HPLC法测定阿加曲班原料药的含量及异构体比例

Determination of Argatroban and the Ratio of Isomer by HPLC

目的建立测定阿加曲班原料药含量及异构体比例的高效液相色谱(HPLC)法。方法采用Agilent ZORBAX SB-C18(100 mm×4.6 mm,1.8μm)色谱柱,以0.1%醋酸铵溶液(用冰醋酸调节p H值至5.5)-甲醇-乙腈(70∶15∶15,V/V)为流动相,检测波长为260 nm。结果阿加曲班的线性范围为159.7~372.6μg·m L^(-1)(r=1.000,n=5),定量限为0.3μg·m L^(-1)。结论本法简便准确,专属性强,分离度好,可用于测定阿加曲班原料药的含量及异构体比例。

OBJECTIVE To established an HPLC method for the determination of Argatroban and the ratio of isomer.METHODS The sample was separated on an Agilent ZORBAX SB-C18（ 100 mm×4.6 mm,1.8 μm） with the mobile phase consisted of 0.1% ammonium acetate（ adjusted with acetic acid to p H 5.5）-methanol-acetonitrile（ 70 ∶15 ∶15,V/V）. The detection wavelength was set at 260 nm. RESULTS There was a good linear relationship for Argatroban in the range of 159. 7-372. 6 μg·m L^（-1）（ r = 1.000,n = 5）.The limit of quantitation was 0.3 μg·m L^（-1）. CONCLUSION The method is simple,accurate and has strong specialization for Argatroban and the ratio of isomer.