CN104387469A - Technology for separating phycocyanin in spirulina by using polyvinyl ether phosphate through two aqueous phase extraction - Google Patents
Technology for separating phycocyanin in spirulina by using polyvinyl ether phosphate through two aqueous phase extraction Download PDFInfo
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- CN104387469A CN104387469A CN201410683691.XA CN201410683691A CN104387469A CN 104387469 A CN104387469 A CN 104387469A CN 201410683691 A CN201410683691 A CN 201410683691A CN 104387469 A CN104387469 A CN 104387469A
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- polyvinyl ether
- phycocyanins
- phosphoric acid
- spirulina
- acid salt
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G2650/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
- C08G2650/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule characterized by the type of post-polymerisation functionalisation
Abstract
The invention provides a technology for separating phycocyanin in spirulina by using polyvinyl ether phosphate through two aqueous phase extraction, belonging to the technical field of biochemical separation engineering. The technology comprises the following steps: preparing polyvinyl ether phosphate; carrying out two phase extraction on phycocyanin; concentrating and freeze-drying, thereby obtaining phycocyanin powder, wherein the phycocyanin recovery rate can be 70-85%. The technology has the beneficial effects that the method is simple; the steps of a purification process are simplified by enriching and separating the spirulina broken wall liquid by directly using polyvinyl ether phosphate; the selectivity is high; the spectral purity of phycocyanin obtained through separation is A620/A280 which is greater than 2.5; the powder of fresh or dry spirulina can be adopted as a raw material, thus providing a technical method with the effect of solving the problem of high value and large scale utilization of spirulina resources.
Description
Technical field
The invention provides a kind of polyvinyl ether method for production of phosphate salt, for extracting Phycocyanins, C-from breaking wall of spirullina princeps liquid.Belong to biochemical separation engineering technical field.
Background technology
Spirulina phycocyanin is present in the phycobilisome of spirulina.Phycobilisome is that specific to red algae and blue-green algae, a kind of supramolecule catches photopigment complex body.Phycocyanins, C-color and luster is in bright blueness, bright-colored, is the first-selected pure natural pigment of food, senior eye shadow, lipstick.Phycocyanins, C-can also regulate and the multiple important enzyme required for synthesized human metabolism, has significantly anti-oxidant, anti-inflammatory effect, regulates human immune system, strengthens function of immune system.Highly purified Phycocyanins, C-with very strong fluorescence, the pure natural fluorescent reagent made, for clinical diagnose, the research field such as immunochemistry and biomedical engineering.
In China, annual spirulina whole nation ultimate production has reached 7000 tons.Yield of spirulina 1/3rd for direct export, other spirulina is used as healthcare products mainly with the form of algae powder, micro-algae algae sheet and perfusion capsule and is used.The mass-producing development and utilization of spirulina is in the elementary process segment, does not also have deep processed product.In spirulina, the extraction of Phycocyanins, C-is also in the laboratory study stage, is not suitable for the good processing method of suitability for industrialized production.At present, the highly purified Phycocyanins, C-commodity that market is sold are all from external import, expensive, and application is restricted.
Therefore, develop leaching process that is easy, Spirulina phycocyanin fast, improving Spirulina phycocyanin product purity is key issue urgently to be resolved hurrily in the mass-producing exploitation of current Spirulina phycocyanin.It is high that polymkeric substance double water-phase isolated protein has biocompatibility, and operational condition is gentle, is easy to carry out the advantages such as continuous operation.The patented technology of the Bi-aqueous extraction Phycocyanins, C-of the relevant preparation algae Phycocyanins, C-reported, such as Chinese patent CN103880950A adopts expensive ionic liquid to form polymkeric substance-aqueous phase, increase production cost, CN102993297A, CN101891809A etc. adopt commercially available PEG to form polymkeric substance-aqueous phase, be separated the poor selectivity of Phycocyanins, C-, need and to saltout etc. that other extracts means conbined usage and just can obtain high purity product, process is loaded down with trivial details.
The invention provides a kind of polyvinyl ether phosphoric acid salt Phycocyanins, C-to specificity extracting power, by two-phase extraction mode direct separation and purification Phycocyanins, C-from breaking wall of spirullina princeps liquid, without the need to saltout etc. that other extracts means conbined usage, just can obtain high purity phycocyanin.
