摘要
目的探讨超高效液相色谱串联质谱法测定血浆不同类型神经酰胺含量的效果。方法采用美国Waters超高效液相色谱仪和ACQUITY UPLC I-Class/Xevo TQD三重四极杆质谱仪对4种神经酰胺进行定量检测,使用的色谱柱为ACQUITY UPLC BEH C18(2.1×50.0 mm,1.7μm),柱温50℃,流动相A为5 mM乙酸铵水溶液,流动相B为0.1%甲酸乙腈,梯度洗脱,扫描方式为多重反应监测模式。结果Cer 16:0、Cer 18:0、Cer 24:0、Cer 24:1神经酰胺的色谱标准曲线显示出良好的线性关系(相关系数r≥0.99)。4种神经酰胺的低、中、高值质控品的批内和批间变异系数(CV)≤15%、相对偏差(Dev)≤±15%,检验结果均满足性能验证要求。平均回收率均在92.20%~111.58%之间,基质效应在92.69%~98.90%之间。高值质控品后的空白标本的测定值均≤定量下限的20%,残留很少,满足实验要求。结论采用超高效液相色谱串联质谱法同时测定不同类型神经酰胺的方法精密度和准确度高,重现性好,可作为检测血清中不同类型神经酰胺含量的工具。
Objective To establish an analytical method for the determination of different types of ceramide in plasma by high-performance liquid chromatography tandem mass spectrometry.Methods American Waters high-performance liquid chromatograph system was employed for quantitative analysis of four ceramides by using ACQUITY UPLC I-Class/Xevo TQD triple quadrupole mass spectrometer,BEH C18(2.1×50.0 mm,1.7μm)chromatographic column.The column temperature was 50℃.The mobile phase A was 5 mM ammonium acetate aqueous solution and the mobile phase B was 0.1%formic acid acetonitrile.The elution method was gradient elution.The scanning mode was multiple response monitoring mode.Results The chromatographic standard curves of Cer 16:0,Cer 18:0,Cer 24:0 and Cer 24:1 showed a good linear relationship(correlation coefficient r≥0.99).The coefficient of variation(CV)of intra-batch and inter-batch assays of low,medium and high value quality control products of the four ceramides were less than 15%,and the relative deviation(Dev)of intra-batch and inter-batch assays were less than±15%.The test results all met the performance verification requirements.The average recovery rate was 92.20%-111.58%and the matrix effect was 92.69%-98.90%.The measured values of blank samples after high-value quality control products were≤20%of the lower limit of quantification,and the residue was very small,which met the experimental requirements.Conclusion The high-performance liquid chromatography-mass spectrometry method for simultaneous analysis of ceramides has high accuracy and reproducibility,which can be used to quantification of ceramide in plasma.
作者
刘兵
刘岩
李斯琪
李红帅
徐丽斯
刘艳霞
LIU Bing;LIU Yan;LI Si-qi;LI Hong-shuai;XU Li-si;LIU Yan-xia(The Second Cadre Ward,General Hospital of Northern Theater Command,National Center for Clinical Research of Geriatric Diseases,Shenyang 110016,China)
出处
《临床军医杂志》
CAS
2022年第11期1121-1124,1128,共5页
Clinical Journal of Medical Officers
基金
全军保健专项基金项目(19BJZ14)
国家自然科学基金面上项目(82170297)
关键词
神经酰胺
液相色谱法
质谱分析法
三重四极杆质谱仪
Ceramide
Liquid chromatograph
Mass spectrometry
Triple quadrupole mass spectrometer
