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Fabrication Process Research of MC-LR

Author: ChenYongFang
Tutor: WangLiGuo；LiuSiQuan
School: Jinan University
Course: Environmental Science
Keywords: Microcystin - LR Preparation High Performance Liquid Chromatography High performance liquid chromatography - mass spectrometry Characterization
CLC: X524
Type: Master's thesis
Year: 2011
Downloads: 78
Quote: 0
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Abstract


In recent decades, the blue-green algae blooms caused by pollution of freshwater lakes gradually intensified. In recent years China's Yunnan Dianchi Lake, Taihu Lake in Jiangsu outbreak of cyanobacterial blooms increased year by year, release harmful substances microcystin (microcystins, MCs). Microcystin-LR standard as MCs major isomer unconventional indicators of drinking water quality detection standard samples. Microcystin isomer pure extraction, purification and preparation of the research reported in the literature are many, involving solid-phase extraction and high performance liquid chromatography, thin layer chromatography. However, our study extracted from cyanobacteria and preparation of MC-LR has not yet formed a mature method required standard almost all imported from abroad. Natural blue-green algae from Dianchi as raw material, to carry out in-depth research by the blue-green algae extract algae toxin extraction conditions, the three aspects of the algal toxin isomer mixture separation conditions, microcystin-LR preparation conditions, has been MC-LR standard to meet the water quality monitoring requirements. And the sample obtained were characterized by HPLC-MS. Isomers of microcystin extracted from natural cyanobacteria exploration and optimization. Different solvent systems, extraction means, and extraction time on the effects of microcystin extract algae residue microcystins crude extracts of the best conditions: methanol / water (65% v / v) as an extraction solvent at room temperature, magnetic stirring 1.5h, ultrasound 20min crude extracted to obtain a crude extract, and then the impurities of macromolecules as well as fibers and the like was removed by filtration with 0.45μm microfiltration membrane MCs. MCs crude pre-purified by solid phase extraction means, explore the eluent eluent the ratio of the effect of purification by solid phase extraction process, 70% eluent maximum microcystin from solid phase extraction column eluted. Means of the microcystin isomers extracted crude product was purified separation effects of different proportion, equal to the flow of the different types of chromatographic conditions on the MCS isomer separation effect obtained using high performance liquid chromatography (HPLC) microcystin isomers optimal chromatographic separation conditions: mobile phase: acetonitrile / water solution (80% v / v); Column temperature: room temperature; detection wavelength: 238 nm; flow rate: 0.5 mL / min; Injection volume: 2μL. Under these conditions, the the microcystin three major isomer can get good separation. Using preparative high performance liquid chromatography for preparative separation of microcystin major isomer, by exploring different proportion of the current phase and injection volume of preparation effects, best preparative chromatography conditions: mobile phase: acetonitrile / water solution (50% v / v); Column temperature: room temperature; detection wavelength: 238 nm; flow rate: 2 mL / min; injection volume: 100 μl. Collection of 10 peaks corresponding components, each component in high performance liquid chromatography, chromatography and spectroscopy (DPD) microcystin mainly three kinds of isomers of MC-LR, and MC-RR and MC-YR basic qualitative, of which more than 90% purity of MC-LR. HPLC-MS characterization was prepared by MC-LR, the molecular ion peaks and peaks consistent with MC-LR. Finally Microcystis aeruginosa strains obtained from the Chinese Academy of Sciences, Wuhan aquatic laboratory culture of Microcystis aeruginosa, MC-LR standard to the same extract preparation, Preparation and characterization results with natural blue-green algae as raw material The results are consistent. Natural blue-green algae as the starting raw material preparation microcystin-LR method is simple, easy to operate, the product is a single batch prepared a large amount of high purity research Microcystis algae extract and preparation of other isomers of the toxin has a good guiding significance.

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CLC: > Environmental science, safety science > Environmental pollution and its prevention > Water pollution and its control > Lakes and reservoirs
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