HPLC-CAD法测定硫酸新霉素中新霉素B、新霉素C、新霉胺及其他有关物质

Determination of neomycin B, neomycin C, neamine and related substances in neomycin by an HPLC-CAD method

目的建立测定硫酸新霉素中新霉素B、新霉素C、新霉胺及其他有关物质的高效液相色谱-电喷雾检测器(HPLCCAD)方法。方法采用Waters HSS T_3色谱柱(4.6mm×250mm, 5μm),以0.15mol/L三氟乙酸水溶液为流动相,流速1.0mL/min,柱温30℃,电喷雾检测器的雾化温度为45℃。结果新建方法对新霉素B、新霉素C、其他有关物质分离度良好,精密度、重复性和回收率均满足分析要求。新霉素B、新霉胺浓度与峰面积线性关系良好(r>0.995),检出限可达到0.02μg和0.003μg。结论新建方法分离度好、灵敏度高,可以检出更多的杂质,可以满足硫酸新霉素原料有关物质分析的要求。

Objective To establish a high performance liquid chromatography-charged aerosol detector (HPLC-CAD) method for the determination of neomycin B, neomycin C, neamine and related substances in neomycin. Methods A Waters HSS T3 column (4.6mm×250mm, 5μm) was used. An aqueous solution containing 0.15mol/L trifluoroacetate was used as the mobile phase. The flow rate was 1.0mL/min, and the column temperature was 30℃. The spray temperature of CAD was 45℃. Results By using this method, neomycin B, neomycin C and related substances were well separated. The precision, the repeatability, and the recoveries of this method were all acceptable. There was a good linear relationship among the neomycin B, neamine concentration, and the peak area (r>0.995). The detection limit was 0.02μg and 0.003μg. Conclusion The novel method had better resolution and higher sensitivity, which could detect more impurities and meet the content analysis requirements of the related substances in neomycin raw materials.