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Method for fast detecting fish collagen by means of capillary electrophoresis

A capillary electrophoresis and collagen technology, applied in the field of allergen detection, can solve the problems of complicated operation, slow detection speed, low sensitivity and accuracy, etc., and achieve the effect of high sensitivity and accuracy, fast detection and high sensitivity

Inactive Publication Date: 2016-06-15
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, these methods generally have the problems of slow detection speed, complicated and cumbersome operation, and low sensitivity and accuracy.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1. Sample Preparation

[0038] Sample preparation: use the kit to extract the whole protein in the fish skin, and dissolve the whole fish protein in the equilibrium buffer solution with pH 7.5 to obtain the sample solution. Add 3 to 5 times the column volume of equilibrium solution at a flow rate of 1.5ml / min to balance the chromatography column, then add the sample solution to the anion exchange chromatography column at a flow rate of 0.5ml / min, and then use 0.2 to 0.3mol / L eluent elutes the chromatographic column at a rate of 1.0ml / min, and collects the solution corresponding to the breakthrough peak, which is the sample.

[0039] 2. Optimization of capillary electrophoresis analysis conditions

[0040] (1) Capillary selection and pretreatment: use uncoated quartz capillary column (45cm×75μm, P / ACE TM MDQSystem, Beckman Company), the new capillary column was first washed with methanol for 10 min, distilled water for 5 min, 0.1M Na0H solution for 30 min, distilled w...

Embodiment 2

[0048] 1. Sample Preparation

[0049] Sample preparation: use the kit to extract the whole protein in the fish skin, and dissolve the whole fish protein in the equilibrium buffer solution with pH 7.5 to obtain the sample solution. Add 3 to 5 times the column volume of equilibrium solution at a flow rate of 1.5ml / min to balance the chromatography column, then add the sample solution to the anion exchange chromatography column at a flow rate of 0.5ml / min, and then use 0.2 to 0.3mol / L eluent elutes the chromatographic column at a rate of 1.0ml / min, and collects the solution corresponding to the breakthrough peak, which is the sample.

[0050] 2. Optimization of capillary electrophoresis analysis conditions

[0051] (1) Capillary selection and pretreatment: use uncoated quartz capillary column (45cm×75μm, P / ACE TM MDQSystem, Beckman Company), the new capillary column was first washed with methanol for 10 min, distilled water for 5 min, 0.1M Na0H solution for 30 min, distilled w...

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PUM

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Abstract

The invention discloses a method for rapidly detecting fish collagen by capillary electrophoresis, which comprises the following steps: (1) extracting the whole protein in fish skin and dissolving it in an equilibrium buffer solution with a pH of 7-8, and then adding anion exchange In the chromatographic column, the eluent is eluted, and the solution corresponding to the breakthrough peak is collected as the sample; (2) The obtained sample is subjected to capillary zone electrophoresis to obtain an electrophoretic spectrum, and the peak area obtained in the electrophoretic spectrum is substituted into the fish collagen The concentration of fish collagen in the sample solution was calculated from the standard curve regression equation of the protein. The present invention uses capillary electrophoresis to rapidly, qualitatively and quantitatively detect a new type of fish allergen, aquatic product allergen fish collagen. The invention uses the established method to qualitatively and quantitatively analyze fish collagen, which has no literature report, and provides new basis for fast and accurate detection of fish collagen.

Description

technical field [0001] The invention relates to the technical field of allergen detection, in particular to a method for fast, qualitative and quantitative detection of fish collagen by capillary electrophoresis. Background technique [0002] Fish is delicious, nutritious and popular among consumers, but it is also one of the eight most common foods with high allergenic activity announced by the Food and Agriculture Organization of the United Nations. In industrialized countries, the population allergic to fish accounts for about 0.1-0.3% of the total population, and accounts for 2.3%-30.4% of the total population with food allergies. Collagen (Pepsinsolublecollagen, PSC) is a newly discovered fish allergen, which widely exists in fish skin and flesh. [0003] At present, the detection methods for identifying fish allergens mainly include the following: [0004] (1) Histamine release test [0005] The histamine release test uses allergens to stimulate mast cells, causing ...

Claims

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Application Information

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IPC IPC(8): G01N27/447
CPCG01N27/447
Inventor 傅玲琳王彦波赵淑淑
Owner ZHEJIANG GONGSHANG UNIVERSITY
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