Enrichment and characterization method of American ginseng polypeptide

A technology of American ginseng and enrichment, which is applied in the fields of separation analysis, plant omics and biology, can solve the problems such as poor knowledge of American ginseng peptides, and achieve high throughput and sensitivity, high sensitivity, and fast analysis speed

Active Publication Date: 2020-06-09
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because its entire genome has not been sequenced, little is known about the peptides in American ginseng

Method used

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  • Enrichment and characterization method of American ginseng polypeptide
  • Enrichment and characterization method of American ginseng polypeptide
  • Enrichment and characterization method of American ginseng polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1. Enrichment and separation of target peptides

[0046] Crush and dry American ginseng slices 100g, add a volume ratio of 50% ethanol aqueous solution as the extraction solvent, the mass volume ratio of the medicinal material powder to the solvent is 0.125g / ml, mix well; ultrasonic for 30 minutes, separate, collect the solvent, repeat the extraction twice, and combine Extract solution; dry the resulting solution under a gentle nitrogen flow, and then redissolve it in 100mL water; add the extract to 300ml ethanol for alcohol precipitation, and dissolve the precipitate in 40ml water; add it to the suspension at a volume ratio of 1:1 Add 40ml of ethyl acetate, repeat 3 times to obtain the extract containing ethyl acetate and the extract not containing ethyl acetate; take the extract not containing ethyl acetate, add 40ml n-butanol according to the volume ratio of 1:1, Repeat 3 times to obtain the extract containing n-butanol and the extract of the aqueous layer; use the A...

Embodiment 2

[0059] 1. Enrichment and separation of target peptides

[0060] Crush and dry American ginseng slices 100g, add a volume ratio of 30% ethanol aqueous solution as the extraction solvent, the mass volume ratio of the medicinal material powder to the solvent is 0.2g / ml, mix evenly; ultrasonic for 30 minutes, separate, collect the solvent, repeat the extraction twice, and combine Extract solution; dry the resulting solution under a gentle nitrogen flow, and then redissolve it in 200mL water; add the extract to 400ml ethanol for alcohol precipitation, and dissolve the precipitate in 50ml water; add to the suspension at a volume ratio of 1:1 Add 50ml of ethyl acetate, repeat 3 times to obtain the extract containing ethyl acetate and the extract not containing ethyl acetate; take the extract not containing ethyl acetate, add 50ml n-butanol according to volume ratio 1:1, Repeat 3 times to obtain the extract containing n-butanol and the extract of the aqueous layer; use the Amicon Ultr...

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Abstract

The invention discloses an enrichment and characterization method of American ginseng polypeptide. The method comprises steps of firstly, preparing American ginseng extracting solution; adding ethyl acetate into the American ginseng extract liquor to obtain extract liquor containing the ethyl acetate and the extract liquor without the ethyl acetate; adding n-butyl alcohol into the extract liquor without the ethyl acetate to obtain the extract liquor containing the n-butyl alcohol and water-bearing layer extract liquor without the n-butyl alcohol; and carrying out ultrafiltration of the water-containing layer extraction liquid, centrifuging, collecting the supernatant, and carrying out HPLC gradient elution to obtain the eluent which is the enriched American ginseng polypeptide. The methodis advantaged in that the enriched polypeptide is characterized by using the mass spectrometry technology and a proteomics analysis database.

Description

technical field [0001] The invention belongs to the fields of separation analysis, plant omics and biotechnology, and relates to a method for effectively screening and enriching polypeptides from complex components of plants, tracking and detecting them based on advanced mass spectrometry technology, and characterizing target polypeptides through modern mass spectrometry technology. Background technique [0002] Peptide is a general term for different peptides consisting of 20 encoded amino acids in different compositions and arrangements, from dipeptides to complex linear and circular structures. A variety of plant oligopeptides, cyclic peptides, cyclic peptide alkaloids, and sugars have been reported. Peptides, etc., have shown gratifying physiological and pharmacological activities from different angles. Peptides in plants are involved in a variety of physiological processes, including the regulation of growth and development, immunity, estimation of genetic variation, cha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/08G01N30/14
CPCG01N30/02G01N30/06G01N30/08G01N30/14G01N2030/027G01N2030/062
Inventor 张晓哲赵强赵楠程孟春
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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