CN106008664B - Efficient separation and purification method of glutathione - Google Patents
Efficient separation and purification method of glutathione Download PDFInfo
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- CN106008664B CN106008664B CN201610593413.4A CN201610593413A CN106008664B CN 106008664 B CN106008664 B CN 106008664B CN 201610593413 A CN201610593413 A CN 201610593413A CN 106008664 B CN106008664 B CN 106008664B
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- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 title claims abstract description 512
- 229960003180 glutathione Drugs 0.000 title claims abstract description 248
- 108010024636 Glutathione Proteins 0.000 title claims abstract description 199
- 238000000034 method Methods 0.000 title claims abstract description 52
- 238000000746 purification Methods 0.000 title abstract description 15
- 238000000926 separation method Methods 0.000 title abstract description 10
- 239000002244 precipitate Substances 0.000 claims abstract description 41
- 239000011347 resin Substances 0.000 claims abstract description 38
- 229920005989 resin Polymers 0.000 claims abstract description 38
- 238000001556 precipitation Methods 0.000 claims abstract description 30
- 238000002425 crystallisation Methods 0.000 claims abstract description 14
- 230000008025 crystallization Effects 0.000 claims abstract description 14
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000001471 micro-filtration Methods 0.000 claims abstract description 7
- 238000003825 pressing Methods 0.000 claims abstract description 5
- 238000001291 vacuum drying Methods 0.000 claims abstract 2
- 239000000243 solution Substances 0.000 claims description 124
- 239000013078 crystal Substances 0.000 claims description 72
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 64
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 52
- 238000003756 stirring Methods 0.000 claims description 45
- 239000006228 supernatant Substances 0.000 claims description 39
- 239000000725 suspension Substances 0.000 claims description 36
- 239000012043 crude product Substances 0.000 claims description 31
- 239000012452 mother liquor Substances 0.000 claims description 26
- 238000005406 washing Methods 0.000 claims description 26
- 238000011068 loading method Methods 0.000 claims description 23
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 22
- 239000000706 filtrate Substances 0.000 claims description 22
- 230000001376 precipitating effect Effects 0.000 claims description 22
- 239000007864 aqueous solution Substances 0.000 claims description 19
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims description 17
- 239000003729 cation exchange resin Substances 0.000 claims description 16
- 239000002253 acid Substances 0.000 claims description 15
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims description 14
- BERDEBHAJNAUOM-UHFFFAOYSA-N copper(I) oxide Inorganic materials [Cu]O[Cu] BERDEBHAJNAUOM-UHFFFAOYSA-N 0.000 claims description 14
- KRFJLUBVMFXRPN-UHFFFAOYSA-N cuprous oxide Chemical compound [O-2].[Cu+].[Cu+] KRFJLUBVMFXRPN-UHFFFAOYSA-N 0.000 claims description 14
- 229940112669 cuprous oxide Drugs 0.000 claims description 14
- 238000001035 drying Methods 0.000 claims description 14
- 238000001179 sorption measurement Methods 0.000 claims description 14
- 239000012528 membrane Substances 0.000 claims description 13
- 238000000967 suction filtration Methods 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 11
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 11
- 229910017604 nitric acid Inorganic materials 0.000 claims description 11
- 239000003463 adsorbent Substances 0.000 claims description 9
- 238000003795 desorption Methods 0.000 claims description 9
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 claims description 8
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 229910000037 hydrogen sulfide Inorganic materials 0.000 claims description 8
- UYJXRRSPUVSSMN-UHFFFAOYSA-P ammonium sulfide Chemical compound [NH4+].[NH4+].[S-2] UYJXRRSPUVSSMN-UHFFFAOYSA-P 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 6
- DPLVEEXVKBWGHE-UHFFFAOYSA-N potassium sulfide Chemical compound [S-2].[K+].[K+] DPLVEEXVKBWGHE-UHFFFAOYSA-N 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 229910052979 sodium sulfide Inorganic materials 0.000 claims description 4
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 claims description 4
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 3
- RFHCUXJIPZTRBX-GEMLJDPKSA-N CCO.OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O Chemical compound CCO.OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RFHCUXJIPZTRBX-GEMLJDPKSA-N 0.000 claims description 3
- 239000000908 ammonium hydroxide Substances 0.000 claims description 3
- 150000001875 compounds Chemical class 0.000 claims description 3
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 3
- 239000004744 fabric Substances 0.000 claims description 3
- 239000012141 concentrate Substances 0.000 claims description 2
- 239000012535 impurity Substances 0.000 abstract description 15
- 239000010949 copper Substances 0.000 abstract description 13
- 150000001879 copper Chemical class 0.000 abstract description 7
- 238000000605 extraction Methods 0.000 abstract description 6
- 238000009776 industrial production Methods 0.000 abstract description 5
