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Oyster endogenous peptide extraction and identification method

An identification method and oyster technology, applied in the field of food science, can solve problems such as oyster detection and identification

Active Publication Date: 2019-10-18
山东佛坤投资有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no way to detect and identify the endogenous peptides of oysters by means of high-resolution mass spectrometry.

Method used

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  • Oyster endogenous peptide extraction and identification method
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  • Oyster endogenous peptide extraction and identification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 A method for extracting and identifying endogenous peptides from oysters based on peptidomics technology:

[0040] (1) Sample information:

[0041] The oysters were collected from a farm in Rongcheng City, Shandong Province. They were transported to the laboratory on the same day under ice storage conditions, and the fresh oysters were processed.

[0042] (2) Sample pretreatment steps:

[0043] (1) Shell the oysters, take the edible parts, freeze them in liquid nitrogen, and homogenize them. Weigh 5 g of the homogenized oyster samples into a powder state, heat at 95°C for 10 minutes, add 25ml of ammonium bicarbonate solution, and heat at 4°C at 15,000 r / Centrifuge for 15 min, collect the supernatant, and obtain the crude peptide extract;

[0044] (2) Add 125 µL of 1 mol / L Dithiothreotol (DTT) to the above protein solution, shake in a water bath at 60°C for 30 min, and then cool to room temperature;

[0045] (3) Take 1250 µL of 1 mol / L freshly prepared Iodo...

Embodiment 2

[0061] Example 2 A method for extracting and identifying endogenous peptides from oysters based on peptidomics technology

[0062] (1) Sample information:

[0063] The oysters were collected from a farm in Rongcheng City, Shandong Province. They were transported to the laboratory on the same day under ice storage conditions, and the fresh oysters were processed.

[0064] (2) Sample pretreatment steps:

[0065] (1) Shell the oysters, take the edible parts, freeze them in liquid nitrogen, and homogenize them. Weigh 5 g of oyster samples that are homogenized into powder, heat at 95°C for 10 minutes, and add 10ml of ammonium bicarbonate solution at 4°C, 15000 r / min. Centrifuge for 15 min, collect the supernatant, and obtain the crude peptide extract;

[0066] (2) Add 50 µL of 1 mol / L Dithiothreotol (DTT) to the above protein solution, shake in a water bath at 60°C for 30 min, and then cool to room temperature;

[0067] (3) Take 500 µL of 1 mol / L freshly prepared iodoacetamide (...

Embodiment 3

[0081] Example 3 A method for extracting and identifying endogenous peptides from oysters based on peptidomics technology:

[0082] (1) Sample information:

[0083] The oysters were collected from a farm in Rongcheng City, Shandong Province. They were transported to the laboratory on the same day under ice storage conditions, and the fresh oysters were processed.

[0084] (2) Sample pretreatment steps:

[0085] (1) Shell the oysters, take the edible parts, freeze them in liquid nitrogen, and homogenize them. Weigh 20 g of the homogenized oyster samples into a powder state, heat at 95°C for 10 minutes, and add 20ml of ammonium bicarbonate solution at 4°C, 15000 r / min. Centrifuge for 15 min, collect the supernatant, and obtain the crude peptide extract;

[0086] (2) Add 100 µL of 1 mol / L Dithiothreotol (DTT) to the above protein solution, shake in a water bath at 60°C for 30 min, and then cool to room temperature;

[0087] (3) Take 1000 µL of 1 mol / L freshly prepared iodoacet...

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Abstract

The invention mainly discloses an oyster endogenous peptide extraction and identification method. The method comprises the steps of: pre-processing an oyster according to a peptidomics technology, andextracting endogenous peptide in the oyster, so that polypeptide filtrate to be detected is obtained; measuring the obtained polypeptide filtrate through an ultra-high phase liquid chromatography-mass spectrography; performing online database comparison on the obtained mass spectrum data, and identifying a peptide fragment; and, verifying the correctness of a peptide fragment sequence and the applicability for conventional liquid quality analysis by synthesizing a part of the peptide fragment and using a multi-reaction monitoring analysis method established by liquid chromatography-triple quadrupole mass spectrometry. The oyster endogenous peptide extraction and identification method is provided in the invention in a targeted manner; furthermore, multiple oyster endogenous peptides are separated and identified by utilization of a mass-spectrometric technology at the first time; an oyster natural substance database is completed; and basis is provided for further researching the oysterendogenous peptide.

Description

technical field [0001] The invention belongs to the field of food science, and in particular relates to a method for identifying oyster endogenous peptides based on mass spectrometry. Background technique [0002] Oysters belong to the phylum Molluscs, the class Phyllobranches, the order Oysters, and the family Oysteridae. It is the largest cultured shellfish in the world and one of the four largest cultured shellfish in my country. It is rich in resources and has a high economic status. Oyster meat is smooth, delicious and nutritious, known as "ocean milk". At the same time, it is also a natural health food with high nutritional value. Historically, oysters are an ancient food in my country, and our ancestors recognized the nutritional and medicinal value of oysters very early. In the Tang Dynasty, it was recorded that oyster meat can promote body fluid, relieve fatigue and nourish. Li Shizhen's "Compendium of Materia Medica" in the Ming Dynasty records that oyster meat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72
CPCG01N30/02G01N30/06G01N30/72G01N2030/062
Inventor 李兆杰陈李品张晓梅张鸿伟仇文峰唐庆娟薛长湖
Owner 山东佛坤投资有限公司
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