HPLC-PAD法测定硫酸依替米星氯化钠注射液中有关物质

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.