论文总字数：20448字

摘 要

敌草隆作为一种在水稻田广泛施用的除草剂，近期研究表明过量使用不仅对作物产生药害，而且对哺乳生物具有急慢性毒性效应和生理生态毒性，对食品安全产生威胁。植物激素茉莉酸在提高植物防御及解毒能力中发挥重要作用。因此，建立一种稳定、灵敏的敌草隆定量方法对研究其茉莉酸处理下的水稻代谢具有非常重要的科研意义。本课题以水稻为研究对象，建立水稻体内敌草隆的液相检测方法，并探讨茉莉酸处理下，体内敌草隆的变化情况。具体结果如下：

1. 敌草隆液相条件的优化。本文从检测波长和流动相选择两方面入手，通过紫外全波长扫描确定敌草隆的最大吸收波长为251 nm；以分离度gt;2.0且达到基线分离为标准，确定流动相比例为甲醇：水=65：35（v/v）；根据敌草隆物化性质，色谱柱选用C18反相柱；
2. 复杂环境中敌草隆前处理方法的优化。本文选择水稻叶、根和培养液为三种基质，从提取试剂和时间、富集净化方法以及洗脱体积等方面入手，通过加标回收实验，确定前处理方法，具体为：15mL乙腈提取三次，每次提取时间为20分钟，旋干后用20 mL超纯水（含千分之五的甲醇）复溶，复溶液过SPE C18小柱浓缩净化，用3 mL洗脱，收集洗脱液，过0.22μm 后进液相检测，该方法的回收率在87%~105%。
3. 用0.125和0.25 mg/L敌草隆处理10天苗期的水稻幼苗，暴露6天后测定水稻植株和其培养液中敌草隆的积累量和残留量。结果显示，0.25mg/L处理的水稻体内敌草隆积累量明显高于0.125mg/L处理组。但两个浓度处理的生物转移因子（BCF）无显著差异，说明水稻对不同浓度的敌草隆的转移能力没有差异。

关键词：茉莉酸 敌草隆 液相检测 水稻

Establishment of Diuron-Detection Liquid Phase in Rice and Its Accumulation in Rice and Its Medium

Abstract

Diuron, a herbicide widely used in rice fields, has recently shown that excessive use not only produces phytotoxicity to crops, but also has acute and chronic toxicity effects and physiological and ecological toxicity to mammals, and poses a threat to food safety. The phytohormone jasmonate plays an important role in enhancing plant defense and detoxification. Therefore, the establishment of a stable and sensitive quantitative method of diuron has important scientific significance for studying the rice metabolism under the treatment of jasmonic acid. In this study, rice was taken as the object of study to establish a liquid-phase detection method for jasmonic acid in rice, and to investigate the changes of diuron in the body treated with jasmonic acid. The specific results are as follows:

Rice is an annual grass plant and is one of the most important food crops in China. The population that uses rice as a staple food accounts for about 65% of the country's population. It can be seen that the safe production of rice is related to the country’s lifeline and is also the basis for the country’s sustainable development. In this study, rice seedlings were used as the research object to investigate how rice seedlings were tested under different concentrations of diuron to detect the residues of diuron in rice and culture fluids to reflect the effects of diuron on rice seedlings and their physiological activities. The effect of diuron on the dose-effect relationship between pesticide stress and rice toxicity. The effect of diuron on antioxidant enzyme activities in rice was further investigated, and the response mechanism of rice to diuron-poisoning was initially explored.

1. Optimization of the liquid phase condition of diuron. In this paper, starting from the detection wavelength and the choice of mobile phase, the maximum absorption wavelength of diuron was determined to be 251 nm by UV full-wavelength scanning; the flow ratio was determined to be methanol:water = 65:35 (v/v); Based on the physicochemical properties of diuron, the column was selected using a C18 reversed-phase column;
2. Optimization of pretreatment methods for diuron in a complex environment. In this study, rice leaves, roots and broth were selected as the three substrates, starting from extraction reagents and time, enrichment and purification methods, and elution volume, etc., and the pretreatment method was determined by adding the standard recovery experiments. Specifically, 15 mL of acetonitrile was extracted three times. Each extraction time is 20 minutes. After spin-drying, it is reconstituted with 20 mL of ultra-pure water (containing 5/1000 of methanol). The complex solution is concentrated and purified by SPE C18 cartridge and eluted with 3 mL to collect the eluent. After 0.22μm late liquid phase detection, the recovery rate of this method is 87% ~ 105%.
3. The seedlings of rice seedlings were treated with 0.125 and 0.25 mg/L diuron for 10 days. After 6 days of exposure, the accumulation and residual amounts of diuron in rice plants and their culture fluids were measured. The results showed that the accumulation of diuron in 0.25 mg/L treated rice was significantly higher than that in the 0.125 mg/L treatment group. However, there was no significant difference in BCF between the two concentrations, indicating that there was no difference in the ability of rice to transfer to different concentrations of diuron.

Key Words：Jasmonate；Quantitative methods；Exogenous stress；Rice

目 录

摘要………………………………………………………………………………………………I

ABSTRACT…………………………………………………………………………………… II

第一章 文献综述 1

1.1敌草隆概况 1

1.1.1 敌草隆的基本性质 1

1.1.2 敌草隆的毒性 1

1.1.3 敌草隆的应用及作用机理 1

1.1.4 敌草隆对环境的影响 2

1.1.5 敌草隆检测方法 2

1.2 高效液相色谱 3

1.2.1 概念 3

1.2.2 应用范围及优点 3

1.3 水稻概况 3

1.3.1 水稻的简介 3

1.3.2 水稻农药污染概况 4

1.4 研究与目的 4

第二章 实验材料及方法 5

2.1 材料 5

2.2 主要仪器 5

2.3实验方法 6

2.3.1水稻种子萌发及幼苗的培养和处理 6

2.3.2敌草隆的配制 7

2.3.3水稻幼苗加药处理 8

2.3.3液相色谱条件 8

2.3.4添加回收 9

2.3.5水稻的农药残留测定 10

第三章 结果与分析 11

3.1 敌草隆液相检测方法确定 11

3.1.1 检测波长的选择 11

3.1.2 标准曲线的绘制和检出限的计算 11

3.1.3 样品前处理方法的优化 12

3.1.4 液相条件的优化及样品检出限和定量限的计算 15

3.1.5不同浓度、不同介质的加标回收 16

3.2水稻及其培养液中的敌草隆积累量和残留量的研究 19

3.3分析及展望 20

参考文献 22

致谢 25

文献综述

1.1敌草隆概况

1.1.1 敌草隆的基本性质

Diuron是敌草隆的英文名，C₉H₁₀Cl₂N₂O是它的分子式，233.09 g/mol是它的分子量，化学结构式为：

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