- 238000005265 energy consumption Methods 0.000 abstract description 2
- 238000005119 centrifugation Methods 0.000 abstract 1
- 238000004925 denaturation Methods 0.000 abstract 1
- 230000036425 denaturation Effects 0.000 abstract 1
- 238000007865 diluting Methods 0.000 abstract 1
- 229910052739 hydrogen Inorganic materials 0.000 description 13
- 238000010828 elution Methods 0.000 description 11
- 239000002245 particle Substances 0.000 description 11
- 238000000921 elemental analysis Methods 0.000 description 10
- 238000001914 filtration Methods 0.000 description 10
- 238000004128 high performance liquid chromatography Methods 0.000 description 10
- 238000005259 measurement Methods 0.000 description 10
- 238000000855 fermentation Methods 0.000 description 6
- 230000004151 fermentation Effects 0.000 description 6
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- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 238000001042 affinity chromatography Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000005342 ion exchange Methods 0.000 description 2
- 239000000693 micelle Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000011085 pressure filtration Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 125000003396 thiol group Chemical group [H]S* 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 239000004201 L-cysteine Substances 0.000 description 1
- 235000013878 L-cysteine Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
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- 238000010364 biochemical engineering Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
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- 210000004027 cell Anatomy 0.000 description 1
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- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
- C07K5/0215—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
Abstract
The invention provides a high-efficiency separation and purification method of glutathione, which mainly comprises the following steps: preparing GSCu precipitate, diluting, adding sulfide to generate Cu2S precipitation and replacement of glutathione, centrifugation, filter pressing or microfiltration, resin purification, reduced pressure concentration, crystallization and vacuum drying. The high-efficiency separation and purification method of the glutathione effectively combines a copper salt precipitation method and a resin purification method, reduces the loss and denaturation of the glutathione in the extraction process to the maximum extent, and eliminates the interference of impurities as much as possible; simple operation, easy control, short treatment period and low energy consumption, and is particularly suitable for industrial production. Therefore, the method for efficiently separating and purifying the glutathione provided by the invention has good application prospect and market potential.
Description
Technical Field
The invention belongs to the field of biochemical engineering, and particularly relates to a high-efficiency separation and purification method of glutathione.
Background
Glutathione (GSH), collectively known as γ -L-glutamic acid-L-cysteinylglycine, is a bioactive tripeptide compound containing sulfhydryl groups, which is formed by condensing L-glutamic acid, L-cysteine and glycine, and is one of the important non-protein sulfhydryl compounds in the body.
Glutathione is ubiquitous in animal and plant cells and microorganisms, and is particularly abundant in yeast, wheat germ, and human and animal liver, kidney, red blood cells, and eye lens. It exists mainly in two forms of reduction type (GSH) and oxidation type (GSSH), wherein, the reduced glutathione exists in the body in large quantity and plays a main role, and the glutathione mentioned in the invention refers to the reduced glutathione. Glutathione has multiple physiological functions, particularly has the most prominent effects of resisting oxidation and integrating detoxification, and plays an important role in the processes of cell proliferation, apoptosis and fibroblast. With the continuous and deep research, the application of glutathione in clinical medicine, health products, food additives, animal feeds and the like is attracting wide attention, and the market prospect demand is increasing day by day.
However, the domestic glutathione production process is not mature enough, the development bottleneck is mainly low in fermentation level, and the yield of the subsequent purification process is low, so that the obtained product has low purity, poor quality and high cost. The method for solving the problems has very important significance for realizing large-scale industrialization of GSH in China, changing the embarrassing situation that GSH raw materials depend on import and promoting the development of medical industry, health care industry and food industry in China, and has good economic benefit and social benefit.
With the optimization of fermentation process conditions and the development of various methods for strain breeding, the fermentation method has become the most common method for producing glutathione at home and abroad, however, the failure to effectively obtain glutathione from fermentation liquor with low cost, high yield and high purity is always an obstacle to the industrialization of glutathione, and is also one of the research hotspots increasingly.
