CN108827922A - 基于多重仿生识别的农药残留可视化快速检测技术 - Google Patents
基于多重仿生识别的农药残留可视化快速检测技术 Download PDFInfo
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Abstract
本发明涉及农药残留检测技术领域,尤其是基于多重仿生识别的农药残留可视化快速检测技术。该检测方法包括以下步骤:(1)在农药提取和富集净化阶段利用优化后的萃取溶剂、脱水剂、除杂剂和除色素对农产品中的农药残留实现提取和净化;(2)在富集筛选阶段,利用合成的有机磷类特异性分子印迹聚合物对有机磷农药进行了筛选和富集;(3)在响应识别阶段,利用合成的光学探针可在碱催化下与有机磷酸酯类农药发生磷酰化反应,实现荧光增强和显色反应的高灵敏度二元响应。本发明通过对有机磷农药快速分析过程中农药提取、富集净化、特异性识别筛选、高灵敏度可视化响应等步骤分别研发了新对策,保证了检测的高灵敏度和高准确性。
Description
技术领域
本发明涉及农药残留检测技术领域,尤其是基于多重仿生识别的农药残留可视化快速检测技术。
背景技术
有机磷农药大多属于磷酸酯类或酰胺类化合物,可用于杀菌、杀虫和除草,是全球使用最广泛的一类农药。但是施用农药中对靶生物起效的只有不到 1%,其余部分则通过降雨、沉降和径流等造成水体污染或者可通过直接喷洒或从土壤中吸收进入农作物中,形成在了食物中的农药残留。据统计,我国每年农药中毒的人数占世界同类事故中毒人数的50%,每年中毒人数超过1000万 (包括生活性中毒),其中急性有机磷农药中毒(AcuteOrganophosphorus Pesticide Poisoning。AOPP)约占70%-90%,平均病死率3%~8%。
因此,亟需快速识别与检测技术来及时发现食品中的有机磷农药。
发明内容
本发明要解决的技术问题是:为了解决现有的检测方法无法快速识别的不足,本发明提供了一种基于多重仿生识别的农药残留可视化快速检测技术,通过对有机磷农药快速分析过程中农药提取、富集净化、特异性识别筛选、高灵敏度可视化响应等步骤分别研发了新对策,保证了检测的高灵敏度和高准确性。
本发明解决其技术问题所采用的技术方案是:
一种基于多重仿生识别的农药残留可视化快速检测技术,该检测方法包括以下步骤:
(1)在农药提取和富集净化阶段利用优化后的萃取溶剂、脱水剂、除杂剂和除色素对农产品中的农药残留实现提取和净化,去除农产品中基质对检测结果的干扰;
(2)在富集筛选阶段,利用合成的有机磷类特异性分子印迹聚合物对有机磷农药进行了筛选和富集,保证只有且足量的有机磷类农药在接下来的识别阶段能与识别元件进行接触;
(3)在响应识别阶段,利用合成的光学探针可在碱催化下与有机磷酸酯类农药发生磷酰化反应,实现荧光增强和显色反应的高灵敏度二元响应。
具体地,所述农药提取方法为,取20mL乙腈作为萃取溶剂,2.5g无水硫酸镁作为脱水剂,500mg C18和500mg PSA作为除杂吸附剂,200mg多壁碳纳米管作为除色素剂对蔬菜中有机磷农药进行提取。
具体地,所述有机磷类特异性分子印迹聚合物的合成方法为,取甲基丙烯酸作为功能单体、三唑磷作为模板分子,二甲基丙烯酸乙二醇酯作为交联剂,按照1:6:12的比例进行组装,以三氯甲烷作为制孔剂,乙腈作为溶剂,在无氧条件下采用溶剂热沉淀聚合的方法合成了有机磷类特异性分子印迹聚合物。
具体地,所述光学探针为,有机磷酸酯特异性可视化响应探针(Z)-3-(1H-苯并[d]咪唑-2-基)-2-羟基-5-甲基苯甲醛肟。
本发明的有益效果是:本发明提供了一种基于多重仿生识别的农药残留可视化快速检测技术,通过对有机磷农药快速分析过程中农药提取、富集净化、特异性识别筛选、高灵敏度可视化响应等步骤分别研发了新对策,保证了检测的高灵敏度和高准确性。
附图说明
下面结合附图和实施例对本发明进一步说明。
图1是本发明的荧光法曲线图;
图2是本发明的比色法曲线图;
具体实施方式
现在结合附图对本发明作进一步详细的说明。
图1是本发明的荧光法曲线图,图2是本发明的比色法曲线图。
(1)本申请首先在农药的提取过程中,优选20mL乙腈作为萃取溶剂,2.5g 无水硫酸镁作为脱水剂,500mg C18和500mg PSA作为除杂吸附剂,200mg多壁碳纳米管作为除色素剂对蔬菜中有机磷农药进行提取,提取回收率在90%以上。
