摘要
目的建立HPLC-MS法测定大鼠血浆中利福布汀浓度的方法,并对大鼠灌胃给药利福布汀后,研究利福布汀在大鼠体内的药动学。方法采用HPLC-MS法测定血浆中利福布汀的浓度。色谱柱:Waters Xbridge-C_(18)(100 mm×3.0mm,3.5μm),流动相:乙腈-水(含0.1%甲酸)=70:30;流速:0.4 ml·min^(-1),分析时间5 min,柱温:25℃,进样量:5μl,质谱条件:采用ESI源负离子模式,毛细管电压3500 V,干燥气流速10 L·min^(-1),雾化气压力35 psi,干燥气温度350℃。选择离子模式(SIM)下,利福布汀的监测离子为[M-H]^-m/z845.4和内标A3的监测离子为[M-H]^-m/z 509.3,碎片电压100 eV。结果利福布汀在0.15~30μg·ml^(-1)浓度范围内线性关系良好(r=0.9987);最低定量限为0.15μg·ml^(-1);低、中、高浓度的提取回收率为86.42%~95.80%;日内、日间精密度RSD均<11.5%。其药动学参数分别为:C_(max)(2.08±0.783)μg·ml^(-1)、T_(max)(2.20±1.247)h、t_(1/2)(12.58±4.909)h、CLz/F(1.15±0.098)L·h·kg^(-1)、Vz/F(20.31±7.276)L·kg^(-1)、AUC_(0-∞)(26.32±2.144)μg·h·ml^(-1)、MRT_(0-∞)(18.29±5.05)h、VRT_(0-∞)(379.94±195.84)h^2。结论该方法简便、准确、可靠,可用于测定大鼠血浆中利福布汀的浓度,并应用于其药代动力学研究。单次口服利福布汀后,药物吸收迅速,消除缓慢,能快速达到峰浓度,且能长时间维持较高浓度。
Objective To establish an HPLC-MS method for the determination of rifabutin in rat plasma and apply it to study the pharmacokinetics of rifabutin after oral administration. Methods Rifabutin was prepared by adding methanol for protein precipitation, and the extraction solution was analyzed by HPLC-MS. The separation of the sample was performed on Waters Xbridge-C18 （100 mm × 3.0 mm ,3.5 μm）. The mobile phase consisted of acetonitrile and 0.1% formic acid aqueous solution 70： 30（ V/V ） at a flow rate of 0.4 ml .rain-1. The column temperature was 25℃ and the injection volume was 5 μl. MS conditions were as follows：The ESI source was in the negative ion mode ,the capillary voltage was 3500 eV,the flow rate of drying gas was 10 L ·min^-1 ,the atomizing air pressure was 35 psi,and drying temperature was 350℃. Results The linear range was 0.15 -30.0 μg·ml^-1 （ r = 0. 9987）. The lowest detectable limit was 0.15 μg ·ml^-1. The recovery of the method was 101.9% - 103.2% ,and the extraction recovery was 86.42% -95.80%. RSD of inter- and intra-day precisions was both less than 11.5%. The pharmacokinetic parameters were： C max（2. 08 ±0. 783） μg ·ml^-1, T max（2. 20 ± 1. 247） h, t 1/2（ 12.58 ± 4. 909） h,CLz/F（1.15 ±0.098） L·h ·kg^-1 ,Vz/F（20.31 ±7. 276） L·kg^-1 ,AUC（0-∞） （26.32 ±2.144） μg·h · ml^-l, MRT（0-∞）（ 18.29 ± 5.05 ） h ,VRT（0-∞）（379.94 ± 195.84） h^2. Conclusion This method is quick, simple and reliable, which can be used to determine rifabutin in rat plasma and in its pharmacokinetic study. After a single oral administration of rifabutin, drug absorption is quick, elimination is slow, peak concentration soon follows;and a high concentration can be maintained for a long time.
出处
《解放军药学学报》
CAS
CSCD
2015年第2期106-109,125,共5页
Pharmaceutical Journal of Chinese People's Liberation Army
基金
国家重大专项项目
No.2013ZX10003009-004
