CN104001180A - Preparation method of alga natural antioxidant - Google Patents
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Abstract
The invention discloses a preparation method of an alga natural antioxidant. The preparation method is as follows: beating and crushing fresh alga, extracting with hot water, centrifuging, drying, crushing, sieving to obtain alga residue, further using a biological enzyme method for broking wall to obtain wall-broken alga residue dry powder; using food grade absolute ethanol for extraction at room temperature, collecting an extracting solution, and performing decompression concentration and vacuum drying to obtain the natural algae antioxidant product. The natural algae antioxidant product is paste, and comprises 5.2 to 13.7 by mass% of total phenol, 3.6 to 8.4 by mass% of total carotenoids, 0.5 to 1.3 by mass% of terpenoids and 0.9 to 2.6 by mass% of sterol compounds. The preparation method adopts the alga residue as a raw material to prepare the natural antioxidant with strong antioxidant activity, realizes the comprehensive utilization of resources, is in simple, efficient, green and environmentally friendly in the production process, and is suitable for industrial production; the natural algae antioxidant product is strong in antioxidant capacity, natural and green, and can replace traditional chemical synthesized antioxidants to use in food industry.
Description
Technical field
The present invention relates to the preparation technology field of the Natural antioxidant, be specifically related to a kind of preparation method of marine alga natural antioxidant.
Background technology
Research shows, oxidative stress is not only that the degenerative disease such as cancer, aging, cardiovascular disease occur, the common pathologic basis of development, is also the main cause that causes or accelerate food spoilage.Traditional antioxidants mainly contains the chemical substances such as tert-butyl hydroquinone (TBHQ), dibenzylatiooluene (BHT), Butylated hydroxyanisole (BHA).Current research discovery, there is potential harm to health in these chemical antioxidants, in European and American developed countries, has forbidden using in infant and infant foods.Therefore, the application of the alternative traditional antioxidants of the antioxidant of exploitation natural green in food industry become urgent need.
In Sargassum, there is a large amount of Natural Antioxidants.Polyphenols is the important antioxidant in Sargassum, its content can account for the 1-14% of Sargassum dry weight, and seeweed polyphenol and terrestrial plant polyphenol structurally have relatively big difference, terrestrial plant polyphenol is mainly the derivant of gallic acid and ellagic acid, seeweed polyphenol is the polymer (Holdt & Kraan, 2011) of phloroglucinol.Studies show that, brown algae polyphenols is being had outstanding performance aspect the multiple biological activitys such as antioxidation.The relatively discovery of the antioxidation evaluation methodology by chemical system and cell system, the antioxidant activity of brown algae polyphenols will be much better than vitamin C and vitamin E (Yotsu-Yamashita
et al., 2013).Fucoxanthin (fucoxanthin) is another critical function active substance in Sargassum, and content can account for the more than 0.5% of Sargassum dry weight.Fucoxanthin also claims Brown algae flavin, fucoxanthine, belongs to phylloxanthin class, and its content accounts for the more than 60% of tang carotene total amount (Yan little Jun etc., 2001).Chen Wenjia etc. (2012) measure the removing ability of hydrogen peroxide, hydroxyl radical free radical, DPPH free radical by fucoxanthin, prove that its non-oxidizability is apparently higher than vitamin C; Ha etc. (2011) study discovery, and fucoxanthin can effectively improve rat plasma glutathion peroxidase and catalase contains activity, reduce the oxidative stress of high fat diet induction.In addition, studies show that, the lipid in Sargassum, terpenoid, halogenide, sulfur-containing compound etc. also have good anti-oxidation function.Therefore, utilize Sargassum to extract Natural Antioxidants and there is huge exploitation value.
Yet, to the exploitation of Sargassum, be mainly Sargassum polysaccharides at present, the antioxidant such as seeweed polyphenol, fucoxanthin are but processed as cheap feedstuff or bio-feritlizer along with Sargassum slag, have caused the significant wastage of resource.Utilize and in Sargassum polysaccharides processing byproduct-Sargassum slag, obtain the Natural antioxidant and still belong to blank.Therefore, from Sargassum slag, extract Natural Antioxidants, there is very important economic worth and realistic meaning.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of extracting Natural Antioxidants from processing byproduct-Sargassum slag of extraction Sargassum polysaccharides.
