小鼠血浆及尿液中多种氨基酸的UPLC-MS检测方法的建立

A Variety of Amino Acid Detection Methods Based on UPLC-MS Technology and Its Application in Mouse Plasma and Urine

目的:采用超高液相色谱-串联质谱(ultra-performance liquid chromatography-tandem mass spectrometry,UPLC-MS)建立16种氨基酸的定量方法,并对小鼠血浆以及尿液样本实现准确定量分析。方法:使用UPLC-MS技术,液相以20 mmol·L^(-1)甲酸铵水溶液(A)以及乙腈(含15%20 mmol·L^(-1)甲酸铵水溶液)(B)为流动相进行梯度洗脱,质谱以多反应监测模式(multiple reaction monitoring,MRM)进行检测,以专属性、检测限、定量限、线性、准确度、精密度、残留效应以及稳定性等方法学指标评价此氨基酸定量方法,并应用于小鼠血浆以及尿液的氨基酸定量。结果:通过对氨基酸标准品的方法学验证,此检测方法符合方法学验证的相关标准。对小鼠的血浆以及尿液中氨基酸的准确定量结果表明,此方法可同时检测小鼠血浆以及尿液中的16种氨基酸并进行准确定量。结论:应用UPLC-MS技术,建立了能对小鼠血浆以及尿液中16种氨基酸准确定量的非衍生化方法,具有良好的精密度以及准确性等,为准确定量小鼠血浆以及尿液中的氨基酸提供了方法参考。

Objective:Ultra-performance liquid chromatography-tandem mass spectrom-etry(UPLC-MS)was used to establish quantitative methods for 16 amino acids.Accurate quantitative analysis of plasma and urine samples was achieved.Methods:Using UPLC-MS technology,the liquid phase was gradient elution with 20 mmol·L^(-1)ammonium formate a-queous solution(A)and acetonitrile(containing 15%20 mmol·L^(-1)ammonium formate aque-ous solution)(B)as the mobile phase,and the mass spectrometry was detected in multiple re action monitoring mode(MRM).The specificity,limit of detection,limit of quantification,linearity,accuracy,precision,residual effect and stability were evaluated.The method was applied to the quantification of amino acids in plasma and urine of mice.Results:The results showed that the method met the relevant standards of methodology verification.The results of accurate quantification of amino acids in plasma and urine of mice showed that this meth-od could simultaneously detect and quantify 16 amino acids in plasma and urine of mice.Con-clusion:A non-derivatized method for accurate quantification of 16 amino acids in mouse plasma and urine has been established by UPLC-MS technology with good precision and ac-curacy,which provides a reference for accurate quantification of amino acids in mouse plasma and urine.