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目的:建立大鼠血浆中青藤碱的高效液相色谱测定方法。方法:以茶碱为内标,血浆样品经二氯甲烷萃取;采用Diamonsil C18色谱柱(200mm×4.6mm,5μm);流动相:甲醇-30mmol.L-1磷酸二氢钾-三乙胺(40:60:0.05);流速:1.0mL.min-1;柱温:35℃;检测波长:263nm。结果:血浆中内源性物质不干扰青藤碱和内标的测定,且在0.2～50.0mg.L-1范围内线性关系良好(r=0.9985),定量下限为0.2mg.L-1;该方法的日内精密度、日间精密度和准确度均符合要求,低、中、高浓度的回收率分别为73.1%、81.1%、85.9%。结论:本方法简便灵敏,干扰小,可用于血浆中青藤碱浓度的测定以及药动学研究。
Objective: To establish an HPLC method for the determination of sinomenine in rat plasma. METHODS: Theophylline was used as the internal standard and the plasma samples were extracted with dichloromethane. Diamonsil C18 column (200mm×4.6mm, 5μm) was used. Mobile phase: methanol-30mmol.L-1 potassium dihydrogen phosphate-triethylamine ( 40:60:0.05); flow rate: 1.0 mL.min-1; column temperature: 35°C; detection wavelength: 263 nm. RESULTS: Endogenous substances in plasma did not interfere with the determination of sinomenine and internal standards, and the linear relationship was good in the range of 0.2-50.0 mg.L-1 (r=0.9985), and the lower limit of quantification was 0.2 mg.L-1; The intra-day precision, inter-day precision and accuracy of the method met the requirements. The recoveries for the low, medium and high concentrations were 73.1%, 81.1% and 85.9%, respectively. Conclusion: This method is simple and sensitive, with little interference, and can be used for the determination of plasma sinomenine concentration and pharmacokinetic study.