CN101250232B - Extraction technique of sargassum fusiform active polysaccharides - Google Patents
Extraction technique of sargassum fusiform active polysaccharides Download PDFInfo
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Abstract
The invention relates to an extraction method of sargassum fusiform active polysaccharide, which uses four-step extraction method as combined ultrasonic extraction, water extraction, alkali extraction, hot water extraction and ultrasonic technique, thereby improving sargassum fusiform polysaccharide extraction rate and avoiding damage on sargassum fusiform activity in polysaccharide extraction. Besides adding the character of polysaccharide on extract, the prepared sargassum fusiform active polysaccharide has high oxidation resistance, which is suitable for cosmetic technical field. In addition, the prepared sargassum fusiform polysaccharide uses active carbon adsorption and AB-8 macroporous adsorption resin to combine decolor and de-fishy, thereby resolving the key techniques as decolor and de-fishy of sea phaeophyceae polysaccharide as sargassum fusiform or the like, to improve polysaccharide purity, appearance quality and confirm the high oxidation resistance of de-fishy and decolor polysaccharide.
Description
Technical field
The present invention relates to biochemical field, specifically refer to a kind of extraction process of Sargassum fusiforme active polysaccharide.
Background technology
Sargassum fusiforme (Sargassum fusiforme (Harv1) Setchell) is the Sargassaceae plant, claims extra large barley, deer horn point etc. again, is a kind of perennial, warm temperate zone marine alga, belongs to phaeophyta, Sargassaceae, Sargassum.Mainly distribute, reach the Lezhou Peninsula in the south, its distribution is all arranged North gets Liaodong Peninsula, coastal at most with Zhejiang.Great mass of data studies show that, the activeconstituents Hijiki polysaccharide that extracts in the marine alga (S1 fusiform e polysaccharides, SFPS), except being widely used in food, healthcare industry, have good anticancer and hypertensive effect, also add as cosmetic material, have more good skin-care effect, it is very thirsty to alleviate skin, recovers water balance, and reaches the effect of the potent skin that compacts of preserving moisture by bioactivation.Hijiki polysaccharide has important use to be worth in healthcare products, medicine and cosmetic industry.Facial mask and face cream skin care product that it mainly applies to anti-oxidant function have the Dermatochalasia of preventing and wrinkle and produce, and improve UV-induced skin pigment precipitation, effects such as desalination senile plaque and other pigmented spots.
In recent years, the extraction of polysaccharide and purification process have had development rapidly, and new method also continues to bring out, and comprises HPLC method, enzyme process, membrane separation process etc.But because each side, these methods are not widely used.At present neutral hot water extraction or acidleach formulations of adopting more.Various nutritive ingredient major parts based on Hijiki polysaccharide are water soluble substances, therefore can utilize the method for hot water lixiviate to be extracted; This extracting method simple possible, easy to operate, not high to equipment requirements, but its production time is long, and polysaccharide yield is not high, and waste is serious.The acidleach formulation is the ad hoc structure relevant with immunocompetence in the destructible polysaccharide then, and two kinds of methods all have its weak point.Adopt the method for enzymolysis and extraction polysaccharide that report was also once arranged, but when the Sargassum fusiforme lixiviate, add cellulase and polygalacturonase, can fully destroy the weave construction of Sargassum fusiforme, can improve polysaccharide extract rate, compare, have certain advantage with the above two.Its effective constituent difference that is contained of the resulting extract of different process, so its Application Areas and effect are also inequality.Above-mentioned technology is intended to improve the extraction yield of Hijiki polysaccharide, has not considered the influence of extraction process to the extraction active polysaccharide, so in the leaching process, and some useful activeconstituentss are more or less destroyed or weeded out.
Summary of the invention
Technical problem to be solved by this invention is the situation at prior art, and a kind of extraction process with Sargassum fusiforme active polysaccharide of high antioxidant is provided.