In the prior art, the reported methods for purifying and separating GSH from fermentation broth mainly include: copper salt method, ion exchange method, affinity chromatography, aqueous two-phase extraction, reverse micelle extraction, etc. Wherein, the copper salt method is used as a traditional extraction method, has strong selectivity, high yield, easy operation and strong practical value; however, impurities are easily introduced during the operation, resulting in low product purity. Ion exchange is also a common means for purifying GSH, but the cost of the used filler is high, the service life is short, the production period is long, and the wastewater treatment capacity is large, so that the method is often limited in industrial production. Among them, affinity chromatography is an effective method for separating GSH, but the mercury-containing resin is poor in stability and easy to leak, so that it is not suitable for application in the field of food and medicine. The use of aqueous two-phase extraction for the separation of GSH, although more efficient, has diminished technical advantages due to the expensive aqueous two-phase composition and high production costs. Among them, reverse micelle extraction of glutathione is a method with great development potential, but the industrial large-scale application of the glutathione is not mature, and the method is still in the laboratory research stage at present.
Therefore, the prior art has not provided a method for separating and purifying glutathione, which is suitable for industrial production.
Disclosure of Invention
Aiming at the defects and shortcomings in the prior art, in order to obtain a method for separating and purifying glutathione, which is suitable for industrial production, the inventor develops a method for separating and purifying glutathione from escherichia coli fermentation liquor, skillfully and effectively combines a copper salt precipitation method and a resin purification method, extracts glutathione liquid through the copper salt precipitation method, removes impurities such as pigments and the like through ion exchange resin to improve the purity, and then concentrates and crystallizes to obtain pure glutathione crystals.
Therefore, the technical scheme provided by the invention is as follows:
a method for efficiently separating and purifying glutathione comprises the following steps:
(1) adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate; adding pure water into the GSCu precipitate, and fully stirring to ensure that the homogenate wet weight of the GSCu precipitate suspension is 300-500 g/L;
(2) adding sulfide into the GSCu precipitation suspension, and stirring for reaction to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor;
(3) centrifuging the glutathione mother liquor to obtain supernatant containing glutathione;
(4) carrying out filter pressing or microfiltration on the supernatant liquid for removing particle impurities;
(5) loading the filtrate obtained in the step (4) on cation exchange resin or macroporous adsorption resin, washing and balancing, and eluting with desorption solution to obtain glutathione solution;
(6) concentrating the glutathione solution under reduced pressure of-0.07 Mpa to 0Mpa at 55 to 65 ℃ to prepare a glutathione concentrated solution;
(7) crystallizing the glutathione concentrated solution by adopting an ethanol solution to obtain a glutathione crystal crude product;
(8) and drying the glutathione crystal crude product in vacuum to obtain a pure glutathione crystal.
Preferably, in the step (2) of the method for efficiently separating and purifying glutathione, the stirring reaction is carried out at a stirring speed of 150-250 rpm for 45-60 min.
Preferably, in the method for efficiently separating and purifying glutathione, the sulfide is selected from any one of the following compounds: hydrogen sulfide, sodium sulfide, potassium sulfide and ammonium sulfide.
Preferably, in the step (4) of the method for efficiently separating and purifying glutathione, the aperture of the filter cloth or the filter membrane used for pressure filtration or microfiltration is 0.45 μm.
Preferably, in the method for efficiently separating and purifying glutathione, the cation exchange resin is selected from any one of the following: d001 macroporous strong acid styrene cation exchange resin, HD-8 macroporous strong acid resin, D-61 macroporous strong acid styrene cation exchange resin; the macroporous adsorption resin is selected from any one of the following: SP207 macroporous adsorbent resin, XDA-1 macroporous adsorbent resin, DA201-C macroporous adsorbent resin.
Preferably, the step (3) of the method for efficiently separating and purifying glutathione further comprises: and adjusting the pH value of the supernatant to 2-4 by using hydrochloric acid or phosphoric acid or nitric acid.
Preferably, in the step (5) of the method for efficiently separating and purifying glutathione, the desorption solution used when cation exchange resin is used is ammonium hydroxide aqueous solution, sodium chloride aqueous solution or hydrochloric acid; when the macroporous adsorption resin is adopted, the desorption solution correspondingly used is ethanol water solution.
Preferably, the concentration of the glutathione concentrated solution in the step (6) of the method for efficiently separating and purifying glutathione is 380-420 g/L.
