HPLC测定磷酸二甲啡烷原料药中的三个异构体杂质

目的 建立磷酸二甲啡烷原料药中3个异构体杂质的定量分析方法。方法 流动相为正己烷-异丙醇-甲醇-三氟乙酸-二乙胺(850∶100∶50∶1∶0.5),流速为1.0 mL·min^-1,采用CHIRALCEL^® OZ-H(4.6 mm×250 mm,5 μm)色谱柱,检测波长为215 nm,柱温35 ℃,进样量20 μL,以主成分自身对照法计算异构体含量。结果 磷酸二甲啡烷主峰与对映异构体杂质M分离度大于1.5、杂质G与杂质I之间分离度大于1.0杂质,M、G、I分别在(0.518 0~1.554 0) (0.508 0~1.524 0) (0.507 5~1.522 5) μg·mL^-1内线性关系良好(相关系数r分别为0.995 0、0.999 5、0.999 5);相对主成分的校正因子分别为1.19、0.99、1.03;加样回收率分别为112.3%、88.3%、102.9%(n=9);定量限为0.508 5、0.526 0、0.507 5 μg·mL^-1;检测限为0.152 6、0.157 8、0.169 2 μg·mL^-1。3批样品结果异构体杂质M最大值为0.017%,杂质G、I均小于检测限。结论 该方法简便,准确度,重复性良好,可用于检测磷酸二甲啡烷合成工艺中产生的3个异构体杂质。

Determination of Three Isomer Impurities in Dimemorfan Phosphate by HPLC

OBJECTIVE To establish a quantitative analysis method for isomer impurities in dimemorfan phosphate APIs. METHODS The mobile phase was n-hexane-iso-propanol-methanol-trifluoroacetic acid-diethylamine (850∶100∶50∶1∶0.5), the flow rate was 1.0 mL·min^-1, and CHIRALCEL^® OZ-H column(4.6 mm×250 mm, 5 μm)was adopted. The column temperature was maintained at 35 ℃, the sample injection quantity was 20 μL, and the detection wavelength was set at 215 nm. The isomer content was calculated by the principal component self-control method. RESULTS The resolution between the peak of dimemorfan phosphate and the impurity M was good (greater than 1.5), and that between impurity G and I was greater than 1.0. The linear ranges of impurity M, G and I were (0.518 0-1.554 0), (0.508 0-1.524 0), (0.507 5-1.522 5) μg·mL^-1 (correlation coefficients of 0.995 0, 0.999 5 and 0.999 5), the correction factors of relative principal components were 1.19, 0.99 and 1.03, the recoveries of samples were 112.3%, 88.3%, and 102.9% (n=9), the quantitative limits were 0.508 5, 0.526 0, 0.507 5 μg·mL^-1, and the detection limits were 0.152 6, 0.157 8, and 0.169 2 μg·mL^-1, respectively. The precision RSDs were less than 2.0%. The analysis results of three batches of samples showed that the maximum content of impurity M was 0.017%, and the contents of impurity G and I were less than the detection limits. CONCLUSION The method is simple, accurate and repeatable. It can be used to detect three isomer impurities produced in the synthesis process of dimemorfan phosphate.