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目的分别建立人血清和尿液中液相色谱-串联质谱法同时测定伏立诺他及其代谢物4-苯胺基-4-氧代丁酸(M2)的方法。方法血清样品经甲醇沉淀蛋白后取上清液进样分析,以Amethyst C_(18)-P(150mm×2.1mm,5μm)为分析柱,流动相为2%的甲酸(含5mmol·L~(-1)醋酸铵水溶液)-甲醇(55:45,V/V);尿液样品以流动相稀释后进样分析,以Hedera ODS-2(150mm×2.1mm,5μm)为分析柱,流动相为2%的甲酸(含30mmol·L~(-1)醋酸铵水溶液)-甲醇(55:45,V/V)。质谱采用气动辅助电喷雾离子化和正离子多反应监测,定量分析的反应离子对分别为m/z 265.2→232.2(伏立诺他)、m/z 194.1→176.1(M2)和m/z 180.1→110.1(内标非那西丁)。结果伏立诺他血药浓度在2.004～1503μg·L~(-1)范围内线性关系良好,平均回收率大于98.3%,M2血药浓度在5.015～5015μg·L~(-1)范围内线性关系良好,平均回收率大于96.8%;伏立诺他尿药浓度在0.03006～20.04mg·L~(-1)范围内线性关系良好,平均回收率大于96.8%,M2尿药浓度在1.003～300.9mg·L~(-1)范围内线性关系良好,平均回收率大于96.5%。尿样测定中,由伏立诺他的另一代谢物O-葡萄糖醛酸苷发生源内裂解而对伏立诺他的测定造成的干扰通过色谱分离得到解决。结论建立的两个方法快速、简便,可应用于人体样本测定。
OBJECTIVE To establish a method for simultaneous determination of vorinostat and its metabolite 4-anilino-4-oxobutyric acid (M2) in human serum and urine by liquid chromatography-tandem mass spectrometry. Methods Serum samples were precipitated with methanol and the supernatant was injected into the sample. Amethyst C_ (18) -P (150 mm × 2.1 mm, 5 μm) was used as the analytical column. The mobile phase consisted of 2% formic acid with 5 mmol·L ~ -1) ammonium acetate in water) -methanol (55:45, V / V); the sample of urine was diluted by mobile phase and injected into Hepra ODS-2 (150mm × 2.1mm, 5μm) 2% formic acid (30 mmol·L -1 ammonium acetate aqueous solution) -methanol (55:45, V / V). The mass spectra were monitored by pneumatically assisted electrospray ionization and positive ion multiple reaction monitoring. The corresponding ion pairs were m / z 265.2 → 232.2 (vorinostat), m / z 194.1 → 176.1 (m 2) and m / z 180.1 → 110.1 (internal standard phenacetin). Results The linear range of vorinostat blood plasma concentration ranged from 2.004 to 1503μg · L ~ (-1), the average recovery was over 98.3%, and the linear range of plasma concentration of M2 was 5.015 ~ 5015μg · L ~ (-1) The average recovery rate was more than 96.8%. The average concentration of vorinostat in the range of 0.03006 ~ 20.04 mg · L ~ (-1) was good, with the average recovery of more than 96.8% and the concentration of M2 urinary drug in the range of 1.003 ~ 300.9 The linear relationship was good in the range of mg · L ~ (-1) with the average recovery of more than 96.5%. In a urine sample assay, interference with the determination of vorinostat caused by in-source lysis of another metabolite of vorinostat O-glucuronide was resolved by chromatographic separation. Conclusion The two established methods are fast and simple and can be applied to the determination of human samples.