温通膏中补骨脂素、异补骨脂素皮肤在体微透析方法的建立

Establishment of in Vivo Skin Microdialysis Method for Psoralen and Isopsoralen in Wentong Ointment

目的建立温通膏中补骨脂素、异补骨脂素的皮肤在体微透析方法。方法以补骨脂素、异补骨脂素回收率为指标,采用增量法考察不同灌流液流速对补骨脂素、异补骨脂素探针体外回收率的影响,确定补骨脂素、异补骨脂素灌流液的流速和取样间隔;利用增量法和减量法观察灌流液流速、浓度对探针体外回收率和损失率的影响,研究反透析法是否可应用于探针体内回收率的计算;将探针植入大鼠皮下,通过减量法考察探针在12 h内体内回收率的稳定性。结果补骨脂素、异补骨脂素探针体外回收率随着灌流液流速的增加,呈现下降的趋势,结合实际操作的时间和采样体积,选择灌流液流速为1μL·min-1,每次采样间隔为20 min;灌流液流速保持一定时,不同浓度的补骨脂素、异补骨脂素下探针体外回收率变化不大,且探针体外回收率与损失率近似相等,表明反透析法可以应用于探针体内回收率的计算;探针植入皮下在12 h内回收率保持相对稳定。结论所建立的补骨脂素、异补骨脂素的皮肤微透析方法可用于温通膏给药后大鼠皮肤药动学研究。

Objective To establish an in vivo skin microdialysis method for psoralen and isopsoralen in Wentong ointment.Methods Using the recovery rates(R)of psoralen and isopsoralen as indicators,the incremental method was used to investigate the effects of different perfusion fluid flow rates on the in vivo R of psoralen and isopsoralen probes,combined with specific experimental operations and relevant factors to determine the flow rate and sampling interval of psoralen and isopsoralen perfusate.The increment method and decrement method were used to explore the influence of perfusate flow rate and concentration on the recovery rate(R)and loss rate(L)of the probe in vitro to study whether the anti-dialysis method can be applied to the calculation of R in the probe.The probe was implanted beneath the skin of the rat,and the stability of the R in the body of the probe within 12 h was investigated by the decrement method.Results The in vitro R of psoralen and isopsoralen probes showed a downward trend with the increase of the flow rate of the perfusion fluid.Combined with the actual operation time and sampling volume,the perfusion fluid flow rate was determined to be 1μL·min-1 and the sampling interval was determined to be 20 min.When the flow rate of the perfusate remained constant,the external R of the probe under different concentrations of psoralen and isopsoralen did not change much,and the external R and L of the probe were approximately equal,indicating that the reverse dialysis method can be used for calculation of R in the probe.R remained relatively stable within 12 hours after the probe was implanted under the skin.Conclusion The established skin microdialysis method of psoralen and isopsoralen in Wentong ointment can be used to study the skin pharmacokinetics of rats after administration.