For achieving the above object, the invention provides a kind of preparation method of marine alga natural antioxidant, it is characterized in that, comprise the steps,
The preparation of Sargassum slag: the making beating of fresh Sargassum is to without obvious bulk thing, 1:5 adds after deionized water 80 ℃ to extract 1 h by volume, centrifugal, preferred 3000 r/min, 5 min, taking precipitate, obtains Sargassum slag;
Optional, can also comprise and continue to add deionized water in Sargassum slag, repeat abovementioned steps, collect Sargassum slag;
The preparation of Sargassum slag dry powder: Sargassum slag drying, pulverize, sieve and make Sargassum slag dry powder;
The biological enzymolysis breaking cellular wall of Sargassum slag dry powder: add deionized water after described Sargassum slag dry powder and compound enzymic preparation are mixed, enzymolysis certain hour, centrifugal under room temperature, preferred 3000 r/min, 5 min, collecting precipitation thing, in baking oven, 80 ℃ of dry 6 h, obtain the Sargassum slag dry powder after breaking cellular wall;
Organic solvent lixiviate: add dehydrated alcohol in the Sargassum slag dry powder after described breaking cellular wall, under room temperature, lixiviate certain hour, centrifugal, preferred 3000 r/min, 5 min, collect respectively supernatant and Sargassum precipitate;
Optional, can also comprise and continue to add dehydrated alcohol in Sargassum precipitate, repeat abovementioned steps, collect respectively supernatant and Sargassum precipitate;
The acquisition of marine alga natural antioxidant: described supernatant is evaporated to dry, obtains marine alga natural antioxidant product through vacuum drying.
Described Sargassum is at least one in Thallus Laminariae (Thallus Eckloniae), Thallus Porphyrae, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch.
Described Sargassum slag is that Sargassum is extracted the residue after polysaccharide.
In the preparation of described Sargassum slag dry powder, 80-100 order sieves.
In the biological enzymolysis breaking cellular wall of described Sargassum slag dry powder, the weight ratio of described seaweed meal and compound enzymic preparation is 1000:5; Described compound enzymic preparation is comprised of pectase, cellulase, and the proportional w/w of preferred group is 1:1;
Optional, the weight/volume that the ratio of seaweed meal and compound enzymic preparation summation and deionized water is 1:15-25;
Optional, add after deionized water enzymolysis 5-8 hour under room temperature.
In described organic solvent lixiviate, add food-grade anhydrous ethanol, under room temperature, the centrifugal precipitation of removing after lixiviate, gets supernatant;
Optional, the weight/volume that the Sargassum slag dry powder after breaking cellular wall and the ratio of food-grade anhydrous ethanol are 1:5-10;
Optional, add after food-grade anhydrous ethanol lixiviate 1-3 hour under room temperature.
In the acquisition of described marine alga natural antioxidant, supernatant is evaporated to dry, through vacuum drying, obtains marine alga natural antioxidant product;
Optional, the temperature of described supernatant concentrating under reduced pressure is 40-60 ℃;
Optional, described vacuum drying temperature is 50-80 ℃;
Optional, the described vacuum drying time is 2-5 hour.
The marine alga natural antioxidative active extractive that described method obtains is paste, and the total phenols containing is 5.2-13.7 quality %, and total carotinoid is 3.6-8.4 quality %, and terpenoid is 0.5-1.3 quality %, and phytosterin compound is 0.9-2.6 quality %.
The present invention also protects the Sargassum antioxidant of either method gained.
Method of the present invention is simple to operate, environment-friendly high-efficiency, be suitable for large-scale production, has fully realized the comprehensive development and utilization of marine algae resource, has improved economic benefit.
The present invention adopts the Sargassum slag extracting after polysaccharide to extract marine alga natural antioxidant, most of glycocalix extraction due to alginic cell wall, the thickness reduction of cell wall, make enzyme shell-broken effect better, cell includes active substance and is able to abundant release, and the extraction ratio of anti-oxidant ingredients is at 4 quality %-6.7 quality %.The main component that Sargassum antioxidant extract now contains has seeweed polyphenol, carotenoid, terpenoid, sterols material.The anti-oxidation active substance obtaining according to method of the present invention and patent application: a kind of enzyme process is prepared the method (application number: the 201310142757.X applying date: 2013-04-24 of Brown algae fucosan and fucoxanthin, open day: 2013-08-07) compare, the active component obtaining is more comprehensive, thereby has realized the maximum using of resource.