Preferably, the step (7) of the method for efficiently separating and purifying glutathione is: adding 50-60% ethanol solution preheated to 55-65 ℃ into the glutathione concentrated solution, then adjusting the pH of the glutathione ethanol solution to 2-4 by using hydrochloric acid or phosphoric acid or nitric acid, slowly stirring at the rotating speed of 150-250 rpm, and slowly separating out crystals; and after full crystallization, carrying out suction filtration, and washing for 2-4 times by using an ethanol solution with the concentration of 65-75% to obtain a glutathione crystal crude product.
Preferably, the step (8) of the method for efficiently separating and purifying glutathione is: and (3) drying the glutathione crystal crude product in vacuum at the temperature of 60 ℃ under the pressure of-0.07 Mpa to 0Mpa to obtain the glutathione crystal with the purity of more than or equal to 98 percent.
The high-efficiency separation and purification method of the glutathione effectively combines a copper salt precipitation method and a resin purification method; in the copper salt precipitation stage, solid-liquid separation is completed by utilizing the characteristic that GSH and cuprous oxide are complexed to generate precipitates, so that most of protein and salt are removed, the GSH is primarily concentrated and purified, and the resin purification of the next step is facilitated; in the purification stage of the resin, because the interference components such as protein, salt and the like are reduced, the adsorption quantity of the resin to the GSH is improved, and the production efficiency is further improved. Therefore, the method for efficiently separating and purifying the glutathione fully utilizes the advantages of the glutathione and the glutathione, and solves the technical problem that the glutathione is not easy to be industrially produced on a large scale; in addition, the separation and purification method is simple to operate, easy to control, short in treatment period and low in energy consumption, so that the production cost is low, and the method is particularly suitable for industrial production. Therefore, the method for efficiently separating and purifying the glutathione provided by the invention has good application prospect and market potential.
Detailed Description
The present invention will be further described with reference to specific embodiments, but the present invention is not limited to the following embodiments.
In a first aspect, the invention provides a method for efficiently separating and purifying glutathione, which comprises the following steps:
(1) adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate; adding pure water into the GSCu precipitate, and fully stirring to ensure that the homogenate wet weight of the GSCu precipitate suspension is 300-500 g/L;
(2) adding sulfide into the GSCu precipitation suspension, and stirring for reaction to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor;
(3) centrifuging the glutathione mother liquor to obtain supernatant containing glutathione;
(4) carrying out filter pressing or microfiltration on the supernatant liquid for removing particle impurities;
(5) loading the filtrate obtained in the step (4) on cation exchange resin or macroporous adsorption resin, washing and balancing, and eluting with desorption solution to obtain glutathione solution;
(6) concentrating the glutathione solution under reduced pressure of-0.07 Mpa to 0Mpa at 55 to 65 ℃ to prepare a glutathione concentrated solution;
(7) crystallizing the glutathione concentrated solution by adopting an ethanol solution to obtain a glutathione crystal crude product;
(8) and drying the glutathione crystal crude product in vacuum to obtain a pure glutathione crystal.
In a preferred embodiment, in the step (2), the stirring reaction is carried out at a stirring speed of 150-250 rpm for 45-60 min.
In a preferred embodiment, the sulfide is selected from any one of: hydrogen sulfide, sodium sulfide, potassium sulfide and ammonium sulfide.
In a preferred embodiment, in the step (4), the filter cloth or the filter membrane used for the pressure filtration or the microfiltration has a pore size of 0.45 μm.
In a preferred embodiment, the cation exchange resin is selected from any one of the following: d001 macroporous strong acid styrene cation exchange resin, HD-8 macroporous strong acid resin, D-61 macroporous strong acid styrene cation exchange resin; the macroporous adsorption resin is selected from any one of the following: SP207 macroporous adsorbent resin, XDA-1 macroporous adsorbent resin, DA201-C macroporous adsorbent resin.
In a preferred embodiment, step (3) further includes: and adjusting the pH value of the supernatant to 2-4 by using hydrochloric acid or phosphoric acid or nitric acid.
In a preferred embodiment, in step (5), the desorption solution used when cation exchange resin is used is aqueous ammonium hydroxide solution, aqueous sodium chloride solution or hydrochloric acid; when the macroporous adsorption resin is adopted, the desorption solution correspondingly used is ethanol water solution.
