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Extraction and application methods of meretrix meretrix linnaeus polysaccharides with antioxidant activity

A technology of clam polysaccharides and extraction methods, which is applied in the direction of organic active ingredients, medical preparations containing active ingredients, applications, etc., can solve the problems that the anti-oxidation effect has not been studied in depth, and achieve simple operation, reduce work intensity, and improve polysaccharides. Yield effect

Active Publication Date: 2014-04-02
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Clam polysaccharide has complex biological activities and functions, but its antioxidant effect has not been studied in depth

Method used

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  • Extraction and application methods of meretrix meretrix linnaeus polysaccharides with antioxidant activity
  • Extraction and application methods of meretrix meretrix linnaeus polysaccharides with antioxidant activity
  • Extraction and application methods of meretrix meretrix linnaeus polysaccharides with antioxidant activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Enzymatic extraction of Clam polysaccharide

[0022] One, plan one

[0023] 1. Beating: Take the fresh clam soft body plus 5 times the quality of deionized water, use high-speed tissue crusher and other homogenizing equipment for pulping, so that the soft body is fully broken.

[0024] 2. Enzymatic extraction: add trypsin, add 2.5% enzyme, stir and hydrolyze at 40°C for 5 h, centrifuge to get the supernatant.

[0025] 3. Concentration: The centrifugal supernatant is concentrated to 1 / 10 of the original volume by vacuum rotary evaporation.

[0026] 4. Alcohol precipitation: Take the concentrated solution, add 3 times the volume of absolute ethanol, leave it at 4 ℃ for 8 hours, centrifuge to collect the precipitate, wash twice with absolute ethanol, and evaporate the ethanol to obtain the crude clam polysaccharide. The yield of polysaccharide is 90 mg / g dry weight.

[0027] Two, plan two

[0028] 1. Beating: Take the fresh clam soft and 3 times the quality of deionized wat...

Embodiment 2

[0032] Example 2 Separation and Purification of Clam Polysaccharide by Column Chromatography

[0033] Using DEAE-52 Cellulose Anion Exchange Column Chromatography to Elute Crude Clam Polysaccharide

[0034] Use 0, 0.1 and 0.3mol / L NaCl as the eluent for stage elution. Load 4 mL of 30 mg / mL Clam polysaccharide solution at a flow rate of 2 mL / min. Each concentration of NaCl solution is eluted with 3 column volumes. , Collect the elution fraction D2 eluted with 0.1mol / L NaCl with the highest sugar content, and collect the fraction D2.

[0035] Using Superdex 200 gel filtration column chromatography to elute the crude polysaccharides of Clams

[0036] Concentrate the collected fraction D2 to a concentration of about 15 mg / mL, use deionized water as the eluent, load 0.5 mL, flow rate 0.3 mL / min, and use Superdex 200 gel filtration column for further separation and purification to obtain two The D21 and D22 components of the polysaccharide peaks were detected as single peaks by the chromat...

Embodiment 3

[0037] Example 3 Antioxidant activity of Clam polysaccharide

[0038] The isolated clam polysaccharides were tested for in vitro antioxidant activity, and the clam polysaccharides were prepared to concentrations of 5, 4, 2, 1, 0.5, 0.25, 0.125 mg / ml, and then the antioxidant model was used to test the in vitro antioxidant activity. The indicators include DPPH scavenging rate, superoxide ion scavenging rate, hydroxyl radical scavenging rate, and total reducing power.

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Abstract

The invention discloses a method for separating and purifying meretrix meretrix linnaeus polysaccharides, relating to the field of biological separation and purification technologies. The method mainly comprises the steps of carrying out added-enzyme assisted hot water extraction, suspended steaming concentration and ethanol precipitation on meretrix meretrix linnaeus soft-bodied homogenate, so as to obtain a crude meretrix meretrix linnaeus polysaccharide product, then, carrying out deproteinization by a sevage method, and gradually separating and purifying the meretrix meretrix linnaeus polysaccharides by adopting DEAE-52 cellulose anion-exchange column chromatography and Superdex 200 gel-filtration column chromatography, wherein the meretrix meretrix linnaeus polysaccharides obtained through purifying are single polysaccharides of two kinds and have the purity over 90%. Thus, the obtained refined meretrix meretrix linnaeus polysaccharide product can serve as raw materials or additives of pharmaceuticals, chemicals, healthcare nutrients and cosmetics; a basis is provided for further researching the meretrix meretrix linnaeus polysaccharides.

Description

Technical field [0001] The invention belongs to the technical field of marine biology, and provides an enzymatic extraction and separation and purification method for the clam polysaccharide with antioxidant activity. The formed clam polysaccharide can be used for the preparation of health care products with antioxidant function. Background technique [0002] Meretrix (Meretrix meretrix Linnaeus) belongs to the phylum Mollusca, Bivalvia, Eubranchidae, Clamidae, and Meretrix. It is one of the main shellfishes traditionally cultured in the tidal flats in my country. Clam polysaccharides mainly exist in the viscera of clams, and have not been widely studied and applied. Modern scientific research has shown that Clam polysaccharide has the functions of clearing heat and dampness, reducing phlegm, dispelling masses, and anti-tumor. It can also play a variety of physiological functions such as hypoglycemic, hypolipidemic, anti-mutation, and anti-aging. In terms of hypoglycemic functio...

Claims

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Application Information

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IPC IPC(8): C08B37/00A23L1/09A61K31/715A61P39/06A23L33/125
Inventor 史劲松
Owner JIANGNAN UNIV
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