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目的:建立高效液相色谱法测定灌胃大鼠血清中甘草苷、异甘草苷、异甘草素、甘草次酸的含量。方法:取大鼠血清样品,采用液液萃取技术,经高效液相色谱进行分离测定。采用Dikma Technologies-C18ODS(200mm×4.6mm,5μm)色谱柱,分别以乙腈-0.01%磷酸0.05mol/L磷酸二氢钾、甲醇-5%醋酸水为流动相,梯度洗脱,柱温30℃。结果:甘草苷血清浓度在0.1355-43.3600μg/mL范围内线性良好;异甘草苷血清浓度在0.0506-5.0600μg/mL范围内线性良好;异甘草素血清浓度在0.02503-1.00100μg/mL范围内线性良好;甘草次酸血清浓度在0.1014-4.0560μg/mL范围内线性良好。结论:该方法专属性强,分析速度快、准确性好,适用于甘草提取物灌胃大鼠的血药浓度测定及药动学研究。
Objective: To establish a HPLC method for the determination of serum glycyrrhizin, isoliquiritin, isoliquiritigenin and glycyrrhetinic acid in gavage rats. Methods: Rat serum samples were taken and separated by liquid-liquid extraction using HPLC. The chromatographic column was Dikma Technologies-C18 ODS (200 mm × 4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.01% phosphoric acid 0.05 mol / L potassium dihydrogen phosphate and methanol-5% . Results: The linearity of glycyrrhizin in the range of 0.1355-43.3600μg / mL was good; the concentration of isoliquiritin was linear in the range of 0.0506-5.0600μg / mL; the concentration of isoliquiritigenin in the range of 0.02503-1.00100μg / mL was linear Good; glycyrrhetinic acid serum concentration in the range of 0.1014-4.0560μg / mL good linearity. Conclusion: The method is specific, rapid, accurate and suitable for the determination of blood concentration and pharmacokinetics of glycyrrhiza extract in rats.