硫酸头孢喹肟乳房注入剂中有关物质的检测方法  

The Detection Methods of the Related Substances in Cefquinome Sulfate Intramammary Infusion

作　　者：刘茂林[1] 刘义明[1] 徐飞[1] 张道康 黄慧丽[1] 李浛 李秀波[1] 

机构地区：[1]中国农业科学院饲料研究所/国家饲料药物基准实验室,北京100081 [2]北京市兽医药品监察所,北京100107

出　　处：《中国农业科学》2016年第15期3054-3062,共9页Scientia Agricultura Sinica

基　　金：国家奶牛产业技术体系北京市创新团队项目;国家公益性行业(农业)科研专项(201303038);中国农业科学院基本科研业务费专项资金项目(2015YWF-ZD-25)

摘　　要：【目的】参考硫酸头孢喹肟注射液兽药质量标准,建立了硫酸头孢喹肟乳房注入剂有关物质的检测方法,建立专属性强、灵敏度高、快速简便的方法,可有效的分析硫酸头孢喹肟乳房注入剂的有关物质,为制剂的质量控制和工艺优化提供依据。【方法】仪器方法按照硫酸头孢喹肟注射液兽药质量标准要求,色谱条件:流动相为高氯酸钠缓冲溶液(精密称取NaClO_4·H_2O_3.45g于1 000 m L水溶解,加入磷酸12 m L,乙腈115 m L,三乙胺调pH3.6),流速为1 m L·min^(-1),检测波长为270 nm,进样量为20μL。采用高效液相色谱法(HPLC)峰面积归一化法计算硫酸头孢喹肟乳房注入剂(干乳期)中有关物质的检测限及其浓度;考察该检查方法精密度及仪器精密度;考察样品主成分及有关物质的稳定性及线性相关性;对样品以适当条件的破坏后对该方法进行专属性考察,例如使用0.2 mol·L^(-1) HCl溶液、0.2 mol·L^(-1) NaOH溶液对硫酸头孢喹肟乳房注入剂进行4 h的破坏;使用10%H_2O_2溶液对硫酸头孢喹肟乳房注入剂进行30 min的破坏;以及90℃高温对硫酸头孢喹肟乳房注入剂进行1 h的破坏。对比样品破坏前后主成分峰面积的降解程度,有关物质含量的变化程度,主成分峰与杂峰之间及有关物质峰与杂质峰之间的分离度,以及杂质峰个数的变化等主要参数。并对硫酸头孢喹肟乳房注入剂(干乳期)三批样品中的有关物质含量进行测定。【结果】硫酸头孢喹肟乳房注入剂经过一系列的梯度稀释后,当样品主成分浓度为7.5μg·m L^(-1)时,有关物质的信噪比≥3,按HPLC峰面积归一化法计算,有关物质浓度为0.05μg·m L^(-1);100μg·m L^(-1)浓度样品进样精密度RSD达到0.8%,方法精密度RSD达到0.6%;主成分在7.5—300μg·m L^(-1)浓度范围内,按HPLC峰面积归一化法计算,有关物质浓度范围为0.05—2μg·m L^(-1),有关物质浓度对峰面积线性回归分析方程为y=45【Objective】 The detection methods of the related substances in cefquinome sulfate intramammary infusion were optimized by referencing the cefquinome sulfate injection in Veterinary Drug Quality Standard Assembly. A specific, sensitive, simple and rapid method was developed, which could be used to analyze the related substance in cefquinome sulfate intramammary infusion, effectively. And it will be useful for its quality control and process optimization. 【Method】 The instrument method was made by the pharmacopoeia requirements. The separation was performed on sodium perchlorate buffer（3.45 g NaClO4·H2O was dissolved in 1 000 mL water, mixed in 12 mL phosphoric acid and 115 mL acetonitrile, and adjusted pH to 3.6 with triethylamine） as the mobile phase at flow rate of 1.0 mL·min^-1. The detection wave length was 270 nm and the injection amount of sample solution was 20 μL. The LOD and concentration of the related substance in cefquinome sulfate were estimated by HPLC area normalization method. The linearity, the RSD of injection precision and method precision were tested. For the specificity test, by using 0.2 mol·L^-1 HCL, 0.2 mol·L^-1Na OH to destroy the drug for 4 h,using 10% H2O2 to destroy the drug for 30 min and 90℃high temperature to destroy the drug for 1 h. The before and after destroyed samples were detected by HPLC after preparation. And the main-substances＇ area, related-substances＇ concentration, and their resolutions were contrasted with each other, and the number of the peak values of the impurities was also contrasted. 【Result】 After a series of dilutions of the cefquinome sulfate intramammary infusion, when the concerntration of the main component was at 7.5 μg·mL^-1, the S/N of the related-substance was no less than 3, and through calculation of the LOD of the related substance was 0.05 μg·mL^-1, and the RSD of injection precision and method precision of 100 μg·mL^-1 were 0.8% and 0.6%, respectively. The calibration curve of the related substances was a liner equat

关 键 词：硫酸头孢喹肟乳房注入剂 有关物质 HPLC法 

分 类 号：S859.79[农业科学—临床兽医学]