高效液相色谱法测定注射用盐酸甲氯芬酯有关物质及质量评价研究

目的：建立注射用盐酸甲氯芬酯有关物质测定的高效液相色谱分析方法，并对药品质量进行评价。方法：采用资生堂MGII C₁₈（4.6 mm×150 mm，5 μm）色谱柱，流动相为0.05 mol·L^-1辛烷磺酸钠溶液（磷酸调节pH至2.5）-乙腈（65：35），流速为1.0 mL·min^-1，柱温为25℃，检测波长为225 nm，进样量10 μL，分析时间为主成分峰保留时间的4倍。结果：考察了有机相比例和缓冲液pH对系统分离情况的影响，根据破坏试验结果，并兼顾分离度要求，确定了有关物质色谱条件并进行方法学验证。盐酸甲氯芬酯检测下限为1 ng，水解杂质（杂质A）、醇解杂质B和C的检测下限分别为0.2、0.7和1.4 ng，新建方法在14批样品中检出现行标准中未能检出的醇解杂质C。结论：方法灵敏度高，专属性强，操作简便，解决了现行标准中盐酸甲氯芬酯有关物质检查法色谱条件不完善等问题，为该系列质量标准的修订提供数据参考，并为药品的科学监管和注射剂的仿制药质量一致性评价提供有力的技术支撑。

Objective: To establish an high performance liquid chromatography method for the determination of related substances of meclofenoxate for injection and evaluate its quality. Methods: The separation was performed on SHISEIDO MGII C₁₈ column (4.6 mm×150 mm, 5 μm) with 0.05 mol·L^-1 sodium octane sulfonate solution (phosphoric acid adjusted to pH 2.5) -acetonitrile (65:35) as the mobile phase at flow rate of 1.0 mL·min^-1. The column temperature was 25, the detection wavelength was 225 nm, the injection volume was 10 μL, and the analysis time was 4 times of the peak retention time of the main component. Results: The effects of organic phase and buffer pH on the separation of the system were investigated. According to the results of the failure tests and the resolution requirements, the chromatographic conditions of the substances were determined and the method validation was carried out. The detection limit of metoclopramide hydrochloride was 1 ng. The detection limits of the impurity A, impurity B and the impurity C were 0.2 ng, 0.7 ng and 1.4 ng, respectively. The new method can effectively detect alcohol-dissolved impurity C which could not be detected in the current standard in 14 batches of samples. Conclusion: The method has high sensitivity, specificity, and simple operation which solves the problems of imperfect chromatographic conditions of the relevant substances in the current standard, and provides data reference for the revision of the series of quality standards. It also provides strong technical support for the scientific supervision of drugs and the consistent evaluation of the quality of generic injections.

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