CN116059691B - Method for extracting active substances through fibroin nanofiber with high beta-sheet content - Google Patents
Method for extracting active substances through fibroin nanofiber with high beta-sheet content Download PDFInfo
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- 108010022355 Fibroins Proteins 0.000 title claims abstract description 95
- 239000002121 nanofiber Substances 0.000 title claims abstract description 93
- 239000013543 active substance Substances 0.000 title claims abstract description 52
- 238000000034 method Methods 0.000 title claims abstract description 36
- 239000011259 mixed solution Substances 0.000 claims abstract description 66
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 32
- 239000000243 solution Substances 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000002904 solvent Substances 0.000 claims abstract description 17
- 230000008569 process Effects 0.000 claims abstract description 14
- 238000000926 separation method Methods 0.000 claims abstract description 10
- 239000007864 aqueous solution Substances 0.000 claims description 33
- 238000000605 extraction Methods 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 6
- 230000008859 change Effects 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 4
- 239000004519 grease Substances 0.000 claims description 3
- 239000006185 dispersion Substances 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 2
- 238000000746 purification Methods 0.000 abstract description 2
- 239000000499 gel Substances 0.000 description 17
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 14
- JUUBCHWRXWPFFH-UHFFFAOYSA-N Hydroxytyrosol Chemical compound OCCC1=CC=C(O)C(O)=C1 JUUBCHWRXWPFFH-UHFFFAOYSA-N 0.000 description 12
- 239000000049 pigment Substances 0.000 description 10
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 7
- RUQCCAGSFPUGSZ-OBWQKADXSA-N Glucoraphanin Natural products C[S@](=O)CCCCC(=NS(=O)(=O)O)S[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RUQCCAGSFPUGSZ-OBWQKADXSA-N 0.000 description 7
- GMMLNKINDDUDCF-JRWRFYLSSA-N [(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] (1e)-5-[(r)-methylsulfinyl]-n-sulfooxypentanimidothioate Chemical compound C[S@@](=O)CCCC\C(=N/OS(O)(=O)=O)S[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GMMLNKINDDUDCF-JRWRFYLSSA-N 0.000 description 7
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 7
- 229960003471 retinol Drugs 0.000 description 7
- 235000020944 retinol Nutrition 0.000 description 7
- 239000011607 retinol Substances 0.000 description 7
- LADGBHLMCUINGV-UHFFFAOYSA-N tricaprin Chemical compound CCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCC)COC(=O)CCCCCCCCC LADGBHLMCUINGV-UHFFFAOYSA-N 0.000 description 7
- 239000004480 active ingredient Substances 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 229940095066 hydroxytyrosol Drugs 0.000 description 6
- 235000003248 hydroxytyrosol Nutrition 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 235000019198 oils Nutrition 0.000 description 6
- AGOYDEPGAOXOCK-KCBOHYOISA-N clarithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@](C)([C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)OC)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 AGOYDEPGAOXOCK-KCBOHYOISA-N 0.000 description 5
- 229960002626 clarithromycin Drugs 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 238000011068 loading method Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- BACYUWVYYTXETD-UHFFFAOYSA-N N-Lauroylsarcosine Chemical group CCCCCCCCCCCC(=O)N(C)CC(O)=O BACYUWVYYTXETD-UHFFFAOYSA-N 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 190000032366 miboplatin Chemical compound 0.000 description 3
- 229950002777 miboplatin Drugs 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108700004121 sarkosyl Proteins 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 229940124350 antibacterial drug Drugs 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 239000000017 hydrogel Substances 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000010491 poppyseed oil Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 241000255789 Bombyx mori Species 0.000 description 1
- 235000011293 Brassica napus Nutrition 0.000 description 1
- 240000008100 Brassica rapa Species 0.000 description 1
- 235000000540 Brassica rapa subsp rapa Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- -1 antibacterial drugs Substances 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- ARQRPTNYUOLOGH-UHFFFAOYSA-N chcl3 chloroform Chemical compound ClC(Cl)Cl.ClC(Cl)Cl ARQRPTNYUOLOGH-UHFFFAOYSA-N 0.000 description 1
- 238000000701 chemical imaging Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 229940115478 isopropyl lauroyl sarcosinate Drugs 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000010399 physical interaction Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- XLCIFRJORZNGEV-UHFFFAOYSA-N propan-2-yl 2-[dodecanoyl(methyl)amino]acetate Chemical compound CCCCCCCCCCCC(=O)N(C)CC(=O)OC(C)C XLCIFRJORZNGEV-UHFFFAOYSA-N 0.000 description 1
