凝胶渗透色谱-固相萃取/高效液相色谱法同时测定水产品中甲基睾酮与己烯雌酚

摘要

建立了凝胶渗透色谱-固相萃取/高效液相色谱法测定水产品中甲基睾酮和己烯雌酚的分析方法.样品经乙醚超声提取和漩涡振荡后以乙酸乙酯-环己烷(1∶1)为流动相进行凝胶渗透色谱净化,再过HLB柱进一步净化,以甲醇-水(72∶28)为流动相,采用高效液相色谱/紫外检测器在254 nm波长下测定,外标法定量.结果显示甲基睾酮和己烯雌酚在0.02～2.0 mg/L质量浓度范围内呈良好的线性关系,线性系数(r)均大于0.999.空白样品在10.0、50.0、200 μg/kg 3个加标水平下的平均回收率为84％～93％,相对标准偏差均小于6.0％.甲基睾酮和己烯雌酚的定量下限分别为6.0 μg/kg和5.0 μg/kg.%A method of high performance liquid chromatography combined with gel permeation chro matography (GPC/HPLC) was developed for the simultaneous analysis of methyltestosterone (MET) and diethylstilbestrol (DES) in aquatic products. The homogenized samples were extracted with dieth-yl ether, then cleaned by GPC using ethyl acetate -cyclohexane(l : l)as mobile phase, follwed by a further HLB solid phase extraction. The targeted compounds were detected by HPLC - ultraviolet detector at the wavelength of 254 nm, using merhanol - water(72: 28) as mobile phase, and quantified by the external standard method. The calibration curves of two compounds were linear in the concentration range of 0. 02 - 2. 0 mg/L with correlation coefficients more than 0. 999. The average recoveries of MET and DES at spiked concentrations of 10. 0, 50. 0, 200 μg/kg ranged from 84% to 93%, with relative standard deviations (RSDs) less than 6. 0% . The LOQs of MET and DES were 6. 0 μg/kg and 5. 0 μg/kg, respectively.