基于HILIC模式的超高效液相色谱-串联质谱法检测牛奶中12种氨基糖苷类药物

Determination of 12 aminoglycoside antibiotics in milk by HILIC-UPLC-MS/MS

利用低密度键合C18柱,在亲水作用色谱(HILIC)模式下,对氨基糖苷类(AGs)抗生素进行分离测定,建立了牛奶中12种AGs类药物的超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。方法采用乙腈、甲酸和水作为流动相,梯度洗脱,40℃柱温条件,整个反应体系中未使用离子对试剂。该方法在250~5000μg/L范围内对庆大霉素和新霉素、150~3000μg/L范围内对安普霉素和巴龙霉素以及在50~1000μg/L范围内对其它6种药物的线性关系良好。方法的检测限为20~100μg/L,定量限为50~250μg/L。20~1000μg/L浓度添加试验表明,样品中的平均添加回收率在60.9%~109.2%之间,日内变异系数在6.8%~15%之间,日间变异系数在3.2%~18%之间。综上所述,本研究建立的AGs类抗生素残留检测方法能够为AGs类抗生素的残留监测提供有效的技术支持。

In the present study,a new Ultra-performance liquid chromatography-tandem mass（ UPLCMS/MS） method utilizing conventional C18 column in hydrophilic interaction chromatography（ HILIC）mode was successfully developed and optimized for the detection of 12 aminoglycoside antibiotics（ AGs） in milk. The common reagents acetonitrile,formic acid and water were used as mobile phase and gradient elution,and 40℃ column temperature was introduced by optimized chromatography and mass spectrometry condition. No ion-pair reagents were used in the method,and all the analytes obtained good retention. Linearity was established over the concentration range of 50-1000 μg/L for spectinomycin（ SPE）, tobramycin（ TOB）, kanamycin（ KANA）, streptomycin（ STR）,dihydrostreptomycin（ DHSTR）,amikacin（ AMK）,150-3000 μg/L for apramycin（ APR） and paromomycin（ PRM）, and 250-5000 μg/L for gentamicin（ GEN） and neomycin（ NEO）,respectively. The correlation coefficients were all 〉0. 98. The limit of detection（ LOD） and limit of quantity（ LOQ） of the method was 20-100 μg/L and 50-250 μg/L,respectively. Fortification at the range of 20-1000μg/L,mean recoveries were 60. 9%-109. 2% with intra-and inter-assay andCVs of 6. 8%-15% and 3. 2%-18% in milk. In summary,the method established in the study was rapid and effective,which could provide effective techniques and enrich the detection system for AGs antibiotics residue monitoring.