栀子苷脑血微透析探针大鼠体内外试验回收率研究

Study on recovery rate of geniposide brain blood microdialysis probe in vitro and in vivo in rats

目的:考察栀子苷脑微透析探针和血微透析探针的大鼠体内外回收率及其稳定性。方法:采用LC-MS/MS法测定大鼠脑、血微透析液中栀子苷的浓度;采用正透析法和反透析法考察不同灌流速度、正透析法考察不同栀子苷浓度(50、200、500、1 000 ng·mL^(-1))及探针使用次数对体外回收率的影响;反透析法考察大鼠体内探针回收率稳定性及灌流速度对回收率的影响,并与体外结果进行比较。结果:栀子苷的脑、血探针体内外回收率均随着灌流速度(0.5、1.0、1.5、2.0、3.0μL·min^(-1))的增加而降低,不同流速下体外正透析法测得的脑、血液探针回收率分别为(48.0±3.4)%、(31.0±2.3)%、(20.4±1.3)%、(17.3±0.8)%、(7.9±0.7)%和(83.0±5.9)%、(56.2±5.2)%、(39.1±3.1)%、(30.4±2.6)%、(26.4±1.9)%;正透析法和反透析法所测得的体外回收率在相同条件下基本一致,且反透析法测得的体内回收率与体外结果基本一致;脑、血探针体内回收率在10 h内的稳定性均良好,平均回收率分别为23.4%和42.6%;脑、血探针回收率与栀子苷的浓度无关;使用不超过3次的探针,经过恢复处理后,仍然能够保持较高的透过率。结论:反透析法能够作为研究栀子苷体内回收率的测定方法,微透析技术能够用于栀子苷脑细胞间液药代动力学、血液药代动力学的同步研究。

Objective： To investigate the recovery rate and stability of geniposide brain and blood microdialysis probe in vitro and in vivo in rats.Methods： The concentrations of geniposide in rats,brain and blood microdialysates were determined by LC-MS/MS.In order to study the effect on recovery in vitro,the different flow rates were investigated by dialysis and reverse dialysis,drug concentrations（50,200,500 and 1 000 ng·mL-1） and the number of probes used were investigated by dialysis.The stability of the rats probe recovery and flow rate on the recovery rate in vivo were determined by reverse dialysis,and the results compared with that of in vitro trials.Results： The in vitro and in vivo recovery rate of brain and blood probe of geniposide were decreased with the increase of flow rate（0.5,1.0,1.5,2.0 and 3.0μL·min-1）.The dialysis recovery of brain and blood probe in vitro were （48.0±3.4）%,（31.0±2.3）%,（20.4±1.3）%,（17.3±0.8）%,（7.9±0.7）% and（83.0±5.9）%, （56.2±5.2）%,（39.1±3.1）%,（30.4±2.6）%,（26.4±1.9）%,respectively.In vitro recovery obtained by dialysis and reverse dialysis were approximately equal under the same condition,and the in vivo recovery determined by reverse dialysis was similar with the in vitro results.The recovery of brain and blood microdialysis probe showed a good stability within 10 h,and the average recovery of brain and blood probe for geniposide were 23.4% and 42.6%,respectively.The recovery rate of the brain and blood probe was not related to the different concentrations of geniposide,and the high transmittance can still be able to be maintained after the recovery of the probe which was used no more than 3 times.Conclusion： The reverse dialysis method can be used to study probe recovery in vivo,and microdialysis technique can be used for the simultaneous study of pharmacokinetic of geniposide in brain cells and blood.