Summary of the invention
The object of this invention is to provide a kind of polyvinyl ether method for production of phosphate salt, for Bi-aqueous extraction Spirulina phycocyanin, obtain highly purified Phycocyanins, C-, be applied to the research and development of natural pigment, protective foods and new drug.
Technical problem to be solved by this invention is to provide a kind of polyvinyl ether method for production of phosphate salt, for extracting and separating Phycocyanins, C-from breaking wall of spirullina princeps liquid.Its method is simple, and fast, cost is low, and be separated Phycocyanins, C-and have higher purity, raw material both can adopt fresh spirulina also can adopt spirulina powder, thus provides a kind of method solving Spirulina phycocyanin resource extentization and utilize.
Technical scheme of the present invention: a kind of isolation technique of polyvinyl ether phosphoric acid salt aqueous two-phase extraction Spirulina phycocyanin, step is:
(1) preparation of bromo polyvinyl ether; (2) the phosphatic preparation of polyvinyl ether; (3) two-phase extraction is from breaking wall of spirullina princeps clear liquid separation and purification Phycocyanins, C-; (4) the Phycocyanins, C-aqueous solution is through concentrating and obtaining Phycocyanins, C-powder after lyophilize.
Described scheme, in step 1, 100g polyoxyethylene glycol is put in round-bottomed flask, phosphorus tribromide is loaded in constant pressure funnel, the weight ratio of phosphorus tribromide and polyoxyethylene glycol is 1:2 ~ 3, under magnetic stirring phosphorus tribromide is slowly instilled in round-bottomed flask, time for adding is 0.5 hour, after dropwising, be warming up to 60 ~ 90 DEG C of reactions 5 ~ 8 hours, slowly instill deionized water in the reactive mixture, until the yellow steam in flask is eliminated, then 500 ~ 1000ml methylene dichloride is added, be transferred in separating funnel, three times are washed with 25% aqueous sodium carbonate of 250ml, collect lower floor's liquid, bromo polyvinyl ether is obtained after methylene dichloride is removed in lower floor's solution distillation.
Described scheme, in step 2, in above-mentioned bromo polyvinyl ether, add anhydrous phosphoric acid salt and 50% isopropanol water solution, between anhydrous sodium phosphate and bromo polyvinyl ether, weight ratio is 1:3 ~ 6, between 50% isopropanol water solution and bromo polyvinyl ether, weight ratio is 1:0.25 ~ 0.5, react 15 ~ 24 hours at 90 ~ 120 DEG C, add 20g gac and stir 2 hours, by filtrate vacuum distilling 1 hour at 80 ~ 100 DEG C after filtration, then in filtrate, add 1000 ~ 2000ml absolute methyl alcohol, filter and obtain polyvinyl ether phosphoric acid salt.
Described scheme, in step 3, the preparation of breaking wall of spirullina princeps clear liquid: adopt water or phosphoric acid buffer to make extraction agent, the concentration of phosphoric acid buffer is 25 ~ 100mmol/L, the solid-to-liquid ratio of algae powder or fresh algae and extracting solution is 1:50 ~ 1:150, after stirring, at being placed in-10 DEG C ~-20 DEG C after freezing certain hour, 37 DEG C of dissolvings, multigelation like this 4-6 time, the broken wall clear liquid of centrifugal acquisition after melting, in broken wall clear liquid, protein concn is 1.0 ~ 5.6mg/ml, and Phycocyanins, C-purity is 0.5 ~ 0.72.
Described scheme, in step 3, adds polyvinyl ether phosphoric acid salt and inorganic salt in breaking wall of spirullina princeps clear liquid, polyvinyl ether phosphoric acid salt: inorganic salt: the weight ratio of broken wall clear liquid is 0.05 ~ 0.25:0.2 ~ 0.35:1, concussion mixing 2 minutes, leave standstill phase-splitting, Phycocyanins, C-is enriched in phase; To be 30000 daltonian ultrafiltration membrance filters through molecular weight cut-off containing Phycocyanins, C-upper, remove polyvinyl ether phosphoric acid salt and inorganic salt, obtain the Phycocyanins, C-aqueous solution, purity be 2.5 ~ 3.2, and protein concentration is 3.15 ~ 5.6mg/ml.
