测定大鼠血浆中士的宁和马钱子碱含量的方法研究

Research Method on Content Determination of Strychnine and Brucine in Rat Plasma

目的:探讨可以同时测定大鼠血浆中士的宁和马钱子碱含量的方法研究。方法:Wistar大鼠血浆样品经液液萃取处理后用Waters Acquity UPLC BEH C18色谱柱分离,流动相为乙腈-酸水(17?83),酸水为0.02 mol·L^(-1)磷酸二氢钾与0.014 8 mol·L^(-1)庚烷磺酸钠等量混合,并用10%磷酸调体系p H为2.8;检测波长为260 nm。结果:Wistar大鼠血浆中杂质不干扰样品的含量测定,士的宁含量在0.067 84-3.392 00μg·m L^(-1),马钱子碱含量在0.052 48-2.624 00μg·m L^(-1),线性良好,日内、日间精密度均小于10%,平均回收率均大于85%,稳定性符合体内药物分析要求。结论:UPLC检测法灵敏度高、精密度好,可应用于通脉丸中士的宁与马钱子碱药物动力学研究。

This study was aimed to establish an effective method for the simultaneous content determination of strychnine and brucine in rat plasma. The Wistar rat plasma samples were processed by liquid-liquid extraction and separated on a UPLC BEH C18 column. The mobile phase was acetonitrile- acid water（17？83）. The acid water was 0.02 mol·L-1. The potassium dihydrogen phosphate and 0.014 8 mol·L-1 sodium heptane sulfonate was mixed with equal amount. The 10% phosphonic acid was used to adjust the system to the p H of 2.8. The detection wavelength was set at 260 nm. The results showed that foreign materials in the Wistar rat plasma did not interfere with the content determination of samples. The calibration curve of strychnine was in good linearity within the range of 0.067 84-3.392 00 μg·m L-1. The calibration curve of brucine was in good linearity with the range of 0.052 48- 2.624 00 μg·m L-1. The intra-and inter-day precisions were less than 10%. The mean extraction recoveries were more than 85%. The stability results met the requirements for biopharmaceutical analysis. It was concluded that the UPLC detection method was sensitive and precise, which can be used in the pharmacokinetics study of strychnine and brucine in Tong-Mai（TM） pill.