基于DNA增强荧光法快速测定养殖用水中孔雀石绿

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英文篇名：Rapid Determination of Malachite Green in Aquaculture Water by DNA Enhanced Fluorescence Method 作者：丁洪流 ; 沈福苗 ; 成玉梁 ; 谢云飞 ; 于航 ; 姚卫蓉 ; 郭亚辉 ; 钱和 英文作者：DING Hong-liu;SHEN Fu-miao;CHENG Yu-liang;XIE Yun-fei;YU Hang;YAO Wei-rong;GUO Ya-hui;QIAN He;The Bureru of Quality and Technical Supervision Comprehensive Inspection and Monitoring Center of Suzhou;School of Food Science and Technology,Jiangnan university; 关键词：DNA ; 荧光光谱 ; 孔雀石绿 ; 快速检测 ; 定量分析 英文关键词：DNA;;Fluorescence spectroscopy;;Malachite green;;Rapid determination;;Quantitative analysis 中文刊名：FXKX 英文刊名：Journal of Analytical Science 机构：苏州市质量技术监督综合检验监测中心;江南大学食品学院; 出版日期：2019-04-20 出版单位：分析科学学报 年：2019 期：v.35 基金：江苏省农业科技自主创新资金(No.CX(16)1028);; 江苏省重点研发计划(No.BE2016664);; 苏州市科技项目(No.SS201734) 语种：中文; 页：FXKX201902016 页数：5 CN：02 ISSN：42-1338/O 分类号：88-92

摘要

本文筛选了由11种富含G碱基的DNA分别与3种缓冲溶液形成的体系,通过优化检测条件,得H22 DNA(5μmol/L)与Tris-HCl缓冲溶液(0.05 mol/L,pH=6.5,100 mmol/L K~+)形成的检测体系,实现了孔雀石绿(MG)的快速检测。进一步探究了体系荧光强度及稳定性,发现DNA浓度增加可显著增加荧光强度,且荧光具有一定的稳定性。此方法对MG检测的线性范围为0.1～6μmol/L,线性相关系数达到0.994,检测限为0.083μmol/L。将本方法用于养殖用水中MG的直接检测,结果未检出;加标准MG配制成0.5、1.0、5.0μmol/L的模拟水样,测得回收率在93.4%～119.3%之间。该方法操作简单快捷,可应用于养殖水体中MG的快速检测。
        By screening the system formed by 11 kinds of DNA and 3 kinds of buffer solutions and optimizing the detection conditions,a system of DNA(H22) and Tris-HCl buffer(0.05 mol/L,pH=6.5,100 mmol/L K~+) was successfully selected to detect malachite green(MG) with a 36-fold increase in fluorescence intensity and rapid detection of malachite green was achieved.The fluorescence intensity and stability of the system were further explored. It was found that increasing the DNA concentration could significantly increase the fluorescence intensity,and the fluorescence had a certain degree of stability.The detection of MG could be realized in the linear range from 0.1 μmol/L to 6.0 μmol/L with the correlation coefficient(R~2) of 0.994,and the actual minimum detectable concentration was 0.083 μm/L.The method was applied to the detection of MG in aquaculture water.No MG was found in this real sample by direct detection.With addition of standard malachite green to form 0.5 μmol/L,1.0 μmol/L and 5.0 μmol/L MG simulation samples,the spiked recoveries of the sample ranged from 93.4% to 119.3%.The method is simple and quick to operate,and can be applied to the rapid detection of MG in aquaculture waters,and has practical significance for screening MG.

引文

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