高效液相色谱-电喷雾串联质谱法测定畜禽粪便中四环素类抗生素

Determination of Tetracyclines Antibiotics Residues in Animal Manure by Liquid Chromatography with Tandem Mass Spectrometry

建立了高效液相色谱～电喷雾串联质谱法（HPLC—MS／MS）测定畜禽粪便中四环素类抗生素的分析方法。样品经甲醇-乙酸-水（体积比6：3：1）溶液提取，OASISHI。B小柱净化后，采用WatersSymmetryC18色谱柱（150×3．9mm，5／lm）进行分离，以0．3％甲酸水溶液-0．3％甲酸乙腈溶液梯度洗脱，电啧雾正离子模式多反应监测（MRM）模式定量分析。在优化的色谱条件下，四环素、土霉素和金霉素在0．05～1．0mg／L范围内具有良好的线性，其检出限分别为0．025、0．084、0．098mg／kg；样品回收率为80％～103％。实验结果表明，该方法较液相色谱法具有更高的准确性和灵敏度，可满足畜禽粪便中四环素类抗生素残留的检测要求。

A method was developed for the determination of tetracyclines antibiotics in animal manure using solid phase extraction followed by liquid chromatography（HPLC） and tandem mass spectrometry （MS/MS）. The mixture of methanol, acetic and water with volume ratio of 6 ： 3 ： 1 was selected as extraction solvent. The analytes were separated on a Waters Symmetry C18 column using 0.3% formic acid in water 0. 3 % formic acid in acetonitrile as mobile phase. Quantitative analysis was performed using multiple reaction- monitoring mode （MRM）. Under the optimal extraction conditions, tetracycline, oxytetracycline, and chlortetracycline were separated with an excellent linear relationship in the range of 0.05-1.0 mg/L. The limits of detection（LOD） were 0. 025,0. 084,0. 098 mg/kg（S/N= 3）. The spiked recoveries of three compounds ranged from 80 % to 103 %. Compared with HPLC method, the proposed method was sensitive and accurate,and could meet the requirement for the determination of tetracyclines antibiotics residue in animal manure.