盐酸小檗碱微透析探针的体内外回收率实验研究

目的：研究微透析探针的体外回收率与灌流速度和盐酸小檗碱浓度之间的关系，确定回收率的校正方法。方法：利用高效液相色谱法测定透析液中盐酸小檗碱含量，采用Agilent Eclipse XDB-C18（4.6 mm×250 mm，5 μm） 色谱柱，流动相为乙腈-0.05 mol·L^-1 磷酸二氢钠溶液（磷酸调节pH 为3.0）（35 : 65），流速1 mL · min^-1，检测波长346 nm，柱温30 ℃。计算不同灌流速度、不同盐酸小檗碱浓度时，正、反透析法测得的探针体外回收率。同时测定固定流速下，不同药物浓度时探针的体内回收率。结果：在体外回收率实验中，将盐酸小檗碱质量浓度定为5.0 μg·mL^-1，灌流速度分别为1、2和3 μL·min^-1 时，透析法测得的回收率依次为（73.6±5.4）%、（60.4±5.8）% 和（35.2±6.3）%；反透析法测得的回收率依次为（70.2±7.0）%、（62.6±4.7）% 和（32.5±8.3）%。在体内回收率实验中，将灌流速度定为2 μL·min^-1，盐酸小檗碱灌流液浓度分别为0.1、0.5 和5.0 μg·mL^-1 时，体内回收率依次为（51.4±3.5）%、（53.2±2.7）% 和（52.6±2.4）%。探针回收率与盐酸小檗碱浓度无关，与流速成反比。体外回收率实验中，透析法与反透析法测得的回收率相近。结论：在用微透析法对盐酸小檗碱生物样品取样时，可采用反透析法对探针回收率进行校正。

Objective: To determine the correction method for microdialysis recovery by studying the relationship between flow rate and berberine hydrochloride concentration and the microdialysis recoveries.Methods: The concentration of berberine hydrochloride in dialysis fluid was detected by HPLC.An Agilent Eclipse XDB-C₁₈ (4.6 mm×250 mm, 5 μm) column was adopted with 1 mL·min^-1 of acetonitrile-0.05 mol·L^-1 sodium dihydrogen phosphate buffer (pH 3.0) as mobile phase.The detection wavelength was 346 nm and the column temperature was 3℃0 .The in vitro recovery of berberine hydrochloride was studied using positive dialysis and retrodialysis methods at different flow rates and concentrations.The in vivo recovery was also analyzed at a certain flow rate but different concentrations.Results: When the concentration of berberine hydrochloride was 5.0 μg·mL^-1 and the flow rates were 1, 2 and 3 μL·min^-1, the in vitro recoveries were (73.6±5.4)%, (60.4±5.8)% and (35.2±6.3)%, respectively, as detected using positive dialysis;while using retrodialysis, the in vitro recoveries were (70.2±7.0)%, (62.6±4.7)% and (32.5±8.3)% , respectively.When the flow rate was 2 μL·min^-1 and the concentrations of berberine hydrochloride were 0.1, 0.5 and 5.0 μg·mL^-1, the in vivo recoveries were (51.4±3.5)%,(53.2±2.7)% and (52.6±2.4)%, respectively.The recoveries were inversely proportional to the flow rates and had nothing to do with the concentration of berberine hydrochloride.The in vitro recoveries detected using positive dialysis and retrodialysis methods were similar.Conclusion: The established retrodialysis method can be used to adjust the in vivo recoveries in the study of berberine hydrochloride in biological samples by microdialysis.