CN114262297A - Preparation method of ergothioneine - Google Patents
Preparation method of ergothioneine Download PDFInfo
- Publication number
- CN114262297A CN114262297A CN202111642632.4A CN202111642632A CN114262297A CN 114262297 A CN114262297 A CN 114262297A CN 202111642632 A CN202111642632 A CN 202111642632A CN 114262297 A CN114262297 A CN 114262297A
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- China
- Prior art keywords
- temperature
- ergothioneine
- preparation
- filtering
- water
- 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.)
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- SSISHJJTAXXQAX-ZETCQYMHSA-N L-ergothioneine Chemical compound C[N+](C)(C)[C@H](C([O-])=O)CC1=CNC(=S)N1 SSISHJJTAXXQAX-ZETCQYMHSA-N 0.000 title claims abstract description 37
- 229940093497 ergothioneine Drugs 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- 238000001914 filtration Methods 0.000 claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 230000008569 process Effects 0.000 claims abstract description 10
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 10
- 239000002244 precipitate Substances 0.000 claims abstract description 9
- 239000000706 filtrate Substances 0.000 claims abstract description 8
- 239000007788 liquid Substances 0.000 claims abstract description 8
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000012528 membrane Substances 0.000 claims abstract description 7
- 238000003756 stirring Methods 0.000 claims abstract description 7
- 239000003480 eluent Substances 0.000 claims abstract description 6
- 238000000605 extraction Methods 0.000 claims abstract description 6
- 230000008014 freezing Effects 0.000 claims abstract description 6
- 238000007710 freezing Methods 0.000 claims abstract description 6
- 238000010411 cooking Methods 0.000 claims abstract description 5
- 239000011347 resin Substances 0.000 claims abstract description 5
- 229920005989 resin Polymers 0.000 claims abstract description 5
- 238000001132 ultrasonic dispersion Methods 0.000 claims abstract description 5
- 238000001816 cooling Methods 0.000 claims abstract description 4
- 238000001179 sorption measurement Methods 0.000 claims abstract description 3
- 238000001694 spray drying Methods 0.000 claims abstract description 3
- 238000004821 distillation Methods 0.000 claims description 4
- 229920006122 polyamide resin Polymers 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 238000002386 leaching Methods 0.000 abstract description 10
- 238000011084 recovery Methods 0.000 abstract description 3
- 239000004480 active ingredient Substances 0.000 abstract description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 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 description 4
- 239000000047 product Substances 0.000 description 4
- PSLIMVZEAPALCD-UHFFFAOYSA-N ethanol;ethoxyethane Chemical compound CCO.CCOCC PSLIMVZEAPALCD-UHFFFAOYSA-N 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 108010024636 Glutathione Proteins 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 229960003180 glutathione Drugs 0.000 description 2
- 208000006820 Arthralgia Diseases 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 208000018937 joint inflammation Diseases 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
- C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D233/84—Sulfur atoms
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention belongs to the field of preparation of active ingredients, and particularly relates to a preparation method of ergothioneine, which comprises the following steps: step 1, adding the mycelium into water, cooking for 20-30min, freezing for 10-20min, then crushing at low temperature, dissolving and filtering to obtain filtrate and precipitate; step 2, putting the precipitate into an ethanol water solution, extracting at constant temperature for 30-60min, performing ultrasonic dispersion for 20-30min, and filtering to obtain an alcohol extract; step 3, combining and stirring the alcohol extract and the water extract, distilling at constant temperature under reduced pressure for 2-3h, then cooling and filtering, distilling at reduced pressure for the second time, and filtering to obtain a concentrated clear solution; and 4, introducing the concentrated clear liquid into an ultrafiltration membrane for ultrafiltration treatment, then carrying out resin exchange adsorption on the filtrate to obtain an eluent, and finally carrying out spray drying on the eluent to obtain the ergothioneine. The invention solves the process difficulty of the existing ergothioneine, greatly improves the extraction amount of the ergothioneine by combining water leaching and alcohol leaching, and achieves the purpose of improving the recovery rate of the ergothioneine.
Description
Technical Field
The invention belongs to the field of preparation of active ingredients, and particularly relates to a preparation method of ergothioneine.
