摘要
目的:建立一种简便、快速、准确、灵敏的高效液相色谱-质谱联用法测定人体内血浆中的阿奇霉素浓度的方法。方法:对18例男性健康志愿受试者进行双交叉试验,随机单剂量口服阿奇霉素干混悬剂(500mg)参比或受试制剂后,按设定时间采集肘静脉血;分取血浆0.2mL 加入内标液(克拉霉素,12.5μg·mL^(-1))50μL,经饱和碳酸钠溶液0.1mL 碱化和乙酸乙酯1mL 萃取处理;采用 HyPurity C_(18)柱(Thermo Hypersil-Keystone,2.1mm×150mm,5μm)色谱柱,柱温40℃,流动相为醋酸铵(10mmol·L^(-1),pH 5.2)-乙腈-甲醇(50:40:10),流速为0.2mL·min^(-1)。HPLC-MS 内标(克拉霉素,[MH]^+,m/z=749)法正离子检测的电喷雾电离方式(ESI)选择离子监测(SIR)离子阿奇霉素([MH]^(2+),m/z=375)血浆浓度。结果:HPLC-MS 法测定血浆中阿奇霉素的最低定量限为3.91μg·L^(-1),线性范围3.91～1000μg·L^(-1)(r=0.9993)。萃取绝对回收率为(72.6±4.6)%～(81.3±5.7)%,日内和日间 RSD 均低于12%。测得口服阿奇霉素干混悬剂受试和参比制剂后的主要药代动力学参数:C_(max)分别为(557.68±110.91)ng·mL^(-1)和(567.90±93.58)ng·mL^(-1);T_(max)分别为(2.00±0.69)h 和(2.00±0.77)h;T_(1/2)分别为(30.95±9.37)h 和(28.94±8.71)h;AUC_(0→120)分别为(5544.31±1852.17)ng·mL^(-1)和(5637.46±1652.44)ng·mL^(-1);相对生物利用度(F)为(98.07±15.51)%。结论:建立的 HPLC-MS 方法灵敏、准确,测定结果可靠;统计分析表明阿奇霉素干混悬剂试验和参比制剂生物等效。
Objective： To establish an accurate and sensitive HPLC - MS method for the determination of azithromycin in plasma and to evaluate its pharmacokinetic parameters and bioavailability in healthy male volunteers after a single oral dose of 500 mg azithromycin or its dry suspensions. Methods： A single oral dose of 500 mg azithromycin or its dry suspensions was given to 18 healthy male volunteers in a two -way cross -over design, the plasma sampies were collected at predetermined time points after the administration, after adding the internal reference （clarithromycin, 12. 5 μg·mL^-1 ） ,0. 2 mL of each plasma sample was alkalized with saturated solution of carbonate azithromycin were determined by HPLC - MS method using HyPutiy Clscolumn（2. 1 mm × 150 mm,5 μm） at 40 ℃, a mixture of NH4Ac（ 10 mmol·L^-1 ,pH 5.2） -acetonitrile -methanol（50： 40： 10） as mobile phase at a flow rate of 0. 2 mL·min^-1, with positive iron ESI detection of azithromycin （ [ MH ]^2±, m/z = 375 ） using clarithromycin （ [MH]^± ,m/z =749） as internal reference. Results：The limit of detection was 3.91 μg·L^-1. A good linearity was obtained in the range of 3.91 -1000 μg·L^-1 （r =0. 9993）. The extraction recoveries were in the range of （72. 6 ±4. 6）% - （81.3 ± 5. 7 ）%, The intra- day and inter- day RSD were both lower than 12%. The main pharmacokinetics parameters after a single oral dose of 500 mg azithromycin or dry suspensions were as follow ： Cmax （ng· mL^-1） ：（557.68 ±110.91） and （567.90 ±93.58） ;Tmax（h） ：（2.00 ±0.69） and （2.00 ±0.77）;T1/2 （h） ： （ 30. 95 ± 9. 37 ） and （ 28.94 ± 8.71 ） ; AUC0→120 （ ng · mL^- 1 ） ： （ 5544. 31 ± 1852. 17 ） and （ 5637. 46 ± 1652. 44）. The relative bioavailability （F） was （ 98.07 ± 15.51 ） %. Condusins： The established HPLC - MS method is sensitive,rapid and accurate. The pharmacokinetic parameters determined is reliable. The statistic results show that azithromycin and its dry suspensions of the test and reference is bioequivalent.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2006年第6期726-729,共4页
Chinese Journal of Pharmaceutical Analysis
