高效液相色谱法测定鸡肉中5种磺胺类药物及乙氧酰胺苯甲酯增效剂残留量

Determination of 5 kinds of sulfonamides and ethopabate residues in chicken by high performance liquid chromatography

中文关键词:  高效液相色谱法  磺胺二甲嘧啶  磺胺间甲氧嘧啶  磺胺甲噁唑  磺胺二甲氧嘧啶  磺胺喹噁啉  乙氧酰胺苯甲酯  抗球虫药

英文关键词:high performance liquid chromatography  sulfamethylamine  sulfamonomethoxine  sulfamethoxazole  sulfadimethoxine  sulfaquinoxaline  ethopabate  coccidiostats

中文摘要:

      目的 建立高效液相色谱法同时检测鸡肉中磺胺二甲嘧啶(sulfamethylamine, SM2)、磺胺间甲氧嘧啶(sulfamonomethoxine, SMM)、磺胺甲噁唑(sulfamethoxazole, SMZ)、磺胺二甲氧嘧啶(sulfadimethoxine, SDM)、磺胺喹噁啉(sulfaquinoxaline, SQ)、乙氧酰胺苯甲酯(ethopabate)的分析方法。方法 样品加入10 mL乙腈提取, 10000 r/min离心10 min, 氮气吹干后, 加入5 mL pH值为7.0的磷酸盐缓冲液溶解, 过HLB固相萃取柱净化。色谱柱为Phenomenex Luna C18 (250 mm×4.6 mm, 5 μm), 流动相为2.2%乙酸水溶液-甲醇乙腈, 等度洗脱, 流速为1.0 mL/min, 进样量为20 μL, 检测波长267 nm。结果 5种磺胺类药物在0.10~1.5 μg/mL呈良好的线性关系, r2均大于0.999, 乙氧酰胺苯甲酯在0.5~10.0 μg/mL呈良好的线性关系, r2均大于0.999。方法中5种磺胺类药物的最低定量限为50 μg/kg, 乙氧酰胺苯甲酯的最低定量限为250 μg/kg。5种磺胺类药物在50~200 μg/kg的浓度添加水平上, 其回收率在60%~100%, 批内、批间的相对标准偏差小于20%, 乙氧酰胺苯甲酯在250~1000 μg/kg的浓度添加水平上, 其回收率在60%~100%, 批内、批间的相对标准偏差小于20%。结论 该方法具有简便快捷、灵敏度高等特点, 适用于鸡肉中5种磺胺类药物和乙氧酰胺苯甲酯的残留检测。

英文摘要:

      Objective To establish a method for determination of sulfamethylamine (SM2)、sulfamonomethoxine (SMM), sulfamethoxazole (SMZ), sulfadimethoxine (SDM), sulfaquinoxaline (SQ) and ethopabate residues in chicken by high performance liquid chromatography (HPLC). Methods Five kinds of sulfonamides and ethopabate were extracted with 10 mL acetonitrile, and centrifuged with 10 min under 10000 r/min, then dissolved by 5 mL phosphate buffered saline after evaporation, and the whole of supernatant was purified with HLB column. The chromatography column was Phenomenex Luna C18 (250 mm×4.6 mm, 5 μm). The mobile phase was methanol-acetonitrile-2.2% acetic acid water (15:15:70, V:V:V) with equivalent elution, the flow rate was 1.0 mL/min, injection volume was 20 μL and detection wavelengths was 267 nm. Results A good linearity of the calibration curve of SM2, SMM, SMZ, SDM, SQ were obtained in the range of 0.1?1.5 μg/mL (r2>0.999), and the limit of quantification were 50 μg/kg. A good linearity of the calibration curve of ethopabate was obtained in the range of 0.5?10.0 μg/mL (r2>0.999), and the limit of quantification was 250 μg/kg. The average recoveries of SM2, SMM, SMZ, SDM, SQ from spiked chicken at 3 concentrations of 50, 100, 200 μg/kg ranged from 60%?100% (n=5) respectively. The average recoveries of ethopabate from spiked chicken at the three concentrations of 250, 500, 1000 μg/kg ranged from 60%?100% (n=5). In day-between days batch variation coefficients were both less than 20%. Conclusion This method is simple, rapid, sensitive and accurate, and can be applied for the determination of SM2, SMM, SMZ, SDM, SQ and ethopabate in chicken.