CN102336725A - Method for extracting fucoxanthine-containing concentrate, product obtained through method, and application of product - Google Patents
Method for extracting fucoxanthine-containing concentrate, product obtained through method, and application of product Download PDFInfo
- Publication number
- CN102336725A CN102336725A CN201110155266XA CN201110155266A CN102336725A CN 102336725 A CN102336725 A CN 102336725A CN 201110155266X A CN201110155266X A CN 201110155266XA CN 201110155266 A CN201110155266 A CN 201110155266A CN 102336725 A CN102336725 A CN 102336725A
- Authority
- CN
- China
- Prior art keywords
- fucoxanthine
- enzyme
- solution
- extracting
- temperature
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention relates to a method for extracting a fucoxanthine-containing concentrate from algae, the fucoxanthine-containing concentrate obtained through the method, and application of the fucoxanthine-containing concentrate. The method comprises the following steps of: pretreating raw materials; performing enzyme treatment; extracting; separating; concentrating and the like. The method overcomes technical defects of low efficiency of extracting fucoxanthine and high consumption of an organic solvent in the prior art, and a fucoxanthine product with high active ingredients is obtained, so the fucoxanthine product has a good application prospect. The raw materials are readily available, the extraction process is simple, and the obtained product has stable content and controllable comprehensive cost, and is suitable for industrial production and lengthening an industrial chain.
Description
[technical field]
The invention belongs to from the technical field of extracted form natural plant effective constituent.More specifically, the present invention relates to a kind ofly contain the method for the enriched material of fucoxanthine, also relate to the enriched material that contains fucoxanthine that adopts aforesaid method to obtain, also relate to the said purposes that contains the enriched material of fucoxanthine from extracting algae.
[background technology]
The marine alga that in the ocean, grows; Particularly biological (comprising sea-tangle (Laminaria japonica), Sargassum fusiforme (Sargassum fusiforme), sargassun (Sargassum), bulk kelp (Macrocystis pyrifera (L.) Ag), wakame (Undaria pinnatifida), Thallus Gracilariae (Gracilaria sp) etc.) body of phaeophyta (Phaeophyta) contains abundant carotinoid compounds; Wherein the fucoxanthine ratio is the highest, is that comparatively ideal extracts the natural resources of fucoxanthine.
Fucoxanthine also is pheophytin, and English name is Fucoxanthin, molecular formula C
42H
58O
6, be a kind of rare oxygen carrotenoid that contains that is present in the phaeophyta biology, half systematic naming method is 3 '-ethanoyl-6 ', 7 '-dideoxy-5,6-epoxy group(ing)-5,5 ', 6,6 ', 7,8-six hydrogen-3,5 '-dihydroxyl-8-carbonyl-β, β-Hu Luobusu.A kind of as xenthophylls, its molecule is made up of a poly alkene chain and two cyclic end groups, in solution He under the unbound state, is revealed as tawny; Fucoxanthine has a plurality of pairs of keys; Have geometrical isomer at occurring in nature, and be main with alltrans, its structural formula is following.
At present, some about the report of fucoxanthine process for extracting.A kind of method of from brown alga, extracting fucoxanthine is disclosed like patent CN100999508A; This method is raw material with the brown alga; Through 60%～100% extraction using alcohol, concentrate, filtering-depositing, macroporous adsorbent resin column chromatography, spraying drying make product; Measure through the HPLC method, the weight content of fucoxanthine is 1%～20% in this product.But present method exists extraction time long, complex process, defective such as it is lower to extract yield, and impurity is many.If can before extracting, carry out broken wall treatment to raw material, promptly help the release of title product, can shorten extraction time greatly again.
Patent CN1706836A discloses a kind of method of from marine alga, separating fucoxanthine, this method be with marine alga with DMSO 99.8MIN. lixiviate in the dark, use ETHYLE ACETATE and (NH then
4)
2SO
4Solution is extracted into the fucoxanthine in the vat liquor in the ETHYLE ACETATE, obtains the fucoxanthine of higher degree at last through 3～4 silica gel column chromatographies.This method and traditional method reduced in comparison extraction time, the yield of fucoxanthine has also improved, but DMSO 99.8MIN. uses as extraction solvent; When carrying out liquid-liquid extraction with ETHYLE ACETATE and ammonium sulfate; Have in mutually residually at ETHYLE ACETATE, and DMSO 99.8MIN. has high boiling point, can not remove through low-temperature reduced-pressure distillatory method; Make that sample can't be dry, and its existence can influence fucoxanthine purifying afterwards; And in this patented claim; Need carry out the silicagel column purifying repeatedly to obtain high-load fucoxanthine product to the enriched material that obtains; A large amount of adsorption losses of target compound have promptly been caused; Increased the complicacy of technology again,, then helped to simplify production technique if can in extraction process, once remove partial impurities.
But, because brown alga belongs to the special physiological structure of marine alga, there are problems such as extraction efficiency is not high, consumption of organic solvent is big from wherein extracting fucoxanthine always, promptly caused the waste of resource, again because of organic solvent residue, influenced the quality of product greatly.In view of above-mentioned these defectives, can't satisfy demand of practical production.Therefore, be necessary to invent a kind of being directed against from marine alga, the particularly technical skill of the biological preparation of phaeophyta fucoxanthine, the inventor has accomplished the present invention finally through lot of experiments.This technology comprises that the pre-treatment, enzyme of raw material content such as handle, extract, separate, concentrate.
