CN103724675A - Method for extracting polysaccharide in algae - Google Patents

Abstract

The invention relates to a method for extracting polysaccharide in algae, and belongs to the field of marine organism medicine. The method is carried out according to the following steps: (1) roguing seaweed, cleaning, drying, and then smashing; (2) weighing 100 g of seaweed powder, adding 85-90% alcohol, performing reflux extraction for 4 to 6 hours, filtering, after medicine residue is dried, adding water, performing ultrasonic extraction for 5 times, filtering for 10 min each time, combining filtrate, condensing at the temperature of 80-90 DEG C until the specific gravity is 1.03, and performing alcohol precipitation for 18 to 24 hours until the alcohol precipitation concentration is 75%; performing centrifugal filtration to obtain precipitate, repeatedly water-dissolving the precipitate, performing alcohol precipitation until no sugars reducing reaction occurs, and then washing by absolute ethyl alcohol and acetone repeatedly so as to obtain polysaccharide crude extract in the algae; (3) performing decoloration on the crude extract through activated carbon, performing deproteinization through a Sevage method, removing micromolecule through a running water dialysis method, performing alcohol precipitation, and performing vacuum drying so as to obtain the extracted polysaccharide in the algae; (4) adding modified starch in the polysaccharide, wherein the added weight is 0.5-1.2 times of that of the polysaccharide, uniformly stirring, using microwave drying to obtain powder-like mixture, sterilizing outward for 15 seconds, sealing, and packaging. The method for extracting the polysaccharide in the algae is simple, convenient and quick, good in repeatability and high in recovery rate.

Description

In a kind of algae, extract the extracting method of polysaccharide

Technical field

The invention belongs to marine organisms field of medicaments, particularly in a kind of algae, extract the extracting method of polysaccharide.

Background technology

Marine alga is the common name of marine products phycophyta.TCM Algae is the dry frond of Sargassaceae Sargassum or Sargassum fusiforme, has softening and resolving hard mass, the function of dissolving phlegm Li Shui.The clinical goiter and tumor that is used for the treatment of, painful and swollen testis, the diseases such as phlegm and retained fluid oedema.Modern study shows, marine alga has the immunological competence of enhancing, suppresses the effects such as virus and antineoplastic activity.In these biological activitys algae contained with it, extract polysaccharide closely related.It is reported that extracting polysaccharide in algae is a kind of irreducibility disaccharide that nature extensively distributes.Its molecular structure is unique, stable chemical nature.In algae, extracting polysaccharide is the efficient biological adsorption agent of a class, and extra large bath polysaccharide has the effects such as reducing blood-fat, anti-oxidant, immunomodulatory, extracts the cold protective medium that polysaccharide derivates can be used as cellular material in algae.These marvellous biological characteristicses of trehalose, make it at aspects such as medicine, food, makeup, agriculturals, have broad application prospects.The exploitation of extracting polysaccharide in algae has become the study hotspot of international coverage.

Summary of the invention

The invention provides the extracting method that extracts polysaccharide in a kind of algae, according to following steps, carry out:

(1) will after marine alga impurity elimination cleaning, drying, pulverize;

(2) take 100g marine algae powder, add 85 ~ 90% alcohol reflux 4 ~ 6 hours, filter, after the dregs of a decoction are dry, add water supersound extraction 5 times, each 10min filters, it is 1.03 that merging filtrate is concentrated into proportion in 80 ~ 90 ℃, alcohol precipitation, and 18 ~ 24h alcohol precipitation concentration is 75%; Centrifuging obtains throw out, and throw out repeatedly water-soluble alcohol is sink to without reducing sugar reaction, then through dehydrated alcohol, washing with acetone repeatedly obtains in algae and extracts polysaccharide crude extract afterwards;

(3) crude extract is through decolorizing with activated carbon, Sevage method deproteinated, and flowing water dialysis method takes off small molecules, alcohol precipitation, vacuum-drying obtains and in algae, extracts polysaccharide.

(4), add treated starch in polysaccharide, the 0.5-1.2 that the quality adding is polysaccharide doubly, after stirring, is used microwave drying, obtains powder mixture, and sterilizing outside, after 15 seconds, packs.

The assay that extracts polysaccharide in algae carries out in accordance with the following methods:

Standard curve making: precision takes in the volumetric flask that dextrose standard sample 12.50mg is dissolved in 100ml, obtains the glucose standardized solution of 0.1250mg/ ml.From standardized solution, draw respectively 0.0,0.4,0.5,0.6,0.7,0.8,0.9,1.0ml, add water to 1ml.Add again 10% phenol solution 1ml, vitriol oil 5ml, shake up rear placement 5min, put boiling water bath and place 15min, take out and be cooled to room temperature.At 490nm, survey its absorbancy.Obtain the regression equation of absorbancy and concentration: A=53186C+ 010108.(?r=?019990)?。

Stability experiment takes: get the polysaccharide soln 0.5ml of concentration known, add water to 1ml, add rapidly phenol solution, the 5ml vitriol oil of 1ml 10%, shake up rear placement 5min, put boiling water bath and place 15min, take out and be cooled to room temperature.At 490nm, survey its absorbancy.In experiment, find that its absorbancy is stable in 0 ~ 25min.

Recovery test: extract polysaccharide soln in the algae of 5 parts of concentration known of absorption, respectively add standard glucose solution, measure absorbancy, by the regression equation calculation rate of recovery in accordance with the law.As calculated, its rate of recovery is more than 95%, and in algae, extracting polysaccharide content extraction yield is 4.26 ~ 4.36%.

