摘要：本文主要用乙酰胆碱酯酶为催化剂，氯化硫代乙酰胆碱为底物，二硫代二硝基苯甲酸为显色剂，采用722可见分光光度计在412 nm波长下测定乙酰胆碱酯酶活性。研究了乙酰胆碱酯酶用量、底物和显色剂浓度对显色反应的影响，并分别对溶液状的乙酰胆碱酯酶和脱乙酰基魔芋葡甘聚糖包埋固定的乙酰胆碱酯酶的活性进行了比较。结果显示酶用量为10 μL、底物用量为5 μL，显色剂用量为50 μL显色效果好。在412 nm的波长下，凝胶状的乙酰胆碱酯酶与溶液状的乙酰胆碱酯酶活性相似。此外，探讨了农药对乙酰胆碱酯酶活性的抑制效果。数据表明，有机磷农药对乙酰胆碱酯酶具有抑制作用，其抑制率与农药的浓度成线性关系。20097
关键词：活性测定；乙酰胆碱酯酶活性；分光光度法；脱乙酰基魔芋葡甘聚糖
Study on the determination of acetylcholinesterase activity by Spectrophotometry
Abstract:This paper is mainly used Acetylcholinesterase as catalyst, Acetylcholine chloride as substrate, 5,5’-Dithio bis-(2-nitrobenzoic acid) as chromogenic agent, using 722 spectrophotometer determination of acetylcholinesterase activity in the 412 nm wavelength. And acetylcholinesterase on solution like and deacetylated konjac glucomannan gel were made from the acetylcholinesterase activity and inhibitory rate comparison. Under certain conditions, organic phosphorus and carbamate pesticide inhibit cholinesterase, the inhibitory rate was positively correlated with the concentration of pesticides。And the different amounts of acetylcholinesterase, substrate and chromogenic agent activity and inhibition rate, determine the amount of enzyme is 10 μL , substrate is 5 μL, chromogenic agent is 50 μL, reduce the cost. Through two kinds of enzyme in the best state, under the wavelength of 412 nm,activity of gel acetylcholinesterase is similar to liquid acetylcholinesterase. Liquid acetylcholinesterase has the higher inhibitory rate.
Keywords: determination of activity ; acetylcholinesterase; spectrophotometry; deacetyled konjac glucomannan
目 录
一 前言 1
1.1 农药残留给人带来的危害 1
1.2 农药残留的一些检验方法 2
1.2.1 色谱法 2
1.2.2 有机磷农药降解酶法 2
1.2.3 生物传感器 2
1.2.4 速测灵法(金属离子催化显色表面皿法) 3
1.2.5 酶抑制分析法 3
1.3 市场上常用农药残留检测仪器 3
1.3.1 速测卡目测法(纸片法)农药残留快速检定仪 3
1.3.2 智能化农药残留综合速测仪 3
1.3.3 全自动大量农残检验仪 4
1.4 酶抑制检验法原理 4
二 试剂和仪器 5
2.1 试剂 5
2.2 仪器 5
2.3实验过程 5
2.3.1脱乙酰基魔芋葡甘聚糖溶胶的配制 5
2.3.2溶液配制 6
2.3.3酶活力的测定 6
2.3.4计算公式 7
三 结果与讨论 8
3.1空白样品吸光度△A0的测定 8
3.2不同农药浓度下的吸光度对比 8
3.3改变试剂用量对吸光度的影响 10
3.3.1改变酶用量对活性的影响 10
3.3.2改变酶用量对抑制率的影响 10
3.3.3改变底物用量对活性的影响 11
3.3.4改变底物用量对抑制率的影响 11
3.3.5改变显色剂用量对活性的影响 12
3.4溶液状酶与凝胶状酶对比 13 分光光度法测定乙酰胆碱酯酶活性的研究:http://www.751com.cn/yixue/lunwen_11735.html