液相色谱串联质谱法检测脂溶性维生素模拟校准品基质的选择和评价  

The Selection and Evaluation of Matrix for the Determination of Fat-solubleVitamin Calibrators by Liquid Chromatography TandemMass Spectrometry

作　　者：李敏 孙江漫 孟祥兆 赵光轮 韩玉霞[1] 于洪远[1] 李会强[2] LI Min;SUN Jiangman;MENG Xiangzhao;ZHAO Guanglun;HAN Yuxia;YU Hongyuan;LI Huiqiang(Beijing Aerospace General Hospital,Beijing 100076,China;School of Laboratory Medicine,Tianjin Medical University,Tianjin 300203,China)

机构地区：[1]北京航天总医院检验科,北京100076 [2]天津医科大学医学检验学院,天津300203

出　　处：《标记免疫分析与临床》2024年第11期2056-2061,2098,共7页Labeled Immunoassays and Clinical Medicine

基　　金：国家自然科学基金面上项目(编号:81772259)。

摘　　要：目的选择和评价液相色谱串联质谱法检测脂溶性维生素标准曲线的模拟基质。方法采用纯甲醇、4%牛血清白蛋白(BSA)作为模拟基质,配制脂溶性维生素的标准曲线。通过线性实验、正确度实验、基质效应实验,对模拟基质进行评价。采用两条标准曲线分别对临床44例血清样本进行定量,依据CLSI EP9-A3文件,对结果进行比对。结果以纯甲醇为基质,维生素A(VA)、25-羟基维生素D 3[25(OH)D 3]、25-羟基维生素D 2[25(OH)D 2]、维生素E(α-VE)对应标准曲线的相关系数分别为0.999、0.997、0.997、0.999,加标回收率为90.0%~110.1%,相对基质效应为-7.40%~7.90%。以4%BSA为基质,VA、25(OH)D 3、25(OH)D 2、α-VE对应标准曲线的相关系数分别为0.999、0.997、0.997、0.999,加标回收率为91.1%~111.2%,相对基质效应为-8.84%~7.60%。两条标准曲线对44例临床样本的定量结果进行线性拟合,VA、25(OH)D、α-VE对应的Passing-Baklok回归方程分别为Y=0.9083 X-3.7026、Y=1.0406 X-3.0114、Y=1.018 X-72.786,医学决定水平处的预估偏移均在可接受范围内。结论纯甲醇和4%BSA均可作为脂溶性维生素校准品的模拟基质,各实验室应评估模拟基质的适用性,以确保临床检测结果的准确性。Objective To select and evaluate a simulated substrate for the determination of standard curves of fat-soluble vitamins by liquid chromatography-tandem mass spectrometry.Methods The standard curve of fat-soluble vitamin was prepared by using pure methanol and 4%bovine serum albumin(BSA)as simulated matrix.The simulated matrix was evaluated by linear experiment,accuracy evaluation experiment and matrix effect mixed experiment.Two standard curves were drawn to quantify serum samples from 44 clinical cases and the results were compared according to CLSI EP9-A3 file.Results Using pure methanol as the substrate,the correlation coefficients of the corresponding standard curves of vitamin A(VA),25-hydroxyvitamin D 3[25(OH)D 3],25-hydroxyvitamin D 2[25(OH)D 2]and vitamin E(α-VE)were 0.999,0.997,and 0.999,respectively,while the recoveries were 90.0%-110.1%.The relative matrix effect ranges from-7.40%to 7.90%.Using 4%BSA as matrix,the correlation coefficients of VA,25(OH)D 3,25(OH)D 2 andα-VE corresponding standard curves were 0.999,0.997,0.997,and 0.999,respectively.The recoveries were 91.1%-111.2%and the relative matrix effects were-8.84%-7.60%.Two standard curves were applied to linearly fit the quantitative results of 44 clinical samples.The developed regression equations of VA,25(OH)D andα-VE were as follows:Y=0.9083 X-3.7026,Y=1.0406 X-3.0114,and Y=1.018 X-72.786,respectively.The estimated deviations at medically determined levels were within acceptable limits.Conclusion Both pure methanol and 4%BSA can be used as simulated substrates for fat-soluble vitamin calibrators.The applicability of simulated substrates should be evaluated in laboratories to ensure the accuracy of clinical test results.

关 键 词：脂溶性维生素 模拟基质 液相色谱串联质谱 

分 类 号：R-331[医药卫生]