Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for detecting amino acids and saccharides in fermentation liquor through gas chromatography-mass spectrometry

A gas chromatography and mass spectrometry detection technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of complicated operation, many interference factors of detection technology, low sensitivity, etc., and achieve simplified operation steps, excellent extraction effect, and good analysis effect. Effect

Inactive Publication Date: 2014-05-14
LIUZHOU LIANHAI TECH
View PDF5 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for detecting amino acids and sugars in fermentation liquid by gas chromatography-mass spectrometry, which can make up for the gap in the current detection method for metabolites in microbial fermentation, and overcome the many interference factors, cumbersome operation, and difficult testing of existing detection technologies. Disadvantages such as high cost, low sensitivity, and narrow detection range

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for detecting amino acids and saccharides in fermentation liquor through gas chromatography-mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Get the yeast fermentation broth.

[0024] (1) Sample preparation of intracellular metabolites: Take 5ml of fermentation broth, centrifuge to obtain the bacteria, wash the bacteria twice with normal saline, dissolve in cold methanol at -40°C, and the dry weight of the biomass reaches 0.2g / ml, and ultrasonically break down to the cells Cleavage to obtain a lysate, extract the lysate at low temperature, the low temperature condition is: extract at -40°C to -50°C for 4 hours, centrifuge to collect 150ul of the extracted lysate supernatant I, add internal standard ribitol solution, and lyse the supernatant I The ratio of the volume to the weight of the internal standard ribitol is 150ul:20μg, and vacuum-dried at room temperature to obtain the intracellular metabolite sample I.

[0025] (2) Preparation of extracellular fluid samples: take fermentation broth and centrifuge to obtain supernatant II, take 300ul supernatant II, add acetonitrile to supernatant II to remove protei...

Embodiment 2

[0029] Get the lactic acid bacteria fermentation broth.

[0030] (1) Sample preparation of intracellular metabolites: Take 5ml of fermentation broth, centrifuge to obtain the bacteria, wash the bacteria three times with normal saline, dissolve in cold methanol at -40°C, and the dry weight of the biomass reaches 1.0g / ml, and ultrasonically break down to the cells Cleavage to obtain a lysate, extract the lysate at low temperature, the low temperature conditions are: extract at -40°C to -50°C for 5 hours, centrifuge to collect 100ul of the extracted lysate supernatant I, add internal standard ribitol solution, and lyse the supernatant I The ratio of the volume to the weight of the internal standard ribitol is 100ul:20μg, and it is vacuum-dried at room temperature to obtain the intracellular metabolite sample I.

[0031] (2) Preparation of extracellular fluid samples: take the fermentation broth, centrifuge to obtain supernatant II, take 350ul of supernatant II, add acetonitrile t...

Embodiment 3

[0036] Take the mold fermentation broth.

[0037] (1) Sample preparation of intracellular metabolites: Take 5ml of fermentation broth, centrifuge to obtain the bacteria, wash the bacteria three times with normal saline, dissolve in cold methanol at -40°C, and the dry weight of the biomass reaches 0.6g / ml, and ultrasonically break down to the cells Cleavage to obtain a lysate, extract the lysate at low temperature, the low temperature condition is: extract at -40°C to -50°C for 3 hours, centrifuge to collect 200ul of the extracted lysate supernatant I, add internal standard ribitol solution, and lyse the supernatant I The ratio of the volume to the weight of the internal standard ribitol is 200ul:20μg, and vacuum-dried at room temperature to obtain the intracellular metabolite sample I.

[0038] (2) Preparation of extracellular fluid samples: take the fermentation broth, centrifuge to obtain supernatant II, take 60ul of supernatant II, add acetonitrile to supernatant II to ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a method for detecting amino acids and saccharides in liquor fermentation liquor through a gas chromatography-mass spectrometry and belongs to the technical field of biochemical detection and measurement. The detection method comprises the steps of (1) taking the fermentation liquor centrifuging at high speed and collecting cells and extracellular fluids respectively; (2) preparing a fermentation liquor sample, namely taking a supernatant liquor obtained in the step 1), adding acetonitrile into the supernatant liquor to remove protein, performing vertex-vibration, then centrifuging for collecting the supernatant liquor III, and adding an internal standard substance ribitol solution to dry at room temperature in vacuum so as to obtain an extracellular fluid sample II; (3) deriving the sample, and adding a saccharide derivating agent and an amino acid derivating agent; (4) performing gas chromatography-mass spectrometry analysis and collecting data. The method for detecting amino acids and saccharides in the fermentation liquor through the gas chromatography-mass spectrometry has the advantages that the method is capable of simply processing the sample, simple and convenient to operate, short in time for detecting amino acids and saccharides, high in sensitivity, high in detection result repeatability and can realize preparation of a plurality of samples.

Description

technical field [0001] The invention belongs to the technical field of biochemical detection and determination, in particular to a method for detecting organic acids and sugars in fermentation broth by gas chromatography-mass spectrometry. Background technique [0002] In terms of using gas chromatography-mass spectrometry to detect amino acids and sugar metabolites, there are related technical reports, such as patent 201210046953.2, which discloses a method for gas chromatography-mass spectrometry detection of urinary organic acids, including the following steps: (1) the urine to be tested (2) add internal standard, use frozen ethanol to process protein precipitation, and dry the sample; (3) carry out methyl silylation derivatization treatment on the above sample; (4) use the above 1- Step 3 to process the standard organic acid to obtain the standard organic acid sample; (5) use gas chromatography-mass spectrometry to detect the standard organic acid sample and the urine sa...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/06
Inventor 陈东梁远雄
Owner LIUZHOU LIANHAI TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com