[summary of the invention]
[technical problem that will solve]
The purpose of this invention is to provide a kind of method that contains the fucoxanthine enriched material from extracting algae.
Another object of the present invention provides a kind of enriched material that contains fucoxanthine.
Another object of the present invention provides a kind of purposes that contains the enriched material of fucoxanthine.
[technical scheme]
The present invention realizes through following technical proposals.
The present invention relates to a kind of method that contains the fucoxanthine enriched material from extracting algae.The step of this method is following:
A, raw materials pretreatment
The impurity and the salinity of water flush away marine alga remained on surface with the marine alga oven dry, are ground into seaweed powder again under the condition of 30～50 ℃ of temperature and lucifuge;
B, enzyme are handled
(1) gets the water of 800～1400 weight parts; And water temperature transferred to 20～50 ℃; Add 0.5～2.0 weight part cellulase and 0.1～0.4 weight part polygalacturonase then; Again with mineral acid or mineral alkali pH regulator to 4～7, at 20～50 ℃ of following activation 20～60min of temperature, so obtain a kind of enzyme treatment solution with its solution;
(2) be sprayed onto 50～100 weight parts to the said enzyme treatment solution of 30～500 weight parts by steps A) in the seaweed powder that obtains, stir again, compacting, sealing, at 20～50 ℃ of following lucifuges insulation 4～96h of temperature, obtain a kind of enzyme and handle seaweed powder;
C, extraction
Taking by weighing 80～120 weight parts by step B) enzyme that obtains handles seaweed powder; Under 2～50 ℃ of conditions of temperature, use 300～800 weight parts, 40～100% hydrophilic organic solvents to carry out refluxing extraction 30～90min; Separation and Extraction liquid then; Leftover materials repeat to extract 3～6 times, merge these extracting solutions;
D, from extracting solution, separate fucoxanthine
With step C) extracting solution that obtains and normal hexane be according to 6: 1～3: 1 mixed of volume ratio, behind the concuss, standing separation 25～35min; Separate lower floor's solution; The residue upper solution repeats this procedure, is colourless mutually until the upper strata normal hexane, merges these lower floor's solution;
Concentrating of E, fucoxanthine component
Pressure-0.06～-condition of 40～55 ℃ of 0.095MPa and temperature under, with step D) lower floor's solution concentration of obtaining is to doing, and obtains containing the enriched material of fucoxanthine.
A preferred embodiment of the invention, described marine alga is selected from sea-tangle, Sargassum fusiforme, sargassun, bulk kelp, wakame or Thallus Gracilariae.
According to another kind of preferred implementation of the present invention, the granularity of said seaweed powder is 20～100 orders.
According to another kind of preferred implementation of the present invention, at step B) in the mineral acid that uses be selected from hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid; Mineral alkali is selected from sodium hydroxide, Pottasium Hydroxide, yellow soda ash, salt of wormwood, sodium hydrogencarbonate or saleratus.
According to another kind of preferred implementation of the present invention, at step B) described in the enzyme work >=4000IU/g of cellulase, the enzyme work >=5000IU/g of described polygalacturonase.
According to another kind of preferred implementation of the present invention, at step C) described in hydrophilic organic solvent be selected from ethanol, methyl alcohol or acetone.
The invention still further relates to the enriched material that adopts said method to obtain, it is characterized in that it contains in the fucoxanthine of enriched material gross weight more than 5%.
The invention still further relates to the purposes of said enriched material in preparation foodstuff additive, functional foodstuff, medicine and fodder additives.
Below the present invention will be described in more detail.
The present invention is during from the extracting algae fucoxanthine, with cellulase and polygalacturonase the marine alga of pulverizing carried out pre-treatment earlier, destroys its cell wall structure; Help the release of activeconstituents, use the water-miscible organic solvent lixiviate again 3～6 times, extracting solution is through liquid-liquid extraction; Remove the impurity such as chlorophyll that exist in the extracting solution, raffinate can obtain having the fucoxanthine condensed cream of certain content through underpressure distillation; This method extraction time is short, and technology is simple, and extraction efficiency is high; The product organic solvent-free that obtains is residual, and comprehensive cost is cheap, for suitability for industrialized production provides the production foundation.
The present invention relates to a kind of method that contains the fucoxanthine enriched material from extracting algae.The step of this method is following:
A, raw materials pretreatment
The impurity and the salinity of water flush away marine alga remained on surface with the marine alga oven dry, are ground into seaweed powder again under the condition of 30～50 ℃ of temperature and lucifuge;
In the present invention, described marine alga is selected from phaeophytas biologies such as sea-tangle, Sargassum fusiforme, sargassun, bulk kelp, wakame or Thallus Gracilariae;
Described marine alga all need be desalted, the pre-treatment of impurity and pulverizing.
In the present invention, because fucoxanthine has the extremely unsettled characteristics of light and heat, therefore, need the marine alga of cleaning is dried under lucifuge and cryogenic situation.
In the present invention, when drying, can use that normally used drying plant carries out drying in the Chinese medicinal materials processing technology, for example the GZX-9146MBE type digital display air dry oven produced of medical apparatus and instruments factory of Shanghai Boxun Industrial Co., Ltd. carries out drying.
