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人体血清中3种脂溶性维生素的液相色谱-串联质谱分析方法研究
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作者单位
刘海培,姜小梅,韩文念,许舒欣,李艳,汪曣,蒋学慧 1.天津大学精密仪器与光电子工程学院2.天津国科医工科技发展有限公司 
基金项目:国家高技术研究发展计划(2014AA022305)
中文摘要:以人体血清中3种脂溶性维生素检测为例,探讨了一种针对人体内源性代谢物的分析方法。采用液相色谱-串联质谱(LC-MS/MS)对人体血清中维生素A、维生素D3和维生素E进行检测,分别通过混合人血清基质加入标准品及内标再扣除内源性物质本底的方法,以及与牛血清白蛋白(BSA)模拟基质加标的方法建立标准曲线进行定量分析。结果表明,采用混合人血清基质所建方法对维生素A、维生素D3和维生素E的检出限分别为4.2、0.8、67.2 ng/mL,定量下限分别为13.7、2.6、221.9 ng/mL。两种方法的线性相关系数均大于0.996;对于实际样品在低、高两个加标浓度下的回收率为90.7%~112%,相对标准偏差(RSD)为1.0%~4.5%,具有良好的准确性和重现性。对40组未知样本的检测结果表明,两种方法无统计学差异。因此,对于人体内源性代谢物采用混合人血清基质扣除本底的方法,可以实现标准品与待测样品基质匹配的准确分析,有利于临床相关疾病的便捷诊断。
中文关键词:脂溶性维生素  液相色谱-串联质谱(LC-MS/MS)  内源性代谢物  人血清基质
 
Study on Analysis Methods of Three Fat-soluble Vitamins in Human Serum by Liquid Chromatography-Tandem Mass Spectrometry
Abstract:Taking the detection of three fat-soluble vitamins in human serum as an example,an analytical method for human endogenous metabolites was explored.A mixed human serum matrix was prepared by adding standard substances and internal standards followed by background subtraction of endogenous substances,and another simulating matrix substrate was made of bovine serum albumin(BSA).They both were detected by liquid chromatography-tandem mass spectrometry(LC-MS/MS).Results showed that the limits of detection for vitamin A,D3 and E in the mixed human serum matrix were 4.2,0.8 and 67.2 ng/mL,while the limits of quantitation were 13.7,2.6 and 221.9 ng/mL,respectively.There existed good linear relationships for the two methods with good accuracy and reproducibility,and their correlation coefficients were both above 0.996.The recoveries for the actual samples at low and high spiked concentrations were between 90.7% and 112%,with relative standard deviations(RSD) of 1.0%-4.5%.There were no statistical differences between the two methods for 40 unknown samples.Therefore,for the endogenous metabolites from human body,the accurate analysis on the matrix matched between the standard sample and the sample to test could be achieved by adding the standard substance and deducting the background from the human serum matrix,which is beneficial to the convenient diagnosis on clinical related diseases.
Key Words:fat-soluble vitamins  liquid chromatography-tandem mass spectrometry(LC-MS/MS)  endogenous metabolite  human serum matrix
引用本文:刘海培,姜小梅,韩文念,许舒欣,李艳,汪曣,蒋学慧.人体血清中3种脂溶性维生素的液相色谱-串联质谱分析方法研究[J].分析测试学报,2019,(6):728-733.
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