免疫亲和柱-高效液相色谱法检测红曲米中的桔霉素

Determination of Citrinin in Red Rice by High Performance Liquid Chromatography with Immunoafinity Clean-up Column

制备抗桔霉素(citrinin,CIT)单克隆抗体免疫亲和柱(IAC),建立了检测红曲米中CIT的免疫亲和柱-高效液相色谱检测方法。采用甲醇-水提取红曲米样品,将提取液用PBS稀释后过免疫亲和柱,甲醇洗脱后以高效液相色谱法检测,激发波长为331 nm,发射波长为500nm,流动相采用乙腈-磷酸溶液(体积比为45:55,pH2.0)。每根抗CIT单克隆抗体免疫亲和柱使用0.5 mL溴化氰活化的4FF琼脂糖凝胶,0.5mg桔霉素单克隆抗体,柱容量为0.35μg。红曲米样品添加CIT标品0.1～0.6 mg/kg,平均回收率为74.2%～87.14%,相对标准偏差为2.85%～13.12%。最低检出限为0.1mg/kg(S/N=3)。

To prepare immunoafinity column of citrinin, high performance liquid chromatography with immunoafinity clean-up column to quantify citrinin in Red rice is described. Methods：samples were extracted with methanol-water and the extract was diluted with methanol-PBS （v ： v=1 ： 10）and applied to a citrinin test immunoafinity column. The column was washed with PBS, and citrinin was eluted, with methanol and quantified by reversed-phase HPLC with fluorometric detection （Ex=331 nm, Em=500 nm） using acetonitrile-H2O （v ： v=45 ： 55, pH2. 0） as mobile phase. Average recoveries of citrinin from Red rice spiked at levels of 0. 1-0. 6 mg/kg ranged from 74. 2% to 87.12%. The detection limit was 0. 1 mg/kg based on a signal to noise ratio of 3 ; 1.