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Detection method for galacto-oligosaccharide based on biological enzyme technology

A technology of galacto-oligosaccharide and detection method, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., can solve the problems of inability to separate and detect galacto-oligosaccharides, poor separation effect, etc., and achieve separation and detection. Simple method and high precision

Inactive Publication Date: 2012-02-15
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is not possible to effectively separate various galacto-oligosaccharides with similar structures and properties and detect the content of each galacto-oligosaccharide
The actual analysis spectrum is not seen in the article, only the detection spectrum of the standard product
Also, galactose and glucose, which have the same molecular weight, do not separate well

Method used

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  • Detection method for galacto-oligosaccharide based on biological enzyme technology
  • Detection method for galacto-oligosaccharide based on biological enzyme technology
  • Detection method for galacto-oligosaccharide based on biological enzyme technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Test results of milk containing galacto-oligosaccharide

[0044] Sample pretreatment: take 10 grams of sample milk, add 5 ml of 1% trichloroacetic acid solution and 2% lead acetate solution, shake well, sonicate for 20 minutes, 4°C, 15000 rpm, centrifuge for 10 minutes, take the supernatant, and pass through 0.22μm fiber Plain membrane and OnGuard II RP column (2.5cc, activated by 10mL methanol and 15mL water), discard the initial 6mL sample and collect 2mL sample for testing.

[0045] Detection conditions: detection temperature and room temperature, flow rate 1mL / min, injection volume 20μL, gradient elution method (see Table 1 for details), 5psi nitrogen detection, four-potential pulse amperometric detector, waveform detection method, and its detection voltage They are E1=+0.10V, E2=-2.00V, E3=+0.60V and E4=-0.10V.

[0046] Reagents: chromatographic grade ultrapure water, 125mmol / L NaOH, 2mol / L sodium acetate.

[0047] Result: see the test result image 3 . The statistical r...

Embodiment 2

[0053] Test results of galactooligosaccharide solution

[0054] Sample pretreatment: Take 5 grams of a galactooligosaccharide solution sample containing 7.2% total sugar, centrifuge at 15000 rpm for 10 minutes, take the supernatant, and pass through a 0.22μm cellulose membrane and OnGuard II RP column (2.5cc, 10mL methanol, 15mL water activation), discard the initial 6mL sample and collect 2mL sample for testing.

[0055] Detection conditions: flow rate 1mL / min, detection temperature room temperature, injection volume 20μL, gradient elution method (see Table 1 for details), 5psi nitrogen detection, integrated pulse ampere four-potential waveform detection method, the detection voltages are: E1 = +0.10V.

[0056] Reagents: chromatographic grade ultrapure water, 125mmol / L NaOH, 2mol / L sodium acetate.

[0057] Result: see the test result Figure 4 .

[0058] The separation and detection results show that the experimental galactooligosaccharide solution has a long reaction time due to the...

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Abstract

The invention provides a detection method for galacto-oligosaccharide based on biological enzyme technology, which belongs to the technical field of food engineering. According to the invention, at first, the method of removing proteins by using trichloroacetic acid and lead acetate is employed for pretreatment of an object to be detected, and membrane filtration is utilized to remove impurities so as to obtain a sample to be detected; then, sodium hydroxide and sodium acetate with different concentration are utilized for ion exchange and gradient elution; finally, a four-potential pulsed amperometric detector is used for detection. The invention is applicable to separation and detection of cow's milk containing galacto-oligosaccharide and a variety of galacto-oligosaccharide components in a galacto-oligosaccharide solution system; employment of ion chromatography, utilization of the methods of ion exchange and gradient elution for separation of galacto-oligosaccharide and cooperativedetection with the four-potential pulsed amperometric detector enable contamination and interference to an analytical column, the detector and the like caused by macro-molecular substances such as proteins, fat and emulsifiers which might exist in the sample to be eliminated, thereby allowing accurate and rapid detection to be achieved.

Description

Technical field [0001] The invention relates to a method in the technical field of food engineering, in particular to a method for detecting galactooligosaccharides based on biological enzyme technology. Background technique [0002] Galacto-oligosaccharide (GOS) is based on lactose, and it is firstly hydrolyzed into half under the action of β-galactosidase (EC3.2.1.23), an enzyme with galactosyl transfer activity. Lactose and glucose, and then galactose is transferred to the galactose base of lactose to produce a class of oligosaccharides containing 2-5 monosaccharide molecules, the molecular formula is (Gal)n-Glu, n=2-4. See the biological enzymatic production process of galactooligosaccharides figure 1 Shown. [0003] In nature, there are traces of galacto-oligosaccharides in animal milk, and a little more in breast milk. There are dozens of confirmed galactooligosaccharides, the structure mainly includes disaccharides (β-D-Gal-(1→6)-D-Glc, β-D-Gal(1→6)-D-Gal, β -D-Gal(1→3)-D...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 张少辉钟瑞强潘彬也余芳
Owner SHANGHAI JIAO TONG UNIV
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