水解衍生-酶联免疫分析在动物肌肉中呋喃唑酮代谢物残留量检测中的应用  

Application of Derivatisation-ELISA in Determining the Residues of Furazolidone Metabolites in Animal Muscles

作　　者：秦燕[1] 朱柳明[1] 庄逸林[2] 林峰[1] 相大鹏[2] 

机构地区：[1]广州出入境检验检疫局食品检验中心,广州510623 [2]汕头出入境检验检疫局技术中心,汕头515031

出　　处：《药物分析杂志》2005年第6期685-688,共4页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:优化并评估了水解衍生-酶联免疫方法在测定动物肌肉中抗生素呋喃唑酮代谢后结合残留物中的应用。方法:样品中的呋喃唑酮代谢物3-氨基-唑烷酮(3-amino-2-oxazolidone,AOZ)的蛋白结合残留物经酸水解释放 AOZ,经2-硝基苯甲醛(2-nitrobenzaldehyde,NBA)衍生生成3{[(2-nitrophenyl)methylene]-amino}-2-oxazolidinone(NPAOZ)后,调 pH 至7.2-7.4,经乙酸乙酯萃取后,10min/4000r·min^(-1)离心取上层氮气流于30℃吹干并用缓冲液溶解,正己烷脱脂后取下层水相进行 NPAOZ 的竞争性酶联免疫吸附分析(Enzyme linked immunosorbent assay,ELISA)。结果:方法定量检测限为200ng·kg^(-1),样本加标水平为1.0μg·kg^(-1)时,回收率在85%以上。实际样本对照实验显示 AOZ 质量浓度>1.0μg·kg^(-1)时,方法与 LC-MS-MS 结果的相对误差<25%。结论:此方法可实现动物肌肉中呋喃唑酮代谢物残留量的高通量筛选。Objective:The potential use of a derivatisation-ELISA method for determining of binding residues of antibiotics furazolidone metabolites in animal muscles was optimized and evaluated.Method:The protein binding residue of furazolidone metabolites(3-amino-2-oxazolidone,AOZ)in samples was released by acid digestion and derived with (2-nitrobenzaldehyde,NBA)to produce3 {[(2-nitrophenyl)methylene]-amino}-2-oxazo- lidinone(NPAOZ),then the pH was adjusted to 7.2-7.4,after extraction with ethyl acetate,the solution was centrifuged for 10 min/4000 r·min^(-1),then the upper organic phase was transferred and evaporated by N_2at 30℃ water bath to dryness.The residue was redissolved with buffer,after removal of lipid components with n-hexane, the NPAOZ in lower phase was tested with competitive ELISA procedure.Results:Limit of quantification of the method was 200 ng·kg^(-1),and the recovery rate was over 85% at the 1.0 μg·kg^(-1) spiked level.The practical test results between ELISA and LC-MS-MS methods for the incurred sample showed the relative errors were un- der 25% when AOZ concentrations were detected over 1.0 μg·kg^(-1).Conclusion:A rapid and effective pretreat- ment procedure suitable for ELISA assay was established.This method maybe used for screening of furazolidone me- tabolites residues in animal muscles.

关 键 词：呋喃唑酮 动物肌肉 代谢物 酶联免疫分析 残留量 衍生 酶联免疫吸附分析 酶联免疫方法 乙酸乙酯萃取 高通量筛选 蛋白结合 质量浓度 对照实验 相对误差 残留物 抗生素 唑烷酮 酸水解 苯甲醛 缓冲液 竞争性 正己烷 检测限 

分 类 号：R573.1[医药卫生—消化系统] R318.08[医药卫生—内科学]