A method to discover and characterize lipid biomarkers in unicellular algae

A biomarker, unicellular algae technology, applied in the field of discovery and identification of unicellular algae lipid biomarkers, can solve the problems of inability to analyze thermally unstable metabolites, limited research scope, etc., to reduce the effect of ion inhibition , The effect of improving analytical throughput and high resolution

Inactive Publication Date: 2011-11-30
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gas chromatography/mass spectrometry (GC-MS) has high sensitivity, but its research scope is limited to the ana

Method used

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  • A method to discover and characterize lipid biomarkers in unicellular algae
  • A method to discover and characterize lipid biomarkers in unicellular algae
  • A method to discover and characterize lipid biomarkers in unicellular algae

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Discovery and Characterization of Lipid Biomarkers of Chlamydomonas nivalis Adaptation to NaCl Stress Environment 1, Chlamydomonas nivalis Cultivation and Harvesting

[0027]After Chlamydomonas nivalis was cultured for 6 days, a sterilized NaCl solution was added to make the final concentration 0.5% (w / v). Continue to cultivate for 0h, 1h, 7h, and 15h to take samples, and set four parallel samples at each time. The algae liquid was centrifuged (5000r / min, 5min), the supernatant was removed, the algae mud was washed three times with pure water, placed in an ultra-low temperature refrigerator for pre-freezing, and freeze-dried to obtain algae powder.

[0028] 2. Extraction of cell lipid compounds

[0029] The algae powder was extracted with methanol-chloroform-water extract. First add 0.025% (w / v, g / ml) antioxidant BHT to 2.5ml of methanol-chloroform-water (1 / 1 / 0.5, v / v / v) extraction solution, and then accurately weigh 10.0mg Algae powder, add the extract, shake with a...

Embodiment 2

[0057] Discovery and Characterization of Lipid Biomarkers of Chlamydomonas nivalis Adaptation to NaCl Stress Environment 1, Chlamydomonas nivalis Cultivation and Harvesting

[0058] After Chlamydomonas nivalis was cultured for 6 days, a sterilized NaCl solution was added to make the final concentration 0.5% (w / v). Continue to cultivate for 0h, 1h, 7h, and 15h to take samples, and set four parallel samples at each time. The algae liquid was centrifuged (5000r / min, 5min), the supernatant was removed, the algae mud was washed three times with pure water, placed in an ultra-low temperature refrigerator for pre-freezing, and freeze-dried to obtain algae powder.

[0059] 2. Extraction of cell lipid compounds

[0060] The algae powder was extracted with methanol-chloroform-water extract. First add 0.05% (w / v, g / ml) antioxidant BHT to 2.5ml of methanol-chloroform-water (1 / 1 / 0.5, v / v / v) extraction solution, and then accurately weigh 10.0mg Algae powder, add the extract, shake with a...

Embodiment 3

[0073] Discovery and Characterization of Lipid Biomarkers of Chlamydomonas nivalis Adaptation to NaCl Stress Environment 1, Chlamydomonas nivalis Cultivation and Harvesting

[0074] After Chlamydomonas nivalis was cultured for 6 days, a sterilized NaCl solution was added to make the final concentration 0.5% (w / v). Continue to cultivate for 0h, 1h, 7h, and 15h to take samples, and set four parallel samples at each time. The algae liquid was centrifuged (5000r / min, 5min), the supernatant was removed, the algae mud was washed three times with pure water, placed in an ultra-low temperature refrigerator for pre-freezing, and freeze-dried to obtain algae powder.

[0075] 2. Extraction of cell lipid compounds

[0076] The algae powder was extracted with methanol-chloroform-water extract. First add 0.05% (w / v, g / ml) antioxidant BHT to 2.5ml of methanol-chloroform-water (1 / 1 / 0.5, v / v / v) extraction solution, and then accurately weigh 10.0mg Algae powder, add the extract, shake with a...

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Abstract

The invention discloses a method for discovering and identifying lipid biomarkers in unicellular algae, comprising the following steps: (1) extraction and antioxidant protection of intracellular lipid compounds; (2) ultra-high performance liquid chromatography/four Grade rod-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) analysis and extraction of lipid compounds; (3) Extract and analyze data with Markerlynx and import it into Simca-p software for orthogonal partial least squares discriminant analysis (OPLS-DA ); (4) Validate the OPLS-DA model (5) Screen out potential lipid biomarkers from the results of the OPLS-DA model; (6) Conduct structural inference and identification of potential lipid biomarkers. The general technical system for discovering and identifying lipid biomarkers in single-cell algae established by the present invention can be radiated and applied to the fields of metabolomic analysis of plants and microorganisms, metabolomic analysis of food safety, and the like.

Description

technical field [0001] The invention belongs to the field of bioanalytical chemistry and relates to a method for discovering and identifying lipid biomarkers of unicellular algae. Background technique [0002] Lipid compounds are small molecular compounds with similar physical and chemical properties. Their existence and abundance are crucial for important physiological activities such as cell metabolism regulation, energy control, and signal transduction. By studying the total lipid extract obtained from unicellular algae, the information of the lipidome (lipidome) can be obtained, including the composition, interaction and function information of all lipid substances in the cell, which reflects To understand the overall changes in lipid metabolites in single-celled algae cells under specific physiological conditions, to understand in more detail the changes in lipid metabolism pathways and networks caused by external interference factors, to discover and identify lipid bio...

Claims

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Application Information

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IPC IPC(8): G01N30/72
Inventor 魏东吕娜
Owner SOUTH CHINA UNIV OF TECH
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