Described scheme, in step 4, to original volume 1/3rd, obtains Phycocyanins, C-powder by Phycocyanins, C-aqueous solution vacuum concentration at 60 DEG C after lyophilize.
Beneficial effect of the present invention: the present invention compared with prior art, mainly has the following advantages: the 1. extraction selectivity of polyvinyl ether phosphoric acid salt to Phycocyanins, C-is high, is applicable to the industrialization production requirements in different scales and place; 2. polyvinyl ether phosphoric acid salt directly can extract and obtain highly purified Phycocyanins, C-from breaking wall of spirullina princeps clear liquid, and extract means without the need to other and coordinate, production cost is low; 3. the raw material preparing Phycocyanins, C-can be the fresh algae of spirulina or dry algae powder, effectively solves the utilization of spirulina data and high-valued problem.
Embodiment
Embodiment 1
(1) preparation of breaking wall of spirullina princeps clear liquid
Get and originate in the spirulina plalensis dry powder that century-old biotechnology company limited is grant in Dongtai city, extraction agent is made with deionized water, the solid-to-liquid ratio of algae powder and deionized water is 1:50, at being placed in-20 DEG C after freezing 4 hours, 37 DEG C melt, multigelation like this 4 times, blue green algae disrupted solution whizzer after thawing under 6000 turns/min centrifugal 10 minutes, getting supernatant liquor is broken wall clear liquid, and in clear liquid, protein concn is 1.0mg/ml, Phycocyanins, C-purity A
620/ A
280for 0.72mg/ml.
(2) preparation of bromo polyvinyl ether
100g polyoxyethylene glycol PEG-1000 (purchased from Hai'an petroleum chemical plant) is put in the round-bottomed flask of 500ml, the molecular-weight average of PEG-1000 is 1000g every mole, 30g phosphorus tribromide (purchased from Huai'an Xin Xin Chemical Co., Ltd.) is loaded in constant pressure funnel, under magnetic stirring phosphorus tribromide is slowly instilled in round-bottomed flask, time for adding is 0.5 hour, after dropwising, be warming up to 60 DEG C of reactions 5 hours, slowly instill deionized water in the reactive mixture, until the yellow steam in flask is eliminated, add 500ml methylene dichloride (purchased from Linzhou City, Zibo City Tian De fine chemistry industry institute), reaction mixture is transferred in separating funnel, with 25% sodium carbonate (purchased from assisting into the industrial chemicals) solution washing three times of 250ml, collect lower floor's liquid distillation and remove methylene dichloride, obtain bromo polyvinyl ether.
(3) the phosphatic preparation of polyvinyl ether
100g bromo polyvinyl ether is put in 1500ml Erlenmeyer flask, add 50% Virahol (purchased from sincerityization light material company limited of the city of Kunshan) aqueous solution of 30g anhydrous sodium phosphate (purchased from Chongqing Chuan Dong Chemical Co., Ltd.) and 400ml, react 15 hours at 90 DEG C, add 20g gac (making the country prosperous gac company limited purchased from Quanzhou City) and stir 2 hours, by filtrate vacuum distilling 1 hour at 80 DEG C after filtration, then in filtrate, add 1000ml absolute methyl alcohol, obtain polyvinyl ether phosphoric acid salt with Büchner funnel suction filtration.
(4) two-phase extraction
0.5g polyvinyl ether phosphoric acid salt and 0.2g anhydrous sodium sulphate (purchased from Nanjing Datang chemical industry limited liability company) is added in 10g breaking wall of spirullina princeps clear liquid, concussion mixing 2 minutes, leave standstill phase-splitting, Phycocyanins, C-is enriched in phase, upper is that 30000 daltonian ultra-filtration membranes (quite that strainer company limited) filter through molecular weight cut-off, removes polyvinyl ether phosphoric acid salt and sodium sulfate, obtains the Phycocyanins, C-aqueous solution, purity is 2.5, and protein concentration is 3.15mg/ml.
