液相色谱串联质谱法测定人血清中可的宁的含量

LC-MS/MS method for determination of continine in human serum

目的建立液相色谱串联质谱法测定人血清中可的宁的含量。方法以稳定同位素化合物(氘3-可的宁)为内标,血清经直接沉淀后进样分析。以XBridge^(TM) Phenyl(2.1 mm×150 mm,5μm)为分析柱,含0.02%甲酸的水溶液-含0.02%甲酸的乙腈溶液(83:17,V/V)为流动相。电喷雾离子源,正离子MRM扫描分析,可的宁和内标离子对分别为m/z 177.0→80.0和m/z 180.5→80.2。结果可的宁在10～2 000μg·L^(-1)范围内线性关系良好,定量下限为10μg·L^(-1)。批内、批间精密度RSD均小于5%,平均提取回收率在77.9%～88.6%范围内。结论本方法灵敏度高、专一性好、操作简单,可用于人血清中可的宁的检测。

AIM To develop a method of LC-MS/MS for determination of continine in human serum. METHODS After adding stable isotope chemical compound （continine-D3） as the internal standard, the analyte was achieved by direct serum protein precipitation and then XBridge^TM Phenyl （2.1 mm× 150 mm, 5 μm） was utilized for the analytical column, with 0.02% formic acid water solution-0.02% formic acid acetonitrile solution （83：17, V/V） as the mobile phase. After analysis by electrospray ionization source with positive ion Multiple Reaction Monitoring （MRM） scans, the outcomings of continine and continine-D3 were m/z 177.0→80.0 and m/z 180.5→80.2, respectively. RESULTS The linear calibration curve of continine showed excellent correlation in the concentration range of 10 - 2 000 μg·L^-1 with the lower limit of quantitation 10 μg ·L^-1. The inter-and intra-run precision （RSD） were less than 5% and the average recovery rate of 77.9% to 88.6%. CONCLUSION The established LC-MS/MS method is shown to be sensitive, accurate, simple and suitable for the determination of continine in human serum.