HPLC测定磷酸二甲啡烷原料药中的三个异构体杂质

Determination of Three Isomer Impurities in Dimemorfan Phosphate by HPLC

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,5μm)色谱柱,检测波长为215 nm,柱温35℃,进样量20μL,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(0.5180~1.5540)(0.5080~1.5240)(0.5075~1.5225)μg·mL^(-1)内线性关系良好(相关系数r分别为0.9950、0.9995、0.9995);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0.5085、0.5260、0.5075μg·mL^(-1);检测限为0.1526、0.1578、0.1692μg·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。

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,5μm)was adopted.The column temperature was maintained at 35℃,the sample injection quantity was 20μL,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(0.5180-1.5540),(0.5080-1.5240),(0.5075-1.5225)μg·mL^(-1)(correlation coefficients of 0.9950,0.9995 and 0.9995),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 0.5085,0.5260,0.5075μg·mL^(-1),and the detection limits were 0.1526,0.1578,and 0.1692μg·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.