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Method for rapidly detecting shrimp tropomyosin through capillary electrophoresis

A technology of tropomyosin and capillary electrophoresis, which is applied in the field of allergen detection, can solve the problems of complex and cumbersome operation, slow detection speed, low sensitivity and low precision, and achieve high analysis speed, fast detection, high sensitivity and precision Effect

Inactive Publication Date: 2016-08-17
ZHEJIANG GONGSHANG UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0007] 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

[0034] 1. Sample Preparation

[0035] The kit is used to extract the whole protein in the shrimp muscle, and the whole shrimp protein is dissolved in the equilibrium buffer solution of 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 0.1 to 0.2mol / L The NaCl eluent was eluted from the chromatographic column at a rate of 1.0ml / min, and the solution corresponding to the breakthrough peak was collected as the sample.

[0036] 2. Optimization of capillary electrophoresis analysis conditions

[0037] (1) Capillary selection and pretreatment: choose an uncoated quartz capillary column, first rinse the new capillary column with methanol for 10 minutes, distilled water for 5 minutes, 0.1M NaOH solution for 30 minutes, distilled water for 5 minutes, and finall...

Embodiment 2

[0045]1. Sample Preparation

[0046] The kit is used to extract the whole protein in the shrimp muscle, and the whole shrimp protein is dissolved in the equilibrium buffer solution of pH 7.5 to obtain the sample liquid. 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 0.1 to 0.2mol / L The NaCl eluent was eluted from the chromatographic column at a rate of 1.0ml / min, and the solution corresponding to the breakthrough peak was collected as the sample.

[0047] 2. Optimization of capillary electrophoresis analysis conditions

[0048] (1) Capillary selection and pretreatment: choose an uncoated quartz capillary column, first rinse the new capillary column with methanol for 10 minutes, distilled water for 5 minutes, 0.1M NaOH solution for 30 minutes, distilled water for 5 minutes, and finally r...

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Abstract

The invention discloses a method for rapidly detecting shrimp tropomyosin through capillary electrophoresis. The method comprises the following steps that 1, holoprotein in shrimp muscle is extracted, dissolved in an equilibration buffer with the pH of 7-8, added into an anion exchange chromatographic column and eluted through an elution solution, and a solution corresponding to a penetrating peak is collected as a sample; 2, the obtained sample is subjected to capillary zone electrophoresis to obtain an electrophoretogram, the peak area in the electrophoretogram is substituted into a standard curvilinear regression equation to calculate the concentration of the tropomyosin in the obtained sample. According to the method for rapidly detecting allergen-shrimp tropomyosin of aquatic products through capillary electrophoresis, holoprotein in shrimp muscle is extracted, added into the anion exchange chromatographic column and eluted through the elution solution, the solution corresponding to the penetrating peak is collected and added into a capillary electrophoresis apparatus, and qualitative and quantitative detection is carried out on the shrimp tropomyosin in the sample solution through capillary zone electrophoresis.

Description

technical field [0001] The invention relates to the technical field of allergen detection, in particular to a qualitative and quantitative detection method for shrimp tropomyosin. Background technique [0002] With the development of aquaculture and international trade, diseases caused by food allergies such as aquatic products are on the rise. About 30% to 40% of the diseases in the world are caused by allergies, and allergies to aquatic products are particularly serious. The main allergen in crustacean aquatic products such as shrimp and crab is tropomyosin (TM). Therefore, establishing a rapid and simple method for detecting tropomyosin can prevent and treat aquatic product allergy diseases, thereby effectively reducing the incidence of aquatic product allergy. [0003] At present, there are mainly the following methods for the detection of food allergens: [0004] (1) Use the principle of antigen-antibody specific reaction for detection, usually using enzyme-linked im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/34G01N27/447
CPCG01N27/447G01N1/34
Inventor 傅玲琳王彦波赵淑淑
Owner ZHEJIANG GONGSHANG UNIVERSITY
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