RP-HPLC梯度洗脱法测定盐酸金霉素有关物质方法研究

Determination of the Related Substances in Chlortetracycline Hydrochloride by RP-HPLC with Gradient Elution

目的 建立准确可靠的RP-HPLC梯度洗脱法用于测定盐酸金霉素原料药有关物质。方法 采用Ultimate XB-C8色谱柱(4.6 mm×250 mm,5μm);以高氯酸-水-二甲基亚砜(16∶934∶525)(pH<2.0)、乙腈、甲醇分别作为流动相A、B、C,进行梯度洗脱;柱温为45℃;检测波长为280 nm;流速为0.8 mL/min。结果 与《中国药典》2020年版相比,检出杂质多2个,盐酸金霉素与11个已知杂质分离度良好。13批样品均检出6个已知杂质,除盐酸四环素、4-差向金霉素外,还检出杂质B、E、J、L,其中杂质B、E为主要其他杂质。结论 该方法适用性好、专属性强,可真实反映盐酸金霉素有关物质状况,为其质量控制提供方法支撑。

Objective To establish a accurate and reliable RP-HPLC method for the determination of the related substances in chlortetracycline hydrochloride.Methods The determination was performed on the Ultimate XB-C8 column(4.6 mm×250 mm,5µm).The mobile phase consisted of the mobile phase A[perchloric acid-water-dimethyl sulfoxide(16∶934∶525)(pH<2.0)]and mobile phase B(methanol)and mobile phase C(acetonitrile)with gradient elution.The column temperature was maintained at 45℃,the detection wavelength was set at 280 nm and the flow rate was 0.8 mL/min.Results Compared with the current quality standard of“Chinese Pharmacopoeia”2020 Edition,two more impurities were detected.The chlortetracycline hydrochloride and eleven known impurities can be completely separated from each other,and six known impurities were detected in the 13 batches of samples.Besides the tetracycline hydrochloride and 4-epichlortetracycline,the impurities B,E,L and J were also detected.And the impurities B and E are the main other impurities.Conclusion The method has good applicability and strong specificity,can truly reflect the status of related substances of the chlortetracycline hydrochloride and provide the methodological support for its quality control.