液相色谱-串联质谱法同时测定人血清与尿液中伏立诺他及其代谢物4-苯胺基-4-氧代丁酸的浓度  

Simultaneous determination of vorinostat and its metabolite 4-anilino-4-oxobutanoic acid in human serum and urine by liquid chromatography-tandem mass spectrometry

作　　者：杜晓琅[1] 朱贺[1] 丁黎[1] 马鹏程[2] 王训强[3] 李文博[1] 罗雪梅[1] 王晶[1] 

机构地区：[1]中国药科大学药物分析教研室,江苏南京210009 [2]中国医学科学院皮肤病研究所,江苏南京210042 [3]江苏正大天晴药业股份有限公司,江苏南京210038

出　　处：《中国新药与临床杂志》2012年第7期385-391,共7页Chinese Journal of New Drugs and Clinical Remedies

摘　　要：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。AIM To establish a LC-MS/MS method for the simultaneous determination of vorinostat and its metabolite 4-anilino-4-oxobutanoic acid （M2） in human serum and urine. METHODS Serum samples were deproteinized using 3 ： 1 methanol/serum, and analyzed by HPLC on a Amethyst C18-P column （150 mm × 2.1 mm, 5 um） with a mobile phase of methanol-5 mmol.L-1 ammonium acetate buffer solution containing 2% formic acid （45 ： 55, V/V）. Urine samples were diluted and analyzed by HPLC on a Hedera ODS-2 C18 column （ 150 mm × 2.1 mm, 5 um） with a mobile phase of methanol-30 mmol .L-1 ammonium acetate buffer solution containing 2% formic acid （45 ： 55, V/V）. An electrospray ionization source was applied and performed in positive ion mode. The ion transitions recorded in multiple reaction monitoring mode were m/z 265.2--232.2 for vorinostat, m/z 194.1-176.1 for M2 and m/z 180.1--110.1 for the internal standard （IS）, respectively. RESULTS The calibration curves were linear over the range of 2.004 - 1 503 ug.L-1 for vorinostat and 5.015 - 5 015 ug.L-1 for M2, respectively, in serum. The average recoveries of vorinostat and M2 in serum were more than 98.3% and 96.8%, respectively. The calibration curves were linear over the range of 0.030 06 - 20.04 mg. L-1 for vorinostat and 1.003 - 300.9 mg. L-1 for M2, respectively, in urine. The average recoveries of vorinostat and M2 in urine were more than 96.8% and 96.5%, respectively. The interference due to decomposition of vorinostat glucuronide in the ionization source of the mass spectrometer was avoided by carrying out the chromatographic separation. CONCLUSION The developed methods can be applied to the pharmacokinetic study and therapeutic drug monitoring of vorinostat and its metabolite.

关 键 词：伏立诺他 4-苯胺基-4-氧代丁酸 色谱法 高压液相 串联质谱法 血清 尿 

分 类 号：R969.1[医药卫生—药理学] R979.1[医药卫生—药学]