CN117244533A - Adsorbent for recycling and pretreatment of acetonitrile waste liquid and preparation method and application thereof - Google Patents
Adsorbent for recycling and pretreatment of acetonitrile waste liquid and preparation method and application thereof Download PDFInfo
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- CN117244533A CN117244533A CN202311306128.6A CN202311306128A CN117244533A CN 117244533 A CN117244533 A CN 117244533A CN 202311306128 A CN202311306128 A CN 202311306128A CN 117244533 A CN117244533 A CN 117244533A
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- adsorbent
- waste liquid
- acetonitrile
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- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 title claims abstract description 333
- 239000007788 liquid Substances 0.000 title claims abstract description 133
- 239000002699 waste material Substances 0.000 title claims abstract description 105
- 239000003463 adsorbent Substances 0.000 title claims abstract description 86
- 238000002360 preparation method Methods 0.000 title abstract description 17
- 238000004064 recycling Methods 0.000 title description 3
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 76
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- 238000011084 recovery Methods 0.000 claims abstract description 37
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- 238000004519 manufacturing process Methods 0.000 claims abstract description 14
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- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 13
- 238000005406 washing Methods 0.000 claims description 12
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 11
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- 238000006243 chemical reaction Methods 0.000 claims description 10
- -1 dichloro isophorone Chemical compound 0.000 claims description 10
- 230000015572 biosynthetic process Effects 0.000 claims description 7
- 238000012986 modification Methods 0.000 claims description 7
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- 238000003786 synthesis reaction Methods 0.000 claims description 7
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- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 claims description 6
- 239000004971 Cross linker Substances 0.000 claims description 5
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 230000004048 modification Effects 0.000 claims description 5
- 238000007781 pre-processing Methods 0.000 claims description 5
- 230000003213 activating effect Effects 0.000 claims description 4
- 230000035484 reaction time Effects 0.000 claims description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 3
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 claims description 3
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 3
- 229920001213 Polysorbate 20 Polymers 0.000 claims description 3
- 125000003158 alcohol group Chemical group 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
- 238000000855 fermentation Methods 0.000 claims description 3
- 230000004151 fermentation Effects 0.000 claims description 3
- 238000011010 flushing procedure Methods 0.000 claims description 3
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims description 3
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims description 3
- 239000012266 salt solution Substances 0.000 claims description 3
- 239000004094 surface-active agent Substances 0.000 claims description 3
- 125000003277 amino group Chemical group 0.000 claims description 2
- 238000001179 sorption measurement Methods 0.000 abstract description 56
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- 239000011159 matrix material Substances 0.000 description 16
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- 150000003839 salts Chemical class 0.000 description 9
- 238000001914 filtration Methods 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- HSJKGGMUJITCBW-UHFFFAOYSA-N 3-hydroxybutanal Chemical compound CC(O)CC=O HSJKGGMUJITCBW-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- HJOVHMDZYOCNQW-UHFFFAOYSA-N isophorone Chemical compound CC1=CC(=O)CC(C)(C)C1 HJOVHMDZYOCNQW-UHFFFAOYSA-N 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
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- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108010059712 Pronase Proteins 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 229920003180 amino resin Polymers 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
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- 238000001976 enzyme digestion Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 235000020793 low-cost food Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- SYSQUGFVNFXIIT-UHFFFAOYSA-N n-[4-(1,3-benzoxazol-2-yl)phenyl]-4-nitrobenzenesulfonamide Chemical class C1=CC([N+](=O)[O-])=CC=C1S(=O)(=O)NC1=CC=C(C=2OC3=CC=CC=C3N=2)C=C1 SYSQUGFVNFXIIT-UHFFFAOYSA-N 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000005373 pervaporation Methods 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000002992 thymic effect Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/265—Synthetic macromolecular compounds modified or post-treated polymers
- B01J20/267—Cross-linked polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/34—Regenerating or reactivating
- B01J20/345—Regenerating or reactivating using a particular desorbing compound or mixture
- B01J20/3475—Regenerating or reactivating using a particular desorbing compound or mixture in the liquid phase
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/28—Treatment of water, waste water, or sewage by sorption
- C02F1/288—Treatment of water, waste water, or sewage by sorption using composite sorbents, e.g. coated, impregnated, multi-layered
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/34—Nature of the water, waste water, sewage or sludge to be treated from industrial activities not provided for in groups C02F2103/12 - C02F2103/32
- C02F2103/343—Nature of the water, waste water, sewage or sludge to be treated from industrial activities not provided for in groups C02F2103/12 - C02F2103/32 from the pharmaceutical industry, e.g. containing antibiotics
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- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Life Sciences & Earth Sciences (AREA)
- Hydrology & Water Resources (AREA)
- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
- Water Supply & Treatment (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the technical field of acetonitrile waste liquid treatment, and in particular relates to an adsorbent for acetonitrile waste liquid recovery pretreatment, a preparation method and application thereof, wherein the adsorbent comprises the following components: an adsorbent containing a functional group, an activator or a cross-linking agent solution, and a protease solution. Compared with the existing adsorbent, the method has the advantages that the regeneration process of the selected adsorbent is simple, the regeneration temperature is 25-35 ℃, the eluent is pure water, and the dosage is low; the invention has stable performance and is suitable for the pretreatment of acetonitrile waste liquid produced in any link in the production process of polypeptide medicaments; the method can greatly remove the residual polypeptide in the acetonitrile waste liquid, degrade the adsorbed polypeptide in the adsorption process, reduce the difficulty of subsequent purification and recovery of the acetonitrile waste liquid, and simultaneously avoid secondary waste liquid pollution.
