串联质谱法研究文拉法辛及其主要代谢物的药动学

Liquid Chromatography Tandem Mass Spectrometry Assay for Simultaneous Determination of Venlafaxine and Its Active Metabolite in Human Plasma and Its Application to a Bioequivalence Study

目的建立同时测定人血浆中文拉法辛(Ven)和O-去甲基文拉法辛(ODV)的串联质谱方法,研究男性健康志愿者单剂量服用盐酸文拉法辛胶囊,原形药文拉法辛和代谢产物O-去甲基文拉法辛体内药动学行为,评价生物等效性。方法 22名健康受试者采用随机分组自身交叉对照试验设计,二周期分别口服受试制剂和参比制剂盐酸文拉法辛胶囊50 mg,用LC-MS-MS联用法同时测定给药后不同时间点血浆中文拉法辛和O-去甲基文拉法辛的经时血药浓度,采用BAPP软件计算其药动学参数和评价生物等效性。结果血浆样品经蛋白沉淀,在选定的色谱/质谱条件下文拉法辛、O-去甲基文拉法辛与内标及血浆杂质分离良好,文拉法辛、O-去甲基文拉法辛分别在1.99～510μg·L-1(r=0.999 7)和1.99～510μg·L-1(r=0.9997)内线性良好;相对回收率在92.2%～105.9%,日内和日间RSD均小于10.5%。受试制剂和参比制剂的文拉法辛主要药动学参数:tmax分别为(2.2±0.7)和(1.9±0.8)h,ρmax分别为(68.90±23.82)和(69.81±23.73)μg·L-1;t1/2分别为(5.0±1.1)和(4.9±1.6)h;AUC0-24分别为(547.91±288.66)和(592.70±330.70)μg·L-1.h。O-去甲基文拉法辛主要药动学参数:tmax分别为(3.8±1.8)和(4.0±1.6)h,ρmax分别为(73.88±21.18)和(73.96±22.09)μg·L-1;t1/2分别为(8.7±1.8)和(8.9±1.9)h;AUC0-48分别为(1 224.41±239.46)和(1 243.53±287.19)μg.h.L-1;文拉法辛、O-去甲基文拉法辛相对生物利用度分别为(107.8±22.0)%和(99.6±10.7)%。结论用LC-MS-MS同时测定血浆中文拉法辛、O-去甲基文拉法辛浓度,样品处理简单快速,杂质无干扰,定量限低,重复性好,准确度高。两制剂的文拉法辛、O-去甲基文拉法辛的主要药动学参数之间无明显差异,经方差分析和检验,证明两制剂生物等效。

OBJECTIVE To establish a liquid chromatography tandem mass spectrometry （LC-MS-MS） method for simultaneous determination of venlafaxine （Ven） and its active metabolite, O-desmethylvenlafaxine （ODV）, in human plasma, and to investigate the pharmaeokinetics and bioequivalence of Ven and ODV in venlafaxine hydrochloride capsules in Chinese healthy volunteers. METH- ODS Twenty-two volunteers took a single oral dose （50 mg） of venlafaxine hydrochloride capsules by 2-way crossover design. The concentrations of Ven and ODV in plasma were determined by HPLC-MS-MS. The pharmacokinetie parameters were calculated by BAPP software. RESULTS Blood samples were deproteinized. The calibration curves of Ven and ODV were linear in the range from 1.99 to 510 μg·L^-1 （ r = 0. 999 7 ） and 1.96 to 501μg·L^-1 （ r = 0. 999 9） , respectively. The relative recovery was 92. 2% - 105.9%. The intra-day and inter-day RSDs were less than 10. 5%. The pharmaeokinetic parameters of test and reference capsules of Ven were as follows ： ρmax were （68. 90 ± 23.8） and （ 69. 81 ± 23.73 ）μg·L^-1·h^-1, tmax were （2. 2 ± 0. 7 ） and （ 1.9 ± 0. 8 ） h, t1/2 were （5.0 ±1.1 ） and （4. 9 ± 1.6） h, AUC0-24 were （547. 91 ± 288.66） and （ 592. 70 ± 330. 70） μg·h^-1·L^-1, respectively; the pharrnacokinetic parameters of test and reference capsules of ODV were as follows： ρmax were （73. 88 ± 21. 18） and （73.96 ±22. 09） μg· L^- 1, tmax were （3.8 ± 1.8） and （4.0 ± 1.6） h, t1/2 were （8.7 ± 1.8） and （8.9 ± 1.9） h, AUC0.48 were （ 1 224.41 ± 239.46） and （1 243.53 ± 287.19） μg·h·L^-1, respectively. The relative bioavalabilities of Ven and ODV in the test capsules were （ 107. 8 ± 22. 0） % and （99.6 ± 10. 7） %, respectively. CONCLUSION The HPLC-MS-MS method for simultaneous determination of Ven and ODV in plasma is proved to be sensitive, accurate and convenient. The reference and test capsules are bioequivalent.