高效液相色谱-电喷雾串联质谱法测定动物饲料中的10种磺胺

Qualification and Quantification of 10 Sulfonamides in Animal Feedstuff by High Performance Liquid Chromatography-Electrospray Tandem Mass Spectrometry

建立了动物饲料中10种常用磺胺(磺胺嘧啶、磺胺吡啶、磺胺甲基嘧啶、磺胺-5-甲氧嘧啶、磺胺二甲基嘧啶、磺胺甲氧哒嗪、磺胺甲基异GFDA4唑、磺胺间甲氧嘧啶、磺胺间二甲氧嘧啶(SDM)和磺胺喹GFDA4啉(SQX))的高效液相色谱(HPLC)-串联质谱检测方法.样品经提取、固相萃取净化、稀释、HPLC分离后进行质谱分析,在多反应监测模式(MRM)下进行特征母-子离子对信号采集.结合保留时间和离子对信息进行定性分析,以共同碎片离子m/z 156进行定量.10种磺胺的定量检测限(S/N=10)为0.5～2.0 μg/kg,在2.0～200 μg/L(SDM和SQX:1.0～100 μg/L)时峰强度与质量浓度的线性关系良好(r>0.999 5).添加水平为1.0 mg/kg时,10种磺胺的平均回收率范围为70% ～92% ,日内相对标准偏差小于10% ,日间相对标准偏差小于15% .结果表明,该法简单、灵敏,特异性强,适用于饲料中多磺胺组分的分析.

The presence of sulfonamide （SA） residues in foods is largely due to the raising of animals with sulfonamide antibiotics added or polluted feedstuff. Because of interference from the matrices, the commonly used immunoassay or chromatographic method is not suitable for the analysis of multi-SAs in feedstuff. A high performance liquid chromatographic-electrospray tandem mass spectrometric （HPLC/ESI-MS-MS） method has been established for the simultaneous determination of multi-SAs including sulfadiazine （ SD ）, sulfapyridine （ SPD ）, sulfamerazine（ SM1 ） , sulfameter （ SM）, sulfamethazine （ SM2 ）, sulfamethoxypyridazine （SMP） , sulfamethoxazole （SMZ）, sulfamonomethoxine （SMM）, sulfadimethoxine （SDM） and sulfaquinoxaline （SQX）. After solvent extraction, solid phase extraction, dilution and reversed-phase HPLC separation, SAs were detected by ESI-MS-MS under multi-reaction monitoring mode. The qualification analysis was done by using retention time and distribution of diagnostic ion pairs,and the quantification was based on the peak intensity of common fragment ion m/z 156. The limits of quantification for l0 SAs were 0. 5 -2.0 μg/kg （S/N = 10）. The correlation coefficient of linear calibration curve was over 0. 999 5 within the SAs concentration range 2.0 - 200 μg/L except for SDM and SQX. At the spiked level of 1.0 mg/kg, the average recoveries for the l0 SAs were between 70% and 92% , the relative standard deviations were under 10% for intra-day and under 15% for inter-day. Routine tests showed the method was fast, sensitive, specific,and practical for the SAs determination in feedstuff.