Method for determining xylooligosaccharide through high-efficiency liquid-phase ion exchange chromatography

A technology of ion exchange chromatography and xylooligosaccharides, which is applied in the field of xylooligosaccharide determination, can solve the problem of inability to distinguish and quantitatively measure different xylosaccharide components, low separation of sugar components, and poor reproducibility of results and other issues to achieve the effect of improving the resolution and detection efficiency

Active Publication Date: 2013-08-21
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Method (1) All xylooligosaccharides were hydrolyzed into xylose by strong acid heating hydrolysis method, and different xylooligosaccharide components could not be distinguished and quantitatively determined; method (2) due to the various sugar components of xylooligosaccharides The molecular structure is similar and the difference in molecular weight is small. It is difficult for the existing thin chromatography and high performance liquid chromatography to realize the direct and effective separation of xylooligosaccharide components with adjacent polymerization degrees, and it is also difficult to separate xylooligosaccharide groups of different xylooligosaccharide groups. Quantitative determination; method (3) needs to derivatize the sugar, the operation is complicated, and the test cost is high
At the same time, due to the lack of standard samples of xylo-oligosaccharides with a higher degree of polymerization such as xyloheptose, it is difficult to qualitatively and quantitatively analyze these xylo-oligosaccharide components in the product
The above-mentioned various xylo-oligosaccharide determination methods all have the disadvantages of cumbersome operation steps, low separation degree of sugar components, poor reproducibility of results or high detection cost;

Method used

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  • Method for determining xylooligosaccharide through high-efficiency liquid-phase ion exchange chromatography
  • Method for determining xylooligosaccharide through high-efficiency liquid-phase ion exchange chromatography
  • Method for determining xylooligosaccharide through high-efficiency liquid-phase ion exchange chromatography

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1 High Performance Liquid Phase Ion Exchange Chromatography System and Chromatographic Conditions

[0039]High performance liquid phase ion exchange chromatography system: American Dionex ICS-3000 ion chromatography system, equipped with double pump (DP) built-in vacuum degassing module, electrochemical detector (ED) and automatic sampler (AS40), operating software of chromatography system Using Chromeleon 6.70 chromatographic workstation, chromatographic conditions: chromatographic column: CarboPacTM PA200 (3×250m) chromatographic column with guard column (3×50mm); column temperature: 30°C; injection volume: 10.0 μL;

[0040] Elution conditions: 500 mmol / L sodium acetate and 100 mmol / L sodium hydroxide were used as eluents for binary gradient elution, the flow rate was 0.3 mL / min, and the concentration gradient of sodium acetate solution elution within 0 to 40 min was 0~120mmol.

[0041] Signal detection: The detection mode of the electrochemical detector is g...

Embodiment 2

[0044] Embodiment 2 Determination of xylo-oligosaccharide standard working equation

[0045] The standard working equation for the determination of xylooligosaccharides: standard products of xylose, xylobiose, xylotriose, xylotetraose, xylopentaose and xylohexaose (produced by Megazyme, Ireland, purity >95%) were prepared to 0.5 ~10mg / L standard solution, using the above-mentioned high performance liquid phase ion exchange chromatography system and chromatographic conditions to determine the standard working equation of xylose to xylose hexose, the measurement results are as follows figure 1 and Figure 4 shown.

[0046] Chromatographic peak retention time RT (min): xylose 4.600, xylobiose 6.667, xylotriose 9.934, xylotetraose 12.684, xylopentaose 15.000, xylohexaose 17.100.

[0047] Standard working equation:

[0048] Xylose A=3.5218c 1 -0.0193, correlation coefficient R 2 =0.9989;

[0049] Xylobiose A=3.0798c 2 -0.1026, correlation coefficient R 2 =0.9998;

[0050] ...

Embodiment 3

[0059] Example 3 Determination of the chromatographic retention time of xyloheptaose and xylooctaose

[0060] Perform linear regression with the polymerization degree value (NDP) of xylose to xylose hexaose and the chromatographic retention time value (RT, min), the results are as follows figure 2 As shown, the linear relationship between RT and NDP can be calculated as:

[0061] RT (min)=2.5785×NDP+1.9726, correlation coefficient R 2 =0.9952.

[0062] The chromatographic retention times of xyloheptaose and xylooctaose were deduced from the linear relationship to be 20.023 min and 22.601 min respectively, which can be used for qualitative analysis and identification of xyloheptaose and xylooctaose components in xylooligosaccharide samples.

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Abstract

The invention discloses a method for determining xylooligosaccharide through high-efficiency liquid-phase ion exchange chromatography, which comprises the following steps: determining a high-efficiency liquid-phase ion exchange chromatography system and chromatography conditions, determining a standard xylooligosaccharide working formula, determining the chromatography reservation time for xyloheptaose and xylooctaose, and analyzing and determining components of xylooligosaccharide. In the method for determining the xylooligosaccharide through the high-efficiency liquid-phase ion exchange chromatography, a method for accurately and quantitatively determining the xylooligosaccharide through the high-efficiency liquid-phase ion exchange chromatography is established for the first time, the CarboPacTMPA200 (3 * 250m) chromatographic column is adopted, the separation degree and the detection efficiency of xylose and various xylooligosaccharide components are improved significantly through binary gradient elution of sodium acetate and sodium hydroxide, the xylooligosaccharide sample, in particular xylobiose, xylotriose, xyloterose, xylopentaose and xylohexaose, can be accurately and quantitatively determined, and the xyloheptaose and xylooctaose can be analyzed qualitatively. Therefore, the method has great significance in detection, evaluation, promotion and application of xylooligosaccharide products.

Description

technical field [0001] The invention relates to a method for measuring xylooligosaccharides, in particular to a method for measuring xylooligosaccharides by using high performance liquid phase ion exchange chromatography. Background technique [0002] Xylooligosaccharide is a general term for a class of oligosaccharides linked by β-1,4-xylosidic bonds with a degree of polymerization of 2 to 8. As a new type of functional oligosaccharide, xylooligosaccharide is a A high-efficiency bifidus factor and immune enhancer for human and animals. It has a variety of health-care and growth-promoting functions such as regulating the intestinal microecology and activating the immune function of the body. It can be widely used in industries such as medicine, food, beverage and feed. [0003] The preparation methods of xylo-oligosaccharides mainly include xylan directional enzymatic degradation, steam explosion and other methods, and the obtained products are all mixtures of various xylo-o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 徐勇余世袁勇强范丽
Owner NANJING FORESTRY UNIV
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