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当前位置: 首页 > 文章 > 奶牛用利福昔明子宫注入剂有关物质反相高效液相色谱检测方法的建立及应用 中国畜牧兽医 2017,44 (4) 1211-1218
Position: Home > Articles > Establishment and Application of Reversed-phase High Performance Liquid Chromatography Method for Detection of Related Substances of Rifaximin Uterine Infusion in Dairy Cows China Animal Husbandry & Veterinary Medicine 2017,44 (4) 1211-1218

奶牛用利福昔明子宫注入剂有关物质反相高效液相色谱检测方法的建立及应用

作  者：
黄慧丽;徐飞;张道康;华伟毅;李秀波;刘义明
单  位：
中国农业科学院饲料研究所国家饲料药物基准实验室
关键词：
利福昔明;子宫注入剂;有关物质;高效液相色谱法;专属性
摘  要：
本试验旨在建立利福昔明有关物质的检测方法。采用反相高效液相色谱法,紫外检测器进行检测,采用WondaSil C18-WR 250mm×4.6mm 5μm色谱柱,柱温:40℃,流动相及其比例为:V(甲醇)∶V(乙腈)∶V(3.16g/L甲酸铵,浓氨水调pH至7.2)=31.5∶31.5∶37,检测波长276nm。采用面积归一化法计算利福昔明子宫注入剂的检测限、定量限及其浓度;考察了方法精密度、线性相关性及有关物质专属性,并通过极端条件的破坏、有关物质的含量、分离度等对主成分峰面积降解程度进行了比较。利福昔明子宫注入剂主成分峰在浓度为40~150μg/mL的范围内,峰面积与浓度呈现良好的线性关系(R2=0.9998),其有关物质浓度对峰面积线性回归方程为y=23 510x+7 182,其进样精密度RSD为0.13%,方法精密度RSD为0.46%,制剂一定时间内经过强酸、强碱、氧化、高温条件破坏后均有不同程度降解,以破坏前的样品主成分含量为100.00%,氧化破坏和高温破坏主成分降解达13%,酸、碱破坏主成分降解在18%左右,主成分峰保留时间在以上4种条件破坏前后一致,且主成分峰与杂质峰之间及杂质峰与杂质峰之间分离度良好(主成分峰与杂质峰之间分离度均大于1.5,杂质峰与杂质峰之间分离度均大于1.2)。本试验建立的利福昔明子宫注入剂有关物质检测方法操作简便,专属性和重复性好,可适用于利福昔明子宫注入剂有关物质的测定。
译  名：
Establishment and Application of Reversed-phase High Performance Liquid Chromatography Method for Detection of Related Substances of Rifaximin Uterine Infusion in Dairy Cows
作  者：
HUANG Hui-li;XU Fei;ZHANG Dao-kang;HUA Wei-yi;LI Xiu-bo;LIU Yi-ming;National Feed Drug Reference Laboratories,Feed Research Institute,Chinese Academy of Agricultural Sciences;
关键词：
rifaximin;;uterine injectants;;related substance;;HPLC;;specificity
摘  要：
The study was aimed to establish a method for detection of rifaximin related substances by reversed-phase high-performance liquid chromatography with UV detector.The instrument methods was WondaSil C18-WR 250 mm ×4.6mm 5μm column,mobile phase was methanol∶acetonitrile∶3.16g/L ammonium formate(concentrated aqueous ammonia adjusted to pH 7.2)=31.5∶31.5∶37,with detection wavelength 276 nm.Area normalization method was used to calculate the limit of detection,limit of quantification and concentration of rifaximin uterine injectant;The precision of the method,linear correlation and related substances specificity were explored.And the degradation degree of the peak area of the principal components was compared by the destruction of the extreme conditions,the content of the related substances and the separationdegree.40 to 150μg/mL concentration of rifaximin,peak area and concentration showed a good linear relationship(R2=0.9998),the related substances concentration on peak area linear regression equation was y=23 510x+7 182,within a certain period of time after preparation of acids,alkalis,oxidation,high temperature degradation damage in varying degrees,in the main ingredient content in the sample before the damage was 100.00%,after oxidative damage and heat damage the main component was 13%,acids,alkalis destruction of the main component degradation was about 18%,the main component peak retention time was consistency before and after the destruction.The retention time and resolution of rifaximin and related-substance were good.(The separation between the main component peak and the impurity peaks were greater than 1.5,the degree of separation between the impurity peak and impurity peaks greater than 1.2).The substance detection methods of rifaximin uterine injectants was simple and had specificity and good repeatability,selective and sensitive,which could be used to analyze the related substance of rifaximin uterine injectants.

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