大黄鱼4种组织中阿维菌素的HPLC-FLD方法的建立

Establishes Method of Avermectin in Four Tissues of Pseudosciaena crocea by High Performance Liquid Chromatography with Fluorescence Detection

建立了大黄鱼肌肉、肝脏、鳃和血液中阿维菌素的高效液相色谱-荧光检测(HPLC-FLD)法.样品用乙腈提取,正己烷脱脂,经碱性氧化铝固相萃取(SPE)柱过滤,1-甲基咪唑和三氟乙酸酐的乙腈溶液在室温下避光衍生化,再经甲醇水解,进行HPLC-FLD测定.结果表明,阿维菌素在0.5～50.0μg/L范围内线性良好,检测限为肌肉0.5μg/kg,其他组织1.0μg/kg.添加5.0～50.0μg/kg时,平均回收率为:肌肉81.2%～87.1%,肝脏68.9%～74.4%,鳃67.6%～80.7%,血液52.4%～60.3%,日内精密度和日间精密度分别为:肌肉4.6%～5.5%、6.1%～6.8%;肝脏5.5%～6.2%、7.6%～8.7%;鳃5.1%～6.3%、6.9%～7.5%;血液7.6%～8.9%、9.7%～11.6%,该方法能满足现行兽药残留分析要求.

In order to understand the avermectin residues in the tissues of Pseudosciaena crocea ,a method for the quantitative deter- mination of this drug in fish muscle, liver, gill and blood sample was established using high performance liquid chromatography with fluorescence detection （HPLC-FLD）. The procedures for sample treatment included extraction by acetonitrile,delipidation by n-hex ane, clean-up by Basic Alumina solid phase extraction cartridge and derivation by 1-methylinidazole and trifluoroacetic anhydride inac etonitrile at room temperature away from light and hydrolysis by methanol. Samples were analyzed by HPLC coupled to a fluores- cence detector. The results indicated that the calibration curves were linear within the working range from 0.5 to 50.0 μg/L. The lim- it of detection of the method for avermectin in muscle was 0.5 μg/kg, and 1.0 μg/kg for the other three tissues. When 5.0-50.0 μg/kg samples were added,the overall recoveries of avermectin in Pseudosciaena crocea muscle,liver,gill and blood sample were re- spectively 81.2%-87.1%, 68.9%-74.4%, 67.6%-80.7%, 52.4%-60.3%. The intra-day and inter-day coefficient variations for muscle,liver,gill and blood sample detection were 4.6%-5.5% and 6.1%-6.8% ,5.5%-6.2% and 7.6%-8.7% ,5.1%-6.3% and 6.9 %-7.5 %, 7.6 %-8. 9 % and 9.7 %-11.6 % ,which meet the current requirements for drug residue analysis. This research provides a reference for the detection of avermectin for the tissues of P. crocea.