HPLC法测定非那雄胺中潜在基因毒性杂质DDQ和DDHQ

Determination of Potential Genotoxic Impurities DDQ and DDHQ in Finasteride by HPLC

目的建立测定非那雄胺中基毒杂质DDQ和DDHQ的HPLC梯度洗脱方法。方法采用GL Science Wondasil C_(18)色谱柱(4.6 mm×150 mm,5μm),以0.04 mol·L^(-1)磷酸二氢钾溶液为流动相A,以甲醇为流动相B,检测波长为220 nm。结果非那雄胺与DDQ和DDHQ之间的分离良好,DDQ在0.06~1.12μg·mL^(-1)浓度范围内,DDHQ在0.06~1.19μg·mL^(-1)浓度范围内,与相应的峰面积呈良好的线性关系(R^(2)>0.999);DDQ和DDHQ的定量限分别为1.12 ng、1.19 n g,检测限分别为0.56 ng、0.60 ng;6批原料药中均未检出DDQ和DDHQ。结论经方法学验证,本方法专属性强,准确度高,适用于非那雄胺中DDQ和DDHQ的测定。

OBJECTIVE To establish an HPLC gradient elution method for the determination of the DDQ and DDHQ of finasteride.METHODS The HPLC method was carried out using a GL Science Wondasil C_(18)(4.6 mm×150 mm,5μm);the mobile phase A was 0.04 mol·L^(-1) potassium dihydrogen phosphate aqueous solution,and the mobile phase B was methanol;the detection wavelength was 220 nm.RESULTS Finasteride,DDQ and DDHQ were separated completely under the above chromatographic conditions.The calibration curve was linear for DDQ and DDHQ in the range of 0.06-1.11μg·mL^(-1)(R^(2)>0.999).The limits of detection and limit of quantification of DDQ were 0.56 ng and 1.12 ng.The limits of detection and limit of quantification of DDHQ were 0.60 ng and 1.19 ng.the impurity was not detected in six batches of finasteride.CONCLUSION The method was applicable for the quality control of DDQ and DDHQ in finasteride through the methodology validation.