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Sample preparation treatment method for excrement and soil metabonomics study

A metabolomics, sample preparation technology, applied in scientific instruments, measuring devices, instruments, etc., can solve problems such as loss of activity, cumbersome process, incomplete metabolome extraction, etc.

Active Publication Date: 2017-09-01
赛思莱(厦门)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current conventional metabolome extraction and processing methods are cumbersome, and the extraction and processing process is prone to pollution and uncertain factors, and is usually accompanied by denaturation of metabolites, loss of activity and low recovery rate of metabolites
Moreover, because a single extraction solution is often used, the extraction of the metabolome is incomplete.

Method used

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  • Sample preparation treatment method for excrement and soil metabonomics study
  • Sample preparation treatment method for excrement and soil metabonomics study
  • Sample preparation treatment method for excrement and soil metabonomics study

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: A method for preparing and processing samples for rat stool metabolomics research

[0036] (1) Sample pretreatment: collect rat feces pellets in a metabolic cage, freeze them overnight in a refrigerator at -80°C, put them in a freeze dryer, dry to constant weight, grind and break with liquid nitrogen, pass through a 100-mesh sieve, and weigh 100 mg of feces Powder, add 1 mL of water-soluble metabolite extract at -20°C (-20°C is to prevent biodegradation of metabolites in the sample), pipette the feces with a pipette, and mix them to obtain the first suspension; The water-soluble metabolite extract is a mixture of methanol, acetonitrile, acetic acid and water in a volume ratio of 1:1:0.1:2.

[0037] (2) Shaking and crushing extraction: adding 2 steel balls with a diameter of 3mm after cleaning and baking at 400°C into the first suspension obtained in step (1), shaking and crushing at low temperature, the oscillation frequency is 80Hz, and the time is 120s;

[0038] ...

Embodiment 2

[0062] Example 2: A method for preparing and processing samples for soil metabolomics research

[0063] (1) Sample pretreatment: Weigh an appropriate amount of soil sample, put it into a glass beaker, freeze it overnight at -80°C, put it in a freeze dryer, dry it to constant weight, add liquid nitrogen to grind and break, and pass through a 100-mesh sieve. Weigh 300 mg of soil powder, add 3 mL of -20°C water-soluble metabolite extract, pipette with a pipette to mix evenly to obtain the first suspension;

[0064] (2) Shaking and crushing extraction: 6 steel balls with a diameter of 3mm after being cleaned and baked at 400°C are added to the first suspension obtained in step (1), and shaken and broken at low temperature, the oscillation frequency is 100Hz, and the time is 90s. Obtain the first suspension after crushing;

[0065] (3) Ultrasonic extraction: the first suspension solution obtained in step (2) is sonicated on ice to further break to dissolve the metabolites. The power of t...

Embodiment 3

[0077] Example 3: A method for preparing and processing samples for neonatal meconium metabolomics research

[0078] (1) Sample pretreatment: Weigh a sample of neonatal meconium, put it in a glass beaker, freeze it overnight at -80°C, put it in a freeze dryer, dry it to a constant weight, add liquid nitrogen to grind and break, and pass through a 100-mesh sieve . Weigh 200 mg of sample powder, add 2 mL of -20°C water-soluble metabolite extract, pipette with a pipette to mix evenly to obtain the first suspension;

[0079] (2) Vibration and crushing extraction: 4 steel balls with a diameter of 3mm after being cleaned and baked at 400°C are added to the first suspension obtained in step (1), and shaken and broken at low temperature, the vibration frequency is 120Hz, and the time is 60s. Obtain the first suspension after crushing;

[0080] (3) Ultrasonic extraction: the first suspension solution obtained in step (2) is sonicated on ice to further break to dissolve the metabolites. The ...

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Abstract

The invention discloses a sample preparation treatment method for excrement and soil metabonomics study. The sample preparation treatment method comprises the following steps: pretreating a sample to obtain first suspension; adding a steel ball and shaking, breaking and extracting to obtain the broken first suspension; performing ultrasonic treatment; centrifuging; collecting first supernate and precipitate; concentrating the first supernate until the first supernate is nearly dry; adding water-soluble re-dissolution liquid and a steel ball, and shaking for re-dissolution; centrifuging at a high speed, wherein obtained second supernate is a water-soluble metabolism extract which can be analyzed directly on a machine; adding a fat-soluble metabolite extracting solution into the collected precipitate, and mixing uniformly to obtain second suspension; shaking, breaking and extracting; performing ultrasonic treatment; centrifuging; collecting a lower-layer solution; blowing to dry the obtained lower-layer solution through a nitrogen blowing way; adding fat-soluble re-dissolution liquid and a steel ball, and shaking for re-dissolution; centrifuging at a high speed, wherein an obtained product is a fat-soluble metabolism extract which can be analyzed directly on the machine. The method disclosed by the invention has the advantages of simple operation, strong stability, comprehensiveness and high recovery rate, and is of great significance to extensive related studies of excrement and soil metabonomics.

Description

Technical field [0001] The invention relates to a method for extracting and processing total metabolites for the purpose of metabolomics analysis in the processing of feces and soil samples. In particular, it relates to a sample preparation and processing method for fecal and soil metabolomics research. Background technique [0002] Metabolomics, as a new technology emerging after genomics and proteomics, is an important component of systems biology. Metabolomics conducts a comprehensive analysis of the qualitative and quantitative changes of metabolites involved in all life activities in the organism at a certain time, thereby revealing the physiological state of the organism. Metabolomics has high-throughput detection and powerful data model analysis capabilities, and has been widely used in many research fields, such as disease diagnosis and mechanism, drug research and development, pollutant toxicology, nutritional food science, botany, and ecology. Using metabolomics, high...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88
CPCG01N30/06G01N30/88
Inventor 张洁王晓飞郭睿
Owner 赛思莱(厦门)科技有限公司
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