色谱

色谱 ›› 2026, Vol. 44 ›› Issue (3): 234-247.DOI: 10.3724/SP.J.1123.2025.08011

• 研究论文 • 上一篇    下一篇

人血清中30种磷脂酰胆碱与溶血磷脂酰胆碱同系物的液相色谱-串联质谱测定及其与冠心病的关联分析

李汶煜, 刘朝阳, 董军, 杨睿悦, 李红霞, 陈文祥, 王思明()   

  1. 北京医院实验研究部 国家老年医学中心，国家卫生健康委老年医学重点实验室，中国医学科学院老年医学研究院，北京 100730
  • 收稿日期:2025-08-15 出版日期:2026-03-08 发布日期:2026-03-12
  • 通讯作者: *Tel：010-85138520，E-mail：wangsiming4446@bjhmoh.cn.
  • 基金资助:
    中央高水平医院临床科研业务费(BJ-2022-125);国家重点研发计划(2021YFE0114200);国家自然科学基金(81501842);北京自然科学基金(7182145)

Determination of 30 homologues of phosphatidylcholines and lysophosphatidylcholines in human serum by liquid chromatography-tandem mass spectrometry and their correlation analysis with coronary artery disease

LI Wenyu, LIU Zhaoyang, DONG Jun, YANG Ruiyue, LI Hongxia, CHEN Wenxiang, WANG Siming()   

  1. Department of Experimental Research，Beijing Hospital，National Center of Gerontology，Key Laboratory of Geriatrics of National Health Commission，Institute of Geriatric Medicine，Chinese Academy of Medical Sciences，Beijing 100730，China
  • Received:2025-08-15 Online:2026-03-08 Published:2026-03-12
  • Supported by:
    National High Level Hospital Clinical Research Funding(BJ-2022-125);National Key R&D Program of China(2021YFE0114200);National Natural Science Foundation of China(81501842);Beijing Natural Science Foundation(7182145)

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摘要：

磷脂酰胆碱和溶血磷脂酰胆碱同系物与冠状动脉粥样硬化密切相关，准确测定其含量可以为临床上冠心病的诊断和预后提供重要依据。本研究建立了一种基于液相色谱-串联质谱的分析方法，仅需10 μL人血清即可同时实现30种磷脂酰胆碱和溶血磷脂酰胆碱同系物的准确测定。采用甲醇-乙腈-甲基叔丁基醚-水作为萃取体系，选用XBridge C18色谱柱，以乙腈-水（1∶1，体积比）混合溶液和异丙醇溶液作为流动相，两相均含有7.5 mmol/L甲酸铵与0.15%（体积分数）甲酸，进行梯度洗脱分离，采用电喷雾离子源，正离子多反应离子检测模式。经验证，该方法线性关系良好，平均线性相关系数≥0.999 7，线性范围为0.125~100 μg/mL，检出限和定量限分别为0.01~1.94 μg/mL和0.03~6.48 μg/mL，回收率为85.4%~114.3%，日内精密度和日间精密度分别不大于4.6%和12.6%。使用本方法测定了临床上110名接受冠状动脉造影术志愿者的血清样本，磷脂酰胆碱同系物平均人群质量浓度为526.80 μg/mL，溶血磷脂酰胆碱同系物平均人群质量浓度为73.67 μg/mL。Spearman相关性检验分析发现，磷脂酰胆碱和溶血磷脂酰胆碱同系物与冠心病严重程度以及相关临床生化、血脂代谢指标存在密切关联，提示其可作为临床冠心病的潜在相关代谢物。本研究针对临床分析需求设计，具有血清用量少、操作简便、响应优良等特点，可高效实现人血清中30种磷脂酰胆碱和溶血磷脂酰胆碱同系物的测定，为探究二者与冠心病的关联及生物标志物转化应用提供了重要参考。

关键词: 磷脂酰胆碱, 溶血磷脂酰胆碱, 动脉粥样硬化, 冠心病, 液相色谱-串联质谱, 脂质组学

Abstract:

Phosphatidylcholine （PC） and lysophosphatidylcholine （LPC） homologues are closely associated with coronary atherosclerosis. Accurate determination of their contents can provide an important basis for the clinical diagnosis and prognosis of coronary artery disease （CAD）. In this study， an analytical method based on liquid chromatography-tandem mass spectrometry was established， which enabled the simultaneous and accurate determination of 30 PC and LPC homologues using only 10 μL of human serum. Methanol-acetonitrile-methyl tert-butyl methyl ether-water was used as the extraction system， and an XBridge C18 column was selected as the stationary phase. The mobile phase consisted of an acetonitrile-water mixture （1∶1， volume ratio） and isopropanol， both containing 7.5 mmol/L ammonium formate and 0.15% （volume ratio） formic acid， and gradient elution was adopted for separation. Detection was performed using an electrospray ionization source in the positive ion mode with multiple reaction monitoring. Method validation results showed that the method exhibited a good linear relationship， with an average linear correlation coefficient of ≥0.999 7 over a linear range of 0.125-100 μg/mL. The limits of detection and limits of quantification were 0.01-1.94 μg/mL and 0.03-6.48 μg/mL， respectively. The recoveries ranged from 85.4% to 114.3%， while the intra-day precision and inter-day precision were no more than 4.6% and 12.6%， respectively. Serum samples from 110 clinical volunteers who underwent coronary angiography were determined using this method. The average population concentration of PC homologues was 526.80 μg/mL， and that of LPC homologues was 73.67 μg/mL. Spearman correlation analysis revealed that PC and LPC homologues were closely correlated with the severity of CAD， as well as with related clinical biochemical and lipid metabolism indicators， suggesting that they could serve as potential CAD-related metabolites in clinical practice. Designed to meet clinical analysis needs， this method features small serum sample volume， simple operation， and excellent response. It can efficiently determine 30 PC and LPC homologues in human serum， providing an important reference for exploring the association between these two lipid classes and CAD， as well as the translational application of related biomarkers.

Key words: phosphatidylcholine, lysophosphatidylcholine, atherosclerosis, coronary artery disease, liquid chromatography-tandem mass spectrometry （LC-MS/MS）, lipidomics

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