Description
Technical Field
The invention belongs to the technical field of acetonitrile waste liquid treatment, and particularly relates to an adsorbent for acetonitrile waste liquid recovery pretreatment and a preparation method and application thereof.
Background
With the rapid development of biotechnology at present, the research and application of the polypeptide have great breakthroughs, and the polypeptide is of great concern in the fields of pharmacy, skin care and food. Because the polypeptide can influence various cell functions in organisms, the polypeptide relates to the fields of cell growth, hormone and nerve, has high medicinal value and specific therapeutic effect in clinic, and many polypeptide medicaments have been successfully used for treating various diseases such as diabetes, cancer, obesity, cardiovascular diseases and the like. The market size of the current global polypeptide drugs exceeds 2000 hundred million yuan, the growth is rapid, and the annual average composite growth rate reaches 7.9%.
However, the polypeptide drug synthesis-purification-modification process is complex, and especially the purification process involves multiple steps of column chromatography, high Performance Liquid Chromatography (HPLC), gel electrophoresis, and the whole production process can generate organic solvent waste liquid, and a large amount of organic solvent is required as a reaction medium, an activator, a dissociating agent, and the like in the synthesis process. Acetonitrile is a common organic solvent in the polypeptide pharmaceutical field, and can provide good conditions for a plurality of links of polypeptide synthesis-purification-modification due to the unique solubility and the reaction activity, so that high yield and high purity of polypeptide synthesis are realized. Therefore, a large amount of acetonitrile solvent is required in the synthesis of polypeptide medicaments, 30-50L of acetonitrile waste liquid is produced per gram of polypeptide medicaments according to statistics, the volume of the produced acetonitrile waste liquid is continuously increased along with the mass production of the polypeptide medicaments, heavy environmental protection pressure is brought to the production, acetonitrile is used as a volatilizable organic compound, the atmospheric environment is polluted, and if the acetonitrile is directly discharged, the water source and soil are seriously damaged.
Considering the universality of acetonitrile used in polypeptide medicine production and the potential influence of acetonitrile waste liquid on the environment, the recovery of acetonitrile becomes an important problem in the polypeptide medicine production process, the adverse influence on the environment can be reduced, and the method has important social value, which is an important medical intermediate solvent, has high price, and the recovery of acetonitrile can greatly reduce the cost and has important economic value.
The acetonitrile waste liquid generally contains 10% -30% of acetonitrile, about 50% -70% of water, and the rest is residual polypeptide and part of organic impurities, polypeptide chains are gradually synthesized in the synthesis process, a large number of unreacted complete polypeptide fragments and free amino acids are in the acetonitrile waste liquid, and besides, during the purification and washing process, a plurality of isomers of target peptides exist, part of carrier proteins, fusion proteins and enzyme digestion residual proteases can be left in the acetonitrile waste liquid, and some polypeptide chains exist in the modification stage. The most common methods used for solvent recovery in the industry at present are rectification, extraction, membrane separation and the like, but the existence of residual polypeptide in acetonitrile waste liquid causes great trouble to the acetonitrile recovery process, and the direct use of the method is easy to block a pipeline and difficult to clean. Patent CN114315641a provides a set of purification flow of acetonitrile waste liquid, firstly, acetonitrile crude product is obtained by azeotropic or rectification mode, then excessive water is removed by adopting a mode of passing through a membrane, but acetonitrile waste liquid produced in the polypeptide production process contains a large amount of residual polypeptide, the separation can not be carried out by directly using a rectifying tower, the pipeline is blocked by the solution, and the subsequent cleaning is also a great difficulty. Patent CN107445863a provides a method for recovering acetonitrile-containing waste liquid generated in biopharmaceutical production, which uses dichloromethane as an extractant to extract the acetonitrile-containing waste liquid, and uses fractionation and rectification treatment of the extract to recover acetonitrile product, but the process introduces new impurities, thereby increasing the difficulty of subsequent acetonitrile purification and recovery. Patent CN1634876A provides a method for separating and recycling acetonitrile solution from production waste liquid of thymic pentapeptide, adopts macroporous resin to adsorb acetonitrile waste liquid, but the regeneration method of the adsorption resin in the process is complex, and needs NaOH solution or Na solution first 2 CO 3 The macroporous resin is washed by the solution, and then is washed by hydrochloric acid solution for the second time, the eluent still contains the residual adsorbed polypeptide, and the eluent also becomes a new waste liquid to wait for treatment. Patent CN111138319a designs a set of acetonitrile recovery device, which comprises a conditioning tank, a pervaporation membrane separator, a first rectifying tower, a second rectifying tower, a third rectifying tower and an oxidation tank, and has complicated steps and high equipment investment. At present, no simple, efficient and low-cost food is availableThe method is used for recovering and treating acetonitrile waste liquid in the production process of polypeptide medicaments.
