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Method for separating wheat gliadin through capillary electrophoresis

A wheat gliadin and capillary electrophoresis technology, which is applied in the direction of material analysis, analysis material, and measuring device by electromagnetic means, can solve the problems of laborious resolution, time-consuming and the like

Active Publication Date: 2017-12-22
INST OF BOTANY CHINESE ACAD OF SCI
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Problems solved by technology

The disadvantage of this method is that it is time consuming, laborious and has low resolution, and requires the use of toxic reagents

Method used

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  • Method for separating wheat gliadin through capillary electrophoresis
  • Method for separating wheat gliadin through capillary electrophoresis
  • Method for separating wheat gliadin through capillary electrophoresis

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Experimental program
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Effect test

Embodiment 1

[0064] Embodiment 1, capillary dynamic coating separation wheat prolamin

[0065] 1. Materials

[0066] Gliadin standard products were purchased from Tokyo Kasei Kogyo Co., Ltd.

[0067] The five wheat varieties are Zhoumai 22, Jimai 22, Liangxing 99, Aikang 58 and Laizhou 953.

[0068] Preparation of gliadin standard substance: Accurately weigh 50 mg of gliadin standard substance, dissolve it in 2 mL, 75% (volume) ethanol aqueous solution, the final concentration is 25 mg / mL.

[0069] Preparation of wheat seed samples: Wheat seeds were crushed directly, baked at 37°C for one week, passed through a 100-mesh sieve, and then extracted according to the following method 1 and method 2:

[0070] Method 1: Accurately weigh 100mg of wheat seed samples, add 1mL of 0.1M sodium chloride aqueous solution respectively, shake and extract at room temperature for 30 minutes, centrifuge (12000r / min) for 10 minutes, discard the supernatant, add 1mL of 0.1M sodium chloride aqueous solution ag...

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Abstract

The invention discloses a method for separating wheat gliadin through capillary electrophoresis. The method comprises the steps as follows: CPVP (cationic polyvinylpyrrolidone) is used as a material, and a coating is prepared on the wall of a capillary; a wheat sample is separated through capillary electrophoresis, and thus, wheat gliadin is separated; the CPVP is cationized polyvinylpyrrolidone with the viscosity average molecular weight of 20,000-25,000; the coating is prepared in the following step: the quartz capillary is washed with an aqueous solution of the CPVP and an operation buffer solution A sequentially. The CPVP is used as the coating material, dynamic modification is performed on a capillary column, the extraction separation condition suitable for wheat gliadin is screened out through optimization, a technological method for stably separating wheat gliadin is established, different varieties of wheat gliadin can be effectively distinguished, and the method becomes a wheat variety fingerprint separation technology.

Description

technical field [0001] The invention relates to a method for separating wheat prolamin by capillary electrophoresis. Background technique [0002] Glamins are the major storage proteins in wheat seed endosperm. In its mature seeds, gliadin accounts for about 40-50% of the total protein. Glamin is a single subunit in structure, and its electrophoretic bands are divided into four types: α, β, γ and ω according to their molecular weight and mobility after electrophoresis on acidic polyacrylamide gel. The composition of gliadin is polymorphic, and there are obvious differences among wheat varieties. The band pattern of its electrophoretic pattern is mainly controlled by genotype. Therefore, the gliadin electrophoretic pattern can be used as a fingerprint for variety identification, seed purity detection, and genetic relationship. analysis etc. [0003] The separation methods of gliadin mainly include acidic polyacrylamide gel electrophoresis, reverse high pressure liquid chro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 张晋丹冯旻
Owner INST OF BOTANY CHINESE ACAD OF SCI
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