摘要
目的:建立硫酸依替米星注射液有关物质及含量的反相高效液相色谱-脉冲安培电化学(HPLC-PAD)法。方法:采用十八烷基硅烷键合硅胶为填充剂(4.6 mm×250 mm,5μm或效能相当的色谱柱),柱温35℃。以0.2 mol·L^-1三氟乙酸溶液(含0.05%五氟丙酸,1.5 g·L^-1无水硫酸钠,0.8%(V/V)的50%氢氧化钠溶液,用50%氢氧化钠溶液调节pH值至3.5)-乙腈(95∶5)为流动相,流速为每分钟1.0 mL。柱后加碱(50%氢氧化钠溶液1→25,流速每分钟0.5 mL)。用积分脉冲安培电化学检测器检测。结果:依替米星在0.075~50μg·mL^-1(r=0.9996)内线性关系良好;单个最大杂质的重复性试验的RSD(n=6)为1.4%,总杂质的重复性试验的RSD(n=6)为1.7%,含量测定的重复性试验的RSD(n=6)为0.8%;方法的检测限(S/N=3)为75 ng·mL^-1,定量限(S/N=10)为250 ng·mL^-1;供试品溶液在12 h内稳定性良好。9批样品用药典方法(反相高效液相色谱-蒸发光散射法)和HPLC-PAD法测定,HPLC-PAD法的总杂和杂质个数结果偏高,其中6批样品HPLC-PAD法的含量结果偏低。结论:经方法学验证,该方法可以用于硫酸依替米星注射液有关物质及含量的测定。
Objective:To establish an HPLC-PAD method for determination of the related substances and assay of etimicin sulfate injection.Methods:The separation was performed on a C 18 column(4.6 mm×250 mm,5μm or equivalent column).The column temperature was set at 35℃.The mobile phase consisted 50 mL acetonitrile and 950 mL of 0.2 mol·L^-1 trifluoroacetic acid solution(contain 0.05%pentafluoropropionic acid,1.5 g·L^-1 sodium sulphate and 0.8%(V/V)of 50%NaOH solution;adjust pH to 3.5 with 50%NaOH solution).The flow rate was 1.0 mL·min^-1.The post column was alkalized with NaOH(50%NaOH 1→25)at a flow rate of 0.5 mL·min^-1.The pulsed amperometric detection(PAD)was used.Results:The linear range of etimicin fell into 0.075-50μg·mL^-1(r=0.9996).The repeatability RSDs(n=6)of the maximum impurity and total impurities were 1.4%and 1.7%respectively,and the repeatability RSD(n=6)of assay was 0.8%.The LOD(S/N=3)and LOQ(S/N=10)were 75 ng·mL^-1 and 250 ng·mL^-1 respectively.The test solution was stable with in 12 h.Nine batches of samples were determined by pharmacopoeia method(RP HPLC-ELSD)and HPLC-PAD method.The contents of total impurities and number of impurities obtained with HPLC-PAD method were higher,but the results of assay of six batches of samples obtained with HPLC-PAD method were lower.Conclusion:The validation results show that the established method is suitable for determination of the related substances and assay of eimicin sulfate injection.
作者
朱晓玥
袁耀佐
张玫
ZHU Xiaoyue;YUAN Yaozuo;ZHANG Mei(Jiangsu Institute for Food and Drug Control,Nanjing 210019,China)
出处
《中国药品标准》
CAS
2019年第6期532-536,共5页
Drug Standards of China