The present invention extracts and in algae, extracts polysaccharide from marine alga, and can according to its have stronger biological adsorption ability, reducing blood-fat, anti-oxidant, strengthen immunoregulatory activity etc., be developed to function food and medicine, or prepared microcapsule.In algae, extract polysaccharide and may become the main drug resource of the mankind.The assay that extracts polysaccharide in algae adopts traditional benzene phenol-sulfuric acid method, and method is easy, and the rate of recovery is high, favorable reproducibility, and data are credible.

In a kind of algae of the present invention, extract the extracting method of polysaccharide, method is easy fast, favorable reproducibility, and the rate of recovery is high.

Embodiment

The present invention marine alga used is that Xiangshan of Zhejiang Province produces, and chemical reagent used is commercial.

embodiment 1

In algae, extract an extracting method for polysaccharide, according to following steps, carry out:

(1) will after marine alga impurity elimination cleaning, drying, pulverize;

(2) take 100g marine algae powder, add 90% alcohol reflux 4 hours, filter, after the dregs of a decoction are dry, add water supersound extraction 5 times, each 10min filters, it is 1.03 that merging filtrate is concentrated into proportion in 80 ~ 90 ℃, alcohol, and 24h alcohol precipitation concentration is 75%; Centrifuging obtains throw out, and throw out repeatedly water-soluble alcohol is sink to without reducing sugar reaction, then through dehydrated alcohol, washing with acetone repeatedly obtains in algae and extracts polysaccharide crude extract afterwards;

(3) crude extract is through decolorizing with activated carbon, Sevage method deproteinated, and flowing water dialysis method takes off small molecules, alcohol precipitation, vacuum-drying obtains and in algae, extracts polysaccharide 4.36g.

(4), add treated starch in polysaccharide, the 0.5-1.2 that the quality adding is polysaccharide doubly, after stirring, is used microwave drying, obtains powder mixture, and sterilizing outside, after 15 seconds, packs.

embodiment 2

In algae, extract an extracting method for polysaccharide, according to following steps, carry out:

(1) will after marine alga impurity elimination cleaning, drying, pulverize;

(2) take 100g marine algae powder, add 70% alcohol reflux 4 hours, filter, after the dregs of a decoction are dry, add water supersound extraction 5 times, each 10min filters, it is 1.03 that merging filtrate is concentrated into proportion in 80 ℃, alcohol precipitation, and 18 ~ 24h alcohol precipitation concentration is 75%; Centrifuging obtains throw out, and throw out repeatedly water-soluble alcohol is sink to without reducing sugar reaction, then through dehydrated alcohol, washing with acetone repeatedly obtains in algae and extracts polysaccharide crude extract afterwards;

(3) crude extract is through decolorizing with activated carbon, Sevage method deproteinated, and flowing water dialysis method takes off small molecules, alcohol precipitation, vacuum-drying obtains and in algae, extracts polysaccharide 4.34g.

(4), add treated starch in polysaccharide, the 0.5-1.2 that the quality adding is polysaccharide doubly, after stirring, is used microwave drying, obtains powder mixture, and sterilizing outside, after 15 seconds, packs.

 

embodiment 3

In algae, extract an extracting method for polysaccharide, according to following steps, carry out:

(1) will after marine alga impurity elimination cleaning, drying, pulverize;

(2) take 100g marine algae powder, add 75% alcohol reflux 5 hours, filter, after the dregs of a decoction are dry, add water supersound extraction 5 times, each 10min filters, it is 1.03 that merging filtrate is concentrated into proportion in 85 ℃, alcohol precipitation, and 18 ~ 24h alcohol precipitation concentration is 75%; Centrifuging obtains throw out, and throw out repeatedly water-soluble alcohol is sink to without reducing sugar reaction, then through dehydrated alcohol, washing with acetone repeatedly obtains in algae and extracts polysaccharide crude extract afterwards;

(3) crude extract is through decolorizing with activated carbon, Sevage method deproteinated, and flowing water dialysis method takes off small molecules, alcohol precipitation, vacuum-drying obtains and in algae, extracts polysaccharide 4.26g.

(4), add treated starch in polysaccharide, the 0.5-1.2 that the quality adding is polysaccharide doubly, after stirring, is used microwave drying, obtains powder mixture, and sterilizing outside, after 15 seconds, packs.

Claims (1)

1. in algae, extract an extracting method for polysaccharide, it is characterized in that carrying out according to following steps:

(1) will after marine alga impurity elimination cleaning, drying, pulverize;

(2) take 100g marine algae powder, add 85 ~ 90% alcohol reflux 4 ~ 6 hours, filter, after the dregs of a decoction are dry, add water supersound extraction 5 times, each 10min filters, it is 1.03 that merging filtrate is concentrated into proportion in 80 ~ 90 ℃, alcohol precipitation, and 18 ~ 24h alcohol precipitation concentration is 75%; Centrifuging obtains throw out, and throw out repeatedly water-soluble alcohol is sink to without reducing sugar reaction, then through dehydrated alcohol, washing with acetone repeatedly obtains in algae and extracts polysaccharide crude extract afterwards;

(3) crude extract is through decolorizing with activated carbon, Sevage method deproteinated, and flowing water dialysis method takes off small molecules, alcohol precipitation, vacuum-drying obtains and in algae, extracts polysaccharide;

(4), add treated starch in polysaccharide, the 0.5-1.2 that the quality adding is polysaccharide doubly, after stirring, is used microwave drying, obtains powder mixture, and sterilizing outside, after 15 seconds, packs.