In a preferred embodiment, the concentration of the glutathione concentrated solution in the step (6) is 380-420 g/L.
In a preferred embodiment, step (7) is: adding 50-60% ethanol solution preheated to 55-65 ℃ into the glutathione concentrated solution, then adjusting the pH of the glutathione ethanol solution to 2-4 by using hydrochloric acid or phosphoric acid or nitric acid, slowly stirring at the rotating speed of 150-250 rpm, and slowly separating out crystals; and after full crystallization, carrying out suction filtration, and washing for 2-4 times by using an ethanol solution with the concentration of 65-75% to obtain a glutathione crystal crude product.
In a preferred embodiment, step (8) is: and (3) drying the glutathione crystal crude product in vacuum at the temperature of 60 ℃ under the pressure of-0.07 Mpa to 0Mpa to obtain the glutathione crystal with the purity of more than or equal to 98 percent.
The following examples of glutathione separation and purification are all carried out according to the technical scheme provided by the invention, wherein the reagents or raw materials used are all obtained from public commercial sources if no specific description is provided, and the steps are all conventional processing operations if no specific description is provided.
Example 1
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, introducing hydrogen sulfide into the GSCu precipitation suspension, and stirring at 250rpm for 1 hour to react to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding hydrochloric acid into the supernatant to adjust the pH value to 3, filtering the supernatant with a 0.45-micron filter membrane to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on HD-8 macroporous strong acid resin with the column volume of 1.5m multiplied by 20cm and the loading flow rate of 50mL/min, washing and balancing, and eluting with 1.5% hydrochloric acid solution with the elution flow rate of 100mL/min to obtain GSH solution; concentrating the GSH solution under-0.07 Mpa at 65 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 50% ethanol solution preheated to 55 ℃ into the glutathione concentrated solution, then adjusting the pH to 3 by using hydrochloric acid, slowly stirring at the rotating speed of 200 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 2 times by using an ethanol solution with the concentration of 65 percent to obtain a glutathione crystal crude product; finally, the crude glutathione crystals are dried under vacuum at 60 ℃ under a pressure of-0.07 Mpa174g of glutathione crystals are obtained, the purity is 98.6 percent and the total yield is 72.5 percent by High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: 39.11 percent of C, 5.54 percent of H and 31.25 percent of O.
Example 2
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, introducing hydrogen sulfide into the GSCu precipitation suspension, and stirring at the rotating speed of 200 rpm for 50 minutes to react to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding phosphoric acid into the supernatant to adjust the pH value to 3, filtering the supernatant by using a filter membrane of 0.45 mu m to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on D001 macroporous strong-acid styrene cation exchange resin with the column volume of 1.5m multiplied by 20cm and the loading flow rate of 50mL/min, washing and balancing, and eluting by using a 2.0% hydrochloric acid solution with the elution flow rate of 100mL/min to obtain a GSH solution; concentrating the GSH solution under-0.05 Mpa at 60 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 55% ethanol solution preheated to 60 ℃ into the glutathione concentrated solution, then adjusting the pH to 3 by using hydrochloric acid, slowly stirring at the rotating speed of 200 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 3 times by using an ethanol solution with the concentration of 70% to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product in vacuum at the temperature of 60 ℃ under the pressure of-0.07 Mpa to obtain 170.6 g of glutathione crystals, wherein the purity is 98.4 percent and the total yield is 71.1 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: 39.07 percent of C, 5.57 percent of H and 31.26 percent of O.
Example 3
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, introducing hydrogen sulfide into the GSCu precipitation suspension, and stirring and reacting at the rotating speed of 150 rpm for 45 minutes to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding dilute nitric acid into the supernatant to adjust the pH value to 2, filtering the supernatant by using a filter membrane of 0.45 mu m to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on SP207 macroporous adsorption resin, wherein the column volume is 1.5m multiplied by 20cm, the loading flow rate is 50mL/min, and after washing and balancing, eluting by using 20% ethanol aqueous solution, and the elution flow rate is 100mL/min to obtain a GSH solution; concentrating the GSH solution under-0.03 Mpa at 65 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 60% ethanol solution preheated to 65 ℃ into the glutathione concentrated solution, then adjusting the pH to 2 by using phosphoric acid, slowly stirring at the rotating speed of 250rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 4 times by using an ethanol solution with the concentration of 75% to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product under the pressure of-0.01 Mpa at the temperature of 60 ℃ in vacuum to obtain 178.6 g of glutathione crystals, wherein the purity is 98.9 percent and the total yield is 74.4 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: c39.09%, H5.59%, O31.23%.