It is the Natural antioxidant that raw material is prepared strong anti-oxidation activity that the present invention adopts by-product-Sargassum slag of Sargassum processing enterprise processing Sargassum polysaccharides, realizes the comprehensive utilization of resource; Production technology is simple, efficient, and environmental protection, is suitable for suitability for industrialized production; The oxidation resistance of product is strong, and green natural can replace the use of the synthetic antioxidant of traditional chemical in food industry.
The Brown algae cell wall that this invention is degraded hard by enzyme wall breaking technology, makes to be present in the abundant strippings of antioxidant content such as intracellular seeweed polyphenol, fucoxanthin, has realized the quick preparation of marine alga natural antioxidant.
Advantage of the present invention and beneficial effect are:
1, China's marine algae resource is abundant, after Sargassum processing enterprise extracts the polysaccharide in Sargassum, Sargassum slag is processed as cheap feedstuff or bio-feritlizer, the antioxidant such as the polyphenol in Sargassum, fucoxanthin do not obtain processing and utilization well, cause the significant wastage of resource, therefore Natural Antioxidants in Sargassum slag is carried out to industrialization development utilization, there is significant economic benefit and social benefit;
2, simple (Fig. 1 is shown in technological process), environment-friendly high-efficiency of technique of the present invention, be suitable for scale operation, can be applicable to suitability for industrialized production.
Embodiments of the invention, provide and have utilized the Sargassums such as Thallus Laminariae (Thallus Eckloniae) to prepare the method for the Natural antioxidant.In embodiment 1,100 g kelp residue dry powder, finally obtain 6.5 g antioxidant extracts; In embodiment 2,100 g Thallus Porphyrae slag dry powder, finally obtain 6.7 g antioxidant extracts; In embodiment 3,100 g Thallus Laminariae slag dry powder, finally obtain 5.2 g antioxidant extracts; In embodiment 4,100 g Sargassum fusiforme (Harv.) Setch slag dry powder, finally obtain 4.9 g antioxidant extracts.
When Sargassum being adopted to cellulase treatment breaking cellular wall by step of the present invention, make the fully stripping of antioxidant content in alginic cell, thereby significantly improve extraction efficiency.Experiment be take kelp residue dry powder as raw material, the shell-broken effect of the compound formulation of pectase, cellulase and composition thereof has been carried out to comparative study, Kelp Powder (80 order) is fixed as 1000:5 (w/w) with the ratio of enzyme, under room temperature, be hydrolyzed 5 h, the best proportioning that judges enzyme with the yield of ethanol extraction, result is as shown in table 1.Experimental result discovery, after using enzyme broken wall treatment, the yield of ethanol extraction all obviously improves, and uses separately pectase, cellulase neither as uses the shell-broken effect of these two kinds of enzymes simultaneously.Meanwhile, test finds, the proportioning of pectase and cellulase is 1:1(w/w) time shell-broken effect and other proportioning there were significant differences statistically.Simultaneously, by changing the ratio of pectase and cellulase, find the shell-broken effect of the compound enzyme of various proportionings, no matter be the proportion that improves pectase or cellulase, effect does not all have pectase and cellulase by 1:1(w/w) shell-broken effect during proportioning is good, so pectase and cellulase optimal proportion are 1:1(w/w).
The affect table of table 1 enzyme breaking cellular wall on kelp residue ethanol extraction yield
2, through analysis of experiments, find, the anti-oxidation active substance content of the marine alga natural antioxidant that the present invention obtains is high, is applicable to the raw material of the industries such as food antioxidant and functional food and medicine.The component analysis of Sargassum antioxidant extract is as table 2:
The component analysis of the different Sargassum antioxidant extracts of table 2
3, the marine alga natural antioxidant that the present invention obtains there is the feature that antioxidant activity is high, the effectively free radical (as Fig. 2) in scavenger cell.As can be seen from Figure 2, the marine alga natural antioxidant that the present invention obtains to the maximal clearance of free radical in cell at 42%-54%, the maximal clearance of vitamin E is 35%, illustrate that marine alga natural antioxidative cell antioxidant activity is better than vitamin E, can be used for the exploitation of food antioxidant and anti-oxidation functional food.
