Method for determining content of binding-state glutamine in protein or protein hydrolysate

A technology of glutamine and determination method, applied in the field of analysis, can solve the problems of high cost, complicated operation, slow speed and the like, and achieves the effect of flexible and convenient application

Inactive Publication Date: 2014-06-18
郑州新威营养技术有限公司
View PDF1 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the slow speed, high cost and cumbersome operation of the existing glutamine content determination method, and the whole determination process requires alanyl-glutamine dipeptide and glycyl-glutamine dipeptide as standard products. Disadvantages, provide a simple, rapid, highly sensitive method for determining the content of bound glutamine in protein or its hydrolyzate

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. Materials and Reagents

[0022] LC600 Binary High Pressure Gradient High Performance Liquid Chromatography System Beijing Labtech Co., Ltd.

[0023] Nitrogen Blowing Apparatus Beijing Shuaien Technology Co., Ltd.

[0024] Wheat hydrolyzed protein (30.0% glutamine content) produced by Zhengzhou Xinwei Nutrition Technology Co., Ltd.

[0025] Chemical reagents: all analytically pure

[0026] 2 Preparation of reagents:

[0027] 2.1 50 μ mol / mL pyridine solution: Dissolve 0.25 mL pyridine in 50 mL distilled water.

[0028] 2.2 10 mg / mL BTI solution: Dissolve 0.125 g of BTI in 12.5 mL of acetonitrile.

[0029] 3 Determination method

[0030] (1) Dissolution of samples: Dissolve 0.50 g of protein or its hydrolyzate in 100.0 mL of distilled water, stir in a water bath at 45°C for 5 hours to fully swell.

[0031] (2) BTI protection treatment of samples: Take 1.0 mL of the sample solution from step (1), add 1.5 mL of BTI solution and 0.1 mL of pyridine solution, and keep...

Embodiment 2

[0043] 1. Materials and Reagents

[0044] LC600 Binary High Pressure Gradient High Performance Liquid Chromatography System Beijing Labtech Co., Ltd.

[0045] Nitrogen Blowing Apparatus Beijing Shuaien Technology Co., Ltd.

[0046] Wheat hydrolyzed protein (30.0% glutamine content) produced by Zhengzhou Xinwei Nutrition Technology Co., Ltd.

[0047] Chemical reagents: all analytically pure

[0048] 2 Preparation of reagents:

[0049] 2.1 50 μ mol / mL pyridine solution: Dissolve 0.25 mL pyridine in 50 mL distilled water.

[0050] 2.2 10 mg / mL BTI solution: Dissolve 0.125 g of BTI in 12.5 mL of acetonitrile.

[0051] 3 Determination method

[0052] (1) Dissolution of samples: Dissolve 0.50 g of protein or its hydrolyzate in 100.0 mL of distilled water, stir in a water bath at 50°C for 6 hours to fully swell.

[0053](2) BTI protection treatment of samples: Take 2.0 mL of the sample solution from step (1), add 2 mL of BTI solution and 0.3 mL of pyridine solution, and keep wa...

Embodiment 3

[0065] 1. Materials and Reagents

[0066] LC600 Binary High Pressure Gradient High Performance Liquid Chromatography System Beijing Labtech Co., Ltd.

[0067] Nitrogen Blowing Apparatus Beijing Shuaien Technology Co., Ltd.

[0068] Wheat hydrolyzed protein (30.0% glutamine content) produced by Zhengzhou Xinwei Nutrition Technology Co., Ltd.

[0069] Chemical reagents: all analytically pure

[0070] 2 Preparation of reagents:

[0071] 2.1 50 μ mol / mL pyridine solution: Dissolve 0.25 mL pyridine in 50 mL distilled water.

[0072] 2.2 10 mg / mL BTI solution: Dissolve 0.125 g of BTI in 12.5 mL of acetonitrile.

[0073] 3 Determination method

[0074] (1) Dissolution of samples: Dissolve 0.50 g of protein or its hydrolyzate in 100.0 mL of distilled water, stir in a water bath at 55°C for 8 hours to fully swell.

[0075] (2) BTI protection treatment of samples: Take 3.0 mL of the sample solution from step (1), add 4.5 mL of BTI solution and 0.5 mL of pyridine solution, and keep...

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

No PUM Login to view more

Abstract

The invention discloses a method for determining the content of binding-state glutamine in protein or protein hydrolysate, belonging to the analysis technology field. In the conventional amino acid analysis, glutamine in protein or peptide molecule is converted into glutamic acid after being hydrolyzed by hydrochloric acid, only the total content of glutamic acid is displayed in an analysis result, but the content of the glutamine in the protein cannot be displayed. Based on the characteristic that binding-state glutamine in protein or peptide molecule cannot be hydrolyzed by hydrochloric acid after reaction with a special reagent (bis(trifluoacetoxy) iodobenzene) (BTI), the method is characterized in that the content of glutamic acid is determined through acid hydrolysis under the protection of the BTI reagent, and finally the content of the binding-state glutamine in protein or peptide molecule can be worked out according to the difference between the content of glutamic acid in the sample under BTI protection and the content of glutamic acid in a sample without BTI protection. The method comprises the steps of preparation of a sample to be tested, BTI protection treatment, acidolysis, derivation, amino acid chromatographic column separation, control sample preparation, and content determination. The method is simple and rapid, has high sensitivity and can timely guide the proteolysis industrial production.

Description

technical field [0001] The invention relates to a method for measuring the content of bound glutamine in protein and its hydrolyzate, which belongs to the field of analysis technology. Background technique [0002] Glutamine is an important amino acid in protein. Glutamine is the amidation product of glutamic acid, which plays an important role in the metabolism of organisms, especially animals. [0003] At present, the analysis of amino acids in proteins adopts the acid hydrolysis method, that is, the protein is hydrolyzed into free amino acids by hydrochloric acid, and then chromatographic techniques (ion exchange or reversed-phase chromatography) are used to separate and determine the content of amino acids. However, during acid hydrolysis, glutamine and aspartic acid in protein are hydrolyzed into glutamic acid and aspartic acid respectively, and these two amino acids are also inherent amino acids in protein. Glutamic acid determined by chromatography is actually protein...

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 Applications(China)
IPC IPC(8): G01N30/02
Inventor 王章存张子峰张新武王许东
Owner 郑州新威营养技术有限公司
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