【作者】 陈建伟；

【导师】 杨明敏；

【作者基本信息】 南京农业大学 ， 应用化学， 2009， 硕士

【摘要】 孔雀石绿、结晶紫等属三苯甲烷类化学物质,许多国家曾将其作为水产养殖业的杀菌剂,用于控制鱼和鱼苗真菌的生长,控制高发的水霉病、原虫病等。但是,孔雀石绿等进入人类或动物机体后,可产生致癌、致畸、致突变等副作用,因此,近年来水产品的国际商贸中,孔雀石绿、结晶紫属类药物成为必检并限制极严的一项指标。测定水产品中孔雀石绿等的方法有多种,其中常见的方法有：液相色谱法、液质联用法和气质联用法等。目前上述方法前处理都较复杂,并且因为水产品种类繁多,基体复杂,对实验条件要求苛刻,因此建立一种简捷测定孔雀石绿染料的分析方法,并应用于实际,具有十分重要的现实意义。本论文共分两个部分：第一部分：本部分描述了一种设计孔雀石绿浊点萃取过程的系统方法,实验目的是计算在染料萃取操作条件下(温度和染料浓度一定)所使用的表面活性剂系列的浓度。在设计计算过程中,本系统讨论了以下几个参数：(1)一定温度下,染料在含有表面活性剂的溶液中的吸附等温线参数；(2)一定温度下,随着表面活性剂和染料的浓度变化,凝聚相体积和稀释相浓度的变化。由聚乙二醇(PEG)系列表面活性剂和孔雀石绿染料组成的实验系统,而这种浊点萃取的过程是可以采用兰谬尔吸附等温式来处理的,故实验数据采用兰谬尔吸附等温式来处理。不同的参数随表面活性剂系列的不同显示出规律性。利用该实验设计方法,我们得出,当染料理论吸附率是90%时,PEG系列各表面活性剂所需要加入的浓度。我们把此方法用于水样和虾样的检测验证,结果满意。第二部分：本部分研究了利用浊点萃取方法富集虾肉中孔雀石绿和结晶紫,建立了用高效液相色谱(二极管阵列检测器)同时测定虾肉中孔雀石绿和结晶紫残留的方法。研究了表面活性剂、盐、平衡时间及平衡温度对浊点萃取效率的影响。该分析方法应用于虾中两种物质的残留分析。在虾肉中分别添加三个不同浓度的孔雀石绿和结晶紫,添加回收率范围分别为75.5-80.2%(孔雀石绿)和81.4-89.6%(结晶紫),相对标准偏差(RSD)分别为2.3-3.1%(孔雀石绿)和1.5-3.3%(结晶紫)。该方法的检测限分别为孔雀石绿2.0μg kg-1、结晶紫1.0μg kg-1,可满足我国和欧盟对虾肉中孔雀石绿和结晶紫的检测要求。更多还原

【Abstract】 Malachite green (MG) and Crystal violet (CV) are triphenylmethane dyes, which was used as a fungicide in aquaculture because it can effectively control gillrot and some other diseases caused by helminthes. Being cheap and easy available, it was widely used in aquaculture industry. Malachite green when discharged into receiving streams will affect the aquatic life and causes detrimental effects in liver, gill, kidney, intestine, gonads and pituitary gonadotrophic cells. In recent years, however, as much more attention has been paid to food safety and sanitation by international community, and many negative effects have been disclosed, such as high residual, great toxicity, carcinogenicity, teratogenicity, mutagenesis and so on, Malachite green has been exciting extensive solicitude of the public. Determination of Malachite green and Crystal violet is a must in today’s international trade.The chief ways to determinate Malachite green are:HPLC, LC-MS and GC-MS. All the above ways need complicated pretreatment. In addition, the wide varieties of aquatic products make the experiment conditions more harshly. Thus, it has very important realistic significance to develop a simple method for the determination of malachite green and apply to practice.The thesis includes two parts:In chapter I:A systematic method is outlined to design Cloud point extraction process. The aim is to calculate the concentration of non-ionic surfactant required for a desired extraction of dye at various operating conditions (feed dye concentration and temperature). In the course of design calculations, information about the following two characteristics of this system are required:(1) the adsorption isotherm of the dye in the surfactant solution at various operating temperature and (2) quantification of the variation of the fractional coacervate phase volume with the feed surfactant, dye concentration and the operating temperature. In case of PEGs and the dye system (Malachite green), a Langmuir type adsorption isotherm is fitted using the experimental data. Correlations are developed for the variation of isotherm parameters with temperature. From the experimental data, the variations of m and n with feed dye concentration and temperature are evaluated. Using the developed correlations and proposed design method, we also calculated the optimal PEG concentrations for MG recovery of 90%and tested the reliability of the proposed procedure to real samples. The developed method will be of immense help for scale up of cloud point extraction process.In chapter II:A cloud point extraction (CPE) method was used to separate and concentrate malachite green (MG) and crystal violet (CV) in shrimp samples followed by HPLC-diode-array detector (DAD) analysis was developed. The effects of surfactant, salt additive, equilibration time and the equilibration temperature on extraction efficiency were studied. The method was applied to determine residues in shrimp samples. Under optimum conditions, recoveries of spiked shrimp sample, relative standard deviations (RSD) and the limit of detection (LOD) of MG and CV were 75.5-80.2%,2.3-3.1%and 2ug kg-1 and 81.4-89.6%,1.5-3.3%and 1μg kg-1, respectively. The results obtained by the proposed method were suitable for the requirements of China and EU.更多还原