免疫亲和柱净化-反相高效液相色谱法检测乳与乳制品中黄曲霉毒素M1  

Immunoaffinity column-reversed phase high performance liquid chromatography for detection of aflatoxin M1 in milk and milk products

作　　者：牛军小 苏军[2] 徐晓枫[2] 李伟[2] 单美娜[2] 蒲云霞[2] 

机构地区：[1]内蒙古妇幼保健院,内蒙古呼和浩特010020 [2]内蒙古自治区疾病预防控制中心,内蒙古呼和浩特010030

出　　处：《中国卫生检验杂志》2014年第7期940-941,945,共3页Chinese Journal of Health Laboratory Technology

摘　　要：目的建立免疫亲和柱净化-反相高效液相色谱法（RP-HPLC）测定乳和乳制品中的黄曲霉毒素M1的方法。方法乳及乳制品中黄曲霉毒素M1经包被特异性M1抗体的免疫亲和柱净化后,采用反相C18高效液相色谱柱进行分离,以乙腈-水（15∶85）为流动相,采用等度法洗脱,荧光检测器检测,激发波长365 nm,发射波长435 nm。结果乳及乳制品中的黄曲霉毒素M1在C18高效液相色谱柱上得到很好的分离,保留时间为6.482 min。方法的检出限0.005,定量限0.015μg/kg,相对标准偏差为3.3%-4.6%,平均回收率为80.1%-88.9%之间,相关系数r〉0.9999。结论采用包被特异性M1抗体的免疫亲和柱净化乳及乳制品中的黄曲霉毒素M1,可以有效的去除样本中杂质的干扰,提高了黄曲霉毒素M1检测的灵敏度及准确度,该法适合乳与乳制品中黄曲霉毒素M1测定。Objective To establish RP-HPLC method for determination of the aflatoxin M1 in milk and milk products based on immunoaffinity column purification. Methods The aflatoxin M1 in milk and milk products was purified by immunoaffinity column coated by specific M1 antibody,separated on reversed-phase C18HPLC column with acetonitrile-water（ 15∶ 85） as mobile phase for isocratic elution,then detected by fluorescence detector. The excitation wavelength was 365 nm and the emission wavelength was 435 nm. Results The aflatoxin M1 in milk and milk products was separated well on C18HPLC column. The retention time was 6. 482 min. The minimum detection limit of the method was 0. 005 μg / kg,and the limit of quantification was 0. 015 μg / kg,with RSD of 3. 3% - 4. 6%. The average recovery was 80. 1% - 88. 9%,and the correlation coefficient was more than 0. 9999. Conclusion The immunoaffinity column coated with specific M1 antibody can effectively remove the interference of impurity in purification of aflatoxin M1 in milk and milk products,thus improving the sensitivity and accuracy of aflatoxin M1 detection. The method is suitable for testing aflatoxin M1 in milk and milk products.

关 键 词：乳和乳制品 黄曲霉毒素M1 高效液相色谱法 

分 类 号：O657.72[理学—分析化学]