Accompanying drawing explanation
Fig. 1 is process chart of the present invention.
Fig. 2 is the effective removing figure of the marine alga natural antioxidant that obtains of the present invention to free radical in cell.
The specific embodiment
Describe embodiments of the invention below in detail, the example of described embodiment is shown in the drawings, and wherein same or similar label represents same or similar element or has the element of identical or similar functions from start to finish.Below by the embodiment being described with reference to the drawings, be exemplary, be intended to for explaining the present invention, and can not be interpreted as limitation of the present invention.Unreceipted concrete technology or condition person in embodiment, according to the described technology of the document in this area or condition or carry out according to product description.The unreceipted person of production firm of agents useful for same or instrument, being can be by the conventional products of commercial acquisition.
Following examples compound enzyme used is 1:1(w/w) pectase and cellulase compound enzyme.
embodiment 1:thallus Laminariae (Thallus Eckloniae) is prepared marine alga natural antioxidant
(1) get fresh Thallus Laminariae (Thallus Eckloniae) 500 g, shears shreds and with beater making beating, adds 2.5 L deionized waters afterwards, stirs;
80 ℃ are extracted 1 h, adopt tubular type continuous centrifuge centrifugal (3000 r/min, 5 min), and taking precipitate obtains Sargassum slag 356.9 g; Sargassum slag drying, pulverizing, cross 80 mesh sieves and make Sargassum slag dry powder 75.8 g;
(2) get above-mentioned kelp residue dry powder 50 g and 0.25 g compound enzyme and fully mix, add deionized water 754 mL, stir.Under 20 ℃ of room temperatures, react after 5 h, adopt tubular type continuous centrifuge centrifugal (per minute 3000 turn, 5 minutes), get kelp residue.This step adopts cellulase treatment breaking cellular wall to Sargassum slag, makes the abundant strippings of polyphenoils mass-energy such as polyphenol in Sargassum, fucoxanthin, thereby significantly improves extraction efficiency.Extraction ratio by ethanol extraction judges that enzyme is to the execution of alginic cell wall (in Table 1).As can be seen from Table 1, the extraction ratio that enzyme is processed front ethanol extraction is 1.52 ± 0.36 g/100 g, when being undertaken after enzyme processing by step 2 pair Sargassum slag, the extraction ratio of ethanol extraction significantly improves, when the proportioning of pectase and cellulase is 1:1 (w/w), the extraction rate reached of ethanol extraction is to 6.53 ± 0.13 g/100 g, and effect is very remarkable;
(3) above-mentioned kelp residue is put into baking oven dry 6 h at 80 ℃, obtains Sargassum slag dry powder 45 g after breaking cellular wall;
(4) in the Sargassum slag dry powder after described breaking cellular wall, add the food-grade anhydrous ethanol of 225 mL, lixiviate 2 h under 20 ℃ of room temperatures, centrifugal (per minute 3000 turn, 5 minutes), collects respectively supernatant and Sargassum precipitate;
(5) repeating step 4, the supernatant that combining step 4 and step 5 obtain;
(6) above-mentioned supernatant is evaporated to dryly at 60 ℃, then adopts vacuum drying in 70 ℃ of dry 3 h, obtains marine alga natural antioxidant product 3.29 g.
embodiment 2:sargassum fusiforme (Harv.) Setch is prepared marine alga natural antioxidant
(1) get Sargassum fusiforme (Harv.) Setch 500 g, shears shreds and with beater making beating, adds 2.5 L deionized waters afterwards, stirs.
