Numerous studies have revealed a close correlation between the levels of apolipoproteins (Apos) (including lipoprotein(a) [Lp(a)]) and an increased risk of cardiovascular disease in recent decades. However, clinically, lipid profiling remains limited to the conventional plasma levels of cholesterol, triglyceride, ApoA1, and ApoB, which brings the necessity to quantify more apolipoproteins in human plasma. In this study, we simultaneously quantified 13 apolipoproteins and Lp(a) in 5 μL of human plasma using the LC-MS/MS platform. A method was developed for the precise detection of Lp(a), ApoA1, A2, A5, B, C1, C2, C3, D, E, H, L1, M, and J. Suitable peptides were selected and optimized to achieve clear separation of each peak. Method validation consisting of linearity, sensitivity, accuracy and precision, recovery, and matrix effects was evaluated. The intra-day CV ranged from 0.58% to 14.2% and the inter-day CV ranged from 0.51% to 13.3%. The recovery rates ranged from 89.8% to 113.7%, while matrix effects ranged from 85.4% to 113.9% for all apolipoproteins and Lp(a). Stability tests demonstrated that these apolipoproteins remained stable for 3 days at 4 °C and 7 days at −20 °C. This validated method was successfully applied to human plasma samples obtained from 45 volunteers. The quantitative results of ApoA1, ApoB, and Lp(a) exhibited a close correlation with the results from the immunity transmission turbidity assay. Collectively, we developed a robust assay that can be used for high-throughput quantification of apolipoproteins and Lp(a) simultaneously for investigating related risk factors in patients with dyslipidemia.

中文翻译：

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对人血浆中 13 种载脂蛋白和脂蛋白 (a) 进行同步定量 LC-MS/MS 分析

近几十年来，大量研究表明载脂蛋白 (Apos)（包括脂蛋白 (a) [Lp(a)]）水平与心血管疾病风险增加之间存在密切相关。然而，临床上，血脂分析仍然仅限于胆固醇、甘油三酯、ApoA1 和 ApoB 的常规血浆水平，这使得有必要对人血浆中更多的载脂蛋白进行定量。在本研究中，我们使用 LC-MS/MS 平台同时定量了 5 μL 人血浆中的 13 种载脂蛋白和 Lp(a)。开发了Lp(a)、ApoA1、A2、A5、B、C1、C2、C3、D、E、H、L1、M和J的精确检测方法。选择合适的肽并进行优化，以实现清晰的检测结果。每个峰的分离。评估了方法验证，包括线性、灵敏度、准确度和精密度、回收率以及基质效应。日内CV范围为0.58%至14.2%，日间CV范围为0.51%至13.3%。所有载脂蛋白和 Lp(a) 的回收率范围为 89.8% 至 113.7%，而基质效应范围为 85.4% 至 113.9%。稳定性测试表明，这些载脂蛋白在 4 °C 下保持稳定 3 天，在 -20 °C 下保持稳定 7 天。这种经过验证的方法已成功应用于从 45 名志愿者获得的人类血浆样本。 ApoA1、ApoB、Lp(a)的定量结果与免疫透射比浊法结果密切相关。总的来说，我们开发了一种强大的检测方法，可同时对载脂蛋白和 Lp(a) 进行高通量定量，以调查血脂异常患者的相关危险因素。