高效液相色谱法检测乳品饮料中四环素类抗生素

Analysis of Tetracycline Antibiotics in Dairy Beverage by High Performance Liquid Chromatography

建立一种用固相萃取-高效液相色谱法测定乳品饮料中土霉素、四环素、金霉素、强力霉素的检测方法。样品前处理用乙腈沉淀蛋白后,采用HLB固相萃取小柱进行净化和富集,氮气吹干后用流动相复溶上机检测。采用Waters RP C18色谱柱(4.6 mm×250 mm,5μm)分离,以乙腈∶甲醇∶0.01mol/L草酸溶液(18∶5∶77)作为流动相,用PDA检测器检测,外标法峰面积定量。结果表明,4种四环素类抗生素在0.10μg/m L^1.00μg/m L浓度范围内线性良好,相关系数r2为0.994~0.999,检出限均为2.0μg/kg^5.0μg/kg,定量限均为6.0μg/kg^15.0μg/kg,加标回收率达到77.6%~90.6%。

A rapid and effective method was established for the determination of oxytetracycline, tetracycline, chlortetracycline and doxycycline in dairy beverage by solid-phase extraction coupled with high performance liquid chromatography detection method. After the sample was treated with acetonitrile,it was concentrated and purified by solid phase extraction column.The elution liquid was dried under nitrogen, and detected after re- dissolution with mobile phase. The separation of target compound was performed on a Waters RP C18 chromato- graphic column （4.6 mm×250 mm, 5μm）using acetonitrile ： methanol ： 0.01 mol/L oxalic acid solution （ 18 ： 5 ： 77 ）as mobile phase, with PDA detector and external standard method peak area quantification. The linear range of 4 tetracycline antibiotics components was in the range of 0.10 μg/mL^-1.00 μg/mL with a correlation coefficient of 0.994-0.999. The detection limit was 2.0μg/kg-5.0μg/kg and the quantitative limit was 6.0 μg/kg- 15.0 μg/kg.The recovery rate was 77.6 %-90.6 %.