基于UHPLC/Q-TOF-MS技术的人血浆磷脂类成分快速分析及质谱裂解规律初探  

Rapid analysis and determination of the fragmentation regularity of phospholipids in human plasma based on UHPLC/Q-TOF-MS

作　　者：钟询龙[1] 王若伦[1] 陈莉诗 钟艳梅[2] ZHONG Xun-long;WANG Ruo-lun;CHEN Li-shi;ZHONG Yan-mei(Department of Pharmacy,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China;New Drug Research and Development Center of Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广州医科大学附属第二医院药学部,广东广州510260 [2]广东药科大学新药研发中心,广东广州510006

出　　处：《药学学报》2022年第10期3214-3222,共9页Acta Pharmaceutica Sinica

基　　金：广东省中医药局面上项目(20221242,20212118);广州市中医药和中西医结合科技项目(20222A011017);广州市卫生健康科技一般引导项目(20191A011085)。

摘　　要：采用超高效液相色谱-四级杆飞行时间质谱联用技术(UHPLC/Q-TOF-MS)对人血浆磷脂类成分进行快速分析并推测其质谱裂解规律。本文采用Acquity UHPLC^(TM) BEH C18色谱柱(50 mm×2.1 mm,1.7μm),以含10 mmol·L^(-1)甲酸铵和0.1%甲酸的水溶液(A)与含10 mmol·L^(-1)甲酸铵和0.1%甲酸的乙腈-异丙醇(体积比为1∶1)有机溶液(B)作为流动相进行梯度洗脱,流速0.3 mL·min^(-1),柱温50℃,ESI源正、负离子模式下采集质谱数据。根据Masslynx 4.1软件计算的精确相对分子质量和元素组成,并通过与对照品、二级质谱碎片离子以及脂质数据库的比对,共鉴定出82个血浆脂质成分,其中包括14个溶血磷脂酰胆碱(LysoPCs)、39个磷脂酰胆碱(PCs)、17个鞘磷脂(SMs)、7个神经酰胺(Cers)、4个磷脂酰乙醇胺(PEs)、1个磷脂酰肌醇(PI)。本实验建立了一种简便高效、快捷、稳定的分析方法,可对人血浆磷脂类成分进行较为全面地定性分析,同时总结了主要磷脂类成分的质谱裂解规律,为进一步以血浆磷脂作为研究对象的代谢组学研究奠定基础。本实验获得广州医科大学附属第二医院临床研究与应用伦理委员会的审查和批准(批准号:2020-hs-07)。Ultra-high performance liquid chromatography-tandem quadrupole time-of-flight mass spectrometry(UHPLC/Q-TOF-MS)was applied to rapidly identify the phospholipids in human plasma and explore the mass spectrometric fragmentation pattern.An acquity UHPLC^(TM) BEH C18 column(50 mm×2.1 mm,1.7μm)was utilized and eluted with a gradient system;the mobile phase consisted of 10 mmol·L^(-1) ammonium formate aqueous solution-0.1% formic acid aqueous solution(A)and acetonitrile-isopropanol(1∶1)organic solution(B)containing 10 mmol·L^(-1) ammonium formate-0.1% formic acid.The flow rate was 0.3 mL·min^(-1) and the column temperature was set at 50℃.An electrospray ionization(ESI)source was used to collect mass spectra in positive and negative ion mode.Based on the precise relative molecular weight and elemental composition calculated by Masslynx 4.1 software,comparison with references,and secondary mass spectrometry fragment ions and lipid databases,a total of 82 plasma lipids were identified,including 14 lysophosphatidylcholines(LysoPCs),39 phosphatidylcholines(PCs),17 sphingomyelins(SMs),7 ceramides(Cers),4 phosphatidylethanolamines(PEs),and 1 phosphatidylinositol(PI).A simple,efficient,fast and stable analytical method was established in this study for the qualitative analysis of phospholipids in human plasma,and the fragmentation regularity of the main phospholipids was determined.This work provides a good foundation for further metabolomics studies of plasma phospholipids.This study was approved by the Second Affiliated Hospital of Guangzhou Medical University Clinical Research and Application Institutional Review Board Approval(No.2020-hs-07).

关 键 词：血浆磷脂 UHPLC/Q-TOF-MS 磷脂酰胆碱 鞘磷脂 神经酰胺 裂解规律 

分 类 号：R917[医药卫生—药物分析学]