- 239000001054 red pigment Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000009210 therapy by ultrasound Methods 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/04—Solvent extraction of solutions which are liquid
- B01D11/0492—Applications, solvents used
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/68—Purification; separation; Use of additives, e.g. for stabilisation
- C07C37/70—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment
- C07C37/72—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment by liquid-liquid treatment
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C403/00—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone
- C07C403/06—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by singly-bound oxygen atoms
- C07C403/08—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by singly-bound oxygen atoms by hydroxy groups
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F15/00—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table
- C07F15/0006—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table compounds of the platinum group
- C07F15/0086—Platinum compounds
- C07F15/0093—Platinum compounds without a metal-carbon linkage
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/08—Hetero rings containing eight or more ring members, e.g. erythromycins
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0096—Purification; Precipitation; Filtration
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2601/00—Systems containing only non-condensed rings
- C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
- C07C2601/16—Systems containing only non-condensed rings with a six-membered ring the ring being unsaturated
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Cosmetics (AREA)
Abstract
The invention relates to the technical field of active substance separation and purification, in particular to a method for extracting active substances by high beta-sheet content fibroin nanofiber, which is characterized by comprising the following steps: (1) Uniformly dispersing a hydrophobic active substance in a hydrophobic solvent to obtain a first mixed solution; (2) Adding the fibroin nanofiber with high beta-sheet content into water to obtain a second mixed solution; (3) Adding the first mixed solution into the second mixed solution, keeping the first mixed solution at the upper layer of the second mixed solution in the adding process, and standing; (4) And when the concentration of the first mixed solution is constant, obtaining a treatment solution at the upper layer, and removing the treatment solution to obtain a target product. The invention can directly realize the water phase dispersion and use of the hydrophobic active substance, and has the advantages of easy removal of hydrophobic solvent and simple operation process.
Description
Technical Field
The invention relates to the technical field of active substance separation and purification, in particular to a method for extracting active substances through fibroin nanofibers with high beta-sheet content.
Background
The active substances such as fat-soluble pigments, vitamins, growth factors, antitumor drugs and the like have wide application prospects in various fields such as chemical imaging, cosmetics, tissue repair, tumor treatment and the like, most of the active substances are hydrophobic active substances, the active substances cannot be directly dispersed in water, and how to solve the problem that the dispersion and transfer of the active substances in an aqueous phase environment become bottlenecks limiting the practical application of the active substances.
With the development of nanotechnology, various kinds of nanocarriers have been successfully developed and used for loading and transferring of fat-soluble substances. The silk fibroin has good biocompatibility and adjustable degradation speed, has the capability of stabilizing active substances and promoting the dispersion/loading/controlled release of the active substances, and is a universal carrier for various types of active substances. The structure of the silk fibroin can be further optimized through modification to form different forms such as nano particles, microspheres, nano fibers, hydrogel and the like, so that the delivery requirements of different active substances are better met, and the active is ensured and meanwhile, the silk fibroin has a designable release behavior. At present, researchers have developed fibroin nanofibers (SFN) with stable structure and high beta-sheet content, the concentration can be regulated to form solution or gel, and active substances are loaded in a physical adsorption mode.
