高效液相色谱法测定硝基呋喃类药物代谢物及其在对虾体内的代谢

Determination of Nitrofuran Metabolites by High Performance Liquid Chromatography and Their Metabolism in Shrimp

建立高效液相色谱法测定对虾中胺基脲(semicarbazide,SEM)、1-氨基-2-内酰脲(1-aminohydantoin,AHD)、5-甲基吗啉-3-氨基-2-唑烷基酮(5-morpholinomethyl-3-amino-2-oxazolidone,AMOZ)和3-氨基-2-恶唑基酮(3-amino-2-oxazolidone,AOZ)4种硝基呋喃类药物代谢物。采用含100 g/L三氯乙酸的甲醇-水(50∶50,V/V)溶液提取,邻氯苯甲醛衍生,经乙酸乙酯萃取、异辛烷净化,以0.05 mol/L乙酸铵-乙腈(30∶70,V/V)为流动相,反相高效液相色谱分析,外标法定量。结果表明,4种待测物在0.1~50 nmol/m L浓度范围内线性良好,检出限分别为SEM 0.75μg/kg、AHD 1.2μg/kg、AMOZ 2.0μg/kg、AOZ 1.0μg/kg。停药后4种硝基呋喃类药物代谢物在对虾肌肉内富集最高含量分别为51.5、35.8、79.3μg/kg和127μg/kg,代谢物残留低于0.5μg/kg所需代谢时间为SEM 21 d、AHD 15 d、AMOZ 17 d和AOZ 23 d。

A high performance liquid chromatography(HPLC) method for the determination of nitrofuran metabolites in shrimp has been established. After being extracted by 100 g/L trichloroacetic acid in methanol-water(50:50, V/V), the four metabolites semicarbazide(SEM), 1-aminohydantoin(AHD), 5-morpholinomethyl-3-amino-2-oxazolidone(AMOZ) and 3-amino-2-oxazolidone(AOZ), were reacted with 2-chlorobenzaldehyde. The analytes were separated in a C18 column with 0.05 mol/L ammonium acetate-acetonitrile(30:70, V/V) as the mobile phase and detected by a UV detector. SEM, AHD, AMOZ and AOZ were quantified by external standard method. The response signals were all linearly proportional to concentrations of the four metabolites in the range of 0.1–50 nmol/m L, and the limits of detection(LODs) were calculated as 0.75, 1.2, 2.0 and 1.0 μg/kg, respectively. The maximum values of the four metabolites accumulated in shrimp muscle were found to be 51.5, 35.8, 79.3 and 127 μg/kg after the discontinuation of the drugs, and the elimination life turns out to be 21, 15, 17 and 23 days in this study, respectively.