平衡透析法测定羊耳菊提取物中9个成分的血浆蛋白结合率

为了测定民族药羊耳菊提取物中9个成分的血浆蛋白结合率。首先采用平衡透析法测定提取物中9个成分血浆蛋白结合率,生物样本用甲醇沉淀蛋白法进行处理,再以葛根素为内标,利用超高效液相色谱-质谱仪串联测定血浆及缓冲溶液中的绿原酸、新绿原酸、隐绿原酸、东莨菪苷、1,3-O-二咖啡酰基奎宁酸、3,4-O-二咖啡酰基奎宁酸、3,5-O-二咖啡酰基奎宁酸和4,5-O-二咖啡酰基奎宁酸等9个成分的浓度。结果发现在所研究浓度范围内,9个成分呈现良好的线性关系(r≥0.999),准确度、精密度、提取回收率和稳定性等均符合生物样品测定要求,且并无内源性杂质干扰,表明该文所建立的方法稳定可靠,可用于羊耳菊药材中9种成分在人和大鼠血浆中的血浆蛋白结合率测定。9种成分在人和大鼠血浆中的平均蛋白结合率分别在(41.07±0.046)%～(94.95±0.008)%,(37.66±0.043)%～(97.46±0.013)%。结果表明羊耳菊提取物中9个成分在人和大鼠中的血浆蛋白结合率存在差异。

Determination of plasma protein binding rates of nine compounds of Inula cappa extraction based on method of equilibrium dialysis

To determine the plasma protein binding rate of the nine compounds in Inula cappa extraction by the method of equilibrium dialysis. The proteins in plasma samples were precipitated by methanol, and the ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for determination of the concentrations of the nine active compounds, namely chlorogenic acid, scopolin, neochlorogenic acid, cryptochlorogenic acid, 1,3-O-dicaffeoylquinic acid, galuteolin, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, with the internal standard of puerarin. We found that all components have a good linearity(r≥0.999), and accuracy, precision, extraction recovery and stability conformed to the requirements of determination, without endogenous compounds disturbing within the range of optimum concentration. This suggested that the method was stable and reliable, and could be used for the determination of the plasma protein binding rates of the nine active compounds in rat and human plasma of I. cappa. The plasma protein binding rates of the nine active compounds in rat and human plasma respectively were(41.07±0.046)%-(94.95±0.008)%, and(37.66±0.043)%-(97.46±0.013)%. According to the results, there were differences in the plasma protein binding rates of the nine compounds in I. cappa extraction between rat and human.