UPLC-PDA法测定甘草浸膏中甘草苷和甘草酸的含量

Determination of Liquiritin and Glycyrrhizic Acidin Extractum Glycyrrhizae by UPLC-PDA

建立超高效液相色谱(UPLC)测定甘草浸膏中主成分甘草苷和甘草酸的含量。采用反相高效液相色谱法对甘草苷和甘草酸进行色谱分离和快速定量测定。以ACQUITY UPLC^TM T 3色谱柱(2.1 mm×100 mm,1.8μm)为分离柱,柱温:35℃;以乙腈-水(0.1%冰乙酸+5 mmol/L醋酸铵)进行梯度洗脱;流速:0.35 mL/min;进样量:1μL;检测波长为232 nm。通过光谱图、保留时间和峰面积参数对甘草苷和甘草酸进行定性定量检测。甘草苷在1~20μg/mL范围内线性关系良好(R^2>0.999),方法检出限为0.5μg/mL,定量限为1μg/mL;甘草酸在10~200μg/mL范围内线性良好(R^2>0.999),方法检出限为5μg/mL,定量限为10μg/mL,完全满足检测需求。该方法色谱分离较好,分析速度较快,前处理简单,适用于甘草浸膏中甘草苷和甘草酸的定性定量检测。

This study aims to establish a high performance liquid chromatography(UPLC-PDA)test method for the determination of liquiritin and glycyrrhizic acid in extractum glycyrrhizae.UPLC was used to isolate and quantitatively determine liquiritin and glycyrrhizic acid.ACQUITY UPLC^TM T 3 chromatographic column(2.1 mm×100 mm,1.8 mm)was used as the separation column,column temperature 35℃;gradient elution was performed with acetonitrile-water(0.1%glacial acetic acid+5 mmol/L ammonium acetate)solution as mobile phase;The flow rate was 0.35 mL/min,the injection volume was 1μL,and the wavelength was 232 nm.In this paper,the quantitative and qualitative determination of liquiritin and glycyrrhizic acid was carried out by spectrogram,retention time and peak area parameters.In the range of 1~20μg/mL of liquiritin,the linear relationship was good(R^2>0.999);the limit of detection was 0.5μg/mL,and the limit of quantitation was 1μg/mL;In the range of 10~200μg/mL of glycyrrhizic acid,the linear relationship was good(R^2>0.999);The limit of detection was 5μg/mL(S/N>3),and the limit of quantitation was 10μg/mL(S/N>10),which fully met the requirements.The test method was suitable for the qualitative and quantitative determination of liquiritin and glycyrrhizic acid in extractum glycyrrhizae for its good separation,quick analysis and easy sample treatment performance.