Background
Ergothioneine is an important active substance in the body, but cannot be synthesized by the animal body per se and can only be taken in from food. Foreign research shows that the glutathione can play an antioxidant role in organisms, has various physiological functions of eliminating free radicals, maintaining DNA biosynthesis, normal cell growth, cellular immunity and the like, can be used for developing outdoor skin care products, protective cosmetics and ophthalmic products, can also be used as an ideal substitute of glutathione, and has gradually attracted more attention. It has been experimentally confirmed that human and animal ingestion of ergothioneine is safe, and dietary supplements developed in the united states using ergothioneine as a raw material for alleviating joint pain and inflammation have been marketed.
The ergothioneine is obtained mainly by chemical synthesis and separation from natural raw materials. However, chemically synthesized ergothioneine has limited its application in the fields of food, medicine and health care products due to the presence of other impurities such as by-products. For the methods using natural sources, most of the methods reported in the patent literature use alumina columns for separation, followed by preparative separation using high performance liquid chromatography. The existing natural raw material extraction method has the problems that the ergothioneine extract obtained by the existing extraction method has low content, generally only about 1-5%, high-content products are difficult to obtain, and large-scale production is difficult to realize.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a preparation method of ergothioneine, which solves the process difficulty of the existing ergothioneine, and greatly improves the extraction amount of the ergothioneine by combining water leaching and alcohol leaching so as to achieve the aim of improving the recovery rate of the ergothioneine.
In order to achieve the technical purpose, the technical scheme of the invention is as follows:
a preparation method of ergothioneine comprises the following steps:
step 1, adding the mycelium into water, cooking for 20-30min, freezing for 10-20min, then crushing at low temperature, dissolving and filtering to obtain filtrate and precipitate; the mass ratio of the mycelium to the water is 5:4-6, the cooking temperature is 100-120 ℃, and the freezing temperature is below-10 ℃; the low-temperature crushing temperature is below 0 ℃;
step 2, putting the precipitate into an ethanol water solution, extracting at constant temperature for 30-60min, performing ultrasonic dispersion for 20-30min, and filtering to obtain an alcohol extract; extracting at constant temperature of 50-60 deg.C, ultrasonic frequency of ultrasonic dispersion of 50-80kHz, temperature of 50-60 deg.C, and filtering temperature of 40-50 deg.C; the volume of the ethanol in the ethanol water solution accounts for 40-60%;
step 3, combining and stirring the alcohol extract and the water extract, distilling at constant temperature under reduced pressure for 2-3h, then cooling and filtering, distilling at reduced pressure for the second time, and filtering to obtain a concentrated clear liquid, wherein the temperature of constant-temperature reduced-pressure distillation is 70-80 ℃, the pressure is 80-90% of the atmospheric pressure, the temperature of cooling and filtering is 5-10 ℃, the pressure of secondary reduced-pressure distillation is 70-80% of the atmospheric pressure, the temperature is 90-100 ℃, and the concentrated clear liquid is 10-30% of the volume of the liquid before concentration;
and 4, introducing the concentrated clear liquid into an ultrafiltration membrane for ultrafiltration treatment, then carrying out resin exchange adsorption on the filtrate to obtain an eluent, and finally carrying out spray drying on the eluent to obtain the ergothioneine. The ultrafiltration membrane adopts a 1-10kDa ultrafiltration membrane, the pore diameter is 100-300nm, and the resin adopts polyamide resin.
The purification method of the ergothioneine comprises the following steps: 1, adding ergothioneine into ether-ethanol solution, stirring uniformly, and then evaporating at constant temperature to remove ether to obtain ethanol solution, wherein the concentration of ergothioneine in the ether-ethanol solution is 20-50g/L, the volume ratio of ether to ethanol in the ether-ethanol solution is 3:4-7, the stirring speed for stirring uniformly is 1000-2000r/min, and the temperature for constant temperature evaporation is 40-50 ℃; a2, filtering the ethanol solution to obtain a filtrate, standing and aging the filtrate overnight, and filtering to obtain a precipitate; a3, dissolving the precipitate in ethanol water solution, stirring uniformly until the precipitate is completely dissolved, then aging twice overnight, filtering to obtain recrystallization, namely the purified ergothioneine, wherein the ethanol volume of the ethanol water solution accounts for 60-80%.
From the above description, it can be seen that the present invention has the following advantages:
1. the invention solves the process difficulty of the existing ergothioneine, greatly improves the extraction amount of the ergothioneine by combining water leaching and alcohol leaching, and achieves the purpose of improving the recovery rate of the ergothioneine.
2. The invention coagulates protein and other materials by combining water leaching with high-temperature leaching to release ergothioneine, and simultaneously thins mycelium by combining freezing solidification with low-temperature crushing to form exposed surfaces of secondary leaching, thereby providing conditions for alcohol leaching.