Example 4
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (Containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, introducing hydrogen sulfide into the GSCu precipitation suspension, and stirring and reacting at the rotating speed of 200 rpm for 60 minutes to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding phosphoric acid into the supernatant to adjust the pH value to 2, filtering the supernatant by using a filter membrane of 0.45 mu m to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on SP207 macroporous adsorption resin, wherein the column volume is 1.5m multiplied by 20cm, the loading flow rate is 50mL/min, and after washing and balancing, eluting by using 20% ethanol aqueous solution, and the elution flow rate is 100mL/min to obtain a GSH solution; concentrating the GSH solution under-0.07 Mpa at 65 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 60% ethanol solution preheated to 55 ℃ into the glutathione concentrated solution, then adjusting the pH to 3 by using phosphoric acid, slowly stirring at the rotating speed of 200 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 4 times by using an ethanol solution with the concentration of 65 percent to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product under the pressure of-0.05 Mpa at the temperature of 60 ℃ in vacuum to obtain 164.4 g of glutathione crystals, wherein the purity is 98.6 percent and the total yield is 68.5 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: 39.07% of C, 5.59% of H and 31.25% of O.
Example 5
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, adding sodium sulfide into the GSCu precipitation suspension, and stirring and reacting at the rotating speed of 200 rpm for 60 minutes to generate Cu2S precipitation and replacement of glutathionePeptide to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding dilute nitric acid into the supernatant to adjust the pH to 2, filtering the supernatant with a 0.45-micrometer filter membrane to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on HD-8 macroporous strong acid resin, wherein the column volume is 1.5m multiplied by 20cm, the loading flow rate is 50mL/min, and after washing and balancing, eluting with 1.5% hydrochloric acid solution at the elution flow rate of 100mL/min to obtain GSH solution; concentrating the GSH solution under-0.06 Mpa at 55 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 55% ethanol solution preheated to 55 ℃ into the glutathione concentrated solution, then adjusting the pH to 4 by using hydrochloric acid, slowly stirring at the rotating speed of 150 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 2 times by using an ethanol solution with the concentration of 70% to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product under the pressure of-0.04 Mpa at the temperature of 60 ℃ in vacuum to obtain 153.8 g of glutathione crystals, wherein the purity is 98.2 percent and the total yield is 64.1 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: 39.10% of C, 5.56% of H and 31.26% of O.
Example 6
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, adding potassium sulfide into the GSCu precipitation suspension, and stirring and reacting at the rotating speed of 250rpm for 45 minutes to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding hydrochloric acid into the supernatant to adjust pH to 4, filtering the supernatant with 0.45 μm filter membrane to remove particulate impurities, collecting filtrate 3.5L, loading onto HD-8 macroporous strong acid resin,the column volume is 1.5m multiplied by 20cm, the sample loading flow rate is 50mL/min, after washing balance, 2.5% hydrochloric acid solution is used for elution, the elution flow rate is 100mL/min, and GSH solution is obtained; concentrating the GSH solution under 0Mpa at 65 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 55% ethanol solution preheated to 60 ℃ into the glutathione concentrated solution, then adjusting the pH to 3 by using hydrochloric acid, slowly stirring at the rotating speed of 200 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 3 times by using an ethanol solution with the concentration of 65% to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product under the pressure of-0.04 Mpa at the temperature of 60 ℃ in vacuum to obtain 150.5g of glutathione crystals, wherein the purity is 98.4 percent and the total yield is 62.7 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: c39.09%, H5.57% and O31.25%.
Example 7
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, adding ammonium sulfide into the GSCu precipitation suspension, and stirring at 150 rpm for 50 minutes to react to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding dilute nitric acid into the supernatant to adjust the pH value to 2, filtering the supernatant by using a filter membrane of 0.45 mu m to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on SP207 macroporous adsorption resin, wherein the column volume is 1.5m multiplied by 20cm, the loading flow rate is 50mL/min, and after washing and balancing, eluting by using 20% ethanol aqueous solution, and the elution flow rate is 100mL/min to obtain a GSH solution; concentrating the GSH solution under-0.02 Mpa at 60 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding glutathione to the glutathioneAdding 50% ethanol solution preheated to 55 deg.C into the peptide concentrated solution, adjusting pH to 3 with dilute nitric acid, slowly stirring at 150 rpm, and slowly precipitating crystal; after full crystallization, carrying out suction filtration, and washing for 2 times by using an ethanol solution with the concentration of 75% to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product under the pressure of-0.06 Mpa at the temperature of 60 ℃ in vacuum to obtain 153.1 g of glutathione crystals, wherein the purity is 98.4 percent and the total yield is 63.8 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: c39.09%, H5.59%, O31.22%.
