Ultra-high performance liquid chromatography method for identifying high molecular weight glutenin subunits in wheat and application thereof

A glutenin subunit and ultra-high performance liquid phase technology, which is applied in the field of identification of high molecular weight glutenin subunits of wheat, can solve the problem that subunits with similar hydrophobicity cannot be separated, the requirements for pre-processing of samples are very high, and the mass spectrometer is expensive. problems, to achieve the effect of short separation time, less sample consumption and high resolution

Inactive Publication Date: 2013-04-24
CAPITAL NORMAL UNIVERSITY
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-performance capillary electrophoresis (HPCE) separation is fast and reproducible, but some subunits with similar molecular weights cannot be distinguished, such as Glu-1Dy10 and Glu-1Dy12 (Gao et al., 2010) (Gao LY, Ma WJ, Chen J, Wang K, Li J, Wang SL, Bekes F, Appels R, Yan YM (2010) Characterization and Comparative Analysis of Wheat High Molecular Weight Glutenin Subunits by SDS-PAGE, RP-HPLC, HPCE, and MALDI-TOF-MS. J Agri Food Chem 58: 2777–2786.)
Matrix-assisted laser desorption ionization / time-of-flight mass spectrometry (MALD I-TOF-MS) has the characteristics of simple operation, sample saving, short measurement time, sensitivity, rapidity, accuracy and wide range of measurement mass. Polymer application has special advantages; however, the requirements for sample pre-treatment are very high, and mass spectrometers are extremely expensive, making it difficult to be widely used
Although reversed-phase high-performance liquid chromatography (RP-HPLC) can perform quantitative analysis of proteins and overcome some weaknesses of SDS-PAGE, it cannot separate subunits with similar hydrophobicity (Dong et al., 2009) (Dong K, Hu CY , Wang AL, Cai MH, Yan YM (2009) Characterization of HMW glutenin subunits in bread and tetraploid wheats by reversed-phase high-performance liquid chromatography. Cereal Res Commun 37: 65-72.)

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

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Ultra-high performance liquid chromatography method for identifying high molecular weight glutenin subunits in wheat and application thereof
  • Ultra-high performance liquid chromatography method for identifying high molecular weight glutenin subunits in wheat and application thereof
  • Ultra-high performance liquid chromatography method for identifying high molecular weight glutenin subunits in wheat and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Identification of wheat high molecular weight glutenin subunits by ultra-high performance liquid chromatography

[0047] 1. Material sampling

[0048] The seeds of wheat variety Zhengmai 9023 were used as test materials. The seeds were randomly selected, ground with liquid nitrogen, and 20-30 mg was weighed for gluten extraction and ultra-high performance liquid chromatography analysis.

[0049] 2. Sample preparation

[0050] Extraction method for high molecular weight glutenin subunits (HMW-GS):

[0051] 1) Take the wheat seeds of the above varieties, grind them with liquid nitrogen, weigh 30 mg of powder, put them into a 1.5 ml centrifuge tube, add 1 ml of 70% ethanol by volume, vortex for 45 min, centrifuge at 13000 rpm for 10 min, remove supernatant;

[0052] 2) Add 1 ml of 55% volume percent isopropanol to the precipitate in step 1), bathe in water at 65°C for 30 minutes (manually shake three times during the period), centrifuge at 13000 rpm for 10 mi...

Embodiment 2

[0085] Example 2, identification of wheat high molecular weight glutenin subunits by ultra-high performance liquid chromatography

[0086] 1. Material sampling

[0087] The seeds of wheat varieties Hortag, wheat varieties Imbross, and Spelt wheat varieties Spelt 80 and Spelt 127 were used as test materials respectively, and the seeds were randomly selected, and 20-30 mg were weighed after grinding with liquid nitrogen for gluten extraction and super-high-efficiency Liquid chromatography analysis.

[0088] 2. Sample preparation

[0089] With embodiment 1.

[0090] 3. Ultra-high performance liquid chromatography (UPLC)

[0091] Instruments and equipment: Ultra-high performance liquid chromatography system produced by Waters (equipped with software for system operation and data processing). The column used was: ACQUITY UPLC? BEH 300 C18 1.7 μm, 2.1 mm x 50 mm.

[0092] Reagents used:

[0093] Solution A: 0.1% by volume of trifluoroacetic acid (TFA) aqueous solution;

[009...

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 the rapid identification of high molecular weight glutenin subunits in wheat by utilizing the ultra-high performance liquid chromatography technology. By adopting the method, the rapid and accurate identification of the high molecular weight glutenin subunits in wheat seed storage proteins can be realized; and mobile phase conditions for ultra-high performanceliquid chromatography, added ingredients, chromatographic condition parameters and a chromatographic column washing procedure are optimized. By adopting the method, the composition of the high molecular weight subunits in different wheat varieties can be fast identified, and the high molecular weight glutenin subunits which can not be separated and identified by the HPLC (high performance liquid chromatography) method can be accurately identified. By adopting the method, fast and accurate identification and screening can be performed on wheat quality and hybrid early generation high-quality storage protein subunits.

Description

technical field [0001] The invention relates to a method for identifying wheat high-molecular-weight glutenin subunits, in particular to a method for quickly identifying wheat high-molecular-weight glutenin subunits through ultra-high performance liquid chromatography (UPLC) technology. Background technique [0002] The quality of wheat is affected by many factors, such as growth environment, agronomic measures and internal genotype, etc. These factors determine the content and quality of wheat protein. Among them, protein content is a quantitative trait, which is mainly affected by environmental and agronomic measures; while protein quality is a qualitative trait, which is mainly controlled by genotype. The composition, content and ratio of different subunits of glutenin are closely related to the processing quality of wheat flour, especially the high molecular weight glutenin subunits, whose different coding sites, different subunit compositions and different expression le...

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 Patents(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 晏月明刘婉李小辉
Owner CAPITAL NORMAL UNIVERSITY
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