HPLC法检查苯甲酸阿格列汀原料药的有关物质

Determination of Related Substances in Alogliptin Benzoate Raw Materials by HPLC

目的：建立检查苯甲酸阿格列汀原料药中有关物质的方法。方法：采用高效液相色谱梯度洗脱法。色谱柱为C18,流动相A为0.2%磷酸溶液（pH 3.0）,流动相B为乙腈,以1.0 ml/min流速进行梯度洗脱,检测波长为225 nm,进样量为20μl。采用不加校正因子主成分自身对照法检查特定杂质F-1的量。结果：苯甲酸阿格列汀与F-1及其他未知杂质均分离良好,前二者检查质量浓度线性范围分别为0.187 2~0.748 8、0.184 5~0.738 0μg/ml（r=0.999 7、0.999 9）;F-1的检测限和定量限分别为2.5、7.5 ng,平均回收率为100.9%（RSD=1.10%,n=3）;3批样品中F-1量均≤0.06%。结论：建立的方法灵敏快速、结果准确可靠,可作为苯甲酸阿格列汀原料药的有关物质检查方法。

OBJECTIVE： To establish a method for the determination of related substances in Alogliptin benzoate （AB） raw materials. METHODS： HPLC-gradient elutior method was adopted. The determination was performed on C18 column with mobile phase A consisted of 0.2% phosphoric acid solution （pH adjusted to 3.0） and mobile phase B consisted of acetonitrile at the flow rate of 1.0 ml/min （gradient elution）. The detection wavelength was set at 225 nm and sample size was 20 μl. The content of impu- rity F-1 was determined by the self-control method without correction factor. RESULTS： AB, F-1 and other impurities were completely separated. The linear range was 0.187 2-0.748 8 μg/ml for AB and 0.184 5-0.738 0 μg/ml for F-1 （r=0.999 7, 0.999 9）. The limits of detection and quantification were 2.5 ng and 7.5 ng for F-1. The average recovery was 100.9% （RSD=1.10%, n= 3）; the content of F-1 in 3 batches of samples were ≤0.06%. CONCLUSIONS： The method is sensitive, rapid, accurate and reliable, and can be used for the determination of related substance in AB raw materials.