酶联免疫法测定水产品中呋喃唑酮代谢物AOZ的残留

Enzyme linked immunosorbent assay(ELISA) for the quantitative analysis of furazolidone residues in aquatic products

运用酶联免疫法对水产品中呋喃唑酮代谢物残留测定的方法进行研究。结果表明，稀释倍数对回收率影响显著，而乙酸乙酯提取次数、光照条件和放置时间对测定结果无显著影响。向样品中分别添加0．60、2．00、6．00μg·kg^-1 3个浓度水平的呋喃唑酮代谢物时，平均回收率分别为99．2％、96．0％和100．0％；批内变异系数为1．76％～12．57％，批间变异系数为5．43％～8．58％，最低检测限为0．3μg·k^-1。该方法灵敏度高，重复性较好，适用于水产品中呋喃唑酮代谢物残留的筛选。

A method of ELISA for the determination of furazolidone metabolites residues in aquatic products is established in this paper. There were no literatures about determination of AOZ residues in aquatic products by Enzyme linked lmmunosorbent assay （ELISA） at home. Samples were derivatived with 2-nitrobenzaldehyde and extracted with ethyl acetate. Then they were purified with n-hexane and determinatod by ELISA. In the paper, effects of different disposal conditions including dilution factors, extraction times, illumination conditions and shelf time to recovery were researched. By determination on Eriocheir sinensis, Micropterus salmoides and Procambarus clarkii spiked with AOZ, recovery, precision and detection limit of the method were researched by single laboratory validation approach as well as between laboratory comparisons. Results show that there is a remarkable effect of dilution factors on the recovery while no such effect is displayed for extraction times by ethyl acetate, illumination conditions and shelf time. So the best disposal condition is that dilution factor and extraction time are both one. The average recoveries are 99.2% ,96.0%and 100.0% respectively when the samples are spiked with 0.60 μg·kg^-1,2.00 μg·kg^-1 and 6.00 μg·kg^-1 of AOZ. The detection limit is 0.3 ·μg·kg^-1 . The relative standard deviations（n= 3） for intra-assay are 1.76% - 12.57% and coefficients of variation for interassay are 5.43% - 8.58%. Three positive samples analyzed by ELISA are confirmed by LC-MS and no false positive samples are found. The method is sensitive with a fairly good reproducibility and suitable for the determination of furazolidone metabolites（AOZ） in aquatic products.