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目的:建立同位素稀释超高效液相色谱串联质谱(ID-UPLC-MSMS)同时测定血清中5种脂溶性维生素的方法，以满足临床检测多种脂溶性维生素的需要。方法:选取2019年4月至2019年8月间上海市第十人民医院的血清样本并通过萃取法提取脂溶性维生素，采用ID-UPLC-MSMS进行检测，参照美国临床和实验室标准协会(CLSI)的相关文件，对该方法进行性能验证。结果:提出并成功验证了ID-UPLC-MSMS快速检测血清中多种脂溶性维生素的方法，该方法的检测线性范围分别为维生素A(VA)：25～2 500 μg/L，25-羟基维生素Dn 2[25(OH)Dn 2]: 2～200 μg/L, 25-羟基维生素Dn 3[25(OH)Dn 3]：2～200 μg/L，维生素E(VE)：0.25～50 mg/L，维生素K1(VK1)：0.1～20 μg/L。5种分析物的批内和批间精密度标准偏差均在±15%以内，25(OH)Dn 2和25(OH)Dn 3标准品测试结果的正确度为96.44%～102.37%。n 结论:采用ID-UPLC-MSMS法同时测定5种脂溶性维生素，方法学性能满意，结果准确可靠，适合临床样本分析。“,”Objective:To establish an analytical method for the simultaneous determination of five fat-soluble vitamins in serum using isotope dilution ultra high performance liquid chromatography-tandem mass spectrometry (ID-UPLC-MSMS).Methods:Fat-soluble vitamins were obtained from serum samples which collected from Shanghai Tenth People′s Hospital between April 2019 and August 2019 by the extraction method, and were detected by ID-UPLC-MSMS. The performance of the method was verified by referring to the relevant documents of the Clinical and Laboratory Standards Institute (CLSI).Results:The ID-UPLC-MSMS method for the rapid detection of various fat-soluble vitamins in serum was proposed and successfully verified. The linear range of the method: vitamin A: 25-2 500 μg/L, 25(OH)D n 2: 2-200 μg/L, 25(OH)D n 3: 2-200 μg/L, vitamin E: 0.25-50 mg/L, vitamin K1: 0.1-20 μg/L. The intra- and inter-assay precision standard deviations of the five analytes were within ± 15%, and the accuracy of the test results of the 25(OH)D n 2 and 25(OH)Dn 3 standards was 96.44%-102.37%.n Conclusion:The performance of ID-UPLC-MSMS method for the simultaneous determination of five fat-soluble vitamins is satisying, and the result is accurate and reliable, which suggested it can be used for the clinical sample.