Efficient cotton metabonomics sample preparation method

A metabolomics, cotton technology, applied in the field of cotton metabolomics analysis

Pending Publication Date: 2021-07-30
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies of the prior art, the present invention provides an efficient sample preparation and detection method

Method used

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  • Efficient cotton metabonomics sample preparation method
  • Efficient cotton metabonomics sample preparation method
  • Efficient cotton metabonomics sample preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Randomly select a cotton variety planted in a field, take a sample of the second leaf of the plant at the budding stage for metabolomics analysis, and test the effect of the patented method on sample pretreatment and detection. The specific steps are as follows

[0027] 1) Take the second leaf sample of cotton in the bud-boll stage, and quickly immerse it in liquid nitrogen to save it for later use;

[0028] 2), put the frozen sample in step 1) into a vacuum freeze-drying apparatus for freeze-drying. The parameters of the vacuum freeze-drying apparatus are set to 0.025 standard atmospheric pressure, minus 50 degrees Celsius, and the freeze-drying time is Twenty-four hours to obtain a dry sample;

[0029] 3), accurately weigh 100 mg of the dried sample obtained in step 2), put 100 mg of the dried sample into a centrifuge tube with a volume of 2 ml, then add steel balls, and then grind it at a frequency of 60 Hz ninety seconds;

[0030] 4) Mix and shake metha...

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Abstract

The invention relates to the technical field of cotton metabonomics analysis, and discloses an efficient cotton metabonomics sample preparation method which is characterized by comprising the following steps: 1) taking a cotton specific tissue sample, and quickly immersing the cotton specific tissue sample in liquid nitrogen for preservation to prepare a frozen sample; according to the sample preparation and detection method disclosed by the invention, the weighing of the sample is more convenient and accurate through vacuum freeze drying, and the stability of metabolites in the sample is also ensured; formic acid is added into the extracting solution, the oxidation reaction of metabolites in the extraction reaction process is weakened by a method for reducing the pH value of the extracting solution, and the stability of the sample extraction process is further maintained; a relatively appropriate chromatographic gradient is optimized through multiple tests, and the chromatographic method can be used for compressing the time of the peak appearance stage of fewer metabolites, reducing the analysis duration, improving the separation efficiency, prolonging the peak appearance time period of more metabolites and reducing the ion inhibition effect in mass spectrometry.

Description

technical field [0001] The invention relates to the field of cotton metabolomics analysis, in particular to an efficient cotton metabolomics sample preparation and detection method. Background technique [0002] Metabolic analysis is an important analytical method for revealing cell activities in organisms. Through qualitative and quantitative analysis of metabolites in organisms, it helps to understand the dynamics of biological growth and development, analyze the genetic basis of metabolism, and thus understand the essence of life activities. Cotton It is an important fiber crop and oil crop. Its abundant secondary metabolites have been widely reported to be involved in cotton fiber development and stress response processes2-4. Using metabolomics to analyze the synthesis and genetic basis of cotton secondary metabolites is important for improving cotton fiber. There are broad prospects for quality improvement and stress resistance. At present, there are relatively few stud...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/72G01N1/42G01N30/86
CPCG01N30/02G01N30/06G01N30/34G01N30/7233G01N1/42G01N30/8679
Inventor 金双侠张献龙斯欢孙艺文
Owner HUAZHONG AGRI UNIV
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