目的: 建立HPLC法研究呋塞米及其注射液的杂质谱,同时检测和分析来源于国内外多家企业的产品,评价该产品的杂质控制现状,以及注射液的主要降解杂质与处方、包装材料的相关性。方法: 采用YMC Hydrosphere C18(250 mm×4.6 mm,3 μm)色谱柱,以0.05%三氟乙酸溶液(pH 2.23)-甲醇-乙腈为流动相,梯度洗脱,流速1.0 mL·min-1,检测波长238 nm和277 nm,柱温30 ℃。计算各杂质与呋塞米的相对保留时间(RRT),已知杂质采用加校正因子(f)的主成分自身对照法计算,未知杂质为不加校正因子的主成分自身对照法。结果: 建立了呋塞米及其注射液中11个已知杂质和1个潜在基因毒杂质的检测方法,各杂质分离度均符合要求,通过强制降解试验归属了各杂质来源。2~3家企业注射液中降解杂质C和降解杂质G的检出量比ICH的鉴定限度大0.2%,5家企业注射液中潜在基因毒杂质糠醛的检出量远高于参比制剂。结论: 建立的HPLC方法可用于呋塞米及其注射液中杂质的快速检测分析,国内只有2家企业生产的注射液杂质控制水平基本与参比制剂一致,其他企业需优化处方和包装材料,本研究可为提高呋塞米注射液的安全性和仿制药质量一致性评价研究提供参考。
Objective: To establish an HPLC method for determination of the impurity profile in furosemide and its injection. At the same time,applying this method to detect and analyze products from many domestic and foreign enterprises,to evaluate the status quo of impurity control in API and injection,and the correlation of major degraded impurities in injection with prescription and packaging materials. Methods: A YMC Hydrosphere C18 column(250 mm×4.6 mm,3 μm)was used,and the mobile phase was 0.05% TFA solution (pH 2.23)-methanol-acetonitrile at the flow rate of 1.0 mL·min-1,gradient elution. Detection wavelength was set at 238 nm and 277 nm and column temperature was 30 ℃. Impurity reference was used for localization and the relative retention time of each impurity was calculated. The known impurities were calculated using the principal component self-control method with correction factor,the unknown impurities were determined by principal component without correction factor. Results: The method for the determination of 11 kinds of known impurities, the potential genotoxic impurity in furosemide, its injection was established,the separation degree of all impurities met the requirements,and the sources of the impurities were identified by forced degradation test. The detected quantity of degraded impurity C and degraded impurity G in the injections produced by 2 to 3 companies exceeded the identification limit in ICH by 0.2%, the amount of potential genotoxic impurity furfural detected in the injection of 5 enterprises was much higher than that of the reference preparation,other impurities were the same as the reference preparation. Conclusion: The established HPLC method can be used for rapid detection and analysis of impurities in furosemide and its injection. The impurity control level of injection produced by only 2 domestic enterprises is basically consistent with the reference preparation,others need to optimize prescription and packaging materials. This study can provide reference for improving the safety of furosemide injection and evaluating the quality consistency of generic drugs.
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