高效液相色谱法测定白龙解郁颗粒中橙皮苷和甘草苷的含量

Determination of hesperidin and liquiritin in Bailong Jieyu granule by high-performance liquid chromatography

目的采用高效液相色谱法(HPLC)测定白龙解郁颗粒中橙皮苷和甘草苷的含量。方法以十八烷基硅烷键合硅胶为填充剂,使用Agilent SB-ZOBAX C18柱(250 mm×4.6 mm,5μm)行HPLC。橙皮苷的测定:乙腈-0.1%磷酸溶液(22∶78)为流动相,柱温30℃,检测波长为283nm,进样量10μL;甘草苷的测定:乙腈-0.1%磷酸溶液(18∶82)为流动相,柱温30℃,检测波长为276nm,进样量10μL。结果 HPLC测定橙皮苷和甘草苷的精密度、重复性、稳定性的范围均符合相关规定;橙皮苷在35.64~178.20μg/mL范围内线性关系良好(r=0.999 8),甘草苷在13.55~216.80μg/mL范围内线性关系良好(r=0.999 9);加样回收率分别为:橙皮苷99.30%,RSD为1.0%(n=9);甘草苷98.60%,RSD为1.5%(n=9)。结论 HPLC快捷、简单、稳定可靠,可用于白龙解郁颗粒的质量控制。

Objective To determine the contents of the hesperidin and liquiritin in Bailong Jieyu granule with high-performance liquid chromatography（HPLC）.Methods HPLC was performed with octadecylsilane chemically bonded silica as filler using the Agilent SB-ZOBAX C18 Column（250 mm×4.6 mm,5μm）.Hesperidin was detected with the mobile phase consisting of acetonitrile and 0.1% of phosphoric acid solution（22∶78）,with column temperature being 30 ℃,the detection wavelength being 283 nm,and the injection volume being 10μL.Liquiritin was determined with acetonitrile-0.1% of phosphoric acid solution（18∶82）as mobile phase,with column temperature being30 ℃,the detection wavelength being 276 nm,and the injection volume being 10μL.Results Accuracy,repeatability and stability of HPLC for determination of hesperidin and liquiritin were in line with the relevant requirements.The linear relationship of hesperidin was good within the range of 35.64-178.20μg/mL（r=0.999 8）,with the average recovery rate being 99.30%,and relative standard deviation（RSD）being 1.0%（n=9）.The good linear range of liquiritin was 13.55-216.80μg/mL（r=0.999 9）,with the average recovery rate being 98.60% and RSD being 1.5%（n=9）.Conclusion HPLC method was fast,simple,stable and reliable,and it can be used for the quality control of Bailong Jieyu granule.