(5) lyophilize obtains Phycocyanins, C-powder
The Phycocyanins, C-aqueous solution vacuum concentration at 60 DEG C obtained in abovementioned steps (4), to original volume 1/3rd, is obtained Phycocyanins, C-powder product after lyophilize.Phycocyanins, C-purity A in product
620/ A
280be 2.5, protein recovery is 80%.
Embodiment 2
(1) preparation of breaking wall of spirullina princeps liquid
Get and originate in the fresh spirulina that Jiangsu great Feng grants century-old bio tech ltd, make extraction agent with 50mmol/L phosphoric acid buffer, the solid-to-liquid ratio of spirulina and deionized water is 1:100, and at being placed in-10 DEG C after freezing 4 hours, 37 DEG C melt, multigelation like this 5 times.Blue green algae disrupted solution whizzer after thawing under 6000 turns/min centrifugal 10 minutes, getting supernatant liquor is broken wall clear liquid, and in clear liquid, protein concn is 3.1mg/ml, Phycocyanins, C-purity A
620/ A
280be 0.5.
(2) preparation of bromo polyvinyl ether
100g polyoxyethylene glycol PEG-3000 (purchased from Hai'an petroleum chemical plant) is put in the round-bottomed flask of 1000ml, the molecular-weight average of PEG-3000 is 3000g every mole, 50g phosphorus tribromide (purchased from Huai'an Xin Xin Chemical Co., Ltd.) is loaded in constant pressure funnel, under magnetic stirring phosphorus tribromide is slowly instilled in round-bottomed flask, time for adding is 0.5 hour, after dropwising, be warming up to 80 DEG C of reactions 7 hours, slowly instill deionized water in the reactive mixture, until the yellow steam in flask is eliminated, then 800ml methylene dichloride (purchased from Linzhou City, Zibo City Tian De fine chemistry industry institute) is added in the reactive mixture, be transferred in separating funnel, with 25% sodium carbonate (purchased from assisting into the industrial chemicals) solution washing three times of 250ml, collect lower floor's liquid, bromo polyvinyl ether is obtained after methylene dichloride is removed in lower floor's solution distillation.
(3) the phosphatic preparation of polyvinyl ether
In 2000ml Erlenmeyer flask, add 50% Virahol (purchased from sincerityization light material company limited of the city of Kunshan) aqueous solution of 100g bromo polyvinyl ether, 20g anhydrous phosphoric acid potassium (purchased from Chongqing Chuan Dong Chemical Co., Ltd.), 250ml, react 20 hours at 110 DEG C, then in reactant, add 20g gac (making the country prosperous gac company limited purchased from Quanzhou City) stir 2 hours, by filtrate vacuum distilling 1 hour at 90 DEG C after filtration, then in filtrate, add 1500ml absolute methyl alcohol, obtain polyvinyl ether phosphoric acid salt with Büchner funnel suction filtration.
(4) two-phase extraction
1.5g polyvinyl ether phosphoric acid salt and 3g anhydrous slufuric acid ammonium (purchased from Nanjing Datang chemical industry limited liability company) is added in 10g breaking wall of spirullina princeps clear liquid, concussion mixing 2 minutes, leave standstill phase-splitting, Phycocyanins, C-is enriched in phase, by upper be that 30000 daltonian ultra-filtration membranes (quite you strainer company limited) filter through molecular weight cut-off, remove polyvinyl ether phosphoric acid salt and ammonium sulfate, obtain the Phycocyanins, C-aqueous solution, purity is 3.1, and protein concentration is 4.8mg/ml.
(5) lyophilize obtains Phycocyanins, C-powder
The Phycocyanins, C-aqueous solution vacuum concentration at 60 DEG C obtained in abovementioned steps (4), to original volume 1/3rd, is obtained Phycocyanins, C-powder product after lyophilize.Phycocyanins, C-purity A in product
620/ A
280be 3.1, protein recovery 65%.