80 ℃ are extracted 1 h, adopt tubular type continuous centrifuge centrifugal (3000 r/min, 5 min), and taking precipitate obtains Sargassum slag 385.7 g; Sargassum slag drying, pulverizing, cross 80 mesh sieves and make Sargassum slag dry powder 81.4 g;
(2) get above-mentioned Sargassum slag dry powder 50 g and 0.25 g compound enzyme fully mixes, add deionized water 1005 mL, stir.Under 20 ℃ of room temperatures, react after 5 h, adopt tubular type continuous centrifuge centrifugal (per minute 3000 turn, 5 minutes), get Sargassum slag;
(3) above-mentioned Sargassum slag is put into baking oven dry 6 h at 80 ℃, obtains Sargassum slag dry powder 46 g after breaking cellular wall;
(4) in the Sargassum slag dry powder after described breaking cellular wall, add the food-grade anhydrous ethanol of 368 mL, lixiviate 2 h under 20 ℃ of room temperatures, centrifugal (per minute 3000 turn, 5 minutes), collects respectively supernatant and Sargassum precipitate;
(5) repeating step 4, the supernatant that combining step 4 and step 5 obtain;
(6) above-mentioned supernatant is evaporated to dryly at 60 ℃, then adopts vacuum drying in 70 ℃ of dry 3 h, obtains marine alga natural antioxidant product 2.47 g.
embodiment 3:thallus Laminariae is prepared marine alga natural antioxidant
(1) get fresh Thallus Laminariae 500 g, shears shreds and with beater making beating, adds 2.5 L deionized waters afterwards, stirs;
80 ℃ are extracted 1 h, adopt tubular type continuous centrifuge centrifugal (3000 r/min, 5 min), and taking precipitate obtains Sargassum slag 322.5 g; Sargassum slag drying, pulverizing, cross 80 mesh sieves and make Sargassum slag dry powder 65.2 g;
(2) get above-mentioned Sargassum slag dry powder 50 g and 0.25 g compound enzyme fully mixes, add deionized water 1256 mL, stir.Under 20 ℃ of room temperatures, react after 5 h, adopt tubular type continuous centrifuge centrifugal (per minute 3000 turn, 5 minutes), get Sargassum slag;
(3) above-mentioned Sargassum slag is put into baking oven dry 6 h at 80 ℃, obtains Sargassum slag dry powder 43 g after breaking cellular wall;
(4) in the Sargassum slag dry powder after described breaking cellular wall, add the food-grade anhydrous ethanol of 430 mL, lixiviate 2 h under 20 ℃ of room temperatures, centrifugal (per minute 3000 turn, 5 minutes), collects respectively supernatant and Sargassum precipitate;
(5) repeating step 4, the supernatant that combining step 4 and step 5 obtain;
(6) above-mentioned supernatant is evaporated to dryly at 60 ℃, then adopts vacuum drying in 70 ℃ of dry 3 h, obtains marine alga natural antioxidant product 2.66 g.
embodiment 4:thallus Porphyrae is prepared marine alga natural antioxidant
(1) get fresh laver 500 g, shears shreds and with beater making beating, adds 2.5 L deionized waters afterwards, stirs;
80 ℃ are extracted 1 h, adopt tubular type continuous centrifuge centrifugal (3000 r/min, 5 min), and taking precipitate obtains Sargassum slag 349.7 g; Sargassum slag drying, pulverizing, cross 80 mesh sieves and make Sargassum slag dry powder 70.6 g;
(2) get above-mentioned Sargassum slag dry powder 50 g and 0.25 g compound enzyme fully mixes, add deionized water 1005 mL, stir.Under 20 ℃ of room temperatures, react after 5 h, adopt tubular type continuous centrifuge centrifugal (per minute 3000 turn, 5 minutes), get Sargassum slag;
(3) above-mentioned Sargassum slag is put into baking oven dry 6 h at 80 ℃, obtains Sargassum slag dry powder 47 g after breaking cellular wall;
(4) in the Sargassum slag dry powder after described breaking cellular wall, add the food-grade anhydrous ethanol of 376 mL, lixiviate 2 h under 20 ℃ of room temperatures, centrifugal (per minute 3000 turn, 5 minutes), collects respectively supernatant and Sargassum precipitate;
(5) repeating step 4, the supernatant that combining step 4 and step 5 obtain;
(6) above-mentioned supernatant is evaporated to dryly at 60 ℃, then adopts vacuum drying in 70 ℃ of dry 3 h, obtains marine alga natural antioxidant product 3.38 g.
