中国临床药学杂志

目的建立用于检测更昔洛韦在家兔房水内药物浓度的HPLC法;并研究更昔洛韦滴眼液在家兔眼内的药动学行为。方法房水样品用20%(V/V)高氯酸沉淀蛋白后取上清液直接进样,HPLC法测定。色谱柱使用LiChrospher C18柱(150 mm×4.6 mm,5μm),流动相为乙腈-0.05 mol.L-1醋酸铵水溶液(0.4∶99.6,V/V)。流速为1.0 mL.min-1,柱温为30℃,检测波长为254 nm。结果房水样品中药物浓度为40.0~400.0μg.L-1时呈良好的线性关系,方程为ρ=0.009 5A-2.410 3,r=0.999 0,(n=6)。低、中、高3种浓度的方法学平均回收率为97.0%、98.7%和102.4%。低、中、高3种浓度的绝对回收率分别为95.0%、98.1%、99.4%。样品溶液在反复冻融条件下,含量基本保持不变,显示更昔洛韦在此条件下稳定性良好。日间和日内RSD均<5%。最低检测限(LOD)为20.0μg.L-1。家兔药动学研究表明,市售更昔洛韦滴眼液药-时曲线符合二室模型一级吸收。结论该方法具有专属性强、灵敏度高、重现性好,适合于更昔洛韦在家兔房水内的药物浓度测定及其在眼内的药动学研究。

AIM To develop an analytical HPLC method for the determination of ganciclovir concentration in rabbit aqueous humor and to study the ocular pharmacokinetics of its eyedrops.METHODS Proteinaceous material in rabbit aqueous humor was precipitated by 20%(V/V) perchloric acid.An aliquot of the supernatant was analyzed by HPLC.The chromatography was performed on a reversed-phase encapped column(LiChrospher C18) at 30℃ with a mobile phase consisting of acetonitrile in 0.05 mol·L-1 ammonium acetate(pH=6.5,0.4∶99.6,V/V).The flow rate was 1.0 mL·min-1.Ganciclovir was monitored with UV detector at 254 nm.The parameters were calculated by 3P87 program.RESULTS Good linearity between concentration of ganciclovir(ρ,μg·L-1)in aqueous humor and peak area(A) was obtained for ganciclovir(ρ=0.009 5A-2.410 3,r=0.999 0,n=6) in the range from 40.0 to 400.0 μg·L-1.The lower limit of detection(LOD) of the method was 20.0 μg·L-1,indicating that the method was adequate for sensitive pharmacokinetic studies.The average recoveries were determined as 97.0%, 98.7%,and 102.4% at a concentration of 40.0,200.0,400.0 μg·L-1.The absolute recovery was determined as 95.0%,98.1%,and 99.4% at a concentration of 40.0,200.0,400.0 μg·L-1.The stability of ganciclovir samples was good in freeze-thawing condition.Within-day and inter-day coefficient of variation was lower than 5%.To study its pharmacokinetic bioavailability,the eyedrop formulation of ganciclovir was given to 40 healthy rabbits.According to 3P87 program,the concentration-time curve was fit for two-compartment open model and the major parameters were as follows: t12α(1.95±0.78) h,t12β(26.46±8.84 )h,AUC0→48(10 184±343 7)μg·h·L-1,tpeak(1.500±0.40)h,ρmax(258.80±67.60)μg·L-1.CONCLUSION This method is a selective,sensitive and reproducible method for the determination of the ganciclovir in rabbit aqueous humor and is suitable for the pharamacokinetic study of ganciclovir in ophthalmic system.