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Rapid detection method of fish parvalbumin by capillary electrophoresis

A capillary electrophoresis and parvalbumin technology, applied in the field of allergen detection, can solve problems such as difficult measurement, and achieve the effects of high sensitivity and precision, high analysis speed, and high accuracy and sensitivity

Inactive Publication Date: 2016-08-31
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many factors affecting the release of histamine in this experiment, and the measurement is difficult

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Sample Preparation

[0036] Sample preparation: use the kit to extract the whole protein in the fish muscle, 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 rate of 0.5ml / min, and then use 0.04 to 0.14mol / min The eluent of L elutes the chromatographic column, and the solution corresponding to the breakthrough peak is collected, which is the sample.

[0037] 2. Optimization of capillary electrophoresis analysis conditions

[0038] (1) Capillary selection and pretreatment: use uncoated quartz capillary column (45cm×75μm, P / ACE TM MDQ System, Beckman Company), the new capillary column was first rinsed with methanol for 10 min, distilled water for 5 min, 0.1M Na0H solution for 30 min, distilled water for...

Embodiment 2

[0046] 1. Sample Preparation

[0047] Sample preparation: use the kit to extract the whole protein in the fish muscle, 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 rate of 0.5ml / min, and then use 0.04 to 0.14mol / min The eluent of L elutes the chromatographic column, and the solution corresponding to the breakthrough peak is collected, which is the sample.

[0048] 2. Optimization of capillary electrophoresis analysis conditions

[0049] (1) Capillary selection and pretreatment: use uncoated quartz capillary column (45cm×75μm, P / ACE TM MDQ System, Beckman Company), the new capillary column was first rinsed with methanol for 10 min, distilled water for 5 min, 0.1M Na0H solution for 30 min, distilled water for...

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Abstract

The invention discloses a method for detecting fish parvalbumin by capillary electrophoresis. The method comprises the following steps: (1) extracting holoprotein from fish-muscle and dissolving in an equilibrium buffer with pH being 7-8, loading into an anion exchange chromatographic column, eluting with an eluant and collecting a solution corresponding to a flow-through peak so as to obtain a sample; and (2) carrying out capillary zone electrophoresis on the sample to obtain substituting peak area obtained in electrophoretogram into a standard curve regression equation of fish parvalbumin and calculating to obtain concentration of fish parvalbumin in the sample solution. The method of the invention is a method for rapid, qualitative and quantitative detection of aquatic product allergen fish parvalbumin by capillary electrophoresis. The method for qualitative and quantitative analysis of fish parvalbumin has been reported in the literature, and provides new basis for rapid and accurate detection of fish parvalbumin.

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] Aquatic products are very popular among people because of their delicious taste and rich nutritional value. Among them, fish animals are one of the eight types of allergic foods. There are many cases of allergic reactions caused by eating fish animals every year, and severe cases even endanger people. life. Parvalbumin (Pav) exists in large quantities in the white muscle of lower vertebrates and is the main allergen of fish animals. [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 them to release histamine and other active mediators, and then measures the releas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
CPCG01N27/447G01N2550/00
Inventor 王彦波傅玲琳周瑾茹
Owner ZHEJIANG GONGSHANG UNIVERSITY
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