The present invention solves the problems of the technologies described above the technical scheme that is adopted: the extraction process of this Sargassum fusiforme active polysaccharide is characterized in that comprising the steps:
The Sargassum fusiforme frond is prepared: reject invalid, fibrous root, and clean with flushing with clean water, be dried to water content≤12%, preferably, and can be 50-60 ℃ of drying, pulverize with pulverizer dry back, crosses 80 mesh sieves then and obtain dry powder;
The dry powder that step a is obtained adds water normal temperature lixiviate 2-4hr, and wherein the mass ratio of dry powder and water is 1: 30-40; Handled 10-15 minute with 800-1200W supersonic cell crusher ice bath, centrifugal settling obtains supernatant liquor one and throw out; Described throw out water under 70-80 ℃ of temperature is carried 1-2hr, and centrifugal settling gets supernatant liquor two and throw out again;
Throw out is carried 1-2h with 1-3% carbonic acid soda, the centrifugal supernatant liquor three that gets;
Merge supernatant liquor one, two, three, regulate its pH value to 7.0, reduced vacuum concentrates, concentrated solution Sevage method Deproteinization 2-4 time, and supernatant liquor 5-10: centrifugal after the 1 granulated active carbon absorption, get supernatant liquor, filter, collect filtrate;
Filtrate is used 4-6: the decolouring of 1 AB-8 macroporous adsorbent resin, centrifugal again or filter after get filtrate, add the ethanol sedimentation of volume fraction 65%-70%, throw out is used 1 successively: the acetone of 5-10, absolute ethanol washing after-filtration, collecting precipitation thing;
The throw out reduced vacuum drying of collecting is obtained the Sargassum fusiforme active polysaccharide, and preferably, described reduced vacuum concentrates and can carry out under 0.1Mpa, 50-60 ℃ condition.
In the above-mentioned Sargassum fusiforme active polysaccharide extracting method, the centrifugation rate in the steps d is 10000 rev/mins, and centrifugation time is 10 minutes; Centrifugation rate in other step is 4000-5000 rev/min, and centrifugation time is 10-15 minute.
Compared with prior art, this product is no raw meat, and is light yellow; This technology combines Sargassum fusiforme pharmacognosy principle, has used the four step extracting method that physics and chemistry combines, and combines promptly that water is carried, alkali is carried, hot water is carried, ultrasonic technology; Not only improve the extraction yield of Hijiki polysaccharide, prevent in the polysaccharide leaching process, and guarantee that the active polysaccharide structure is not destroyed, and has the high anti-oxidation activity the bioactive destruction of Sargassum fusiforme.Sargassum fusiforme Crude polysaccharides extraction yield is about 8-9%, and thick product is about 11-13%, stable yield; Product is handled through decolouring and deodorization technology, has solved ocean phaeophyta polysaccharide such as Sargassum fusiforme and has gone raw meat and decolouring gordian technique, effectively improves purity of polysaccharide, exterior quality, and guarantees to take off the high antioxidant of raw meat decolouring back polysaccharide.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail.
Sargassum fusiforme is rejected invalid, fibrous root, clean with flushing with clean water., pulverize to water content 10% at 55 ℃ of blowing-type constant temperature oven inner dryings, cross 80 mesh sieves then and obtain dry powder with pulverizer;
Get above-mentioned dry powder 50 grams and add 1500 milliliters of normal temperature lixiviates of water 2h, handled 11 minutes with 900W ultrasonic wave ice bath, with the said extracted thing with 4500 rev/mins centrifugal 10 minutes, obtain supernatant liquor one and throw out; Throw out water under 80 ℃ of temperature is carried 1 hour, and the same then method is centrifugal, obtains supernatant liquor two and throw out; Throw out is carried 1 hour with 2% carbonic acid soda again, centrifugal with method, obtain supernatant liquor three and throw out.
Merge the above-mentioned gained supernatant liquor one that respectively goes on foot; two; three; regulate its pH value to 7.0; at 0.1Mpa; 60 ℃ of following reduced vacuum concentrate; concentrated solution Sevage method Deproteinization 4 times; after using granulated active carbon absorption in 10: 1 then; with 10000 rev/mins speed centrifugal 10 minutes; behind the gained supernatant liquid filtering; with 4: the decolouring of 1AB-8 macroporous adsorbent resin; get filtrate after centrifugal for 4500 rev/mins, add the ethanol sedimentation of volume fraction 65%, throw out is used successively 1: 7 acetone; absolute ethanol washing; filter, taking precipitate is at 0.1Mpa; 60 ℃ of reduced vacuum dryings obtain Hijiki polysaccharide 6.8 grams.
Calculate the extraction yield of Sargassum fusiforme active polysaccharide by following formula:
Reducing sugar content (g) in total reducing sugar (g)-sample in polysaccharide content (g)=sample
Polysaccharide extract rate (%)=polysaccharide content (g)/Sargassum fusiforme dry powder quality (g) * 100%
Above-mentioned resulting Hijiki polysaccharide is detected (Harbin University of Commerce's journal, 2006 the 1st phases, Hijiki polysaccharide extraction and assay through the sulfuric acid phynol method; " Biochemistry Experiment method and technology ", Science Press, reducing sugar and total sugar content are fixed), as calculated, the extraction yield of Hijiki polysaccharide is 8.53% in the present embodiment.Be compared to the 6-7% in the 9-10% of Enzymatic Extraction technology and microwave extraction technology, the extraction process by water, the extraction rate reached of polysaccharide of the present invention is to 8-9%, and the product activity is good, has high antioxygenic property.
