Method for quickly identifying sugar

A recognition method and monosaccharide technology, applied in the field of analytical chemistry, can solve problems such as the inability to determine the composition of sugar and monosaccharide residues, and the inability to analyze accurately

Inactive Publication Date: 2016-06-22
CHANGCHUN UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It solves the problem that in the existing technology for identifying oligosaccharide isomers, it is impossible to analyze the actual mixed sugar samples simply, qu

Method used

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  • Method for quickly identifying sugar
  • Method for quickly identifying sugar
  • Method for quickly identifying sugar

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (1) Preparation of standard products

[0058] Sucrose, lactose, kojibiose, nigerbiose, maltose, isomaltose, sophorose, laminaribiose, cellobiose, gentiobiose, glucose, galactose were mixed with 50% methanol-water (v / v) , mannose and fructose standard substances were prepared into a solution with a concentration of 2 μM.

[0059] (2) IM-MS detection of standard products

[0060] The prepared standard was directly measured by ESI-IM-MS, and the injection speed of the peristaltic pump was 5 μL / min. ESI ion source conditions are as follows: positive ion mode, capillary voltage: 2.8kV, sample cone voltage: 50V, extraction cone voltage: 4V, source temperature: 120°C, desolvation temperature: 350°C, cone gas flow : 30L / h. IM-MS conditions: T-wave rate: 550-2500m / s, T-wave height: 40V, drift (nitrogen) flow rate: 90mL / min.

[0061] see results figure 1 , comparing the ion mobility mass spectra of monosaccharide and disaccharide isomers, according to t D , found that only ...

Embodiment 2

[0063] (1) Preparation of calibrators and standard substances to be tested

[0064] Calibrator (sucrose, lactose, cellobiose, maltose, melibiose, trehalose, melezitose, raffinose and maltotriose) and the standard substance to be measured (koji disaccharide, aspergillus niger, isomaltose, sophorose, laminaribiose, gentiobiose, glucose, galactose, mannose and fructose) were prepared into a solution with a concentration of 2 μM.

[0065] (2) Ammonia-methanol solution preparation

[0066] Take 500 μL of 28% NH 3 ·H 2 O, was added to 9.5mL methanol and shaken fully to obtain

[0067] Ammonia-methanol solution with a concentration of 0.82mol / L.

[0068] (3) Preparation of PMP derivative solution

[0069] Weigh 60.67mg of PMP and put it in a 10mL volumetric flask, fully dissolve it with ammonia-methanol solution, adjust to volume, fully oscillate, and prepare a concentration of 0.0349mol / L, and store it in a 4°C refrigerator for later use.

[0070] (4) PMP derivatization proces...

Embodiment 3

[0085] The PHN derivatization process is the same as in Example 2, and the IM-MS detection conditions of the derivatized products are the same as in Example 1.

[0086] see results image 3 , comparing the ion mobility mass spectra of monosaccharide and disaccharide isomer PHN derivatized products, according to t D , found that Aspergillus nigerbiose PHN derivatives (t D =4.34ms), laminaribiose PHN derivatives (t D =4.45ms) and cellobiose PHN derivatives (t D =5.15ms) can be distinguished from other disaccharide isomer PHN derivatives, galactose PHN derivatives (t D =2.71ms) can be distinguished from other monosaccharide isomers PHN derivatives, and other monosaccharide and disaccharide isomers cannot be distinguished.

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Abstract

The invention relates to a method of combining PMP (1-Phenyl-3-Methyl-5-Pyrazolone) derivatization and IM-MS/MS (Ion Mobility Tandem Mass Spectrometry), which solves the problem that an actually mixed sugar sample cannot be analyzed easily, quickly and accurately in the prior art for identifying oligosaccharide isomers, and meanwhile, solves the problem that the compositions of monosaccharide residues of the sugar cannot be determined by utilizing a mass spectroscopy means. According to the method for quickly identifying the sugar, the effects on identifying sugar isomers by taking PMP and PHN (Phenylhydrazine) as derivatization reagents and combining the IM-MS/MS method are compared, the CCS (Collision Cross Section) value of a derivatization product is calculated theoretically for further proving that mixed extracts of the sugar can be qualitatively identified successfully by combining the PMP derivatization and the IM-MS/MS method, and meanwhile, the connection, structures and compositions of the sugar can be identified.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and relates to a method for identifying sugars by PMP (1-phenyl-3-methyl-5-pyrazolone) derivatization combined with ion mobility tandem mass spectrometry (IM-MS / MS), specifically , a method involving PMP derivatization coupled with ion mobility tandem mass spectrometry for the identification of sugar structures in natural product extracts. Background technique [0002] Oligosaccharides are important information substances in organisms. Sugars play an important role in various biological processes, such as anticoagulant activity, immune regulation, anti-tumor, hypoglycemic, inhibition of pathogenic microorganisms, anti-oxidation, anti-inflammatory activities, etc. (X . Zhang. Curr. Med. Chem. 2006, 13, 1141-1147; S. Hakomori. Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 225-232.). The connection, configuration, and composition of monosaccharide residues of sugars have always been the difficulties i...

Claims

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Application Information

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IPC IPC(8): G01N27/68G01N1/28
CPCG01N1/28G01N27/68
Inventor 杨洪梅石磊苏蕊连文慧李金英刘淑莹
Owner CHANGCHUN UNIV OF CHINESE MEDICINE
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