However, loading the active substances by the above-described method has the following disadvantages: (1) Because of its aqueous dispersion or hydrogel form, the loaded active substance needs to be water-soluble or water-dispersible, whereas the hydrophobic active substance is difficult to dissolve or disperse in water; (2) The silk fibroin nanofiber can reduce the oil-water interfacial tension, and can coat hydrophobic active substances in an oil-water system in an emulsifying mode, however, the hydrophobic solvent is difficult to remove by coating the active substances by the method.
Disclosure of Invention
Aiming at the defects existing in the prior art, the invention aims to provide a method for extracting active substances through fibroin nanofibers with high beta-sheet content, so that hydrophobic active substances can be dispersed in water, and the method has the advantage of easy removal of hydrophobic solvents.
The technical aim of the invention is achieved by the following technical scheme, namely a method for extracting active substances through fibroin nanofibers with high beta-sheet content, which comprises the following steps:
(1) Uniformly dispersing a hydrophobic active substance in a hydrophobic solvent to obtain a first mixed solution;
(2) Adding the fibroin nanofiber with high beta-sheet content into water to obtain a second mixed solution;
(3) Adding the first mixed solution into the second mixed solution, keeping the first mixed solution at the upper layer of the second mixed solution in the adding process, and standing;
(4) And when the concentration of the first mixed solution is constant, obtaining a treatment solution at the upper layer, and removing the treatment solution to obtain a target product.
In one embodiment, the hydrophobic active is an oil-soluble active or an oil-dispersible active.
In one embodiment, the hydrophobic solvent is a water-immiscible grease or a nonpolar organic solvent.
In one embodiment, the high beta-sheet content fibroin nanofibers have a length of 20-4000nm and a diameter of 5-30nm.
In one embodiment, the high β -sheet content fibroin nanofibers have a β -sheet content of greater than or equal to 30%.
In one embodiment, the second mixed solution comprises any one of a high beta-sheet content fibroin nanofiber aqueous solution and a high beta-sheet content fibroin nanofiber gel.
In one embodiment, the concentration of the high β -sheet content fibroin nanofibers in the second mixed liquor is less than or equal to 10 wt%.
In one embodiment, in the step (4), the consistency of the concentration of the first mixed solution is determined by observing a color change or chemical analysis.
In one embodiment, in the step (4), the treatment liquid is removed by a liquid separation or extraction method.
The method for extracting the active substances through the fibroin nanofiber with high beta-sheet content has the following beneficial effects:
firstly, the invention directly realizes the water phase dispersion and use of the hydrophobic active substances by utilizing the excellent adsorption capacity of the special structural silk proteins on the fat-soluble substances through a similar extraction method, does not involve complex carrier preparation and hydrophobic solvent removal processes, can recycle the hydrophobic solvent, has simple and controllable method, is easy to prepare stably, and has excellent industrial transformation prospect;
secondly, the invention realizes the dispersion of oil-soluble medicines through physical interaction, can realize the transfer of different active ingredients such as pigments, vitamins, antibacterial medicines, antitumor medicines and the like, can solve the problem of water phase application of active substances in multiple fields, and has excellent universality;
thirdly, the silk protein adopted by the invention has excellent biocompatibility, combines mild preparation conditions, can effectively keep the activity of the active ingredients, can avoid side effects of the carrier, and greatly improves the applicability of various active ingredients in the fields of cosmetics, medical treatment and the like.
Drawings
Fig. 1 is a flow chart of the steps of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are intended to facilitate the understanding of the present invention without any limitation thereto. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, definitions, will control.
The term "prepared from …" as used herein is synonymous with "comprising". The terms "comprising," "including," "having," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, step, method, article, or apparatus.
The conjunction "consisting of …" excludes any unspecified element, step or component. If used in a claim, such phrase will cause the claim to be closed, such that it does not include materials other than those described, except for conventional impurities associated therewith. When the phrase "consisting of …" appears in a clause of the claim body, rather than immediately following the subject, it is limited to only the elements described in that clause; other elements are not excluded from the stated claims as a whole.
