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 determination 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-hexane,clean-up by Basic Alumina solid phase extraction cartridge and derivation by 1-methylinidazole and trifluoroacetic anhydride inacetonitrile at room temperature away from light and hydrolysis by methanol.Samples were analyzed by HPLC coupled to a fluorescence detector.The results indicated that the calibration curves were linear within the working range from 0.5 to 50.0 μg/L.The limit 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 respectively 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.国家公益性(农业)行业专项(200903029

Observation on the Life History of Cryptocaryon irritans in Eastern Sea of Fujian and Research on Antiparasitic Effect of Acetic Acid

近年来,由刺激隐核虫(CryPTOCAryOn IrrITAnS)感染引发的“白点病“为闽东大黄鱼(PSEudOSCIAEnA CrOCEA)养殖业带来连续重大损失.大量针对该病的研究工作已经开展,但仍未找到治疗“白点病“安全、有效的方法.本实验从福建省福鼎市八尺门海域取得一种寄生虫,通过形态学及分子生物学相关鉴定方法,证明了所取寄生虫为刺激隐核虫.利用乙酸对大黄鱼幼鱼进行药物安全性评价,同时开展杀灭不同生活史阶段刺激隐核虫的时间效应与剂量效应实验.结果显示:刺激隐核虫包囊对乙酸的耐受力显著强于幼虫,乙酸在杀灭幼虫的同时,对大黄鱼幼鱼也有一定的刺激作用,因此在实际用药中应严格控制剂量.根据实验结果,建议在生产中防治“白点病“时,要以刺激隐核虫幼虫为目标,使用157.4Mg/l乙酸,累计处理2H,可以有效杀灭全部幼虫,又不会对鱼体造成危害,是一种治疗“白点病“的安全用药策略.Cryptocaryon irritans,a holotrichous marine ciliate protozoan that causing "white spot disease",is one of the most devastating parasites of marine fish in Fujian Province.In the present study,a strain of C.irritans-like parasite was isolated from Pseudosciaena crocea aquiculture areas in eastern Fujian province and then identified.The morphological features and biochemical characteristics of the parasites were identical to those C.irritans strains which had been reported in other places,this fact suggested that the sampled pathogen was C.irritans.In addition,the acute toxicity of acetic acid to juvenile P.crocea and C.irritans was studied.The results showed that C.irritans theronts were more sensitive to acetic acid than tomonts.Meanwhile,the susceptibility of acetic acid to juvenile P.crocea was an important factor which was considered in the process of killing C.irritans.The results indicated that dealt "white spot disease" infected P.crocea with 157.4 mg/L acetic acid for 2 h could kill C.irritans theronts effectively as well as unharmful to the hosts,it was a safe drug-using strategy for preventing and curing "white spot disease".公益性(农业)行业专项(200903029); 国家科技支撑计划项目(2011BAD13B08); 广西科技攻关计划项目(桂科攻1222013-2