Example 8
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, introducing hydrogen sulfide into the GSCu precipitation suspension, and stirring and reacting at the rotating speed of 150 rpm for 60 minutes to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding dilute nitric acid into the supernatant to adjust the pH value to 3, filtering the supernatant by using a filter membrane of 0.45 mu m to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on HD-8 macroporous strong acid resin, wherein the column volume is 1.5m multiplied by 20cm, the loading flow rate is 50mL/min, and after washing and balancing, eluting by using 1.5% hydrochloric acid and 0.3mol/L sodium chloride aqueous solution, and the elution flow rate is 100mL/min to obtain a GSH solution; concentrating the GSH solution under-0.07 Mpa at 65 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 60% ethanol solution preheated to 60 ℃ into the glutathione concentrated solution, then adjusting the pH to 3 by using hydrochloric acid, slowly stirring at the rotating speed of 150 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 2 times by using an ethanol solution with the concentration of 65 percent to obtain a glutathione crystal crude product; finally, at-0.0And (3) drying the glutathione crystal crude product under the pressure of 5Mpa at the temperature of 60 ℃ in vacuum to obtain 178.8 g of glutathione crystal, wherein the purity is 98.7 percent and the total yield is 74.5 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: 39.06% of C, 5.56% of H and 31.27% of O.
Example 9
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, adding ammonium sulfide into the GSCu precipitation suspension, and stirring and reacting at the rotating speed of 250rpm for 60 minutes to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding hydrochloric acid into the supernatant to adjust the pH value to 3, filtering the supernatant with a 0.45-micron filter membrane to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on HD-8 macroporous strong acid resin with the column volume of 1.5m multiplied by 20cm and the loading flow rate of 50mL/min, washing and balancing, and eluting with 1.5% hydrochloric acid solution with the elution flow rate of 100mL/min to obtain GSH solution; concentrating the GSH solution under 0Mpa at 65 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 50% ethanol solution preheated to 60 ℃ into the glutathione concentrated solution, then adjusting the pH to 3 by using hydrochloric acid, slowly stirring at the rotating speed of 200 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 3 times by using an ethanol solution with the concentration of 75% to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product in vacuum at the temperature of 60 ℃ under the pressure of-0.02 Mpa to obtain 134.8g of glutathione crystals, wherein the purity is 98.5 percent and the total yield is 56.2 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: c39.09%, H5.59%, O31.23%.
Example 10
Adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate, and separating and purifying according to the following steps:
taking 600g of GSCu precipitate (containing 240g of GSH), adding pure water, fully stirring, and fixing the volume to 4L to obtain GSCu precipitate suspension; transferring the GSCu precipitation suspension into another container, adding ammonium sulfide into the GSCu precipitation suspension, and stirring and reacting at the rotating speed of 200 rpm for 60 minutes to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor; centrifuging the glutathione mother liquor by a centrifuge with the rotating speed of 6000 rpm to obtain supernatant containing glutathione; adding hydrochloric acid into the supernatant to adjust the pH value to 2, filtering the supernatant with a 0.45-micron filter membrane to remove particle impurities, collecting 3.5L of filtrate, loading the filtrate on SP207 macroporous adsorption resin, wherein the column volume is 1.5m multiplied by 20cm, the loading flow rate is 50mL/min, and after washing and balancing, eluting with 1.5% hydrochloric acid solution at the elution flow rate of 100mL/min to obtain GSH solution; concentrating the GSH solution under-0.07 Mpa at 65 deg.C under reduced pressure to reach GSH concentration of 400g/L to obtain glutathione concentrated solution; adding 50% ethanol solution preheated to 55 ℃ into the glutathione concentrated solution, then adjusting the pH to 3 by using phosphoric acid, slowly stirring at the rotating speed of 200 rpm, and slowly precipitating crystals; after full crystallization, carrying out suction filtration, and washing for 2 times by using an ethanol solution with the concentration of 65 percent to obtain a glutathione crystal crude product; and finally, drying the glutathione crystal crude product in vacuum at the temperature of 60 ℃ under the pressure of-0.07 Mpa to obtain 134.8g of glutathione crystals, wherein the purity is 98.6 percent and the total yield is 56.2 percent through High Performance Liquid Chromatography (HPLC).