When an equivalent, concentration, or other value or parameter is expressed as a range, preferred range, or a range bounded by a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when ranges of "1 to 5" are disclosed, the described ranges should be construed to include ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a numerical range is described herein, unless otherwise indicated, the range is intended to include its endpoints and all integers and fractions within the range.
The present invention will be described in detail below with reference to the accompanying drawings and examples.
Referring to fig. 1, a method for extracting active substances through high beta-sheet content fibroin nanofibers comprises the following steps:
(1) Uniformly dispersing a hydrophobic active substance in a hydrophobic solvent to obtain a first mixed solution;
(2) Adding the fibroin nanofiber with high beta-sheet content into water to obtain a second mixed solution;
(3) Adding the first mixed solution into the second mixed solution, keeping the first mixed solution at the upper layer of the second mixed solution in the adding process, and standing;
in actual operation, the adding speed of the first mixed solution needs to be controlled, the first mixed solution and the second mixed solution are prevented from being mixed, if the two mixed solutions are mixed, the high beta-folding content fibroin nanofiber has an emulsification-like effect, so that the emulsification reaction is easy to occur on the contact surface of the two solutions, the contact surface of the two solutions is not clear in limit, and the two solutions are difficult to separate;
(4) When the concentration of the first mixed solution is constant, the treatment solution at the upper layer is obtained, and the treatment solution is removed, so that the target product is obtained.
In actual operation, when the concentration of the first mixed solution is constant, the treatment solution of the upper layer is a hydrophobic solvent, but there are cases where the treatment solution is a hydrophobic solvent and a small amount of an unloaded hydrophobic active substance; the lower layer solution is a mixed solution of the second mixed solution and the hydrophobic active substance, and the target product is obtained after the treatment solution is removed.
Through the steps, the hydrophobic active substances can be distributed in an oil-free system by utilizing the excellent adsorption capacity of the special structural silk proteins on the fat-soluble substances and the method for directly extracting the hydrophobic active substances, so that the hydrophobic solvent is easy to directly remove and recycle, the operation process is simple, the stable preparation is easy, and the method has excellent industrial conversion prospect; the transfer of different active ingredients such as pigment, vitamin, antibacterial drugs, antitumor drugs and the like can be realized, and the problem of water phase application of active substances in multiple fields can be solved; in addition, the preparation conditions are mild, the activity of the active ingredients can be maintained, the side effects of the carrier are avoided, and the applicability of the active ingredients in multiple fields is improved.
In particular, the hydrophobic active is an oil-soluble active or an oil-dispersible active.
In particular, the hydrophobic solvent is a water-immiscible grease or a nonpolar organic solvent.
Specifically, the length of the fibroin nanofiber with high beta-sheet content is 20-4000nm, and the diameter is 5-30nm.
Specifically, the content of beta-sheet in the fibroin nanofiber with high beta-sheet content is more than or equal to 30%.
Specifically, the second mixed solution comprises any one of a fibroin nanofiber aqueous solution with high beta-sheet content and a fibroin nanofiber gel with high beta-sheet content.
Specifically, the concentration of the fibroin nanofiber with high beta-sheet content in the second mixed solution is less than or equal to 10w% wt.
Specifically, in the step (4), the consistency of the concentration of the first mixed solution is judged by observing the color change or chemical analysis.
Specifically, in the step (4), the treatment liquid is removed by a liquid separation or extraction method.
