极性切换方法

摘要 几种氨基酸、核苷酸代谢物、有机酸和胆碱相关代谢物已被确定为心血管疾病 (CVD) 的可能危险因素。然而，目前还没有一种方法可以快速处理大量样品，同时量化代谢物浓度。在这里，我们开发了一种使用液相色谱-串联质谱 (LC-MS/MS) 量化 CVD 代谢组学风险因素的简单而可靠的方法，并使用来自血管造影确定的冠状动脉疾病 (CAD) 患者的血清样本评估其疗效。血清样品与同位素标记的内标混合，用甲醇萃取，并通过亲水相互作用 LC 进行分离。MS 检测在正负电离模式下同时进行，并结合实时极性切换，从而显着提高了监测能力并缩短了总运行时间。总共在 4 分钟内分析了 20 种代谢物。平均线性相关系数高于 0.999，平均分析回收率为 101.9%（范围：90.1–110.9%）。运行内和总变异系数 (CV) 分别为 1.02-6.11% 和 1.25-9.41%。几种代谢物与传统的 CVD 风险因素显着相关。多变量logistic回归分析显示色氨酸和次黄嘌呤与CAD呈负相关，而三甲胺-N-氧化物与CAD呈正相关。本研究开发的方法简单、精确、准确、