Embodiment 3
(1) preparation of breaking wall of spirullina princeps liquid
Get the spirulina powder originating in spirulina base, Lijiang, yunnan Cheng Hai source, make extraction agent with 100mmol/L phosphoric acid buffer, the solid-to-liquid ratio of spirulina and deionized water is 1:150, and at being placed in-20 DEG C after freezing 4 hours, 37 DEG C melt, multigelation like this 6 times.Blue green algae disrupted solution whizzer after thawing under 6000 turns/min centrifugal 10 minutes, getting supernatant liquor is broken wall clear liquid, the Phycocyanins, C-purity A in clear liquid
620/ A
280be 0.72, shell-broken liquid protein concn is 5.6mg/ml mg/ml.
(2) preparation of bromo polyvinyl ether
In the round-bottomed flask of 2000ml, put into 100g polyoxyethylene glycol PEG-4000 (purchased from Hai'an petroleum chemical plant), the molecular-weight average of PEG-4000 is 4000g every mole, 30g phosphorus tribromide (purchased from Huai'an Xin Xin Chemical Co., Ltd.) is loaded in constant pressure funnel, under magnetic stirring phosphorus tribromide is slowly instilled in round-bottomed flask, time for adding is 0.5 hour, after dropwising, be warming up to 90 DEG C of reactions 8 hours, slowly instill deionized water in the reactive mixture, until the yellow steam in flask is eliminated, then 1000ml methylene dichloride (purchased from Linzhou City, Zibo City Tian De fine chemistry industry institute) is added in the reactive mixture, be transferred in separating funnel, three times are washed with 25% aqueous sodium carbonate of 250ml, collect lower floor's liquid, bromo polyvinyl ether is obtained after methylene dichloride is removed in lower floor's solution distillation.
(3) the phosphatic preparation of polyvinyl ether
In 3000ml Erlenmeyer flask, add 50% Virahol (purchased from sincerityization light material company limited of the city of Kunshan) aqueous solution of 100g bromo polyvinyl ether, 15g anhydrous phosphoric acid potassium (purchased from Chongqing Chuan Dong Chemical Co., Ltd.), 200ml, react 24 hours at 120 DEG C, then in reactant, add 20g gac (making the country prosperous gac company limited purchased from Quanzhou City) stir 2 hours, by filtrate vacuum distilling 1 hour at 100 DEG C after filtration, then in filtrate, add 2000ml absolute methyl alcohol, obtain polyvinyl ether phosphoric acid salt with Büchner funnel suction filtration.
(4) two-phase extraction
2.5g polyvinyl ether phosphoric acid salt and 3.5g is added Wu Shui Liu Suan Magnesium (purchased from Nanjing Datang chemical industry limited liability company) in 10g breaking wall of spirullina princeps clear liquid, concussion mixing 2 minutes, leave standstill phase-splitting, Phycocyanins, C-is enriched in phase, by upper be that 30000 daltonian ultra-filtration membranes (quite you strainer company limited) filter through molecular weight cut-off, remove polyvinyl ether phosphoric acid salt and Liu Suan Magnesium, obtain the Phycocyanins, C-aqueous solution, purity is 3.2, and protein concentration is 5.6mg/ml.
(5) lyophilize obtains Phycocyanins, C-powder
The Phycocyanins, C-aqueous solution vacuum concentration at 60 DEG C obtained in abovementioned steps (4), to original volume 1/3rd, is obtained Phycocyanins, C-powder product after lyophilize.Phycocyanins, C-purity A in product
620/ A
280be 2.8, protein recovery 75%.
Claims (4)
1. an isolation technique for polyvinyl ether phosphoric acid salt aqueous two-phase extraction Spirulina phycocyanin, is characterized in that step is: the phosphatic preparation of polyvinyl ether; Two-phase extraction Phycocyanins, C-; Through concentrating and obtaining Phycocyanins, C-powder after lyophilize.