The performance test of Sargassum fusiforme active polysaccharide antioxidation in vitro
The DPPH oxidation-resistance detects the method with reference to " biological chemistry and biophysics progress " 2006 the 6th phases of periodical " estimating the plant resistance of oxidation with removing organic free radical DPPH method ", utilize the absorption honeybee of the feature red-purple group of DPPH solution, with spectrophotometry, add OD after the liquid glucose
514The decline that absorbs represents that it eliminates ability to organic free radical.DPPH dissolves with small amount of ethanol earlier, and is mixed with 1 * 10 with 10% ethanol
-4Mol/L keeps in Dark Place.Get 2mLDPPH solution, add the 2mL facial mask and extract the solution thorough mixing.Measure its OD at 514nm (maximum absorption wave strong point) after 30 minutes
514To remove DPPH50% is IC
50The gram number is the oxidation-resistance index.Clearance rate P calculates available following formula:
P＝[A
0-(A
i-A
j)]/A
0
A in the formula
0The absorbancy of=2mL DPPH solution+2mL solvent; A
i=2mL DPPH solution+2mL sample liquid absorbancy; A
jThe absorbancy of=2mL sample+2mL solvent.Its test result is as shown in table 1.
Table 1: the comparison of the Hijiki polysaccharide of Different Extraction Method and vitamins C, E
The result shows: the external clearance rates for trial of this technology ultrasonic extraction Hijiki polysaccharide DPPH free radical, the oxidation-resistance effect is better than that traditional water is carried, enzyme is carried, microwave extract method, compare with vitamins C, E, its DPPH IC50 is support one's family 2 times and more than 3 times of vitamin-E of C.Promptly every gram Hijiki polysaccharide of this technology extraction can be removed external DPPH IC
50The time free radical 0.025g.
Claims (4)
1. the extraction process of a Sargassum fusiforme active polysaccharide is characterized in that comprising the steps:
A) the Sargassum fusiforme frond is prepared: reject invalid, fibrous root, and clean with flushing with clean water, be dried to water content≤12%, pulverize with pulverizer, cross 80 mesh sieves then and obtain dry powder;
B) dry powder that step a is obtained adds water normal temperature lixiviate 2-4hr, and wherein the mass ratio of dry powder and water is 1: 30-40; Handled 10-15 minute with 800-1200W supersonic cell crusher ice bath, centrifugal settling obtains supernatant liquor one and throw out; Described throw out water under 70-80 ℃ of temperature is carried 1-2hr, and centrifugal settling gets supernatant liquor two and throw out again;
C) throw out is carried 1-2h with 1-3% carbonic acid soda, the centrifugal supernatant liquor three that gets;
Merge supernatant liquor one, two, three, regulate its pH value to 7.0, reduced vacuum concentrates, concentrated solution Sevage method Deproteinization 2-4 time, and supernatant liquor 5-10: centrifugal after the 1 granulated active carbon absorption, get supernatant liquor, filter, collect filtrate;
D) filtrate is used 4-6: the decolouring of 1 AB-8 macroporous adsorbent resin, centrifugal again or filter after get filtrate, add the ethanol sedimentation of volume fraction 65%-70%, throw out is used 1 successively: the acetone of 5-10, absolute ethanol washing after-filtration, collecting precipitation thing;
E) the throw out reduced vacuum drying of collecting is obtained the Sargassum fusiforme active polysaccharide.
2. the extraction process of Sargassum fusiforme active polysaccharide according to claim 1 is characterized in that: the drying temperature among the step a is 50-60 ℃.
3. the extraction process of Sargassum fusiforme active polysaccharide according to claim 1 is characterized in that: described reduced vacuum is concentrated under 0.1Mpa, 50-60 ℃ the condition carries out.
4. the extraction process of Sargassum fusiforme active polysaccharide according to claim 1 is characterized in that: the centrifugation rate in the steps d is 10000 rev/mins, and centrifugation time is 10 minutes; Centrifugation rate in other step is 4000-5000 rev/min, and centrifugation time is 10-15 minute.
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