建立了中空纤维液相微萃取(HFLPME)耦合高效液相色谱法(HPLC)用于测定血浆和尿液中大黄素及其代谢物的浓度,比较了中药有效成分大黄素在不同性别大鼠体内的吸收和代谢能力,阐述了大黄素在体内的代谢和转化过程.本实验以聚偏氟乙烯纤维为溶剂载体,正辛醇为萃取溶剂,对血浆和尿液样品进行HFLPME处理,萃取后挥干有机溶剂,用50μL甲醇溶解,进行HPLC测定.在优化的微萃取条件下,血浆和尿液样品中大黄素及其代谢物标准曲线线性良好(相关系数(r)大于0.9960)；检出限为0.1～3.0μg/L;富集倍数为12.2～26.3；日内、日间精密度(以相对标准偏差(RSD)计)小于11.0％；血浆和尿液中代谢物的平均回收率为97.9％～103％.HFLPME操作简单,富集倍数高,能有效去除生物样品中复杂基体的干扰,适用于复杂样品中微量、痕量成分分析物的分析测定.%Hollow fiber liquid phase microextraction (HFLPME) coupled with high performance liquid chromatography (HPLC) has been developed to analyze the emodin and its metabolites in plasma and urine samples. The abilities of the absorption and metabolism for the active components in traditional Chinese medicines between the male and female rats were compared, and the biological metabolism and transmutation of the analyte were detailed discussed. Emodin and its metabolites in plasma and urine samples were extracted into n-octanol (acceptor) in hollow fiber. The acceptor phase was dried and dissolved by 50μL methanol and then analyzed by HPLC. Under the optimal conditions, the linearities of the analytes were all very good in biological samples (r >0. 996 0), the detection limits of the analytes were within the ranges of 0. 1 -3. 0 μg/L. The enrichment factors were 12. 2 to 26. 3. The relative standard deviations for intra-day and inter-day precision were lower than 11. 0%. The average recoveries of the analytes in plasma and urine samples were all in the range of 97. 9% to 103%. HFLPME-HPLC can eliminate interference from complex biological samples, improve the sensitivity and reduce the detection limit, thus this method is suitable for the determination of trace compounds in complex sample.
展开▼