(2)本申请通过优化实验,优选甲基丙烯酸(MAA)作为功能单体、三唑磷作为模板分子,二甲基丙烯酸乙二醇酯(EGDMA)作为交联剂,按照1:6:12 的比例进行组装,以三氯甲烷作为制孔剂,乙腈作为溶剂,在无氧条件下采用溶剂热沉淀聚合的方法合成了有机磷类特异性分子印迹聚合物,能够特异性识别筛选甲胺磷、对硫磷、三唑磷等有机磷类农药。
(3)本申请合成了(Z)-3-(1H-苯并[d]咪唑-2-基)-2-羟基-5-甲基苯甲醛肟作为有机磷酸酯特异性可视化响应探针。此探针可在碱催化下与有机磷酸酯类农药发生磷酰化反应,导致探针母环上的电子密度发生改变,从而实现荧光增强和显色反应的高灵敏度二元响应机制,识别浓度可低至纳摩尔级。
(4)本申请将上述分技术串联耦合,在农药提取和富集净化阶段利用优化后的萃取溶剂、脱水剂、除杂剂和除色素对农产品中的农药残留实现高效提取和净化,有效去除了农产品中基质对检测结果的干扰;在富集筛选阶段,利用合成的有机磷类特异性分子印迹聚合物对有机磷农药进行了筛选和富集,保证只有且足量的有机磷类农药在接下来的识别阶段能与识别元件进行接触;在响应识别阶段,利用合成的光学探针可在碱催化下与有机磷酸酯类农药发生磷酰化反应,实现荧光增强和显色反应的高灵敏度二元响应。
实施例1:以甲胺磷为有机磷农药代表,配制0.05ppm、0.1ppm、0.2ppm、 0.5ppm、1ppm五个浓度的甲醇10%标准溶液作为模拟样品,分别经过本研究中上述的基质分散萃取、分子印迹萃取筛选、探针荧光及比色识别反应,记录不同浓度在365纳米波长激发下415纳米发射波长的荧光强度(Intensity),以及 570nm下的可见光吸光度值(OD),并分别制作标准标准曲线,结果表明在 0.05ppm-1ppm之间本方法具有良好的线性范围和检测灵敏度。如附图1所示,线性相关系数为0.9762(荧光法),如附图2所示,线性相关系数为0.9142(比色法),检测灵敏度为0.05ppm。
实施例2:以黄瓜为检测对象,将其切碎成小块,分别称取2g并添加0.2ppm,0.5ppm,1ppm三个不同浓度的甲胺磷,作为模拟实际基质样品,分别经过本研究中上述的基质分散萃取、分子印迹萃取筛选、探针荧光及比色识别反应,记录不同浓度在365纳米波长激发下415纳米发射波长的荧光强度(Intensity),以及570nm下的可见光吸光度值(OD),根据此前制作的标准曲线进行校正。结果表明,三个不同浓度的回收率分别为82.55%(荧光法),91.32%(荧光法) 和103%(比色法)。说明此方法可用于实际农产品中有机磷农药的快速检测。
以上述依据本发明的理想实施例为启示,通过上述的说明内容,相关工作人员完全可以在不偏离本项发明技术思想的范围内,进行多样的变更以及修改。本项发明的技术性范围并不局限于说明书上的内容,必须要根据权利要求范围来确定其技术性范围。
Claims (4)
1.一种基于多重仿生识别的农药残留可视化快速检测技术,其特征在于,该检测方法包括以下步骤:
(1)在农药提取和富集净化阶段利用优化后的萃取溶剂、脱水剂、除杂剂和除色素对农产品中的农药残留实现提取和净化,去除农产品中基质对检测结果的干扰;
(2)在富集筛选阶段,利用合成的有机磷类特异性分子印迹聚合物对有机磷农药进行了筛选和富集,保证只有且足量的有机磷类农药在接下来的识别阶段能与识别元件进行接触;
(3)在响应识别阶段,利用合成的光学探针可在碱催化下与有机磷酸酯类农药发生磷酰化反应,实现荧光增强和显色反应的高灵敏度二元响应。
2.根据权利要求1所述的基于多重仿生识别的农药残留可视化快速检测技术,其特征在于:所述农药提取方法为,取20mL乙腈作为萃取溶剂,2.5g无水硫酸镁作为脱水剂,500mgC18和500mg PSA作为除杂吸附剂,200mg多壁碳纳米管作为除色素剂对蔬菜中有机磷农药进行提取。
3.根据权利要求1所述的基于多重仿生识别的农药残留可视化快速检测技术,其特征在于:所述有机磷类特异性分子印迹聚合物的合成方法为,取甲基丙烯酸作为功能单体、三唑磷作为模板分子,二甲基丙烯酸乙二醇酯作为交联剂,按照1:6:12的比例进行组装,以三氯甲烷作为制孔剂,乙腈作为溶剂,在无氧条件下采用溶剂热沉淀聚合的方法合成了有机磷类特异性分子印迹聚合物。
4.根据权利要求1所述的基于多重仿生识别的农药残留可视化快速检测技术,其特征在于:所述光学探针为,有机磷酸酯特异性可视化响应探针(Z)-3-(1H-苯并[d]咪唑-2-基)-2-羟基-5-甲基苯甲醛肟。
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