The following are specific examples:
example 1
The natural plant pigment dispersed by the high beta-sheet content fibroin nanofiber extraction oil comprises the following steps:
(1) Uniformly dispersing 0.25g of glucoraphanin in 5g of caprylic/capric triglyceride to obtain a first mixed solution;
(2) Preparing 0.5% high beta-sheet content fibroin nanofiber aqueous solution;
(3) Taking 5g of 0.5% high beta-sheet content fibroin nanofiber aqueous solution, slowly adding 5.25g of first mixed solution into 5g of high beta-sheet content fibroin nanofiber aqueous solution, keeping the first mixed solution at the upper layer of the high beta-sheet content fibroin nanofiber aqueous solution in the adding process, standing, wherein the red pigment can be observed to be transferred from the upper layer to the lower layer in the standing process, and the interface position has the darkest color;
(4) When the color of the upper layer solution is completely transferred to the lower layer, the upper layer is caprylic capric triglyceride, the lower layer is the high beta-folding content fibroin nanofiber aqueous solution containing the glucoraphanin, and the upper layer caprylic capric triglyceride is removed by a liquid separation or extraction method to obtain the high beta-folding content fibroin nanofiber aqueous solution containing the glucoraphanin.
The first mixed solution and the aqueous solution of the fibroin nanofiber with high beta-sheet content in the embodiment have the following change conditions:
| observation period | The first mixed solution is just added into silkworm with high beta-sheet content Upper layer of silk fibroin nanofiber aqueous solution | When the concentration of the first mixed solution is constant | Pouring the container with constant concentration of the first mixed solution 90° |
| Observation of phenomena | The upper layer is oil dispersed red natural pigment, and the lower layer Is fibroin nanofiber water with high beta-sheet content Solution | Migration of color to underlying high beta-sheet Fibroin-containing nanofiber water In the solution, the color is concentrated in oil water Interface(s) | Clear oil normally flows and high beta-adsorption of pigment Aqueous solution of folded fibroin-containing nanofibers Gel formation |
Examples using coloured actives show that the actives migrate slowly into aqueous fibroin nanofiber solutions with high beta-sheet content and highest concentration at the oil-water interface. After loading the active, the texture of the aqueous solution of high β -sheet content fibroin nanofibers may change, typically with reduced flowability, indicating that the active is bound to the high β -sheet content fibroin nanofibers, rather than simply migrating into the water without interacting with the high β -sheet content fibroin nanofibers.
Comparative example 1
On the basis of example 1, 5g of 0.5% high beta-sheet content fibroin nanofiber aqueous solution was replaced with 5g of deionized water, as follows:
(1) Uniformly dispersing 0.25g of glucoraphanin into 5g of caprylic-capric triglyceride to obtain a first mixed solution;
(2) 5g of deionized water was taken, the first mixture was added to deionized water, and left to stand without color transfer to the lower layer.
The experiment was performed on the aqueous solution of the fibroin nanofiber with high beta-sheet content containing glucoraphanin obtained in example 1 and the mixed solution containing glucoraphanin obtained in comparative example 1, and the steps are as follows:
the two solutions are respectively placed in a dialysis bag, the bag opening is fastened, and then the two solutions are respectively placed in water, so that the phenomenon is observed.
The two solutions were changed as follows:
| experimental objects | Comparative example 1 solution | The solution obtained in example 1 |
| Phenomenon (1) | The dialysis bag contains an aqueous solution with pigments migrating to the outside | No migration of pigments to the outside is observed in the dialysis bag |
Therefore, the fibroin nanofiber with high beta-sheet content can be proved to adsorb oil to disperse pigment, so that the pigment can not be freely transferred in water.
Example 2
The oil-soluble active substance retinol is extracted by the fibroin nanofiber with high beta-sheet content, and the steps are as follows:
(1) Uniformly dispersing 0.25g of retinol in 5g of caprylic/capric triglyceride to obtain a first mixed solution;
(2) Preparing 2% high beta-sheet content fibroin nanofiber gel, performing ultrasonic treatment on the gel for 10min, converting the gel into a solution, and reducing the length of the high beta-sheet content fibroin nanofiber from 2000nm to 100nm;
(3) Taking 5g of 2% high beta-sheet content fibroin nanofiber aqueous solution, slowly adding 5.25g of first mixed solution into the high beta-sheet content fibroin nanofiber aqueous solution, keeping the first mixed solution at the upper layer of the high beta-sheet content fibroin nanofiber aqueous solution in the adding process, standing, wherein the transfer of light yellow retinol from the upper layer to the lower layer can be observed in the standing process, and the color of an interface position is the deepest;
(4) After standing for 24 hours, the colors of the upper layer and the lower layer are not changed any more, the upper layer is caprylic capric triglyceride, the lower layer is a high beta-folding content fibroin nanofiber aqueous solution containing retinol, and the upper layer caprylic capric triglyceride is removed by a liquid separation or extraction method to obtain the high beta-folding content fibroin nanofiber aqueous solution containing retinol active substances.