In the present invention, described pulverizing is to use in the Chinese medicinal materials processing technology normally used disintegrating apparatus that marine alga is carried out high speed shear and pulverizes, and is crushed to 20～100 orders.
Among the present invention, employed kibbler can be the normally used kibbler of selling in the market in the Chinese medicinal materials processing technology, for example the food masher produced of the new friendly machinofacture in Jiangyin City ltd.Typically use kibbler raw material pulverizing is arrived certain particle size, sieve with the standard medicine then, collect 20～100 purpose seaweed powders.
B, enzyme are handled
(1) gets 800～1400 weight parts waters and water temperature transferred to 20～50 ℃; Add 0.5～2.0 weight part cellulase and 0.1～0.4 weight part polygalacturonase then; Again with mineral acid or mineral alkali pH regulator to 4～7 with its solution; At 20～50 ℃ of following activation 20～60min of temperature, so obtain a kind of enzyme treatment solution.
Cellulase (EC 3.2.1.4) is a kind of important enzyme product, is a kind of prozyme, mainly is made up of circumscribed beta-glucanase, inscribe beta-glucanase and beta-glucosidase etc.Usually, cellulase is divided into three types: C1 enzyme, Cx enzyme and beta-glucosidase enzyme.The C1 enzyme is at first to the acting enzyme of Mierocrystalline cellulose, destroys the crystalline texture of cellulose chain.The Cx enzyme is Mierocrystalline cellulose, the decomposition β-1 that acts on through the C1 enzyme activation, the cellulase of 4-glycosidic link.Beta-glucosidase enzyme can be decomposed into glucose with cellobiose, procellose and other low molecule cellodextrins.
In the present invention, described cellulase for example is Pangbo Bioengineering Co Ltd, Nanning's production of cellulose enzyme.
According to the present invention, if the enzyme work of said cellulase is lower than 4000IU/g, then can be not high because of enzyme activity; And need to use a large amount of cellulases to guarantee that the raw material cell wall broken wall is abundant; Therefore the enzyme work of said cellulase should be higher than 4000IU/g, lives in view of the enzyme of the cellulase of selling on the market, and the enzyme work of said cellulase is 4000～10000IU/g; Preferably 5000～9000IU/g, more preferably 6000～8000IU/g.
Wherein, enzyme work is defined as 1g enzyme powder or 1mL enzyme liquid under 40 ℃ of conditions with pH4.6, and the enzyme amount that 1h decomposing soluble starch produces 1mg glucose is 1 enzyme unit that lives.
Described polygalacturonase is a multienzyme complex of decompose pectin, generally includes protopectinase, pectinesterase lytic enzyme, pectinesterase.Combined action through them makes pectin substance be able to decompose fully.Natural pectin substance changes into the pectin of water dissolvable under the protopectinase effect; Pectin is generated pectic acid by pectin methyl esters lytic enzyme catalytic elimination methyl esters group; Pectic acid generates galacturonic acid through pectic acid hydrolase and the degraded of pectate lyase class.
In the present invention, described polygalacturonase for example is the polygalacturonase that Pangbo Bioengineering Co Ltd, Nanning produces.
According to the present invention; If the enzyme work of said polygalacturonase is lower than 5000IU/g; Then can be not high because of enzyme activity; And need to use a large amount of polygalacturonases to guarantee that the whole hydrolysis of the pectin that raw material is contained are better discharged to guarantee target compound, therefore, the enzyme work of said polygalacturonase should be higher than 5000IU/g; Seeing that the enzyme of the polygalacturonase of selling is lived on the market, therefore the enzyme work of said polygalacturonase is 5000～12000IU/g, preferably 6000～10000IU/g, more preferably 8000～9000IU/g.
Preferably, with 800～1400 weight parts waters dissolving 0.8～1.8 weight part cellulase and 0.15～0.35 weight part polygalacturonase; Preferred, with 800～1400 weight parts waters dissolving 1.0～1.6 weight part cellulases and 0.2～0.3 weight part polygalacturonase.
In this step, the mineral acid of use is selected from hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid, preferably is selected from hydrochloric acid, sulfuric acid or phosphoric acid; More preferably be selected from hydrochloric acid or phosphoric acid.
In this step; The mineral alkali that uses is selected from sodium hydroxide, Pottasium Hydroxide, yellow soda ash, salt of wormwood, sodium hydrogencarbonate or saleratus; Preferably be selected from sodium hydroxide, Pottasium Hydroxide, yellow soda ash, sodium hydrogencarbonate or saleratus, more preferably be selected from sodium hydroxide, yellow soda ash, sodium hydrogencarbonate or saleratus.
In this step; Cellulase and polygalacturonase are enzyme common in the plant materials, and it can guarantee that the pH scope of enzyme activity is 4～7, in order in reaction process, to guarantee the maximum vigor of enzyme; The pH of reaction solution can not be too high; Can not be low excessively, therefore, need be with mineral acid or mineral alkali pH regulator to 4～7 with its enzyme solution.Too high or too low pH can make enzyme suffer irreversible breaking because of the stability that influences enzyme, thereby influences reaction effect.
(2) be sprayed onto 50～100 weight parts to the said enzyme treatment solution of 30～500 weight parts by steps A) in the seaweed powder that obtains, stir again, compacting, sealing, at 20～50 ℃ of following lucifuges insulation 4～96h of temperature, obtain a kind of enzyme and handle seaweed powder.
