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A pretreatment method for determining intracellular microcystins in natural water bodies

A microcystin and pretreatment technology, applied in the field of environmental protection, can solve problems such as heavy workload, low precision, and poor adaptability, and achieve the effects of improving efficiency, avoiding algae residues, and improving utilization

Inactive Publication Date: 2016-09-14
CHINESE RES ACAD OF ENVIRONMENTAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a pretreatment method for determining intracellular microcystins in natural water, so as to solve the problems of large workload, low accuracy and poor adaptability in the background technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The pretreatment of the three intracellular microcystins in the water of Yangcheng Lake in July included the following steps:

[0026] 1) The water sample is not turbid after mixing, and Microcystis is not seen in the process of microscopic examination. Take 3L water sample and filter it.

[0027] 2) The filter membrane was frozen at -20°C for 24 hours and thawed, then shredded into 10mL centrifuge tubes, added 5% acetic acid, vortexed and mixed for 1min, and then crushed intermittently for 10min by an ultrasonic cell disruptor in an ice bath, every 5min Vortex for 0.5 min at 50% power. Centrifuge at 4°C for 12 min at 8000 r / min, and collect the supernatant. Repeat the centrifugation twice, add 5% acetic acid before each centrifugation, vortex mix for 0.5min, and combine the supernatant.

[0028] 3) Activate the HLB solid-phase extraction cartridge with 15mL of methanol, and then activate with 15mL of pure water when the methanol is close to the upper sieve at a flow ...

Embodiment 2

[0031] Pretreatment methods of three intracellular microcystins in water body of Taihu Lake in July

[0032] 1) The water sample is turbid after mixing, and the presence of Microcystis can be clearly seen in the process of microscopic examination. Take 0.5L water sample and filter it.

[0033] 2) The filter membrane was frozen at -20°C for 24 hours and thawed, then shredded into 10mL centrifuge tubes, added 5% acetic acid, vortexed and mixed for 1min, and then crushed intermittently by an ultrasonic cell disruptor in an ice bath for 15min, every 5min Vortex for 0.5 min at 50% power. Centrifuge at 4°C for 12 min at 8000 r / min, and collect the supernatant. Repeat the centrifugation for 4 more times, add 5% acetic acid before each centrifugation, vortex and mix for 0.5 min, and combine the supernatant.

[0034] 3) Activate the HLB solid-phase extraction cartridge with 15mL of methanol, and then activate with 15mL of pure water when the methanol is close to the upper sieve at a ...

Embodiment 3

[0037] Pretreatment methods of three intracellular microcystins in water body of Taihu Lake in April

[0038] 1) The water sample is turbid after mixing, and Microcystis can be seen in the process of microscopic examination. Take 1L of water sample and filter it.

[0039] 2) The filter membrane was frozen at -20°C for 24 hours and thawed, then shredded into 10mL centrifuge tubes, added 5% acetic acid, vortexed and mixed for 1min, and then crushed intermittently by an ultrasonic cell disruptor in an ice bath for 15min, every 5min Vortex for 0.5 min at 50% power. Centrifuge at 4°C for 12 min at 8000 r / min, and collect the supernatant. Repeat the centrifugation 3 times, add 5% acetic acid before each centrifugation, vortex mix for 0.5min, and combine the supernatant.

[0040]3) Activate the HLB solid-phase extraction cartridge with 15mL of methanol, and then activate it with 15mL of pure water when the methanol is close to the upper sieve, with a flow rate of 5mL / min. Then the...

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Abstract

The invention relates to a pretreatment method for measuring intracellular microcystin in natural water. The method comprises the steps of (1) carrying out microscopic examination before a water sample is filtered, and determining the quantity of needed water according to the turbid degree of the water sample and the detection effect of microcystis; (2) filtering the water sample, carrying out cell disruption after a filter membrane is cut into pieces, rotated in a vortex way and evenly mixed, and carrying out centrifugal separation to obtain supernate; (3) enriching the supernate containing microcystin in a solid-phase extraction column; and (4) eluting the microcystin enriched in the solid-phase extraction column, drying by blowing nitrogen, and making up to volume for testing. The method is simple, high in flexibility and high in operability, and is suitable for the pretreatment in the long-term measurement of intracellular microcystin in natural water at different seasons and in different areas.

Description

technical field [0001] The invention belongs to the technical field of environmental protection, and in particular relates to a water sample pretreatment method for long-term determination of common intracellular microcystins in different natural water bodies in different seasons. Background technique [0002] In recent years, the eutrophication of water bodies has become increasingly serious, resulting in frequent occurrence of cyanobacterial blooms. There are more than 40 species of toxins in freshwater cyanobacteria, among which Microcystins (MCs) are the cyanotoxins with the highest frequency and the most serious damage, which seriously endanger the health of the ecosystem of water bodies. More than 80 isomers of microcystins have been found, the most common are MC-LR, MC-RR and MC-YR, among which MC-LR is the most toxic. [0003] Algal toxins are divided into intracellular algal toxins and extracellular algal toxins. Normally, algal toxins exist in algal cells, but in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28
Inventor 苏婧席北斗魏代春霍守亮纪丹凤王骥
Owner CHINESE RES ACAD OF ENVIRONMENTAL SCI
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