绵羊血浆中多拉菌素的高效液相色谱检测  

Determination of Doramectin in Plasma of Sheep by HPLC with Fluorescence Detection

作　　者：石艳丽[1] 韩彩霞[1] 张子群 李晓云[1] 宋铭忻[1] 李巍[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]黑龙江出入境检验检疫局,黑龙江哈尔滨150036

出　　处：《中国兽医杂志》2014年第1期73-75,78,共4页Chinese Journal of Veterinary Medicine

基　　金：哈尔滨市科技创新人才研究专项资金(2006RFQXN016);"十一五"黑龙江省科技攻关计划项目(GA06B202-3)

摘　　要：本试验建立了绵羊血浆中多拉菌素的荧光高效液相色谱（HPLC）检测法。用乙腈作为血浆中蛋白质的沉淀剂和多拉菌素的提取剂，离心后取上清液用旋转真空蒸发器挥干溶剂，用5％乙腈-水溶解残渣后用ODS—C18固相萃取法对提取的多拉菌素进行纯化。用氮气在60℃沙浴中对提取和纯化后的多拉菌素进行干燥。在无水条件下，用三氟乙酸酐和N-甲基咪唑作荧光衍生化试剂，在常温避光条件下对多拉菌素进行荧光衍生化。用荧光HPLC进行检测，检测条件：流动相为无水甲醇，流速1．0mL／min，激发波长365nm，发射波长475nm，进样量20乩，柱温为20℃。结果表明，本方法的检测限为0．1ng／mL，在1－100ng／mL血浆浓度范围内与峰面积呈良好的线性关系；样品的回收率为89．5％～95．67％；不同浓度水平的日内变异系数和日间变异系数分别小于4％和5％。结果显示，本方法样品提取、纯化和衍生化过程准确、灵敏度高、干扰少、重复性好。A novel method was developed to determinate doramectin in plasma of sheep using high-perfonnance liquid chromatography （HPLC） with fluorescence detection. Doramectin in plasma of sheep was extracted using acetonitrile, purified by solid phase extraction on a ODS-C 18 column. and dried in airless box.Subsequently, it was modified with trifluoroacetic anhydride and N methyl glyoxaline at room temperature. After being injected into the HPLC column, the doramectin was detected by fluorescence detector under 365 nm excitation and 475 nm emission wave lengths.The limit of quantitation was 0.1 ng / mL. The method generated good linear calibration graphs within the concentrations from 1 to 100 ng / mL.The mean recovery rates of Doramectin ranged from 89.50% to 95.67%. The coefficient of variability within one day and among several days was smaller than 4% and 5 %, respectively. This study provides a rapid, reliable, and sensitive method for determination of doramectin in plasma of sheep.

关 键 词：多拉菌素 固相萃取 荧光高效液相色谱 

分 类 号：S858.26[农业科学—临床兽医学]