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水质下降、水体环境污染和富营养化问题日趋严重，特别是蓝藻水华问题在淡水湖泊的频繁爆发，已引起全球的关注。在蓝藻死亡后，其细胞裂解，并向环境介质中释放多种不同类型的微囊藻毒素(MCs)，对人类产生危害。因此需要建立快速、灵敏、稳定、高效的方法对环境介质中的微囊藻毒素进行定性、定量分析。同时，需要进一步研究其生物降解机理，对已爆发蓝藻问题的水域进行治理。本论文在比较有关研究蓝藻、微囊藻毒素研究的基础上，进一步验证和探究微囊藻毒素MC-LR的提取、纯化以及分析检测条件。优化了固相萃取(SPE)结合高效液相色谱法(HLPC)分析水中的MC-LR的方法，主要确定了20%的甲醇为淋洗液、含有0.06%三氟乙酸(THF)的纯甲醇为洗脱液的MC-LR的SPE提取条件。并初步探究了漆酶、锰过氧化物酶对不同浓度MC-LR的降解作用。8110
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关键词微囊藻毒素、固相萃取、高效液相色谱、锰过氧化物酶
Title Study on the extraction and biodegradation of microcystins
Abstract
The deterioration of water quality, serious environmental pollution and eutrophication
of water, especially the high frequency of algal bloom in freshwater lakes, has attracted
global attention. After the death of the cyanobacteria, the cells split and release various
types of microcystins (MCs) to the environmental media, which are harmful to human
beings. Therefore, it’s needed to establish a rapid, sensitive, stable and efficient method
for qualitative and quantitative analysis of MCs in environmental media. At the same
time, a further study on its biodegradation mechanism is required, which is used for
controlling algal bloom in lakes.
On the foundation of the studies available on cyanobacteria and MCs, the
extraction, purification and analysis of testing conditions of MC-LR are verified and
explored. A method for analysis of MC-LR in water using SPE and HPLC has been
optimized. Mainly carried out SPE extraction conditions of MC-LR, 20% methanol and
a mix solution of pure methanol and 0.06% trifluoroacetic acid (THF) was used as
eluent. And the degradation of different concentrations of MC-LR by using laccase and
manganese peroxidase is elementarily inquired.
Keyword Microcystins SPE HLPC Manganese peroxidase 目 次
1 绪论 .. 1
1.1 蓝藻水华的现状、危害及防治 . 1
1.1.1 蓝藻水华的现状 1
1.1.2 蓝藻水华的危害 2
1.1.3 蓝藻水华的防治 2
1.2 微囊藻毒素的研究进展 . 3
1.2.1 微囊藻毒素的种类、结构 .. 3
1.2.2 微囊藻毒素的毒理性质 4
1.2.3 微囊藻毒素的分析检测 5
1.2.4 微囊藻毒素的生物降解 5
1.3 漆酶的结构与应用 6
1.4 锰过氧化物的结构与应用 7
1.5 本课题的研究内容 7
2 实验原理 8
2.1 固相萃取(SPE)原理 . 8
2.2 高效液相色谱原理 9
3 实验部分 .. 11
3.1 微囊藻的培养 11
3.1.1 实验材料 . 11
3.1.2 实验方法 . 11
3.2 MC-LR的提取、分离 . 12
3.2.1 实验仪器 12
3.2.2 MC-LR的提取 12
3.3 MC-LR的分析检测 .. 13
3.3.1 实验材料 13