Among the present invention; Use common mechanical stirrer or whipping device with said enzyme treatment solution with by steps A) in the seaweed powder mixing that obtains; Use compact machine equipment that the density of mixing mixture is improved then, seal with plastic material again, so that can reach the lucifuge effect.In the present invention, employed heat-preserving equipment can be the thermostat water bath that in experimentation, often uses, like the DZKW-C type electronic thermostatic water-bath of Beijing Bo Lian medical apparatus corporation, Ltd production.
In this step, preferably use said enzyme treatment solution of 50～300 weight parts and 50～100 weight parts by steps A) seaweed powder that obtains.More preferably use said enzyme treatment solution of 75～200 weight parts and 50～100 weight parts by steps A) seaweed powder that obtains.
In this step, preferably, the mixture of said enzyme treatment solution and seaweed powder is at 25～45 ℃ of following lucifuge insulation 10～85h of temperature.More preferably, the mixture of said enzyme treatment solution and seaweed powder is at 30～40 ℃ of following lucifuge insulation 20～75h of temperature.
C, extraction
Taking by weighing 80～120 weight parts by step B) enzyme that obtains handles seaweed powder; Under 2～50 ℃ of conditions of temperature, use the hydrophilic organic solvent of 300～800 weight parts 40～100% to carry out refluxing extraction 30～90min; Separation and Extraction liquid then, leftover materials repeat to extract 3～6 times, united extraction liquid.
Described hydrophilic organic solvent is selected from ethanol, methyl alcohol or acetone etc.They all are the solvents that very generally uses in the chemical technology field, and are product solds in the market.
Among the present invention; The RE-5210A type Rotary Evaporators that employed extraction element is produced for the prompt experimental installation ltd of being shaken by Shanghai; Capacity is 10L, and vacuum pump is SHZ-D (III) the type circulation ability of swimming vacuum pump of being produced by Yuhua Instrument Co., Ltd., Gongyi City.
D, from extracting solution, separate fucoxanthine
With step C) extracting solution that obtains and normal hexane be according to 6: 1～3: 1 mixed of volume ratio, behind the concuss, standing separation 25～35min; Separate lower floor's solution; The residue upper solution repeats this procedure, is colourless mutually until the upper strata normal hexane, merges these lower floor's solution;
Among the present invention; When carrying out liquid-liquid extraction with normal hexane and described extracting solution, in order to guarantee under the minimum situation of normal hexane consumption that Impurity removals such as chlorophyll is clean, need follow a small amount of principle repeatedly; Said extracting solution and normal hexane are advisable according to 6: 1～3: 1 ratio of volume ratio; Be preferably 5: 1～4: 1, guarantee that promptly removal of impurities is complete, reduce the solvent waste again.
When the present invention is used for industrial amplification production, can adopt the high-speed counter-current extraction equipment to replace described concussion extraction.
Concentrating of E, fucoxanthine component
Pressure-0.06～-condition of 40～55 ℃ of 0.09MPa and temperature under, with step D) lower floor's solution concentration of obtaining is to doing, and obtains containing the fucoxanthine enriched material.
The enriched material that obtains adopts the HPLC method to detect the content of fucoxanthine in a conventional manner.
The invention still further relates to the enriched material that adopts said method to obtain, it is characterized in that it contains in the fucoxanthine of enriched material gross weight more than 5%.
Described enriched material also contains a certain amount of oil-soluble impurities except fucoxanthine.
The invention still further relates to the purposes of said enriched material in preparation foodstuff additive, functional foodstuff, medicine and fodder additives.
Be noted that and use and add the enriched material that the present invention contains fucoxanthine according to the relevant regulations of country in foodstuff additive, functional foodstuff, medicine and feed additive field.
[beneficial effect]
The invention has the beneficial effects as follows:
1, extraction time is short, and it is high to extract fucoxanthine efficient;
Because fucoxanthine is the pigment that is present in the alginic cell,,, be difficult to fucoxanthine is extracted then owing to cell institute inherent constructional feature and osmotic pressure problem if directly marine alga is extracted with the ethanol equal solvent.In the present invention, marine alga is through the pre-treatment of enzyme liquid, and with the cell walls disorganization of stubbornness, the compositions such as carrotenoid that contained in the marine alga exist with the free state form.And fucoxanthine has good solubility in ethanol, and when refluxing lixiviate with ethanol, fucoxanthine can be discharged into rapidly and extract in the solvent, has shortened extraction time greatly, and has improved extraction efficiency.
2, the product organic solvent-free that obtains is residual;
In the present invention, the used solvent of leaching process is an ethanol, and sepn process adopts normal hexane and the mode of extracting the solvent liquid-liquid extraction to remove impurity; The solvent for use kind is few; And both can reach preferably through the mode of regulating alcohol concn and separate, therefore, and in products obtained therefrom; Organic solvent-free residual, product quality is guaranteed.
