柱前衍生超高效液相色谱法测量脂蛋白相关磷脂酶A2含量  

Determination of themass fraction of Lp-PLA2 by pre-column derivatization UPLC

作　　者：崔宏恩 冯速 叶人元 刘阳 方帅[1] 朱茂林 樊晓琳 CUI Hong en;FENG Su;YE Renyuan;LIU Yang;FANG Shuai;ZHU Maolin;FAN Xiaolin(Jiangsu Institute of Metrology,Nanjing 210023,China;Nanjing Vazyme Biotech Co.,Ltd.,Nanjing 210000,China)

机构地区：[1]江苏省计量科学研究院,江苏南京210023 [2]南京诺唯赞生物科技股份有限公司,江苏南京210000

出　　处：《中国计量大学学报》2023年第3期325-331,共7页Journal of China University of Metrology

基　　金：江苏省市场监督管理局科技计划项目(No.KJ185602)。

摘　　要：目的:脂蛋白相关磷脂酶A2(Lp-PLA2)在血液中的含量能够反映动脉粥样硬化及其它心脑血管疾病的进展,故建立Lp-PLA2柱前衍生超高效液相色谱(UPLC)测定方法。方法:Lp-PLA2酸水解后,经柱前衍生剂处理,以γ-氨基丁酸为内标,采用超高效液相色谱法测定其中丝氨酸、丙氨酸、异亮氨酸、赖氨酸4种衍生物的含量,进而推算出Lp-PLA2纯品的含量。测定结果与同位素稀释质谱法相比较,验证方法的可行性。结果:在最优条件下,所建立的柱前衍生超高效液相色谱(UPLC)法测定的Lp-PLA2含量为0.1183 mg/mL,相对标准偏差为2.48%,扩展不确定度为0.0026 mg/mL(k=2),检出限和定量限分别为1.96×10^(-2)mg/mL和6.55×10^(-2)mg/mL。结论:该方法成本较低且溯源链清晰、准确度高,可作为一种补充验证方法用于Lp-PLA2标准物质的定量。Aims:The content of lipoprotein associated phospholipase A2(Lp-PLA2)in blood reflects the progress of atherosclerosis and other cardiovascular and cerebrovascular diseases.A method for the determination of Lp-PLA2 by precolumn derivatization ultra performance liquid chromatography(UPLC)was established.Methods:After Lp-PLA2 acid hydrolysis and pre-column derivatization,the content of four derivatives,i.e.,serine,alanine,isoleucine,and lysine was determined by ultra-high performance liquid chromatography withγ-aminobutyric acid as the internal standard;and the content of Lp-PLA2 was calculated.The feasibility of the proposed method was verified as compared with the result of the isotope dilution mass spectrometry.Results:The Lp-PLA2 content determined by the established pre-column derivatization UPLC method was 0.1183 mg/mL,with a relative standard deviation(RSD)of 2.48%,an expanded uncertainty of 0.0026 mg/mL(k=2),and the detection and quantitation limits of 1.96×10^(-2)mg/mL and 6.55×10^(-2)mg/mL,respectively.Conclusions:This method has low cost,clear traceability chain,and high accuracy,and can be used as a supplementary validation method for the calibration of Lp-PLA2 reference materials.

关 键 词：计量学 脂蛋白相关磷脂酶A2 含量 柱前衍生超高效液相色谱法 

分 类 号：TQ464.7[化学工程—制药化工]