A method for purifying and analyzing breast milk acid oligosaccharides

An analysis method, an acidic technology, applied in the field of breast milk acid oligosaccharide purification and analysis, can solve the problems of tedious time-consuming, low selectivity, etc., and achieve the effect of wide application range, high selectivity, and high throughput

Active Publication Date: 2021-07-09
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the selectivity of the above two methods is low, cumbersome and time-consuming

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
  • A method for purifying and analyzing breast milk acid oligosaccharides
  • A method for purifying and analyzing breast milk acid oligosaccharides
  • A method for purifying and analyzing breast milk acid oligosaccharides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Defat and protein removal process of human milk oligosaccharides

[0032] Take a certain amount of human milk and centrifuge at 4000g for 60min at 4°C to remove the upper layer of lipid. Remove the lower aqueous layer, add 2 times the volume of absolute ethanol and mix well, let stand at 4°C for 12 hours, then centrifuge at 4000g for 60min at 4°C, centrifuge once, and collect the supernatant to obtain the defatted and deproteinized oligosaccharide sample.

[0033] Human milk oligosaccharide purification and analysis process

[0034] The liquid sample of defatted and deproteinized human milk oligosaccharides was taken directly for analysis. The injection volume was 20 μL, the SPE purification column used ClickTE-GSH column, the column specification was 2.1*50mm, the flow rate was 0.2mL / min, 80% acetonitrile water wasocratic elution for 5min, and the eluent was a solution containing acidic oligosaccharides. By switching the valve, the SPE purification column and the HIL...

Embodiment 2

[0037] Defat and protein removal process of human milk oligosaccharides

[0038] The process is the same as in Example 1.

[0039] Human milk oligosaccharide purification and analysis process

[0040] The liquid sample of defatted and deproteinized human milk oligosaccharides was taken directly for analysis. The injection volume was 20 μL, and the HILIC analytical column was used for analysis without SPE purification column. Other conditions are with embodiment 1.

[0041] After mass spectrometry detection, in addition to acidic oligosaccharides, there were also lactose and neutral oligosaccharides, and neutral oligosaccharides and acidic oligosaccharides crossed in the chromatogram, and neutral oligosaccharides seriously interfered with the analysis of acidic oligosaccharides. Therefore, before analysis, it is very important to purify acid oligosaccharides to remove lactose and neutral oligosaccharides.

Embodiment 3

[0043] Defat and protein removal process of goat milk oligosaccharides

[0044] Take a certain amount of goat milk and centrifuge at 2000g for 5min at 4°C to remove the upper layer of lipid. Remove the lower water layer, add 4 times the volume of acetone and mix well. After standing at 4°C for 18 hours, centrifuge at 2000g at 4°C for 10 minutes. After centrifuging twice, collect the supernatant to obtain a liquid sample for degreasing protein and oligosaccharides. Concentrate until dry, set aside.

[0045] Purification and analysis process of goat milk oligosaccharides

[0046]Weigh the defatted deproteinized goat milk oligosaccharide sample, use 50% acetonitrile water to prepare a solution with a concentration of 100 mg / mL, and the injection volume is 100 μL. The SPE purification column uses a chromatographic column with aspartic acid groups bonded to the surface of silica gel. The inner diameter of the chromatographic column is 2.1*100mm, the flow rate is 0.2mL / min, 85% ac...

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

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Login to view more

Abstract

The invention relates to a method for purifying and analyzing breast milk acid oligosaccharides. Its characteristics are: the sample is degreased by low-temperature centrifugation, after adding an organic solvent to precipitate and remove protein, and then remove the interference of lactose and neutral oligosaccharides through an online purification column, the obtained acidic sugar is separated by a hydrophilic column, evaporated light, electrospray , UV or mass spectrometry to obtain information on the structure, composition and content of acidic sugars in breast milk. The method has the characteristics of good selectivity, good stability, simple and controllable operation, and high throughput, and is suitable for rapid and high-throughput analysis of mammalian milk and human milk samples.

Description

technical field [0001] The invention relates to a method for purifying and analyzing breast milk acid oligosaccharides. Its characteristics are: the sample is degreased by low-temperature centrifugation, organic solvent is added to precipitate and remove protein, the acid oligosaccharide is purified by column chromatography in electrostatic repulsion mode, and the interference of lactose and neutral oligosaccharide is removed, and then separated and analyzed by online HILIC column, evaporated light, electrospray , UV or mass spectrometry to obtain structural composition analysis. The method has the characteristics of good selectivity, good stability, simple and controllable operation, and high throughput, and is suitable for rapid analysis of a large number of breast milk samples. [0002] technical background [0003] Oligosaccharides are important components in breast milk and have important physiological functions. It plays a vital role in the normal growth and developme...

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 Patents(China)
IPC IPC(8): G01N30/06G01N30/64G01N30/72G01N30/74
CPCG01N30/06G01N30/64G01N30/72G01N30/74
Inventor 梁鑫淼闫竞宇郭志谋丁俊杰
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap