HPLC测定达比加群酯中的有关物质

Determination of the Related Substances of Dabigatran Etexilate by HPLC

关键词:  达比加群酯  高效液相色谱法  有关物质

Key words:  dabigatran etexilate  HPLC  related substances

摘要:

目的 建立达比加群酯原料药中有关物质的测定方法。方法 采用Agilent SB-C₁₈色谱柱(250 mm×4.6 mm，5 μm)，以乙腈(A)-0.2%的醋酸铵(B，用冰醋酸调节pH值至4.4)为流动相进行梯度洗脱：0~18 min，90%→40%B；18~30 min，40%B。流速为1.0 mL·min^-1，检测波长为340 nm。结果 各杂质与主峰之间的分离度良好。5个已知杂质：杂质A浓度在0.117 0~1.872 μg·mL^-1内与峰面积呈良好的线性关系，r为0.999 7；杂质B浓度在0.126 5~2.024 μg·mL^-1内与峰面积呈良好的线性关系，r为0.999 5；杂质C浓度在0.113 0~1.808 μg·mL^-1内与峰面积呈良好的线性关系，r为0.999 5；杂质D浓度在0.120 5~1.928 μg·mL^-1内与峰面积呈良好的线性关系，r为0.999 9；杂质E浓度在0.123 0~1.968 μg·mL^-1内与峰面积呈良好的线性关系，r为0.999 6；杂质A、杂质B、杂质C、杂质D和杂质E加样回收率的平均值分别为98.75%，98.91%，98.39%，99.0%和99.73%；RSD分别为0.91%，1.09%，1.22%，1.35%和1.18%。结论 本方法简便、准确可靠，适用于达比加群酯中有关物质的控制。

Abstract:

OBJECTIVE To develop a method for determing the content of related substances of dabigatran etexilate. METHODS The condition of detection were: Agilent SB-C₁₈ column (250 mm×4.6 mm, 5 μm), the gradient mobile phase consisted of acetonitrile(A) and 0.2% ammonium acetate (B, adjust with glacial acetic acid to pH 4.4): 0-18 min, 90%→40%B; 18-30 min, 40%B. The flow rate was 1.0 mL·min^-1, the UV detection wavelength was 340 nm. RESULTS The resolution of dabigatran etexilate and relate substances was good. The calibration curves were linear in the range of 0.117 0-1.872 μg·mL^-1 for Impurity A (r=0.999 7); the calibration curves were linear in the range of 0.126 5-2.024 μg·mL^-1 for Impurity B (r=0.999 5); the calibration curves were linear in the range of 0.113 0-1.808 μg·mL^-1 for Impurity C (r=0.999 5); the calibration curves were linear in the range of 0.120 5-1.928 μg·mL^-1 for Impurity D (r=0.999 9); the calibration curves were linear in the range of 0.123?0-1.968 μg·mL^-1 for Impurity E (r=0.999 6). The average recoveries of Impurity A, Impurity B, Impurity C, Impurity D and Impurity E were 98.75%, 98.91%, 98.39%, 99.0% and 99.73% respectively; the RSD were 0.91%, 1.09%, 1.22%, 1.35% and 1.18%, respectively. CONCLUSION The correction factor method is viable to determine the content of related substances of dabigatran etexilate.