替硝唑、达克罗宁和氯己定微透析探针体外回收率及大鼠体内稳定性的研究

Study on the in vitro recovery of microdialysis probe and in vivo stability in rats of tinidazole,dyclonine and chlorhexidine

目的:测定替硝唑、达克罗宁和氯己定的微透析体外回收率,并进行体内回收率稳定性的考察。方法:采用HPLC-MS/MS联用技术建立微透析样品中替硝唑、达克罗宁、氯己定的分析方法。体外回收率的测定采用浓差法(增量法、减量法),考察灌流液、流速、浓度对回收率的影响;体内回收率的测定采用减量法,将探针植入大鼠皮下组织中,考察探针在12 h回收率的稳定性。结果:所建立的HPLC-MS/MS,方法灵敏可靠,在所需浓度范围内线性良好。增量法及减量法测定的回收率一致;回收率随灌流速度的增大而减小,探针的回收率与药物的浓度无关。体内皮下探针对替硝唑、达克罗宁、氯己定的回收率分别为(34.25±2.97)%、(31.20±1.75)%、(25.19±2.52)%,3个药物在皮下探针中12 h回收率的变化基本上保持稳定。结论:本研究提示体内研究的反透析法可作为微透析研究中替硝唑、达克罗宁、氯己定回收率的测定方法,所建立的微透析取样技术可用于3个药物的在体药动学研究。

Objective： To detect the in vitro recovery of tinidazole,dyclonine and chlorhexidine in microdialysis technique and to study its stability in vivo. Methods： HPLC- MS/MS method used for the determination of tinidazole,dyclonine and chlorhexidine in microdialysis samples was established. Comparable in vitro recovery was obtained by different established approaches including recovery estimation by gain and loss method,and the effects of perfusion fluid,flow rate and concentration on the recovery were investigated. Recovery by loss was used to study the in vivo 12 h recovery of tinidazole,dyclonine and chlorhexidine from rat subcutaneous tissue. Results：The HPLC- MS / MS method established was sensitive and reliable. The linearity correlation was good within the required concentration range. The in vitro experiment results showed that the in vitro recoveries of tinidazole,dyclonine and chlorhexidine from the microdialysis samples determined by gain and by loss were comparable at each flow rate; the recovery decreased as the flow rate increased; the recoveries of the three components were independent of concentration over the concentration range investigated. In the present study,the in vivo recoveries of three components were（ 34. 25 ± 2. 97） %,（ 31. 20 ± 1. 75） % and（ 25. 19 ± 2. 52） %,respectively,and were stable over the 12 h study period. Conclusion： Following characterization of tinidazole,dyclonine and chlorhexidine both in vitro and in vivo microdialysis,microdialysis sampling could be used for the pharmacokinetic study of the three components.