牛奶中埃谱利诺菌素(eprinom ectin)荧光高效液相色谱检测方法的建立  

Determination of Eprinomectin in Milk of Cow by HPLC with Fluorescence Detection

作　　者：潘保良[1] 汪明[1] 王玉万[2] 潘贞德[2] 戴晓曦[2] 高凌云[2] 那顺[2] 

机构地区：[1]中国农业大学动物医学院,北京100094 [2]中国农业大学生物技术股份有限公司,北京100094

出　　处：《中国兽医学报》2004年第2期174-176,共3页Chinese Journal of Veterinary Science

基　　金：国家"十五"攻关课题项目 (2 0 0 2 BA5 14 A-17-0 2 )

摘　　要：建立了以多拉菌素 (doramectin,DOR)作为内标检测牛奶中埃谱利诺菌素 (eprinomectin,EPR)的荧光高效液相色谱 (HPL C)检测法。用乙腈作为牛奶中蛋白质的沉淀剂和 EPR的提取剂。用 ODS- C1 8固相萃取法对提取的 EPR进行纯化。用真空干燥箱对提取和纯化后的 EPR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剂 ,加入冰醋酸 ,在加热条件下对干燥后的 EPR进行荧光衍生化。用荧光 HPL C进行检测 ,检测条件 :流动相为 A液(为乙腈与无水甲醇按 1∶ 2的比例配制而成 )与水之比为 98∶ 2 ,流速为 1.0 m L/ m in,激发波长为 36 6 nm,发射波长为 4 6 5 nm,进样量为 10 0 μL,柱温为室温。本方法的检测限为 0 .1μg/ L,在 1～ 4 0 μg/ L 血浆的浓度范围内 ,标准曲线方程为 Y=0 .1787x- 0 .117,R2 =0 .997;样品的回收率为 83.4 0 %～ 10 5 .6 5 % ;变异系数为 6 .32 %～ 9.0 5 %。结果显示 :本方法灵敏度高 ,干扰少 ,重复性好。A novel method by high-performance liquid chromatography(HPLC) with fluorescence detection was developed to determinate eprinomectin(EPR) in milk of cow.Samples were extracted with acetonitrile,cleaned up by solid phase extraction(SPE) on a C_(18) column and dried in airless box.Then EPR was derived with trifluoroacetic anhydride and N-methylimidazole,addition of acetic acid,heat at 65℃.Then the samples were injected into the HPLC column.EPR was detected by fluorescence detector set at 366 nm excitation and 465 nm emission wavelength.The method gave linear calibration graphs(R^2=0.997) over the concentration between 1 and 40 μg/L.The limit of quantification was 0.1 μg/L EPR in milk of cow.The mean recovery ranged between 83.40% and 105.40%.The procedure provides a rapid,reliable and sensitive method for determination of eprinomectin in milk.

关 键 词：埃谱利诺菌素 DOR 内标法 固相萃取 荧光高效液相色谱 牛奶 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程] S859.796[轻工技术与工程—食品科学与工程]