高效液相色谱法考察更昔洛韦在Caco-2细胞上的转运特征  

Study on transport of ganciclovir in Caco-2 cell monolayer model by HPLC

作　　者：黄建耿[1] 姜玲黎[1] 斯陆勤[1] 吴祥根[1] 范昭泽[1] 李高[1] 

机构地区：[1]华中科技大学同济医学院药学院药剂系,湖北武汉430030

出　　处：《中国医院药学杂志》2008年第10期775-778,共4页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金(编号:30572265);湖北省卫生厅科研基金项目(编号:JX2B04)

摘　　要：目的:建立更昔洛韦的高效液相色谱测定法,研究其在Caco-2细胞上的转运特征。方法:采用Eurospher-100C_(18) (250mm×4.6 mm,5μm)色谱柱,流动相为甲醇-水(8:92).流速为1.0mL·min^(-1),检测波长为254 nm,考察介质pH、药物浓度、P-gp抑制剂对更昔洛韦在Caco-2细胞上转运的影响。结果:更昔洛韦在Caco-2细胞单层膜上的转运量具有一定的浓度和时间依赖性,但介质pH对其跨膜转运无显著影响;更昔洛韦在50mg·L^(-1)质量浓度下AP→BL、BL→AP方向转运表观渗透系数分别为(0.59±0.11)×10^(-6)"和(2.0±0.4)×10^(-6)cm·s^(-1)。P-gp抑制剂维拉帕米能够增加更昔洛韦跨膜转运量。结论:更昔洛韦在Caco-2细胞上转运表现为被动扩散且有外转运蛋白参与更昔洛韦跨膜转运。OBJECTIVE To establish an HPLC method for determination of ganciclovir and investigate its transport in Caco- 2 cell monolayer model. METHODS Separation of ganciclovir was carried out by a Eurospher-100 C18 （250 mmx 4. 6 ram, 5 μm） column. The mobile phase consisted of methanol-water （8：92） at the flow rate of 1.0 mL·min^-1. The detective wave- length was set at 254 nm. This HPLC method was applied to determine ganciclovir concentration in Caco-2 cell monolayer transport system under various conditions, including changing ganciclovir concentration and pH of DHank＇s. Furthermore, influence of positive P-gp inhibitor verapamil on ganciciolovir transport was involved in the study. RESULTS Ganciciclovir transport in Caco-2 cell monolayer was time and concentration dependent, but not pH dependent. The apparent permeability coefficient （Papp） of ganciclovir with the concentration of 50μg·mL^-1 transporting from Apical to Basolateral and Basolateral to Apical was （0. 59 ± 0. 11 ） × 10^-6 cm·s^-1 and （2. 0 ± 0. 4） × 10^-6 cm·s^-1 , respectively. The transport of ganciclovir was enhanced by verapamil, CONCLUSION Ganciclovir transport complied with the passive diffusion in Caco-2 cell monolayer model, and furthermore efflux transporters might be involved in the transepithelial transport of ganciclovir.

关 键 词：高效液相色谱法 更昔洛韦 CACO-2细胞 跨膜转运 

分 类 号：R945[医药卫生—微生物与生化药学]