3, technology is simple, and comprehensive cost is cheap, is suitable for suitability for industrialized production
In the present invention, raw material is phaeophyta marine algas such as sea-tangle, Sargassum fusiforme, sargassun, bulk kelp, wakame or Thallus Gracilariae, in China rich in natural resources is arranged; Raw material is prone to obtain, and this technology is simple, after handling through enzyme; Can largely wherein contained fucoxanthine be extracted with ethanol; Separate with n-hexane extraction, the fucoxanthine enriched material that can obtain having using value after concentrating has been avoided the use of a large amount of organic solvents in the prior art again; And obtain simplifying from technology, also practiced thrift production cost to a certain extent.This technology is strong operability aborning, has favorable industrial and amplifies prospect.
[embodiment]
Embodiment 1: the present invention contains the preparation of fucoxanthine enriched material
Implementation step is following:
A, raw materials pretreatment
Water is rinsed sea-tangle top layer remaining impurities and salinity well; And the GZX-9146MBE type air dry oven that places medical apparatus and instruments factory of Shanghai Boxun Industrial Co., Ltd. to produce; In 35 ℃ of oven dry down of temperature, take by weighing the dry sea-tangle of 1kg, be crushed to≤20 orders through high speed shear.
B, enzyme are handled
(1) gets 1000g water and water temperature transferred to 30 ℃; Add the cellulase of 1.0g Pangbo Bioengineering Co Ltd, Nanning production and the polygalacturonase that 0.2g Pangbo Bioengineering Co Ltd, Nanning produces then; Again with hydrochloric acid or sodium hydroxide dilute aqueous soln pH regulator to 6.0 with its solution; At 30 ℃ of following activation 40min of temperature, so obtain a kind of enzyme treatment solution;
(2) be sprayed onto 700g to the said enzyme treatment solution of 1400 grams by steps A) in the sea-tangle powder that obtains; In the 3000mL large beaker, add material on one side; Spray the enzyme treatment solution on one side; Stir again, compacting, with preservative film sealing, lucifuge insulation 60h obtains a kind of enzyme and handles the sea-tangle powder in the thermostat water bath of 30 ℃ of temperature;
C, extraction
Taking by weighing 100g at step B) enzyme that obtains handles the sea-tangle powder; Join in the 10L Rotary Evaporators, under 20 ℃ of conditions of temperature, using 400g concentration is that 80% ethanol carries out refluxing extraction 70min, then separation and Extraction liquid; Leftover materials repeat to extract 5 times again, merge these extracting solutions;
D, from extracting solution, separate fucoxanthine
With step C) extracting solution that obtains mixes with the 400g normal hexane, and behind the concuss, standing separation 30min separates lower floor's solution, and the residue upper solution repeats aforesaid operations, and normal hexane is colourless mutually to the upper strata, merges these lower floor's solution;
Concentrating of E, fucoxanthine component
Under the condition of 40 ℃ of pressure 0.095MPa and temperature, with step D) lower floor's solution concentration of obtaining is to doing, and obtains containing the enriched material of fucoxanthine.Adopting the HPLC method to detect its fucoxanthine content in a usual manner is 6.41%.
Embodiment 2: the present invention contains the preparation of the enriched material of fucoxanthine
Implementation step is following:
A, raw materials pretreatment
Water is rinsed sargassun top layer remaining impurities and salinity well; And the GZX-9146MBE type air dry oven that places medical apparatus and instruments factory of Shanghai Boxun Industrial Co., Ltd. to produce; In 30 ℃ of oven dry down of temperature, take by weighing the dry sargassun of 1kg, be crushed to≤20 orders through high speed shear.
B, enzyme are handled
(1) gets 800g water and water temperature transferred to 40 ℃; Add the cellulase of 0.5g Pangbo Bioengineering Co Ltd, Nanning production and the polygalacturonase that 0.4g Pangbo Bioengineering Co Ltd, Nanning produces then; Again with hydrochloric acid or sodium hydroxide dilute aqueous soln pH regulator to 5.0 with its solution; At 40 ℃ of following activation 50min of temperature, so obtain a kind of enzyme treatment solution;
(2) be sprayed onto 900g to the said enzyme treatment solution of 2700 grams by steps A) in the sargassun powder that obtains; In bucket, add material on one side; Spray the enzyme treatment solution on one side; Stir again, compacting, with preservative film sealing, lucifuge insulation 30h obtains a kind of enzyme and handles the sargassun powder in the thermostat water bath of 40 ℃ of temperature;
C, extraction
Taking by weighing 80g at step B) enzyme that obtains handles the sargassun powder; Join in the 10L Rotary Evaporators, under 50 ℃ of conditions of temperature, using 300g concentration is that 40% ethanol carries out refluxing extraction 30min, then separation and Extraction liquid; Leftover materials repeat to extract 6 times again, merge these extracting solutions;
D, from extracting solution, separate pheophytin
With step C) extracting solution that obtains mixes with the 600g normal hexane, and behind the concuss, standing separation 30min separates lower floor's solution, and the residue upper solution repeats to the upper strata that normal hexane merges these lower floor's solution mutually for colourless;
Concentrating of E, fucoxanthine component
Under the condition of 45 ℃ of pressure 0.095MPa and temperature, with step D) obtain lower floor's solution concentration to doing, obtain containing the enriched material of fucoxanthine.Adopting the HPLC method to detect its fucoxanthine content in a usual manner is 6.93%.
Embodiment 3: the present invention contains the preparation of the enriched material of fucoxanthine
Implementation step is following:
A, raw materials pretreatment
Water is rinsed wakame top layer remaining impurities and salinity well; And the GZX-9146MBE type air dry oven that places medical apparatus and instruments factory of Shanghai Boxun Industrial Co., Ltd. to produce; In 50 ℃ of oven dry down of temperature, take by weighing the 1kg dried undaria pinnatifida, be crushed to≤20 orders through high speed shear.
