HPLC法测定氨基葡萄糖类药物的有关物质

Determination of the Related Substance in Glucosamine by HPLC

目的:建立高效液相色谱法测定氨基葡萄糖类药物的有关物质的方法,为质量标准提高提供参考。方法:采用高效液相色谱-紫外检测法,色谱柱为岛津Inertsil ODS-3柱(150 mm×4.6 mm,3μm),以庚烷磺酸钠溶液(取庚烷磺酸钠0.50 g,溶解至适量水中,加0.5 ml磷酸和1 mol·L-1的氢氧化钾溶液4 ml,加水至1 000 ml)-乙腈(96∶4)为流动相,流量:1.0 ml·min-1,检测波长为195 nm,柱温:30℃,进样量:20μl。以2-甲基吡嗪作为外标对照品,测定氨基葡萄糖类药物的有关物质的限量。结果:2-甲基吡嗪在1.0~10.0μg·ml-1范围内呈良好的线性关系(r=1.000 0),平均回收率为99.0%,RSD为0.8%(n=9)。结论:该法能有效分离氨基葡萄糖与其降解杂质2,5-果糖嗪、2,5-脱氧果糖嗪,用吸收系数相当的2-甲基吡嗪作为外标对照品控制单个杂质和总杂质的限量,方法简便、准确,专属性强,适用于氨基葡萄糖类药物的质量控制。

Objective: To establish an HPLC method for determining the related substance in glucosamine. Methods: HPLC-UV was applied. An Inertsil ODS-3( 150 mm×4.6 mm,3 μm) column was used and the mobile phase was 0.50 g sodium heptanesulfonate dissolved in water,and 0.5 ml phosphoric acid and 4 ml potassium hydroxide solution( 1 mol·L-1) diluted by water to 1 000 ml-acetonitrile( 96 ∶4). The flow rate was 1.0 ml·min-1,and the detection wavelength was set at 195 nm. Results: A good linear range for 2-methylpyrazine was 1.0-10.0 μg·ml-1( r= 1.000 0),and the average recovery was 99.0%( RSD = 0.8%,n = 9). Conclusion: The method can effectively separate glucosamine from its impurities 2,5-fructosazine and 2,5-Deoxyfructosazine. 2-Methylpyrazine with the same absorption coefficient is used as the external reference for the determinations of single impurity and the total impurities,and the method is simple,accurate and reliable,which is suitable for the quantity control of glucosamine.