导　　师： 印春生

学科专业： 083001

授予学位： 硕士

作　　者： ;

机构地区： 上海海洋大学

摘　　要： 抗生素是“在低微浓度即可对某些生物的生命活动有特异抑制作用的化学物质的总称”。近年来随着人类社会的发展，为了满足生活的需要，水产养殖业一度大规模，集中式的扩大，在集约型发展模式下，多种由细菌或微生物引起的疾病给水产养殖带来了巨大的损失。为了防治鱼病减少不必要的损失，抗生素的使用频率越来越高。因此,建立水环境中抗生素类药物的检测方法，无疑是一件重要意义的工作。 化学发光分析是利用化学发光反应原理来测定物质中某种成分的含量的一种痕量分析方法。化学发光分析法背景值比较低、可检测的范围广、分析快速、操作简便，是分析测定微量和痕量物质的非常有效的方法。在工业，医药，生物领域的分析应用越来越广泛。 化学发光反应分为直接发光和间接发光两大类。根据化学发光反应中，某些发光体系的本底发光强度会因为外来无机离子或有机物的加入而增强或减弱的现象，建立了增强化学发光或抑制化学发光两种检测方法。化学发光分析进样方式分为两种：静态注射进样和流动注射进样。 本研究就是通过建立化学发光反应体系，测定水产养殖中常用的3种抗生素——金霉素、土霉素和呋喃西林渔药中相应抗生素的含量。 本论文主要分为两大部分，前一部分为综述，后一部分为研究报告。 综述部分简单介绍了目前全球水产养殖业抗生素的使用情况及其后果，阐述了化学发光分析方法的原理，技术，并总结了化学发光分析法在常用抗生素检测方面的应用。 研究报告主要由五个部分组成： 1）根据金霉素能增强KMnO4-Na2SO3化学发光体系发光强度的原理，建立了静态注射化学发光检测金霉素的方法。在优化的实验条件下，盐酸金霉素溶液浓度与发光强度的方程为ICL=220.31x+18.89，R~2=0.9794，方法的检出限为2.0×10~/(-8/)mg//L。对1.0mg／L盐酸金霉素溶液进行10次平行测定的相对标准偏差RSD为7.13％。用该方法测定盐酸金霉素眼膏中盐酸金霉素的含量，加标回收率分别为96.9/%,98.5/%和102.5/%，平均值为99.3/%。结果较令人满意，表明该方法能应用于对金霉素的测定。 2）根据碱性介质中，盐酸土霉素对铜/(II/)离子-鲁米诺-过氧化氢化学发光体系产生的发光信号能起到明显的抑制作用，并且其抑制程度与盐酸土霉素的浓度线性相关的，建立了静态注射抑制化学发光测定盐酸土霉素的方法。在最适实验条件下，盐酸土霉素在2.0×10~/(-7/)mol//L~1.0×10~/(-5/)mol//L的浓度范围内与其抑制发光程度呈现良好的线性关系，方法的检出限为4.97×10~/(-8/)mol//L。对浓度为1.0×10~/(-5/)mol//L的盐酸土霉素标准溶液进行11次平行测定的相对标准偏差RSD为2.20％。用该方法测定盐酸土霉素可溶性拌料粉中盐酸土霉素的含量，加标回收率在97/%~103/%之间，平均回收率为100.67/%，结果较令人满意。 3）根据呋喃西林能增强H2SO4-KMnO4化学发光体系发光强度的原理，建立了静态注射化学发光测定呋喃西林精粉中呋喃西林含量的方法。在最适条件下，呋喃西林在2.5×10~/(-6/)mol//L~4.0×10~/(-5/)mol//L的浓度范围内与体系发光强度呈现良好的线性关系，线性方程为y=10.269x+5.6963，R~2=0.9887。方法的检出限为1.26×10~/(-6/)mol//L。对2.0×10~/(-5/)mol//L呋喃西林标准溶液进行11次平行测定的相对标准偏差RSD为2.00％。用该方法测定呋喃西林精粉中呋喃西林含量，加标回收率分别为93/%,96/%和92/%，平均值为93.67/%，结果较令人满意。 4）根据甲醛增强OTC-KMnO4-HCl体系化学发光强度的原理,建立了流动注射化学发光法测定盐酸土霉素可溶性拌料粉中土霉素的含量的方法。在最适实验条件下，盐酸土霉素在0.4～60mg//L范围内与发光程度呈现良好的线性关系，其线性方程为y=12.86x-2.13,R~2=0.9906，方法的检出限为0.28mg//L。对10mg//L土霉素标准溶液进行11次平行测定的相对标准偏差RSD为3.4％。用该方法测定盐酸土霉素可溶性拌料粉中盐酸土霉素的含量，加标回收率分别为99.2/%,101.4/%和102.7/%，平均值为101.1/%，结果较令人满意。 5）根据Co~/(2+/)抑制CTC-NaOH-Luminol-KIO4体系化学发光的原理,建立了流动注射抑制化学发光法测定盐酸金霉素可溶性拌料粉中金霉素的含量的方法。在最适实验条件下，盐酸金霉素在1.0×10~/(-7/)～1.0×10~/(-5/)g//L的浓度范围内与其抑制发光程度呈现良好的线性关系，线性方程为I=1.22×107x+1.43,R~2=0.9945。方法的检出限为2.0×10~/(-8/)g//L。对1.0×10~/(-6/)g//L盐酸金霉素标准溶液进行11次平行测定所得相对标准偏差RSD为2.7％。用该方法测定盐酸土霉素可溶性拌料粉中盐酸土霉素的含量，加标回收率分别为98.5/%,101.2/%和100.8/%，平均值为100.2/%，结果较令人满意。该方法可实现渔药中金霉素含量的快速检测。 Antibiotics is the general term for chemicals with the specific inhibition to somebiological activities of life in low concentration. In recent years, with the developmentof society, in order to meet the needs of life, the expansion of aquaculture was oncelarge-scale and centralized. Under the intensive mode of development, a variety ofdiseases caused by bacteria or micro-organisms has brought great losses to aquaculture.In order to control the diseases to make losses unnecessary reduce, frequency ofantibiotics using was increasing. Therefore, the establishment of the detection ofantibiotics in the water environment is undoubtedly an significance work. Chemiluminescence /(CL/) is a trace analysis method that is based on light radiation generated by chemical reaction /(chemiluminescence/) to determine the content of materials. The technology with low background, high sensitivity, wide linear range and simple instruments makes it a sensitive optical detection technology. So that CL technology has become an effective method for micro and trace analysis. The chemiluminescence reaction is divided into direct chemiluminescence andindirect chemiluminescence. In the chemiluminesce reaction, the luminescenceintensity is enhanced or weakening because of the foreign inorganic ions or organiccompounds joining into the CL system. Two detection methods of the enhancedchemiluminescence and inhibited chemiluminescence are established on this. Theinjection of chemiluminescence analysis is divided into two types: static-injection andflow-injection. In this study, the content of three kinds of aquaculture antibiotics/(chlortetracycline,oxytetracycline, nitrofurazone/) in fishery drugs have determined by the establishmentof the chemiluminescence reaction system. This thesis is divided into two main parts, the first part is the