特种黑鸡鸡肉四环素类药残检测及调查研究

A Study on the Detection of Tetracycline Residues in the Endemic Species of Gallus gallus

针对四川乐山特种黑鸡鸡肉中四环素类兽药检测,得到采用Mcllivaine-Na_2EDTA缓冲溶液提取样品,Cleanert~PEP-2小柱净化;高效液相色谱法测定:色谱柱为Eclipse C_(18)(4.6 mm×100 mm,3.5μm);流动相:A相-甲醇,B相-0.01 mol/L草酸水溶液,C相-乙腈;柱温:40℃;进样量:100μL;检测波长:355 nm。方法标准曲线相关系数均大于0.995,四种抗生素最低检出限(LOD,S/N=3)分别为52,24,48和44μg/kg;定量限分别为84,80,88和84μg/kg。加标回收率均大于90%(加标浓度范围40~800μg/kg),RSD%均小于5%,方法准确可靠。以该方法调查养殖黑鸡四环素类抗生素残留,得到5个来自规模化养殖场黑鸡鸡肉样品四环素类兽药残留全低于100μg/kg限量[1];9个来自散养户的黑鸡样品,有6个样品1种四环素类兽药少量超限(超限量低于50%)。

This study focuses on achieving a reliable detection method to detect tetracycline residues in Black Chicken, the endemic and endangered species lives in Leshan, Sichuan Province. Samples are extracted with buffer solution Mcllivaine- Na：EDTA, purified in spin columns Cleanert PEP-2 and detected by HPLC （high performance liquid chromatography）. Parameters are set as follows： chromatographic colunm was Eclipse C18 4.6 mm×100 mm, 3.5 μm; Mobile phase was methanol as phase A, 0.01 mol/L oxalic solution as phase B, acetonitrile as phase C; Column temperature was 40℃; Sample size was 100 p.L; Detection wavelength was 355 nm. All the correlation coefficients of calibration curves are above 0.995. LODs （S/ N=3） of aureomycin, doxycycline, tetracycline and oxytetracycline are 52, 24, 48 and 44 μg/kg, respectively, while LOQs are 84, 80, 88 and 84 μg/kg, respectively. Recovery rates are all above 90% （concentration range of standard liquid： 40-800 μg）, all of RSD% is below 5%, which indicates this is a reliable method. Of all the 14 samples, 5 samples are from large-scale farms and 9 samples are from household raisers. The results show that the residues in the samples from large-scale farms are all lower than the regulated limit of 100 μg/kg, while in 6 samples from household raisers, there is one tetracycline residue above the limit （limit-exceeding proportion lower than 50%）.