HPLC-ELSD法测定硫酸庆大霉素碳酸铋胶囊中庆大霉素C组分及有关物质含量

HPLC-ELSD analysis of the components of gentamicin C and related substances in gentamicin sulfate and bismuth carbonate capsules

目的为硫酸庆大霉素碳酸铋胶囊中庆大霉素C组分及有关物质建立质控方法。方法采用HPLC-ELSD方法,色谱柱:GRACEApollo C18柱(250 mm×4.6 mm,5μm);流动相:0.2 mol/L三氟乙酸溶液-甲醇(96:4,V/V),流速0.6 mL/min;柱温30℃;进样量20μL;Waters低温分流型蒸发光散射检测器,载气流量40 psi,漂移管温度55℃。结果所建方法通过方法学验证,庆大霉素C组分与各杂质间分离完全,庆大霉素、西索米星和小诺霉素分别在0.5~6 mg/mL、25~100μg/mL和25~100μg/mL的浓度范围内,各浓度的对数值与对应色谱峰面积的对数值呈现出良好的线性。采用所建方法对市售16家生产企业43批次硫酸庆大霉素碳酸铋胶囊样品进行测定,获得该制剂产品中各组分及有关物质的数据。结论所建方法是在中国药典方法基础上优化并验证适用于硫酸庆大霉素碳酸铋胶囊C组分及有关物质的含量测定,方法简便准确,专属性强,稳定性好。由获得的实验数据可知,该复方制剂中C组分及有关物质的含量在各厂家间或同厂家不同批次间差异显著,可能存在影响用药安全的潜在隐患,建议在该剂型质量标准中增加相关质控项。

Objective This study aimed to establish a quality control method for the contents of gentamicin C components and related substances in gentamycin sulfate and bismuth carbonate capsules.Methods HPLC-ELSD method was used.The column was GRACEApollo C18(250 mm×4.6 mm,5μm),the mobile phase was 0.2 mol/L trifluoroacetic acid solution:methanol(96:4,V/V),the flow rate was 0.6 mL/min,the column temperature was 30℃,the injection volume was 20μL,the Waters low-temperature diverging evaporative light scattering detector was used,carrier gas flow was 40 psi,and the drift tube temperature was 55℃.Results The method was validated and performed well on the separation of gentamicin C components and other related substances.Good linearities were observed between the logarithmic values of each concentration and the corresponding chromatographic peak area for gentamicin,sisomicin and micronomicin with concentration ranges of 0.5~6 mg/mL,25~100μg/mL and 25~100μg/mL,respectively.Samples of 43 batches of gentamicin sulfate and bismuth carbonate capsules from 16 companies on the market were tested using the method that had already been set up.This gave information about the ingredients and other substances in the preparation product.Conclusion The method was developed and optimized on the basis of Chinese Pharmacopeia with the strengths of convenience,specificity and robustness.It is validated for the determination of the content and related substances of gentamicin C components in gentamycin sulfate and bismuth carbonate capsules.The results showed significant differences in the content and related substances in the capsules from different manufacturers and batches,which indicated substantial risks for medication safety.Therefore,it was suggested that the method be considered in the quality control of such capsules.