反相高效液相色谱法同时测定甘草酸单铵盐原料药主成分及有关物质含量

建立了同时测定甘草酸单铵盐原料药中主成分18α-甘草酸、18β-甘草酸及其有关物质A、有关物质B含量的高效液相色谱法,并用于其质量标准的建立。采用Durashell C₁₈色谱柱(250 mm×4.6 mm, 5 μm),以10 mmol/L高氯酸铵(氨水调节pH 8.20)-甲醇(48:52, v/v)为流动相,流速0.80 mL/min,检测波长254 nm,柱温50 ℃,进样量10 μL。在优化的色谱条件下,18α-甘草酸、18β-甘草酸、有关物质A、有关物质B在0.50~100 mg/L范围内线性关系良好(r>0.9999),检出限分别为0.15、0.10、0.10、0.15 mg/L,平均回收率在97.32%~99.33%之间(n=3),相对标准偏差(RSD)在0.05%~1.06%之间。本方法检测灵敏、重现性好,结果准确可靠,可用于甘草酸单铵盐原料药主成分及有关物质的检测分析,有利于其原料药的质量控制。

Simultaneous determination of principal components and related substances of raw material drug of ammonium glycyrrhizinate by reversed-phase high performance liquid chromatography

An analytical method for the simultaneous determination of 18α-glycyrrhizic acid, 18β-glycyrrhizinic acid, related substances A and B and drug quality standard by reversed-phase high performance liquid chromatography (RP-HPLC) was established. The assay was carried out on a Durashell-C₁₈ column (250 mm×4.6 mm, 5 μm) with 10 mmol/L ammonium perchlorate (the pH value was adjusted to 8.20 with ammonia)-methanol (48:52, v/v) as mobile phase at a flow rate of 0.80 mL/min, and the detection wavelength was set at 254 nm. The column temperature was 50 ℃ and the injection volume was 10 μL. Under the separation conditions, the calibration curves of the analytes showed good linearities within the mass concentrations of 0.50-100 mg/L (r>0.9999). The detection limits for 18α-glycyrrhizic acid, 18β-glycyrrhizinic acid, related substances A and B were 0.15, 0.10, 0.10, 0.15 mg/L, respectively. The average recoveries were between 97.32% and 99.33% (n=3) with the relative standard deviations (RSDs) between 0.05% and 1.06%. The method is sensitive, reproducible, and the results are accurate and reliable. The method can be used for the determination of principal components and related substances of ammonium glycyrrhizinate for the quality control of raw material drug of ammonium glycyrrhizinate.