Claims (8)
1. A preparation method of ergothioneine is characterized in that: the method comprises the following steps:
step 1, adding the mycelium into water, cooking for 20-30min, freezing for 10-20min, then crushing at low temperature, dissolving and filtering to obtain filtrate and precipitate;
step 2, putting the precipitate into an ethanol water solution, extracting at constant temperature for 30-60min, performing ultrasonic dispersion for 20-30min, and filtering to obtain an alcohol extract;
step 3, combining and stirring the alcohol extract and the water extract, distilling at constant temperature under reduced pressure for 2-3h, then cooling and filtering, distilling at reduced pressure for the second time, and filtering to obtain a concentrated clear solution;
and 4, introducing the concentrated clear liquid into an ultrafiltration membrane for ultrafiltration treatment, then carrying out resin exchange adsorption on the filtrate to obtain an eluent, and finally carrying out spray drying on the eluent to obtain the ergothioneine.
2. A process for the preparation of ergothioneine according to claim 1, wherein: the mass ratio of the mycelium to the water in the step 1 is 5:4-6, and the cooking temperature is 100-120 ℃.
3. A process for the preparation of ergothioneine according to claim 1, wherein: the freezing temperature in the step 1 is below-10 ℃; the low-temperature crushing temperature is below 0 ℃.
4. A process for the preparation of ergothioneine according to claim 1, wherein: the temperature of the constant-temperature extraction in the step 2 is 50-60 ℃, the ultrasonic frequency of the ultrasonic dispersion is 50-80kHz, the temperature is 50-60 ℃, and the filtering temperature is 40-50 ℃.
5. A process for the preparation of ergothioneine according to claim 1, wherein: the volume of the ethanol in the ethanol water solution in the step 2 accounts for 40-60%.
6. A process for the preparation of ergothioneine according to claim 1, wherein: the temperature of the constant-temperature reduced-pressure distillation in the step 3 is 70-80 ℃, the pressure is 80-90% of the atmospheric pressure, and the temperature of the temperature reduction filtration is 5-10 ℃.
7. A process for the preparation of ergothioneine according to claim 1, wherein: the pressure of the secondary reduced pressure distillation in the step 3 is 70-80% of the atmospheric pressure, the temperature is 90-100 ℃, and the concentrated clear liquid is 10-30% of the volume of the liquid before concentration.
8. A process for the preparation of ergothioneine according to claim 1, wherein: the ultrafiltration membrane in the step 4 is a 1-10kDa ultrafiltration membrane, the pore diameter is 100-300nm, and the resin is polyamide resin.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111642632.4A CN114262297A (en) | 2021-12-29 | 2021-12-29 | Preparation method of ergothioneine |
| CN202211333738.0A CN115536590A (en) | 2021-12-29 | 2022-10-28 | Preparation method of ergothioneine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111642632.4A CN114262297A (en) | 2021-12-29 | 2021-12-29 | Preparation method of ergothioneine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114262297A true CN114262297A (en) | 2022-04-01 |
Family
ID=80831696
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111642632.4A Pending CN114262297A (en) | 2021-12-29 | 2021-12-29 | Preparation method of ergothioneine |
| CN202211333738.0A Withdrawn CN115536590A (en) | 2021-12-29 | 2022-10-28 | Preparation method of ergothioneine |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211333738.0A Withdrawn CN115536590A (en) | 2021-12-29 | 2022-10-28 | Preparation method of ergothioneine |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN114262297A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114573731A (en) * | 2022-04-24 | 2022-06-03 | 青岛先迈海洋生物科技有限公司 | Extraction process of Chondrus crispus |
| CN118908894A (en) * | 2024-08-23 | 2024-11-08 | 江苏仅三生物科技有限公司 | Preparation method of novel ergothioneine crystal form |
-
2021
- 2021-12-29 CN CN202111642632.4A patent/CN114262297A/en active Pending
-
2022
- 2022-10-28 CN CN202211333738.0A patent/CN115536590A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114573731A (en) * | 2022-04-24 | 2022-06-03 | 青岛先迈海洋生物科技有限公司 | Extraction process of Chondrus crispus |
| CN118908894A (en) * | 2024-08-23 | 2024-11-08 | 江苏仅三生物科技有限公司 | Preparation method of novel ergothioneine crystal form |
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| Publication number | Publication date |
|---|---|
| CN115536590A (en) | 2022-12-30 |
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| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
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Application publication date: 20220401 |