B, enzyme are handled
(1) gets 1200g water and water temperature transferred to 50 ℃; Add the cellulase of 1.5g Pangbo Bioengineering Co Ltd, Nanning production and the polygalacturonase that 0.3g Pangbo Bioengineering Co Ltd, Nanning produces then; Again with hydrochloric acid or sodium hydroxide dilute aqueous soln pH regulator to 7.0 with its solution; At 50 ℃ of following activation 60min of temperature, so obtain a kind of enzyme treatment solution;
(2) be sprayed onto 1000g to the said enzyme treatment solution of 5000 grams by steps A) in the undaria powder that obtains; In bucket, add material on one side; Spray the enzyme treatment solution on one side; Stir again, compacting, with preservative film sealing, lucifuge insulation 4h obtains a kind of enzyme and handles undaria powder in the thermostat water bath of 50 ℃ of temperature;
C, extraction
Taking by weighing 120g at step B) enzyme that obtains handles undaria powder; Join in the 10L Rotary Evaporators, under 2 ℃ of conditions of temperature, using 800g concentration is that 100% ethanol carries out refluxing extraction 90min, then separation and Extraction liquid; Leftover materials repeat to extract 3 times again, united extraction liquid;
D, from extracting solution, separate pheophytin
With step C) extracting solution that obtains mixes with the 500g normal hexane, and behind the concuss, standing separation 30min separates lower floor's solution, and it is colourless mutually that the residue upper solution repeats aforesaid operations to upper strata normal hexane, merges these lower floor's solution;
Concentrating of E, fucoxanthine component
Under the condition of 50 ℃ of pressure 0.095MPa and temperature, with step D) lower floor's solution concentration of obtaining is to doing, and obtains containing the enriched material of fucoxanthine.Adopting the HPLC method to detect its fucoxanthine content in a usual manner is 5.79%.
Embodiment 4: the present invention contains the preparation of the enriched material of fucoxanthine
Implementation step is following:
A, raw materials pretreatment
Water is rinsed sea-tangle top layer remaining impurities and salinity well; And the GZX-9146MBE type air dry oven that places medical apparatus and instruments factory of Shanghai Boxun Industrial Co., Ltd. to produce; In 40 ℃ of oven dry down of temperature, take by weighing the dry sea-tangle of 1kg, be crushed to≤20 orders through high speed shear.
B, enzyme are handled
(1) gets 1400g water and water temperature transferred to 20 ℃; Add the cellulase of 2.0g Pangbo Bioengineering Co Ltd, Nanning production and the polygalacturonase that 0.1g Pangbo Bioengineering Co Ltd, Nanning produces then; Again with hydrochloric acid or sodium hydroxide dilute aqueous soln pH regulator to 4.0 with its solution; At 20 ℃ of following activation 20min of temperature, so obtain a kind of enzyme treatment solution;
(2) be sprayed onto 500g to the said enzyme treatment solution of 300 grams by steps A) in the sea-tangle powder that obtains; In the 3000mL large beaker, add material on one side; Spray the enzyme treatment solution on one side; Stir again, compacting, with preservative film sealing, lucifuge insulation 96h obtains a kind of enzyme and handles the sea-tangle powder in the thermostat water bath of 20 ℃ of temperature;
C, extraction
Taking by weighing 110g at step B) enzyme that obtains handles the sea-tangle powder; Join in the 10L Rotary Evaporators, under 40 ℃ of conditions of temperature, using 600g concentration is that 60% ethanol carries out refluxing extraction 50min, then separation and Extraction liquid; Leftover materials repeat to extract 4 times again, united extraction liquid;
D, from extracting solution, separate pheophytin
With step C) extracting solution that obtains mixes with the 600g normal hexane, and behind the concuss, standing separation 30min separates lower floor's solution, and it is colourless mutually that the residue upper solution repeats aforesaid operations to upper strata normal hexane, merges these lower floor's solution;
Concentrating of E, fucoxanthine component
Under the condition of 55 ℃ of pressure 0.095MPa and temperature, with step D) lower floor's solution concentration of obtaining is to doing, and obtains containing the enriched material of fucoxanthine.Adopting the HPLC method to detect its fucoxanthine content in a usual manner is 5.86%.
