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Method for detecting hydroxyl radicals in earthworm body

A technology of free radicals and earthworms, applied in the direction of analysis by electron paramagnetic resonance, etc., to achieve the effect of easy mastery, simple operation and wide practicability

Inactive Publication Date: 2010-02-03
NANJING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are many studies on the oxidative damage of the body caused by the free radicals produced by earthworms under the exposure stress of various pollutants, and the responses of antioxidant enzymes or the determination of the end products of reactive oxygen toxicity are often used to speculate the body’s oxidative damage. However, direct evidence of free radical production induced by pollutant stress is seldom reported.

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  • Method for detecting hydroxyl radicals in earthworm body
  • Method for detecting hydroxyl radicals in earthworm body
  • Method for detecting hydroxyl radicals in earthworm body

Examples

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Embodiment 1

[0028] Determination of Hydroxyl Free Radicals in Earthworms under Heavy Metal Cd Stress in Example 1

[0029] Select the model organism species of international soil ecotoxicology research—Eiseniafetida, and after domestication for a period of time, select healthy adult worms with a body weight of 0.3-0.4g, basically the same size, and mature rings. After 24 hours, wash the earthworms 2-3 times repeatedly at room temperature with ultrapure water, and dry the water on the surface of the earthworms with filter paper. Earthworms were exposed to soil contaminated with different concentrations of Cd for 14 days. The concentrations of Cd were 0, 0.1, 0.5, 1.0, 5.0, and 10.0 mg / kg soil, with 4 parallels for each concentration group. After the exposure was complete, the earthworms were taken out and washed repeatedly with iced saline, and 0.1 g of earthworms were quickly weighed and placed in a glass homogenizer, and 1 mL of dimethyl sulfoxide was added as a solvent to prepare a PBN ...

Embodiment 2

[0033] Determination of Hydroxyl Free Radicals in Earthworms under Example 2 Tetrabromobisphenol A (TBBPA) Stress

[0034] Select the model organism species of international soil ecotoxicology research—Eiseniafetida, and after domestication for a period of time, select healthy adult worms with a body weight of 0.3-0.4g, basically the same size, and mature rings. After 24 hours, wash the earthworms 2-3 times repeatedly at room temperature with ultrapure water, and dry the water on the surface of the earthworms with filter paper. Earthworms were exposed to soil contaminated with different concentrations of TBBPA for 14 days. After the exposure was complete, the earthworms were taken out and washed repeatedly with iced saline, and 0.1 g of earthworms were quickly weighed and placed in a glass homogenizer, and 1 mL of dimethyl sulfoxide was added as a solvent to prepare a PBN solution with a concentration of 50 mmol / L (the addition process In order to avoid the entry of water, wh...

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Abstract

The invention discloses a method for detecting hydroxyl radicals in an earthworm body. Firstly, a fully exposed earthworm undergoes bowel clearing and is then cleaned. Radical PBN capture is carried out in a deoxygenated hermetical operation box. Finally, EPR technology is used for detection. The method has the advantages that the operation is relatively simple and easy to master and the hydroxylradicals of the earthworm can be detected within a short time; according to a hyperfine structure constant of an EPR spectrogram and the shape analysis of the spectrogram, the method discovers the detected radicals to be the hydroxyl radicals and is the most direct, accurate and effective method at present for detecting the hydroxyl radicals; and the method has a good dose-response relationship. Research results show that the strength of the hydroxyl radical and the exposed concentration are in good dose-response relationship.

Description

technical field [0001] The invention relates to a method for measuring hydroxyl free radicals in earthworms. Background technique [0002] Free radicals are atoms, molecules or groups of atoms containing unpaired electrons, which are produced by mitochondria in the process of electron transfer. In the metabolic process of life activities, various free radicals are constantly produced, including superoxide anion (O 2 · - ), hydroxyl radical ( OH ), hydroperoxyl (HO 2 - ), alkoxy (RO ), alkoperoxy (ROO ), hydrogen peroxide (H 2 o 2 ), nitric oxide (NO), etc. Hydroxyl radicals are the most powerful and important active oxygen radicals. Under normal physiological conditions, there is a dynamic balance mechanism between the production of free radicals and the antioxidant defense system, which is beneficial to the body. When the production of free radicals exceeds the balance mechanism, the body produces oxidative stress. Studies have shown that the cytotoxicity of free ra...

Claims

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Application Information

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IPC IPC(8): G01N24/10
Inventor 王晓蓉薛银刚姜锦林顾雪元吴可
Owner NANJING UNIV
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