Example 3
The hydroxytyrosol is extracted by the fibroin nanofiber with high beta-sheet content, and the steps are as follows:
(1) Dispersing 0.1g of hydroxytyrosol in 5g of isopropyl lauroyl sarcosinate to obtain a first mixed solution;
(2) Preparing 0.5% high beta-sheet content fibroin nanofiber aqueous solution;
(3) Taking 5g of 0.5% high beta-sheet content fibroin nanofiber aqueous solution, slowly adding 5.25g of first mixed solution into the high beta-sheet content fibroin nanofiber aqueous solution, keeping the first mixed solution at the upper layer of the high beta-sheet content fibroin nanofiber aqueous solution in the adding process, standing, and spontaneously transferring hydroxytyrosol to the lower layer;
(4) When the concentration of the first mixed solution in the container is constant, the upper layer solution in the container is lauroyl sarcosine isopropyl ester, the lower layer solution is a high beta-sheet content fibroin nanofiber aqueous solution containing hydroxytyrosol, and the upper layer lauroyl sarcosine isopropyl ester is removed by a liquid separation or extraction method to obtain the high beta-sheet content fibroin nanofiber aqueous solution containing hydroxytyrosol.
Example 4
The oil-soluble drug Miripla (Miriplat) is extracted by the fibroin nanofiber with high beta-sheet content, and the steps are as follows:
(1) Dispersing 0.1g of miboplatin in 5g of poppy seed oil iodized fatty acid ethyl ester to obtain a first mixed solution;
(2) Preparing 1% high beta-sheet content fibroin nanofiber gel;
(3) Taking 5g of 1% high beta-sheet content fibroin nanofiber gel, slowly adding 5.1g of first mixed solution into the high beta-sheet content fibroin nanofiber gel, keeping the first mixed solution at the upper layer of the high beta-sheet content fibroin nanofiber gel in the adding process, and standing;
(4) Standing for 12 hours, transferring 70% of the miboplatin from the upper layer to the high beta-sheet content fibroin nanofiber gel layer, basically balancing, and removing the upper layer oily solvent by a liquid separation or extraction method to obtain the high beta-sheet content fibroin nanofiber gel containing the miboplatin medicine.
Example 5
The oil-soluble antibacterial drug clarithromycin is extracted by the fibroin nanofiber with high beta-sheet content, and the steps are as follows:
(1) Dissolving 0.1g of clarithromycin in 5g of chloroform to obtain a first mixed solution;
(2) Preparing 0.5% high beta-sheet content fibroin nanofiber aqueous solution;
(3) Taking 5g of 0.5% high beta-sheet content fibroin nanofiber aqueous solution, slowly adding 5.1g of first mixed solution into the high beta-sheet content fibroin nanofiber aqueous solution, keeping the first mixed solution at the upper layer of the second mixed solution in the adding process, and standing;
(4) Standing for 48 hours, transferring clarithromycin from the upper layer to a high beta-sheet content fibroin nanofiber solution layer, and removing the upper oily solvent by a liquid separation or extraction method to obtain a high beta-sheet content fibroin nanofiber aqueous solution containing clarithromycin medicine.
