盐酸氯苯胍原料药主成分含量及有关物质的测定

Determination of Principle Components and Related Substances of Raw Material Drug of Robenidine Hydrochloride

为建立测定盐酸氯苯胍原料药中主成分含量及有关物质的超高效液相色谱法,采用ACQUITY UPLC BEH C18柱(1.7μm,2.1mm×100mm),以乙腈-2mL/L磷酸水(V∶V=40∶60)为流动相,检测波长为313nm,进样量10μL。在优化的色谱条件下,主成分与各杂质峰分离度良好,盐酸氯苯胍在15μg/mL～35μg/mL范围内线性关系良好(R2=0.999),低、中、高浓度样品的平均回收率分别为100.85%、100.61%、100.40%,相对标准偏差(RSD)分别为1.50%、0.07%、0.64%。盐酸氯苯胍的主要杂质为保留时间1.612的R1和5.416的R2,以主成分自身对照法计算,暂定单个杂质峰面积不得大于对照溶液主峰面积的0.16倍(0.16%),各杂质峰面积的和不得大于对照溶液主峰面积的0.26倍(0.26%)。该方法分离时间短,检出限低,分离度好,结果准确、可靠,可作为盐酸氯苯胍及其有关物质的检测方法。

An analytical method for the determination of robenidine hydrochloride and related substances by ultra high performance liquid chromatography(UPLC)was established.The assay was carried out on a ACQUITY UPLC BEH C18 column(1.7μm,2.1 mm×100 mm)with acetonitrile-0.2%phosphoric acid aqueous solution(V∶V=40∶60)as mobile phase,and the detection wavelength was set at 313 nm.The injection volumn was 10μL.Under the separation conditions,the calibration curves of analytes showed good linearities within the mass concentration of 15μg/mL-35μg/mL(R2=0.999).The average recoveries of samples with low,medium,and high concentrations were 100.85%,100.61%,100.40%with the relative standard deviation(RSD)were 1.50%,0.07%,0.64%,respectively.The major impurities of robenidine hydrochloride were R1 and R2,whose retention time was 1.612 and 5.416 respectively.The peak area of a single impurity was not more than 0.16 times(0.16%)of the peak area of robenidine hydrochloride in the control solution.The whole peak areas of each impurity were not more than 0.26 times(0.26%)the peak area of robenidine hydrochloride in the control solution.The method can be used for the determination of robenidine hydrochloride for the short separation time,low detection limit,good resolution and accurate and reliable results.