overview,and thesecond part is the research report. In the section of the review, it briefly introduced the usage of the aquaculture inthe global antibiotics with its consequences, and the principles, techniques onchemiluminescence analysis method. Then it summarizes the application of thechemiluminescence method used in the antibiotics detection. The research report consists of five parts: According to the principle that chlortetracycline can sensitize the chemiluminescence system of KMnO4‐Na2SO3, a method with static injection to determine chlortetracycline was set up. Under the best conditions, the chemiluminescence intensity was found to respond linearly to the concentration of chlortetracycline, ICL=220.31x+18.89, R2=.9794, with the detection limit of2.0×10-8mg//L. Precision of the method was tested at the concentration method level of1.0mg//L and the RSD′s /(n=11/) found was7.13％. The proposed was applied to the analysis of chlortetracycline hydrochloride ointment, giving values of recovery in the range96.9/%~102.5/%, and the average recovery was993/%.The results obtained were satisfactory. 2/) It was found that in the alkaline solution, chemiluminescence, produced by thereaction system of copper /(II/) ions-luminol-hydrogen peroxide, was remarkablyinhibited by the presence of oxytetracyline, and the magnitude of decrease ofchemiluminescence intensity was found to be directly proportional to the concentrationof oxytetracyline. Based on these facts, a method of static injection chemiluminescencedetermination of oxytetracyline was proposed. Linearity was kept in the range2.0×10-7mol//L~1.0×10-5mol//L with a detection limit of4.97×10-8mol//L. Precision of themethod was tested at the concentration method level of1.0×10-5mol//L and the RSD′s/(n=11/) found was2.20％. The proposed was applied to the analysis of oxytetracyclinehydrochloride soluble powder, giving values of recovery in the range97/%~103/%, andthe average recovery was100.67/%.The results obtained were satisfactory. 3/) According to the principle that nitrofurazone can sensitize the chemiluminescence system of H2SO4‐KMnO4, a method with static injection to determine nitrofurazone was set up. Under the best conditions, linearity was kept in the range2.5×10~/(‐6/)mol//L~4.0×10~/(‐5/)mol//L /(y=10.269x+5.6963, R~2,=0.9887/)with a detection limit of1.26×10~/(‐6/)mol//L. Precision of the method was tested at the concentration method level of2.0×10~/(‐5/)mol//L and the RSD′s /(n=11/) found was2.00％. The proposed was applied to the analysis of nitrofurazone powder, giving values of recovery in the range92/%~96/%, and the average recovery was93.67/%.The results obtained were satisfactory. 4/) According to the principle that HCHO can sensitize the chemiluminescence system of OTC-KMnO4-HCl, a method with flow injection to determine oxytetracyline was set up. Under the best conditions, linearity was kept in the range0.4~60mg//L /(y=12.86x‐2.13, R~2=0.9906/)with a detection limit of0.28mg//L. Precision of the method was tested at the concentration method level of10mg//L and the RSD′s /(n=11/) found was3.4/%. The proposed was applied to the analysis of oxytetracycline hydrochloridesoluble powder, giving values of recovery in the range99.2/%~102.7/%, and the average recovery was101.1/%.The results obtained were satisfactory. 5/) According to the principle that Co2+can inhibit the chemiluminescencesystem of CTC-NaOH-Luminol-KIO4, a method with flow injection to determinechlortetracycline was set up. Under the best conditions, linearity was kept in the range1.0×10-7~1.0×10~/(-5/)g//L /(y=12.86x-2.13, R~2=0.9906/)with a detection limit of2.0×10~/(-8/)g//L. Precision of the method was tested at the concentration method levelof10mg//L and the RSD′s /(n=11/) found was2.7/%. The proposed was applied to theanalysis of chlortetracycline hydrochloride soluble powder, giving values of recoveryin the range98.5/%~101.2/%, and the average recovery was100.2/%.The resultsobtained were satisfactory.

关 键 词： 渔药 化学发光 金霉素 土霉素 呋喃西林 静态注射 流动注射

分 类 号： [X132]

领　　域： [环境科学与工程]