Claims (8)
1. one kind contains the method for fucoxanthine enriched material from extracting algae, it is characterized in that the step of this method is following:
A, raw materials pretreatment
The impurity and the salinity of water flush away marine alga remained on surface with the marine alga oven dry, are ground into seaweed powder again under the condition of 30～50 ℃ of temperature and lucifuge;
B, enzyme are handled
(1) gets the water of 800～1400 weight parts; And water temperature transferred to 20～50 ℃; Add 0.5～2.0 weight part cellulase and 0.1～0.4 weight part polygalacturonase then; Again with mineral acid or mineral alkali pH regulator to 4～7, at 20～50 ℃ of following activation 20～60min of temperature, so obtain a kind of enzyme treatment solution with its solution;
(2) be sprayed onto 50～100 weight parts to the said enzyme treatment solution of 30～500 weight parts by steps A) in the seaweed powder that obtains, stir again, compacting, sealing, at 20～50 ℃ of following lucifuges insulation 4～96h of temperature, obtain a kind of enzyme and handle seaweed powder;
C, extraction
Taking by weighing 80～120 weight parts by step B) enzyme that obtains handles seaweed powder; Under 2～50 ℃ of conditions of temperature, use 300～800 weight parts, 40～100% hydrophilic organic solvents to carry out refluxing extraction 30～90min; Separation and Extraction liquid then; Leftover materials repeat to extract 3～6 times, merge these extracting solutions;
D, from extracting solution, separate fucoxanthine
With step C) extracting solution that obtains and normal hexane be according to 6: 1～3: 1 mixed of volume ratio, behind the concuss, standing separation 25～35min; Separate lower floor's solution; The residue upper solution repeats this procedure, is colourless mutually until the upper strata normal hexane, merges these lower floor's solution;
Concentrating of E, fucoxanthine component
Pressure-0.06～-condition of 40～55 ℃ of 0.095MPa and temperature under, with step D) lower floor's solution concentration of obtaining is to doing, and obtains containing the enriched material of fucoxanthine.
2. preparation method according to claim 1 is characterized in that described marine alga is selected from sea-tangle, Sargassum fusiforme, sargassun, bulk kelp, wakame or Thallus Gracilariae.
3. method according to claim 1, the granularity that it is characterized in that said seaweed powder is 20～100 orders.
4. method according to claim 1 is characterized in that at step B) in the mineral acid that uses be selected from hydrochloric acid, sulfuric acid, nitric acid or phosphoric acid; Mineral alkali is selected from sodium hydroxide, Pottasium Hydroxide, yellow soda ash, salt of wormwood, sodium hydrogencarbonate or saleratus.
5. method according to claim 1 is characterized in that at step B) described in the enzyme work >=4000IU/g of cellulase, the enzyme work >=5000IU/g of described polygalacturonase.
6. method according to claim 1 is characterized in that at step C) in, described hydrophilic organic solvent is selected from ethanol, methyl alcohol or acetone.
7. the enriched material that obtains according to the said method of each claim in the claim 1～6 is characterized in that it contains in the fucoxanthine of enriched material gross weight more than 5%.
8. according to the purposes of the said enriched material of claim 7 in preparation foodstuff additive, functional foodstuff, medicine and fodder additives.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110155266.XA CN102336725B (en) | 2011-06-10 | 2011-06-10 | Method for extracting fucoxanthine-containing concentrate, product obtained through method, and application of product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110155266.XA CN102336725B (en) | 2011-06-10 | 2011-06-10 | Method for extracting fucoxanthine-containing concentrate, product obtained through method, and application of product |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102336725A true CN102336725A (en) | 2012-02-01 |
| CN102336725B CN102336725B (en) | 2014-09-10 |
Family
ID=45512722
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110155266.XA Active CN102336725B (en) | 2011-06-10 | 2011-06-10 | Method for extracting fucoxanthine-containing concentrate, product obtained through method, and application of product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102336725B (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103012327A (en) * | 2012-12-21 | 2013-04-03 | 国家海洋局第三海洋研究所 | Preparation method of fucoxanthin |
| CN103232552A (en) * | 2013-04-24 | 2013-08-07 | 集美大学 | Method for preparing brown algae fucosan and fucoxanthin in enzymic way |
| JPWO2013161378A1 (en) * | 2012-04-27 | 2015-12-24 | 株式会社カネカ | Method for producing fucoxanthin-containing composition |
| CN105287648A (en) * | 2015-11-05 | 2016-02-03 | 广东医学院 | Seaweed active component extraction and preparation method and application thereof |
| CN106566760A (en) * | 2016-11-08 | 2017-04-19 | 云南爱尔康生物技术有限公司 | Fucoxanthin healthcare wine and preparation method thereof |
| CN107188870A (en) * | 2017-06-29 | 2017-09-22 | 厦门理工学院 | A kind of method that fucoxanthin is extracted from brown alga |
| CN107382912A (en) * | 2017-07-31 | 2017-11-24 | 中国科学院海洋研究所 | A kind of method for improving sargassum crude extract fucoxanthin purity |
| CN107581434A (en) * | 2017-10-27 | 2018-01-16 | 山东省农业科学院生物技术研究中心 | A kind of lactic acid bacteria beverage rich in astaxanthin and preparation method thereof |
| CN110560474A (en) * | 2019-09-05 | 2019-12-13 | 王旭 | Method for removing total chromium pollutants in chromium-polluted soil by water removal |