(1) the phosphatic preparation of polyvinyl ether: put into 100g polyoxyethylene glycol in round-bottomed flask, phosphorus tribromide is loaded in constant pressure funnel, the weight ratio of phosphorus tribromide and polyoxyethylene glycol is 1:2 ~ 3, under magnetic stirring phosphorus tribromide is slowly instilled in round-bottomed flask, time for adding is 0.5 hour, after dropwising, be warming up to 60 ~ 90 DEG C of reactions 5 ~ 8 hours, slowly instill deionized water in the reactive mixture, until the yellow steam in flask is eliminated, then 500 ~ 1000ml methylene dichloride is added in the reactive mixture, be transferred in separating funnel, three times are washed with 25% aqueous sodium carbonate of 250ml, collect lower floor's liquid, bromo polyvinyl ether is obtained after methylene dichloride is removed in lower floor's solution distillation.In bromo polyvinyl ether, add anhydrous phosphoric acid salt and 50% isopropanol water solution, between anhydrous phosphoric acid salt and bromo polyvinyl ether, weight ratio is 1:3 ~ 6, between 50% isopropanol water solution and bromo polyvinyl ether, weight ratio is 1:0.25 ~ 0.5, react 15 ~ 24 hours at 90 ~ 120 DEG C, then in reactant, add 20g gac stir 2 hours, by filtrate vacuum distilling 1 hour at 80 ~ 100 DEG C after filtration, then in filtrate, add 1000 ~ 2000ml absolute methyl alcohol, filter and obtain polyvinyl ether phosphoric acid salt.
(2) two-phase extraction: add polyvinyl ether phosphoric acid salt and inorganic salt in breaking wall of spirullina princeps clear liquid, polyvinyl ether phosphoric acid salt: inorganic salt: the weight ratio of broken wall clear liquid is 0.05 ~ 0.25:0.2 ~ 0.35:1, concussion mixing 2 minutes, leave standstill phase-splitting, Phycocyanins, C-is enriched in phase.
(3) by above-mentioned be 30000 daltonian ultrafiltration membrance filters containing Phycocyanins, C-upper through molecular weight cut-off, remove polyvinyl ether phosphoric acid salt and inorganic salt, obtain the Phycocyanins, C-aqueous solution, purity is 2.5 ~ 3.2, and protein concentration is 3.15 ~ 5.6mg/ml.
(4) Phycocyanins, C-aqueous solution vacuum concentration at 60 DEG C, to original volume 1/3rd, is obtained Phycocyanins, C-powder after lyophilize.
2. polyvinyl ether method for production of phosphate salt according to claim 1, is characterized in that the molecular-weight average of the polyoxyethylene glycol adopted can be 500,1000,2000,3000,4000.
3. polyvinyl ether method for production of phosphate salt according to claim 1, is characterized in that the anhydrous phosphoric acid salt adopted can be anhydrous sodium phosphate, anhydrous phosphoric acid potassium.
4. two-phase extraction method according to claim 1, is characterized in that, for becoming the inorganic salt of phase can for anhydrous sodium sulphate, without water sulphur sour Magnesium, anhydrous slufuric acid ammonium, anhydrous chlorides of rase sodium, anhydrous phosphoric acid potassium.
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| CN105254752A (en) * | 2015-10-20 | 2016-01-20 | 合肥工业大学 | Method for extracting and purifying phycocyanin by means of activated carbon pretreatment and salt fractionation |
| CN106008705A (en) * | 2016-06-22 | 2016-10-12 | 瑞安市智造科技有限公司 | Method for separating and purifying phycocyanin by means of combination of two aqueous phase extraction and ultrasonic waves |
| CN106432481A (en) * | 2016-11-30 | 2017-02-22 | 江南大学 | Separation technology for performing two aqueous phase extraction on spiral seaweed phycocyanin by octadecyl polyvinylether |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105254752A (en) * | 2015-10-20 | 2016-01-20 | 合肥工业大学 | Method for extracting and purifying phycocyanin by means of activated carbon pretreatment and salt fractionation |
| CN105254752B (en) * | 2015-10-20 | 2019-05-14 | 合肥工业大学 | A kind of method of Activated Carbon Pretreatment joint salting out method extraction purification phycocyanin |
| CN106008705A (en) * | 2016-06-22 | 2016-10-12 | 瑞安市智造科技有限公司 | Method for separating and purifying phycocyanin by means of combination of two aqueous phase extraction and ultrasonic waves |
| CN106432481A (en) * | 2016-11-30 | 2017-02-22 | 江南大学 | Separation technology for performing two aqueous phase extraction on spiral seaweed phycocyanin by octadecyl polyvinylether |
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