Although illustrated and described embodiments of the invention above, be understandable that, above-described embodiment is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change above-described embodiment within the scope of the invention in the situation that not departing from principle of the present invention and aim, modification, replacement and modification.
Claims (9)
1. a preparation method for marine alga natural antioxidant, is characterized in that, comprise the steps,
The preparation of Sargassum slag: the making beating of fresh Sargassum is to without obvious bulk thing, 1:5 adds after deionized water 80 ℃ to extract 1 h by volume, centrifugal, preferred 3000 r/min, 5 min, taking precipitate, obtains Sargassum slag;
Optional, can also comprise and continue to add deionized water in Sargassum slag, repeat abovementioned steps, collect Sargassum slag;
The preparation of Sargassum slag dry powder: Sargassum slag drying, pulverize, sieve and make Sargassum slag dry powder;
The biological enzymolysis breaking cellular wall of Sargassum slag dry powder: add deionized water after described Sargassum slag dry powder and compound enzymic preparation are mixed, enzymolysis certain hour, centrifugal under room temperature, preferred 3000 r/min, 5 min, collecting precipitation thing, in baking oven, 80 ℃ of dry 6 h, obtain the Sargassum slag dry powder after breaking cellular wall;
Organic solvent lixiviate: add dehydrated alcohol in the Sargassum slag dry powder after described breaking cellular wall, under room temperature, lixiviate certain hour, centrifugal, preferred 3000 r/min, 5 min, collect respectively supernatant and Sargassum precipitate;
Optional, can also comprise and continue to add dehydrated alcohol in Sargassum precipitate, repeat abovementioned steps, collect respectively supernatant and Sargassum precipitate;
The acquisition of marine alga natural antioxidant: described supernatant is evaporated to dry, obtains marine alga natural antioxidant product through vacuum drying.
2. the preparation method of a kind of marine alga natural antioxidant of claim 1, is characterized in that: described Sargassum is at least one in Thallus Laminariae (Thallus Eckloniae), Thallus Porphyrae, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch.
3. the preparation method of a kind of marine alga natural antioxidant of claim 1, is characterized in that: described Sargassum slag is that Sargassum is extracted the residue after polysaccharide.
4. the preparation method of a kind of marine alga natural antioxidant of claim 1, is characterized in that: in the preparation of described Sargassum slag dry powder, 80-100 order sieves.
5. the preparation method of a kind of marine alga natural antioxidant of claim 1, is characterized in that: in the biological enzymolysis breaking cellular wall of described Sargassum slag dry powder, the weight ratio of described seaweed meal and compound enzymic preparation is 1000:5; Described compound enzymic preparation is comprised of pectase, cellulase, and the proportional w/w of preferred group is 1:1;
Optional, the weight/volume that the ratio of seaweed meal and compound enzymic preparation summation and deionized water is 1:15-25;
Optional, add after deionized water enzymolysis 5-8 hour under room temperature.
6. the preparation method of a kind of marine alga natural antioxidant of claim 1, is characterized in that: in described organic solvent lixiviate, add food-grade anhydrous ethanol, under room temperature, the centrifugal precipitation of removing after lixiviate, gets supernatant;
Optional, the weight/volume that the Sargassum slag dry powder after breaking cellular wall and the ratio of food-grade anhydrous ethanol are 1:5-10;
Optional, add after food-grade anhydrous ethanol lixiviate 1-3 hour under room temperature.
7. the preparation method of a kind of marine alga natural antioxidant of claim 1, is characterized in that: in the acquisition of described marine alga natural antioxidant, supernatant is evaporated to dry, through vacuum drying, obtains marine alga natural antioxidant product;
Optional, the temperature of described supernatant concentrating under reduced pressure is 40-60 ℃;
Optional, described vacuum drying temperature is 50-80 ℃;
Optional, the described vacuum drying time is 2-5 hour.
8. marine alga natural antioxidant product according to claim 7, it is characterized in that: this marine alga natural antioxidative active extractive is paste, the total phenols containing is 5.2-13.7 quality %, total carotinoid is 3.6-8.4 quality %, terpenoid is 0.5-1.3 quality %, and phytosterin compound is 0.9-2.6 quality %.
9. the Sargassum antioxidant of claim 1-8 either method gained.
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