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Method for detecting microcystin in water

A technology of microcystin and detection method, which is applied in measurement devices, instruments, Raman scattering, etc., can solve the problems of expensive equipment, large sample consumption, inability to identify MC species, etc., and achieve the effect of sensitive detection

Inactive Publication Date: 2012-06-20
INST OF CHEM CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The detection methods for microcystins mainly include traditional mouse intraperitoneal injection bioanalysis method, enzyme-linked immunoassay, protein phosphatase inhibition and other biochemical analysis methods, but these methods are complicated to operate, time-consuming and unable to identify the type of MC, etc. Disadvantages, while the commonly used high-performance liquid chromatography (HPLC), sample consumption is large and pollutes the environment
Although capillary electrophoresis and mass spectrometry (CE-MS) require less samples and high separation efficiency, and can also confirm the type of MCs through molecular weight information, this method requires expensive equipment, professional skills, high cost and time-consuming

Method used

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  • Method for detecting microcystin in water

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preparation example Construction

[0025] (1) Preparation of MAb-MPs solution

[0026] Firstly prepare magnetic nanoparticles (select ferric oxide nanoparticles in this embodiment), prepare by hydrothermal method, the specific steps are as follows: 0.45g~0.75g ferric chloride, 0.10g~0.20g trisodium citrate are dissolved in 20ml ~40ml of ethylene glycol, then add 1.0g~2.0g of sodium acetate, stir for 30min, pour the resulting mixture into a 50ml reaction kettle, and react at 200°C for 10h; Wash with ethanol three times, and finally dissolve in water to obtain an aqueous solution of iron ferric oxide nanoparticles with a diameter of 180nm to 250nm. figure 1 as shown in (c);

[0027] Microcystin monoclonal antibodies (MAbs), respectively MC-LR, MC-YR and MC-RR, were linked to the prepared magnetic nanoparticles with EDC-NHS coupling agent, the specific steps are as follows : Mix 20 μL of a molar concentration of 3.0 mM NHS (N-hydroxysuccinimide) and 20 μL of a molar concentration of 3.0 mM EDC (1-(3-dimethylamin...

Embodiment 1

[0037] Embodiment 1, the detection of microcystin MC-LR in water

[0038] (1) Drawing of detection standard curve

[0039] 10 μL of MC-LR standard solutions of different concentrations (concentrations are 0 μg / L, 0.02 μg / L, 0.04 μg / L, 0.06 μg / L, 0.1 μg / L, 0.5 μg / L, 1 μg / L, 1.5 μg / L L, 2μg / L, 2.5μg / L, 3μg / L, 3.5μg / L and 4μg / L) were mixed with 27.5μL of MC-SERS Tags (connected with MC-LR molecule) solution in turn to obtain the incubation solution; solution was added to 20 μL of MAb-MPs (MC-LR monoclonal antibody) solution and shaken for 25 minutes, the magnetic immune complex in the solution was separated with a magnet, the remaining solution was discarded, and the obtained immune complex was washed with PBS Wash twice, then detect its Raman signal with a laser Raman spectrometer, (laser Raman detects the incident light wavelength is 532nm, the power is 2mW, and the spectral detection time is 10s); when using different MC-LR concentrations, the Raman spectrum is 1076cm -1 The...

Embodiment 2

[0042] Embodiment 2, the detection of microcystin MC-YR in water

[0043] (1) Drawing of detection standard curve

[0044] 10 μL of different concentrations of MC-YR standard solution (concentrations are 0 μg / L, 0.02 μg / L, 0.04 μg / L, 0.06 μg / L, 0.1 μg / L, 0.5 μg / L, 1 μg / L, 1.5 μg / L L, 2μg / L, 2.5μg / L, 3μg / L, 3.5μg / L and 4μg / L) were mixed with 27.5μL of MC-SERS Tags (connected with MC-YR molecule) solution in turn to obtain the incubation solution; solution was added to 20 μL MAb-MPs (connected with MC-YR monoclonal antibody) solution and reacted for 25 minutes by shaking, the magnetic immune complex in the solution was separated with a magnet, the remaining solution was discarded, and the obtained immune complex was washed with PBS Wash twice, then detect its Raman signal with a laser Raman spectrometer, (laser Raman detects the incident light wavelength is 532nm, the power is 2mW, and the spectral detection time is 10s); when using different MC-YR concentrations, the Raman spe...

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Abstract

The invention discloses a method for detecting microcystin in water. The method comprises the following steps of: (1) preparing magnetic particles connected with monoclonal antibodies of microcystin; (2) preparing a Raman probe connected with microcystin molecules by Raman enhancement of silver nano particles on micromolecules; (3) drawing a standard detection curve of the microcystin under the determined optimum reaction condition; and (4) determining the concentration of a sample to be determined by comparing detected peak intensity of a specified peak of a Raman spectrum of the sample and the standard curve. In the detection process, laser Raman enhancement and competition immunity are combined, and magnetic separation is applied; and compared with the conventional detection method, the method for detecting the microcystin in water greatly shortens detection time. The method for detecting the microcystin in water can also be used for detecting other kinds of microcystin molecules and small molecules, and a quick and convenient detection way is provided with pollutant in water.

Description

technical field [0001] The invention relates to a method for detecting microcystin in water, belonging to the technical field of microcystin detection. Background technique [0002] The frequent occurrence of harmful cyanobacterial blooms caused by eutrophication of water bodies caused by environmental pollution has become an environmental problem of widespread concern at home and abroad. Microcystins (MCs) are a class of hepatotoxins released by harmful cyanobacteria blooms with strong cancer-promoting effects, and more than 60 isomers have been found. Microcystins are stable in nature, not deactivated after boiling, non-volatile, resistant to pH changes, soluble in water, methanol and acetone. The damage of microcystins to organisms is mainly manifested as liver toxicity and neurotoxicity, and also damages the kidney, adrenal gland, lung and stomach to varying degrees. Cyanobacterial blooms and their toxins have been listed as detection items for microorganisms and organ...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/65
Inventor 江龙张兴华李津茹
Owner INST OF CHEM CHINESE ACAD OF SCI
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