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Method for purifying and separating oligosaccharides

A technology for purification and separation of oligosaccharides, which is applied in the direction of chemical instruments and methods, disaccharides, oligosaccharides, etc., can solve the problems of large amount of solvents, cumbersome operations, and increased experimental steps, so as to achieve simple and controllable experimental operations and a wide range of applications Broad and repeatable effect

Inactive Publication Date: 2011-06-22
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Size exclusion chromatography separates according to the molecular size of the sample, the selectivity is relatively poor, and it is difficult to separate oligosaccharide samples with the same degree of polymerization
Ion-exchange chromatography is based on the difference in the charge of the sample. It will encounter certain problems when separating oligosaccharides with the same charge, and it is difficult to achieve baseline separation.
Although the combination of the above two methods can solve the problem of selectivity to a certain extent [Ballance, S.; Aarstad, O.A.; Aachmann, F.L.; -Braek, G.; Christensen, B.E.Carbohydrate Research 2009, 344, 255], but the coupling technique undoubtedly increases the experimental steps, which may lead to a decrease in the sample recovery rate
However, the operation of thin-layer chromatography is relatively cumbersome, and the amount of solvent is large, so it is not easy to realize automation.

Method used

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  • Method for purifying and separating oligosaccharides
  • Method for purifying and separating oligosaccharides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 50 mg of galactooligosaccharides was dissolved in 1 mL of pure water, and the injection volume was 30 μL.

[0029] A maltose chromatographic column (Click Maltose, 10×100mm, see invention patent 200710010808.8) was used with a column temperature of 30°C and a flow rate of 3.0mL / min. Phase A is water, phase B is acetonitrile, 0-10min, A / B (V / V): 70 / 30→60 / 40, 10-25min, A / B: 60 / 40.

[0030] Post-column split, split volume ratio 5:1, about 1 / 6 of which enters the detector, the detector is evaporative light scattering (ELSD), instrument parameter settings: gas pressure 35psi, drift tube temperature 85°C, gain value 1. A total of 6 different sample peaks were detected; they were galactooligosaccharides with degrees of polymerization of 2, 3, 4, 5, 6, and 7;

[0031] Samples were injected 10 times continuously, and the fractions were collected according to the retention time of the 6 sample peaks. A total of 6 fractions were obtained. The fractions were concentrated to about ...

Embodiment 2

[0033] 50 mg of galactooligosaccharides was dissolved in 1 mL of pure water, and the injection volume was 5 μL.

[0034]A diol-based chromatographic column (Diol, 4.6×150 mm) was used with a column temperature of 30° C. and a flow rate of 1.0 mL / min. Phase A is water, phase B is acetonitrile, 0-10min, A / B (V / V): 80 / 20→70 / 30, 10-25min, A / B: 70 / 30-60 / 40. The detector is evaporative light scattering (ELSD), and the instrument parameter settings are: gas pressure 35psi, drift tube temperature 85°C, gain value 10. A total of 6 different sample peaks were detected; they were galactooligosaccharides with degrees of polymerization of 2, 3, 4, 5, 6, and 7, respectively.

Embodiment 3

[0036] Fructose oligosaccharide 30mg, dissolved in 1mL pure water, injection volume 10μL.

[0037] A maltose chromatographic column (Click Maltose 4.6×100mm, see invention patent 200710010808.8) was used with a column temperature of 30°C and a flow rate of 1.0mL / min. Phase A is water, phase B is acetonitrile, 0-60min, A / B (V / V): 70 / 30→50 / 50, 60-70min, A / B: 50 / 50-40 / 60. The detector is evaporative light scattering (ELSD), and the instrument parameter settings are: gas pressure 35psi, drift tube temperature 85°C, gain value 10. A total of 46 different sample peaks were detected; they were fructooligosaccharides with a degree of polymerization of 5-50.

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Abstract

The invention discloses a method for purifying and separating oligosaccharides. A polar chromatographic column is prepared by taking a polar filler as a chromatographic stationary phase in the chromatographic mode of a hydrophilic interaction chromatography principle, wherein water, acetonitrile and a buffer salt serve as a mobile phase; water serves as a strong elution solvent; and an isocratic or gradient elution condition is used. In the method, parameters such as the type and concentration of the buffer salt, the gradient of the mobile phase, column temperature and the like are optimized, so that separation and preparation of acid, neutral and alkaline oligosaccharides are realized. The method has the characteristics of high stability, large loading amount, easiness, convenience and controllability in operating and the like, and is suitable for separating and purifying different types of oligosaccharides.

Description

technical field [0001] The invention relates to the separation and purification of oligosaccharides, in particular to a method for purification and separation of oligosaccharides. technical background [0002] Sugars are aldehyde or ketone compounds containing polyhydroxyl groups, which are widely distributed in the biological world. According to the composition of molecules, sugars can be divided into monosaccharides, oligosaccharides, polysaccharides, conjugated sugars and derivative sugars. Among them, oligosaccharides refer to low-level polymerized sugars composed of 3-9 monosaccharides linked by glycosidic bonds. Studies have shown that oligosaccharides have various biological and potential pharmacological activities. Sialooligosaccharides can stimulate angiogenesis [West, D.C.; Hampson, I.N.; Arnold, F., Kumar, S. Science, 1985, 228, 1324] and inhibit tumor growth [Zeng, C.; Toole, B.P.; Kinney, S.D.; Kuo, J.W.; Stamenkovic, I.Int.J.Cancer, 1998, 77, 396]; Oligosacch...

Claims

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Application Information

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IPC IPC(8): C07H3/06C07H3/04C07H1/06C08B37/00C08B37/08
Inventor 梁鑫淼傅青郭志谋肖远胜张秀莉
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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