人参皂苷Rg_1脑、血微透析探针体内外回收率研究

Recovery of Ginsenoside Rg_1 by Brain and Blood Microdialysis Probe in Vivo and in Vitro

目的以人参皂苷Rg_1为检测药物,研究脑、血微透析探针的体内外回收率。方法采用正透析法和反透析法进行人参皂苷Rg_1脑、血微透析探针体内、体外回收率的研究,采用LC-MS/MS测定微透析液中人参皂苷Rg_1的浓度,计算探针回收率。结果在流速为1.5μL·min^(-1)时,脑、血探针体内回收率在10 h内保持稳定,平均回收率分别为17.0%和34.4%;恒定流速下,质量浓度(50,200,500,1 000 ng·mL^(-1))对脑、血探针回收率均无影响;恒定浓度下,探针体内外回收率均随着流速(0.5,1.0,1.5,2.0,3.0μL·min^(-1))的增加而减小,脑、血探针的体外正透析法测的回收率分别为(40.6±4.3)%,(23.5±2.3)%,(17.7±0.8)%,(12.2±1.1)%,(8.8±0.6)%和(70.6±3.6)%,(46.0±2.1)%,(32.9±1.6)%,(25.6±0.7)%,(18.2±1.3)%,正、反透析法所测得的探针体外回收率一致,且体内和体外回收率结果也一致;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回收率。结论反透析法能够作为人参皂苷Rg_1脑、血微透析探针体内回收率的测定方法,微透析技术能够用于人参皂苷Rg_1脑细胞间液、血液药动学的同步研究。

OBJECTIVE To establish a measuring method for microdialysis probe recovery of ginsenoside Rg1 and investigate the effects of flow rate, concentration and using times of probe on the recovery in vlvo and in vitro. METHODS Dialysis method and retrodialysis method were used for the study. The concentration of ginsenoside Rg1 in brain and blood dialysate was determined by LC-MS/MS and the probe recovery was calculated. RESULTS The recoveries of brain and blood microdialysis probes showed good stability within 10 h, with average values of 17.0% and 34.4% respectively for ginsenoside Rg1 at 1.5μL·min±（-1）. Concentrations （50,200,500,1 000 ng·mL±（-1）） had no obvious effect on recovery. At the same concentration, the recovery of brain and blood probes for ginsenoside Rg1 decreased with the increase of flow rate （0.5, 1.0, 1.5, 2.0, 3.0 μL·min±（-1） ） in vitro and in vivo. The dialysis recoveries of brain and blood probes in vitro were （40.6 ± 4.3 ） % , （23.5±2.3）%, （17.7 ±0.8）%, （12.2 ± 1.1）%, （8.8 ±0.6）% and （70.6 ± 3.6）%, （46.0 ±2.1）%, （ 32.9 ± 1.6） % , （ 25.6 ± 0.7 ） % , （ 18.2 ± 1.3 ） % , respectively. The recoveries of dialysis and retrodialysis in vitro were approximately equal, and the recovery detected by retrodialysis in vivo was similar with the in vitro results. Probe used for no more than 3 times still kept high transmittance by flushing with 2% heparin sodium and ultrapure water successively. CONCLUSION Retrodialysis method can be used to study brain and blood probe recovery in vivo, and microdialysis can be used for simutaneous pharmacokinetic studies of ±insenoside Rg, in intercelluar fluid and blood.