色谱柱参数对盐酸四环素有关物质HPLC分析的影响研究

摘要：目的 分析色谱柱参数对盐酸四环素有关物质HPLC分析的影响,筛选影响关键杂质分离度以及理论塔板数的主要柱参数。方法 针对《中国药典》2020版中盐酸四环素有关物质检查的色谱条件,使用多个品牌不同型号的色谱柱进行色谱分析,使用SPSS软件进行模型回归,得到柱参数与目标因子分离度及理论塔板数的回归方程,并用实测值进行验证。结果 色谱柱参数C是影响杂质4-差向四环素和土霉素分离度(R12)的重要参数,利用柱参数C可快速预测不同色谱柱的相似度,进而筛选适宜四环素分析的色谱柱;探讨了金霉素理论塔板数(R5N)与柱参数之间的关系,其中参数B2对R5N影响较大。结论 建立柱参数与色谱分析目标因子之间的联系,为盐酸四环素有关物质测定选择合适的色谱柱提供便利与指导。

Study on the influence of chromatographic column parameters on the HPLC analysis of tetracycline hydrochloride related substances

Abstract Objective The influence of chromatographic column parameters on the HPLC determinationof tetracycline hydrochloride related substances was analyzed, and the main parameters that affect the resolutionof impurities and the number of theoretical plates were screened. Methods According to the chromatographicconditions for the determination of tetracycline hydrochloride related substances in Chinese Pharmacopoeia2020 edition, chromatographic analysis was performed using a variety of brands and models of chromatographiccolumns, and model regression was performed by SPSS software. The regression equation about column parameters,resolution between objective factors, and theoretical plate number was obtained and verified by real experiments.Results Column parameter C was an important parameter that affects the resolution (R12) of 4-epitetracyclineand oxytetracycline. Parameter C might replace similarity calculation as a convenient means for column screeningin tetracycline analysis. The relationship between the theoretical plate number (R5N) and column parameterswas discussed and parameter B2 had the greatest impact on R5N. Conclusion Relationship between the columnparameters and the target factors of impurity analysis was established to provide convenience and guidance for theselection of suitable chromatographic columns for determination of tetracycline hydrochloride related