Elemental analysis of the prepared glutathione crystals was as follows: theoretical value: 39.08 percent of C, 5.58 percent of H and 31.24 percent of O; measurement values: 39.07 percent of C, 5.57 percent of H and 31.26 percent of O.
The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered by the present invention.
Claims (8)
1. The method for efficiently separating and purifying the glutathione is characterized by comprising the following steps of:
(1) adding a freshly prepared cuprous oxide solution into a glutathione aqueous solution to prepare a GSCu precipitate; adding pure water into the GSCu precipitate, and fully stirring to ensure that the homogenate wet weight of the GSCu precipitate suspension is 300-500 g/L;
(2) adding sulfide into the GSCu precipitation suspension, and stirring for reaction to generate Cu2S, precipitating and replacing glutathione to prepare glutathione mother liquor;
(3) centrifuging the glutathione mother liquor to obtain supernatant containing glutathione;
(4) carrying out filter pressing or microfiltration on the supernatant;
(5) loading the filtrate obtained in the step (4) on cation exchange resin or macroporous adsorption resin, washing and balancing, and eluting with desorption solution to obtain glutathione solution;
(6) concentrating the glutathione solution under reduced pressure of-0.07 Mpa to 0Mpa at 55 to 65 ℃ to prepare glutathione concentrated solution;
(7) crystallizing the glutathione concentrated solution by adopting an ethanol solution to obtain a glutathione crystal crude product;
(8) vacuum drying the glutathione crystal crude product to obtain a pure glutathione crystal;
wherein, in the step (4), the aperture of the filter cloth or the filter membrane adopted by the filter pressing or the microfiltration is 0.45 μm;
the step (7) is as follows: adding 50-60% ethanol solution preheated to 55-65 ℃ into the glutathione concentrated solution, then adjusting the pH of the glutathione ethanol solution to 2-4 by using hydrochloric acid or phosphoric acid or nitric acid, slowly stirring at the rotating speed of 150-250 rpm, and slowly separating out crystals; and after full crystallization, carrying out suction filtration, and washing for 2-4 times by using an ethanol solution with the concentration of 65-75% to obtain a glutathione crystal crude product.
2. The method for efficiently separating and purifying glutathione according to claim 1, wherein the stirring reaction is performed at a stirring speed of 150 to 250rpm for 45 to 60min in the step (2).
3. The method for efficiently separating and purifying glutathione according to claim 1, wherein the sulfide is selected from any one of the following compounds: hydrogen sulfide, sodium sulfide, potassium sulfide and ammonium sulfide.
4. The method for efficiently separating and purifying glutathione according to claim 1, wherein the cation exchange resin is selected from any one of the following: d001 macroporous strong acid styrene cation exchange resin, HD-8 macroporous strong acid resin, D-61 macroporous strong acid styrene cation exchange resin; the macroporous adsorption resin is selected from any one of the following: SP207 macroporous adsorbent resin, XDA-1 macroporous adsorbent resin, DA201-C macroporous adsorbent resin.
5. The method for efficiently separating and purifying glutathione according to claim 1, wherein the step (3) further comprises: and adjusting the pH value of the supernatant to 2-4 by using hydrochloric acid or phosphoric acid or nitric acid.
6. The method for efficiently separating and purifying glutathione according to claim 1, wherein in the step (5), the desorption solution used when cation exchange resin is used is aqueous ammonium hydroxide solution, aqueous sodium chloride solution or hydrochloric acid; when the macroporous adsorption resin is adopted, the desorption solution correspondingly used is ethanol water solution.
7. The method for efficiently separating and purifying glutathione according to claim 1, wherein the concentration of the glutathione concentrate in the step (6) is 380 to 420 g/L.
8. The method for efficiently separating and purifying glutathione according to claim 1, wherein the step (8) comprises: and (3) drying the glutathione crystal crude product in vacuum at the temperature of 60 ℃ under the pressure of-0.07 Mpa to 0Mpa to obtain the glutathione crystal with the purity of more than or equal to 98 percent.
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