Example 6
The oil-soluble active substance is extracted by the fibroin nanofiber with high beta-sheet content and the gradient material is prepared, and the steps are as follows:
in examples 1-4, the aqueous solution of high beta-sheet content fibroin nanofibers had a consistency after extraction of the actives, formed a gel, and the aqueous solution of high beta-sheet content fibroin nanofibers did not contain an oil agent. The high beta-sheet content fibroin nanofiber gel loaded with the glucoraphanin obtained in example 1 is spread first, and then the high beta-sheet content fibroin nanofiber gel loaded with the retinol obtained in example 2 is spread above the high beta-sheet content fibroin nanofiber gel, and the gradient material containing two layers of insoluble active substances can be obtained without mixing and homogenizing. And the gradient material obtained in example 6 may be subjected to concentration processing by dehydration, heating, or the like.
The concentrations of the raw materials used in examples 1 to 6 and comparative example 1 of the present invention are shown in the following table,
| using raw materials | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 | Example 6 | Comparative example 1 |
| Caprylic capric triglyceride | 5g | 5g | / | / | / | 10g | 5g |
| 0.5% aqueous fibroin nanofiber solution with high beta-sheet content | 5 g | / | 5g | / | 5 g | 5 g | / |
| 1% aqueous fibroin nanofiber solution with high beta-sheet content | / | / | / | 5 g | / | / | / |
| 2% aqueous fibroin nanofiber solution with high beta-sheet content | / | 5 g | / | / | / | 5 g | / |
| Deionized water | / | / | / | / | / | / | 5g |
| Turnip red element | 0.25g | / | / | / | / | 0.25g | 0.25g |
| Retinol | / | 0.25g | / | / | / | 0.25g | / |
| Lauroyl sarcosine isopropyl ester | / | / | 5g | / | / | / | / |
| Hydroxytyrosol | / | / | 0.1g | / | / | / | / |
| Miripla (Miriplat) | / | / | / | 0.1g | / | / | / |
| Iodized poppy seed oil fatty acid ethyl ester | / | / | / | 5g | / | / | / |
| Clarithromycin | / | / | / | / | 0.1g | / | / |
| Chloroform (chloroform) | / | / | / | / | 5g | / | / |
The foregoing examples illustrate only a few embodiments of the invention and are described in detail herein without thereby limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (6)
1. A method for extracting active substances by high beta-sheet content fibroin nanofibers, comprising the steps of:
(1) Uniformly dispersing a hydrophobic active substance in a hydrophobic solvent to obtain a first mixed solution;
(2) Adding the fibroin nanofiber with high beta-sheet content into water to obtain a second mixed solution;
(3) Adding the first mixed solution into the second mixed solution, keeping the first mixed solution at the upper layer of the second mixed solution in the adding process, and standing;
(4) When the concentration of the first mixed solution is constant, obtaining a treatment solution at an upper layer, and removing the treatment solution to obtain a target product;
the hydrophobic solvent is a nonpolar organic solvent, the hydrophobic active substance is an oil-soluble active substance or an oil-dispersible active substance, the length of the high beta-sheet content fibroin nanofiber is 20-4000nm, the diameter is 5-30nm, and the beta-sheet content in the high beta-sheet content fibroin nanofiber is more than or equal to 30%.
2. A method of extracting active substances by high β -sheet content fibroin nanofibers according to claim 1, wherein: the nonpolar organic solvent is water-insoluble grease.
3. A method of extracting active substances by high β -sheet content fibroin nanofibers according to claim 1, wherein: the second mixed solution comprises any one of a fibroin nanofiber aqueous solution with high beta-sheet content and a fibroin nanofiber gel with high beta-sheet content.
4. A method of extracting active substances by high β -sheet content fibroin nanofibers according to claim 1, wherein: the concentration of the fibroin nanofiber with high beta-sheet content in the second mixed solution is less than or equal to 10w% wt.
5. A method of extracting active substances by high β -sheet content fibroin nanofibers according to claim 1, wherein: in the step (4), the consistency of the concentration of the first mixed solution is judged by observing a color change or chemical analysis.
6. A method of extracting active substances by high β -sheet content fibroin nanofibers according to claim 1, wherein: in the step (4), the treatment liquid is removed by a liquid separation or extraction method.
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