氯法拉滨大鼠血药浓度检测及药动学研究

Determination of Clofarabine in rat plasma and the pharmacokinetic research

目的:建立氯法拉滨血药浓度HPLC检测法,研究其在大鼠体内的血浆药动学。方法:色谱条件为Agilent ODS Hypersil C18分析柱(150 mm×4.6 mm,5μm),柱温30℃,紫外检测波长263 nm,流动相为乙腈-水(16∶84),乙酸调pH=4.0,流速1.2 mL/min;氯法拉滨30 mg/kg静脉注射,于给药后0.17、0.33、0.5、0.75、1.0、1.5、2.0、4.0、8.0 h取血,测血药浓度。结果:氯法拉滨与血浆组分分离完全,在0.05～20.0μg/mL浓度范围内线性关系良好(线性方程y=21.758x-0.9155,r=0.999910)。氯法拉滨高、中、低三个浓度的回收率为95%～105%,日内、日间精密度均小于10%。氯法拉滨大鼠体内药动学符合二室模型,主要药动学参数为A=14.20±2.45,B=1.81±0.51,CL=(2.91±0.47)L.h-1.kg-1,AUC(0-t)=(10.22±1.88)mg.L-1.h,t1/2β=(2.03±0.16)h。结论:HPLC紫外法简便、可靠,能够满足氯法拉滨血药浓度监测及药动学研究需要;氯法拉滨在大鼠体内药动学符合二室模型,消除半衰期约为2 h。

AIM： To establish an HPLC method for the determination of Clofarabine concentration in the plasma of rats and calculate the pharmacokinetic parameters. METHODS： All the samples were separated on a Agilent ODS Hypersil C18 analytical column（150 mm×4.6 mm,5 μm） by a mobile phase of acetonitrile-water（16∶84）.The analytical column temperature was 30 ℃,the ultraviolet detection wavelength was at 263 nm,and the flow rate was 1.2 mL/min.The dosage of 30 mg/kg was used for the intravenous injection of Clofarabine before we took blood from each rat at 0.17,0.33,0.5,0.75,1.0,1.5,2.0,4.0 and 8.0 h post administration.The plasma concentration of Clofarabine was measured by HPLC.RESULTS：The chromatographic peak of Clofarabine was separated completely with the impurities in rat plasma.Good linearity was found over the range of 0.05-20.0 μg/mL for Clofarabine determination（the linear equation was y=21.758x-0.9155,r=0.999910）.The recovery of methodology was 95%～105%,the inter-and intra-day precisions were both less than 10%.The concentration time curve of Clofarabine in rat could be fitted to two compartment model,and the major pharmacokinetic parameters were as follows： A=14.20±2.45,B=1.81±0.51,CL =（2.91±0.47） L·h-1·kg-1,AUC（0-t）=（10.22±1.88） mg·L-1·h,t1/2β=（2.03±0.16） h.CONCLUSION： This method is selective and sensitive for Clofarabine determination,and can also be used in pharmacokinetic research.The drug shows us a two-compartment model in rat plasma,and it＇s half-life in plasma of rats was 2 h.