| CN115530372A (en) * | 2022-10-09 | 2022-12-30 | 北京姿美堂生物技术股份有限公司 | Seaweed extract with fat reducing function, preparation method thereof and seaweed extract composition |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1706836A (en) * | 2004-06-11 | 2005-12-14 | 中国科学院海洋研究所 | Process of separating fucoxanthin from algae |
| CN101445496A (en) * | 2008-10-10 | 2009-06-03 | 汤洁 | Method for preparing fucoxanthine by using supercritical CO2 technology |
-
2011
- 2011-06-10 CN CN201110155266.XA patent/CN102336725B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1706836A (en) * | 2004-06-11 | 2005-12-14 | 中国科学院海洋研究所 | Process of separating fucoxanthin from algae |
| CN101445496A (en) * | 2008-10-10 | 2009-06-03 | 汤洁 | Method for preparing fucoxanthine by using supercritical CO2 technology |
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPWO2013161378A1 (en) * | 2012-04-27 | 2015-12-24 | 株式会社カネカ | Method for producing fucoxanthin-containing composition |
| CN103012327A (en) * | 2012-12-21 | 2013-04-03 | 国家海洋局第三海洋研究所 | Preparation method of fucoxanthin |
| CN103012327B (en) * | 2012-12-21 | 2015-05-13 | 国家海洋局第三海洋研究所 | Preparation method of fucoxanthin |
| CN103232552A (en) * | 2013-04-24 | 2013-08-07 | 集美大学 | Method for preparing brown algae fucosan and fucoxanthin in enzymic way |
| CN105287648B (en) * | 2015-11-05 | 2019-02-12 | 广东医学院 | Sea is delivered vegetables the extraction preparation method and applications of active constituent |
| CN105287648A (en) * | 2015-11-05 | 2016-02-03 | 广东医学院 | Seaweed active component extraction and preparation method and application thereof |
| CN106566760A (en) * | 2016-11-08 | 2017-04-19 | 云南爱尔康生物技术有限公司 | Fucoxanthin healthcare wine and preparation method thereof |
| CN107188870A (en) * | 2017-06-29 | 2017-09-22 | 厦门理工学院 | A kind of method that fucoxanthin is extracted from brown alga |
| CN107382912A (en) * | 2017-07-31 | 2017-11-24 | 中国科学院海洋研究所 | A kind of method for improving sargassum crude extract fucoxanthin purity |
| CN107382912B (en) * | 2017-07-31 | 2020-04-03 | 中国科学院海洋研究所 | Method for improving fucoxanthin purity of crude sargassum extract |
| CN107581434A (en) * | 2017-10-27 | 2018-01-16 | 山东省农业科学院生物技术研究中心 | A kind of lactic acid bacteria beverage rich in astaxanthin and preparation method thereof |
| CN110560474A (en) * | 2019-09-05 | 2019-12-13 | 王旭 | Method for removing total chromium pollutants in chromium-polluted soil by water removal |
| CN110560474B (en) * | 2019-09-05 | 2021-05-14 | 王旭 | Method for removing total chromium pollutants in chromium-polluted soil by water removal |
| CN115530372A (en) * | 2022-10-09 | 2022-12-30 | 北京姿美堂生物技术股份有限公司 | Seaweed extract with fat reducing function, preparation method thereof and seaweed extract composition |
| CN115530372B (en) * | 2022-10-09 | 2024-01-30 | 北京姿美堂生物技术股份有限公司 | Seaweed extract with lipid-lowering function, preparation method thereof and seaweed extract composition |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102336725B (en) | 2014-09-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102336725B (en) | Method for extracting fucoxanthine-containing concentrate, product obtained through method, and application of product | |
| CN105193863B (en) | A kind of preparation method of high-purity seeweed polyphenol | |
| CN101961427B (en) | Method for extracting polysaccharide and polyphenol of oil-tea camellia husk | |
| US8871217B2 (en) | Method for producing fucoxanthin | |
| CN102787519A (en) | Method for preparing soluble kelp cellulose | |
| CN102417162B (en) | Recycling method of hydrochloric acid-acetic acid waste liquid | |
| CN105902578B (en) | The extracting method of triterpene compound in a kind of lucidum spore powder | |
| CN101914075B (en) | Method for extracting fucoxanthin from brown algae | |
| CN103804503A (en) | Method of extracting polysaccharide of enteromorpha by enzymolysis in steps | |
| CN103012544B (en) | A kind of method extracting saponin and polysaccharide from tea seed grouts | |
| CN107382912B (en) | Method for improving fucoxanthin purity of crude sargassum extract | |
| CN102807678A (en) | Method for circularly and continuously extracting eucommia gum, eucommia seed oil, eucommia extractum and eucommia resin from eucommia seeds | |
| CN106832045B (en) | A kind of method that gingko episperm extracts pectin and phenolic acid compound simultaneously | |
| CN108014143B (en) | Method for synchronously extracting total triterpenoids and polysaccharides of antrodia camphorata | |
| CN106520154A (en) | Processing method of husk slag of Xanthoceras sorbifolia Bunge and products | |
| CN105925363A (en) | Extracting method of pine seed oil with low acid value and pinolenic acid | |
| CN103204895B (en) | A kind of method of separating ursolic acid from Pericarpium Mali pumilae | |
| CN110903277A (en) | Method for saponifying and hydrolyzing tobacco leaves by ultrasonic alkaline water and extracting nicotine by reduced pressure distillation | |
| CN107488413A (en) | The extracting method of tannin extract in a kind of granatum | |
| CN101486951A (en) | Method for separating oleate, linolic acid, oleate and linoleate | |
| CN102816050A (en) | Method for extracting solanesol by activating waste and defective tobacco by enzyme catalysis | |
| CN101054357A (en) | Method for extracting total carotinoid from turmeric | |
| CN103709136B (en) | Combined type method for extracting schisandrin b from fructus schisandrae | |
| CN114588657B (en) | Extraction method of torreya grandis bark active ingredient | |
| CN107828421A (en) | A kind of preparation method for being sustained olive pomace discarded object soil conditioner |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |