氨糖美辛肠溶片中盐酸氨基葡萄糖有关物质研究

Study on Related Substances of Glucosamine Hydrochloride in Glucosamine Indometacin Enteric - Coated Tablets

目的 建立测定氨糖美辛肠溶片中盐酸氨基葡萄糖有关物质的高效液相色谱法,并分析其影响因素。方法 色谱柱为纳谱ChromCore AQ C18柱(250 mm×4.6 mm,3μm),流动相为水-乙腈(梯度洗脱),流速为1.0 mL/min,检测波长为280 nm,柱温为30℃,进样量为20μL。采用加速稳定性试验和辅料相容性试验,结合生产工艺分析有关物质的影响因素。结果 已知杂质果糖嗪、2,5-脱氧果糖嗪、吡嗪、5-羟甲基糠醛、糠醛、2-甲基吡嗪、吡咯-2-甲醛、5-甲基糠醛质量浓度分别在0.20~3.30μg/mL、0.20~3.00μg/mL、0.25~7.54μg/mL、0.18~7.04μg/mL、0.25~10.10μg/mL、0.25~10.18μg/mL、0.25~10.17μg/mL、0.26~10.50μg/mL范围内与峰面积线性关系良好(R2≥0.999);检测限为0.3~5.0μg/mL,定量限为0.7~20.1μg/mL;精密度、稳定性、重复性试验结果的RSD均小于2.0%;平均加样回收率为98.24%~101.20%,RSD为0.33%~2.19%(n=9)。影响制剂有关物质产生的因素为工艺、温度和时间,影响有关物质含量增长速率的因素除工艺外,还有辅料(主要为羟甲基淀粉钠)和水分。结论 该方法简便、准确、专属性好,可用于氨糖美辛肠溶制剂中盐酸氨基葡萄糖有关物质的检测。建议优化工艺中干燥温度及时间,考察添加辅料的合理性,并控制片剂中的水分,进一步提高制剂质量的均一性和稳定性。

Objective To establish a high-performance liquid chromatography(HPLC)method for the determination of related substances of glucosamine hydrochloride in Glucosamine Indometacin Enteric-Coated Tablets,and to analyze their influencing factors.Methods The chromatographic column was the NanoChrom ChromCore AQ C18 column(250 mm×4.6 mm,3μm),the mobile phase was water-acetonitrile(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was 280 nm,the column temperature was 30℃,and the injection volume was 20μL.The accelerated stability test and excipient compatibility test were used to analyze the influencing factors of related substances based on the production process.Results The linear ranges of known impurities including fructosazine,2,5-deoxyfructosazine,pyrazine,5-hydroxymethylfurfural,furfural,2-methylpyrazine,pyrrole-2-carboxaldehyde,5-methylfurfural were 0.20-3.30μg/mL,0.20-3.00μg/mL,0.25- 7. 54 μg / mL,0. 18 - 7. 04 μg / mL,0. 25 - 10. 10 μg / mL,0. 25 - 10. 18 μg / mL,0. 25 - 10. 17 μg / mL,0. 26 -10. 50 μg / mL (R2 ≥ 0. 999) respectively. The limit of detection was in the range of 0. 3 - 5. 0 μg / mL,the limit ofquantification was in the range of 0. 7 - 20. 1 μg / mL. The RSDs of precision,stability,and repeatability tests were all lower than2. 0%. The average recovery rate of the above known impurities was in the range of 98. 24% - 101. 20%,with RSDs in the rangeof 0. 33% - 2. 19% (n = 9). The factors that affected the generation of related substances in the formulation were process,temperature,and time;the factors that affected the growth rate of related substance content included process,excipients (mainlysodium hydroxymethyl starch) and moisture. Conclusion This method is simple,accurate,and specific,which can be used for thedetection of related substances of glucosamine hydrochloride in Glucosamine Indometacin Enteric - Coated Tablets. We shouldoptimize the drying temperature and time in the process,improve the rationality of adding excipients,and control the moisture inthe tablets to further improve the uniformity and stability of the formulation quality.