高效液相色谱法测定扎那米韦含量及有关物质

HPLC determination of zanamivir content and the related substances

目的:建立测定扎那米韦的含量和有关物质的高效液相方法。方法:采用NH2P-50色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-5 mmol·L^(-1)硫酸溶液(60∶40)[用氨水调节pH至(6.2±0.05)];检测波长为234和210 nm;流速为1.0 mL·min^(-1),柱温为30℃。同时采用液质联用对各杂质进行了结构确证。结果:主药与各杂质、各杂质之间分离度良好,各杂质能有效检出。在100.10~1 001.0μg·mL^(-1)浓度范围内,扎那米韦与其峰面积呈良好的线性关系(r=0.999 9,n=5),最小检测限为25.0 ng·mL^(-1),含量测定及有关物质检查的中间精密度RSD分别为0.24%(n=6)与1.24%(n=6)。结论:本法简便、精密、专属、灵敏,可用于扎那米韦含量和有关物质测定。

Objective： To develop an HPLC method for simultaneously determining zanamivir content and its related substances for quality control. Methods： The method was performed on an Asahipak NH2P-50 column （ 250 mm x 4.6 mm, 5 Ixm） with acetonitrile-5 mmol. L - 1 sulphuric acid （60： 40） [ adjusting pH to （6.2 ± 0.05 ） with ammonia solution ] as the mobile phase. The flow rate was 1.0 mL· min-l. The detection wavelength was 234 and 210 nm, and the column temperature was set at 30 ℃. Meanwhile, the structure of each impurity was confirmed by LCMS. Results： Zanamivir was completely separated from the related substances. The calibration curve of zanamivir was linear （ r = 0. 999 9, n = 5 ） in the range of 100.10 ± 1 001.0 μg·mL^（-1） ; detection limit was 25.0 μg·mL^（-1） the intermediate precisions RSD were 0.24% and 1.24% （n =6） in the content and related substance determination. Conclusion： The method is simple, accurate, specific and sensitive for simultaneously determining the content and its related substances of zanamivir.