Study And Application Of Analysis Of Trace Zearalenone By HPLC

Zearalenone(ZEN,also known as F-2 toxin),the toxin released by Fusarium in the environment,can enter the human body through the food chain or pollute the environment through animal excreta.The analysis of the residual levels of ZEN in animals and their distribution is an important component of the study on the environmental behavior that ZEN migrates from the environment to the organism.Based on the high performance liquid chromatography method with fluorescence detection(HPLC-FLD),various factors including the aspects of sample pretreatment,matrix effects,on-column enrichment,separation of drug residues and pharmacokinetics,have been well investigated,andthus the analysis of trace ZEN and its metabolites in animal samples was established to realize the highly sensitive monitoringof trace ZEN,The pharmacokinetic models of ZEN in animal(rabbit)plasma were built and the distribution of its residues in the tissues of the animal(rabbit)were also studied.The paper mainly includes the followingparts:In chapter 1,literature review.the knowledge about the nature of ZEN and its metabolites,analysis methods and metabolic kinetics for ZEN was reviewed,and then the aims and research scheme of this study were proposed.In chapter 2,a study on the pretreatment of trace ZEN in animal tissues and plasma.To solve the low recovery ratio of ZEN in real samples,effects of various parameters including the extractant concentration,the loading speed of solid phase extraction(SPE),the elution rate,the type of eluent,the amount of eluent,the evaporation temperature under nitrogen and the amount of solvent were investigated systemically.Under the optimal conditions,the pretreatment of ZEN in real samples was established and used for determining trace ZEN residue in animal tissues and plasma.By using HPLC method,trace ZEN could be well detected with the detection limit of ZEN at 1.0 ng/g.The recovery rates of ZEN were in the range of 72.6?75.5%,74.4?91.5%,and 74.2?94.0%for muscle,liver and plasma,respectively.In chapter 3,the study on continuous separation of trace ZEN and its metabolites.The on-column enrichment technique of HPLC-FLD for ZEN was developed after investigating the key parameters such as sample compositions and injection volumes.The sensitivity of ZEN analysis was improved,and the detection limit was 0.2 ng/g.Zearalenone and its metabolites viz.a-zearalcoin,?-zearalcoin,a-zearalenol,??zearalenol and zcaralanone were successfully separated under the optimial conditions.The methods were applied for the analysis of animal tissue and plasma samples with the recovery rates of 62.7?107.9%and 70.7?127.9%,respectively.And the RSD were 3.5?10.3%for muscle and liver samples and 3.1?14.9%for plasma samples.In chapter 4,the study on pharmacokinetic for ZEN.The pharmacokinetic models and the residual distribution data of ZEN in the animal(rabbit)tissues were established.ZEN is mainly metabolized to a-ZOL in rabbit and the ZEN metabolic model belongs to the first-class absorption of two-compartment model,with key parameters including eliminate half life t1/2? =1.02h,clearance rate CLs = 1.18L/h/kg L/h/kg,Cmax = 2263.12?g/L,the curve area AUC = 2293.09?g/L/h and the apparent distribution Volume =15.3 L/kg.