RP-HPLC法测定DHDK原料药中11种有关物质的含量

Determination of eleven related substances in DHDK drug by RP-HPLC method

目的建立RP-HPLC法同时测定DHDK原料药中11种有关物质的含量。方法采用Diamonsil C_(18)(250 mm×4.6 mm,5μm)色谱柱,以乙腈-体积分数0.1%磷酸水为流动相进行梯度洗脱,流速为1.0 mL·min^(-1),检测波长为220 nm,柱温30℃,进样量20μL。结果DHDK与有关物质A~K及未知杂质均能达到良好分离;杂质A在质量浓度0.180~2.000 mg·L^(-1)、杂质B、C和J在质量浓度0.096~2.000 mg·L^(-1)、杂质D、F和H在质量浓度0.144~2.000 mg·L^(-1)、杂质E和G在质量浓度0.060~2.000 mg·L^(-1)、杂质I在质量浓度0.216~4.000 mg·L^(-1)、杂质K在质量浓度0.312~10.000 mg·L^(-1)内与峰面积呈良好的线性关系(r≥0.9990);回收率在92%~108%(RSD≤5.0,n=9)内。结论该方法操作简便、专属性强,能够准确测定DHDK原料药中11种有关物质的含量。

Objective To establish an RP-HPLC method for simultaneous quantification of related su-bstances in DHDK drug.Methods The determination was performed on a Diamonsil C_(18)(250 m m×4.6 mm,5μm)column with mobile phase A consisted of acetonitrile and mobile phase B con-sisted of 0.1%phosphoric acid aqueous solution at the flow rate of 1.0 mL·min^(-1)using gradient e-lution.The detection wavelength was set at 220 nm and sample size was 20μL,and the column t-emperature was set at 30℃.Results DHDK could be effectively separated with the related substan-ces A-K and unknown impurities.The linear range of the related substance A was 0.180-2.000 mg·L^(-1)、the linear range of the related substances B、C、J was 0.096-2.000 mg·L^(-1)、the linear ra-nge of the related substances D、F、H was 0.144-2.000 mg·L^(-1)、the linear range of the related s-ubstances E、G was 0.060-2.000 mg·L^(-1)、the linear range of the related substances I was 0.216-4.000 mg·L^(-1)、the linear range of the related substances K was 0.312-10.000 mg·L^(-1)(r≥0.9990).The recovery was between 92%and 108%(RSD≤5.0,n=9).Conclusion The method is sample and spe-cfic,which can be used for the determination of the related substances in DHDK drug.