HPLC-PAD法测定硫酸依替米星氯化钠注射液中有关物质

Determination of related substances in etimicin sulfate and sodium chloride injection by HPLC- PAD

目的：建立硫酸依替米星氯化钠注射液有关物质的反相高效液相色谱-脉冲安培电化学（HPLC-PAD）检测方法。方法：采用Welch Ultimate LP-C18（4.6 mm×300 mm，5 μm）色谱柱，柱温30 ℃，以0.2 mol·L-1三氟乙酸（用50%氢氧化钠溶液调pH至2.0）-乙腈（92：8）为流动相，流速1.0 mL·min-1，柱后碱溶液为0.5 mol·L-1氢氧化钠溶液，流速0.5 mL·min-1，检测器为脉冲安培电化学检测器，工作电极为金电极（3 mm直径）。结果：依替米星在0.3~30 μg·mL-1（r=0.9985）内线性关系良好；单个最大杂质的重复性试验的RSD（n=6）为0.8%，总杂质的重复性试验的RSD（n=6）为1.3%；方法的检测限（S/N=3）为2.5 ng，定量限（S/N=10）为7.5 ng；供试品溶液在12 h内稳定性良好。结论：该方法经方法学验证，可以用于硫酸依替米星氯化钠注射液有关物质的测定。

Objective： To establish an HPLC-PAD method for the determination of the related substances in etimicin sulfate and sodium chloride injection. Methods： The separation was performed on a Welch Ultimate LP-C18column （4.6 mm×300 mm,5 μm） and detected by pulsed amperometric detection （PAD） on a gold electrode （diameter：3 mm）.The column temperature was set at 30 ℃.The mobile phase consisted of an aqueous solution containing 0.2 mol·L-1 trifluoroacetic acid （pH adjusted to 2.0 with 50% sodium hydroxide solution） and acetonitrile （92：8）,the flow rate was 1.0 mL·min-1.The alkaline solution after column was 0.5 mol·L-1 sodium hydroxide solution at a flow rate of 0.5 mL·min-1. Results： The linear rang of etimicin was 0.3-30 μg·mL-1（r=0.9985）.The repeatability RSDs （n=6） of the maximum impurity and total impurities in a sample were 0.8% and 1.3%,respectively.The LOD （S/N=3） and LOQ （S/N=10） were 2.5 ng and 7.5 ng,respectively.The sample solution was stable within 12 h. Conclusion： The method is simple,accurate and sensitive,and suitable for the determination of the related substances in etimicin sulfate and sodium chloride injection.