高效液相色谱法同时测定盐酸二甲双胍中5种杂质的含量

Simultaneous Determination of Five Impurities in Metformin Hydrochloride by High Performance Liquid Chromatography

目的:借助高效液相色谱法同时测定盐酸二甲双胍中的5种杂质的含量,为盐酸二甲双胍药物中杂质成分分析探索方法。方法:选取盐酸二甲双胍原料药,色谱柱为Ultimate HILIC AmphionⅡ,流动相选取乙腈-25 mmol·L^(-1)磷酸二氢钾,柱温45℃,每次进样量为20μL。检测时所用波长为213 nm,流动相的流速为1.0 mL·min^(-1),对原料药中的5种杂质进行测定,分析杂质含量、试验的重复性、稳定性。结果:经检测,盐酸二甲双胍和杂质甲、乙、丙、丁及戊均能有效分离,且分离度为2.0以上。各项杂质的峰面积RSD分别为0.64%、0.58%、0.52%、0.61%和2.1%,均在3.0%以内;杂质甲、乙、丙、丁和戊检测限分别是11.84μg·L^(-1)、29.36μg·L^(-1)、10.76μg·L^(-1)、10.57μg·L^(-1)和180.26μg·L^(-1),而其定量限分别为39.35μg·L^(-1)、98.79μg·L^(-1)、36.57μg·L^(-1)、35.41μg·L^(-1)和605.78μg·L^(-1)。经分析,这一检测方法及仪器的重复性及稳定性试验结果良好,各项杂质在不同浓度之下,回收率为92.9%~104.3%,平均回收率的RSD在3.0%以内。用该方法对4个厂家生产的盐酸二甲双胍原料药中杂质进行分析,发现4个厂家所生产的盐酸二甲双胍原料药中,杂质的含量均符合相关要求。结论:高效液相色谱法同时测定盐酸二甲双胍中的5种杂质含量的方法可靠、结果准确,可推荐应用。

Objective: The contents of five impurities in metformin hydrochloride were determined simultaneously by high performance liquid chromatography, so as to explore a method for the analysis of impurities in metformin hydrochloride.Method: Metformin hydrochloride API was selected, the chromatcolumn was Ultimate HILIC AmphionⅡ, mobile phase selected acetonitrile-25 mmol·L^(-1)potassium dihydrogen phosphate, column temperature of 45 ℃, each sample intake of 20 μL. The wavelength was 213 nm and the flow rate of the mobile phase was 1.0 mL·min^(-1). Five impurities in the API were determined, and the impurity content and the repeatability of the test were analyzed. Result: Metformin hydrochloride and impurities were effectively separated and separated above 2.0. The peak area RSD of each impurity was 0.64%, 0.58%,0.52%, 0.61% and 2.1%, all within 3.0%;the detection limit of each impurity was 11.84 μg·L^(-1), 29.36 μg·L^(-1), 10.76 μg·L^(-1),10.57 μg·L^(-1), 180.26 μg·L^(-1), and its quantitative limit was 39.35 μg·L^(-1), 98.79 μg·L^(-1), 36.57 μg·L^(-1), 35.41 μg·L^(-1)and 605.78 μg·L^(-1). After analysis, the repeatability and stability test results of this detection method and instrument were good.The recovery rate of various impurities was ranging from 92.9% to 104.3% under different concentrations, and the average recovery rate of RSD was within 3.0%. This method was used to analyze the impurities in metformin hydrochloride API produced by four manufacturers. It was found that the content of impurities in metformin hydrochloride API produced by four manufacturers met the relevant requirements. Conclusion: The simultaneous determination of five impurities in metformin hydrochloride is reliable and recommended need.