HPLC法测定犬血浆中葛根素含量

HPLC Determination of Puerarine in Dog Plasma

目的:建立血浆中葛根素含量测定方法。方法:色谱柱:Waters Symmetry C18柱(150 mm×3.9 mm,5μm);流动相:乙腈-0.0075 mol·L-1磷酸盐缓冲液(pH 2.7)(10:90);测定波长:248 nm;柱温:40℃;流速:1 mL·min-1。血浆样品采用固相萃取处理。并用健康家犬6只,单剂量给予葛根总黄酮片600 mg,进行了血药浓度测定。结果:血浆样品中葛根素无干扰,最低检测限为0.1 ng·mL-1,葛根素在0.0175-2.2μg·mL-1范围内血浆药物浓度与峰面积线性关系良好,r=0.9995,回收率为85.0%-117.1%,日内、日间RSD分别为7.5%-11.6%和4.7%-8.7%。结论:本法样品处理方法简单,无干扰,提取完全,灵敏度高,重现性好,可用于葛根素生物样本分析。

Objective: To establish an HPLC method for determination of puerarine in plasma. Methods: Waters Symmetry C18 column(150 mm×3. 9 mm,5μm) ,mobile phase was acetonitrile -0. 0075 mol·L-1 potassium di-hydrogen phosphate buffer( pH 2. 7 ) (10:90 ) ; flow rate was 1 mL·min-1; puerarine was extracted from dog plasma by solid phase extract (SPE) method and then detected in 248 nm wavelength. Blood samples were obtained at 0, 0.25,1.0,1.5,2.0,3.0,4.0,5.0,7.0,9.0,12 h after a single dose of 600 mg total puerariae flavones(TPF)tablets. Results: Puerarine was saparated completely without interference. The peak area of puerarine showed good linearity within the range between 0. 0175 -2. 2μg·mL-1 ,r =0. 9995. The LOD of puerarine in plasma was 0. 1 ng·mL-1. The recovery was 85. 0% - 117. 1% ,the within - and between - day RSD was 7. 5% -11. 6% and 4. 7% -8. 7% respectively. Conclusion: This method is simple and sensitive with good recovery,and can be used in analyzing puerarin in biological sample.