The design and preparation of the adsorbent are all the time important in research at home and abroad, and the adsorbent such as molecular sieve, silica gel, resin and the like has the advantages of large specific surface area, high pore volume and the like, has good stability in aqueous solution, and has good application prospect of polypeptide adsorption. The covalent coupling mode is used for modifying the adsorbent, and the grafted substance with enzyme activity plays an important role in degradation after polypeptide adsorption, so that the pressure of subsequent acetonitrile purification and the pressure of residual polypeptide adsorption after treatment can be relieved. The invention aims to provide an adsorbent for pretreatment of acetonitrile waste liquid, which is used for adsorbing and degrading residual polypeptide and relieving the difficulty of subsequent purification of acetonitrile waste liquid.
Disclosure of Invention
The invention aims to provide an adsorbent for recovering and preprocessing acetonitrile waste liquid, and a preparation method and application thereof, which can greatly remove residual polypeptide and reduce the difficulty of subsequent purification recovery of acetonitrile waste liquid and residual polypeptide adsorption post-treatment.
The invention aims at realizing the following technical scheme:
the invention provides an adsorbent for recovering and preprocessing acetonitrile waste liquid, which comprises the following components: an adsorbent containing a functional group, an activator or a cross-linking agent solution, and a protease solution.
Further, the adsorbent containing a functional group is an adsorbent containing any one of an amino group, a carboxyl group and an alcohol group.
Further, the adsorbent comprises mesoporous resin, macroporous resin, silica gel and molecular sieve.
Further, the activator or crosslinker solution comprises any one or two of N-hydroxysuccinimide, carboxyl activator, disulfide crosslinker, dichloro isophorone and glutaraldehyde.
Further, the concentration of the protease solution is 1-10mg/mL.
The invention also provides a preparation method of the adsorbent for recovering and preprocessing the acetonitrile waste liquid, which comprises the following steps:
(1) Adding an activating agent or a cross-linking agent solution into the adsorbent containing the functional groups, stirring at room temperature for reaction, and flushing with deionized water until no residual activating agent or cross-linking agent exists after the reaction is completed to obtain a treated adsorbent;
(2) Mixing the treated adsorbent with protease solution, stirring at room temperature for reaction, centrifuging, washing with detergent, and drying at room temperature to obtain adsorbent.
Further, in the step (1), the molar ratio of the adsorbent containing the functional group to the activator or cross-linking agent solution is 1:0.05-0.5, and the stirring reaction time is 0.5-6h.
Further, in the step (2), the solid-to-liquid ratio of the treated adsorbent to the protease solution is 1:1-10, the stirring reaction time is 2-12h, and the detergent is a salt solution or a surfactant comprising Tris-HCl and Tween-20.
The invention also provides application of the adsorbent in recovery pretreatment of acetonitrile waste liquid.
Furthermore, the acetonitrile waste liquid is waste liquid produced in any link of polypeptide synthesis, fermentation, purification and modification.
The invention has the beneficial effects that:
(1) Compared with the existing adsorbent, the adsorbent has the advantages that the regeneration process of the selected adsorbent is simple, the regeneration temperature is 25-35 ℃, the eluent is pure water, and the dosage is low.
(2) The invention has stable performance and is suitable for the pretreatment of acetonitrile waste liquid produced in any link in the production process of polypeptide drugs.
(3) The method can greatly remove the residual polypeptide in the acetonitrile waste liquid, degrade the adsorbed polypeptide in the adsorption process, reduce the difficulty of subsequent purification and recovery of the acetonitrile waste liquid, and simultaneously avoid secondary waste liquid pollution.
(4) The invention can hydrolyze the residual adsorbed polypeptide, and avoid the generation of secondary peptide-containing waste liquid in the subsequent elution process.
Detailed Description
The adsorbent for recovering and pretreating the acetonitrile waste liquid provided by the embodiment of the invention is used for recovering and pretreating the acetonitrile waste liquid in polypeptide drug production, and the specific preparation process comprises the following steps:
(1) Adding an activator or cross-linking agent solution into an adsorbent containing proper functional groups, wherein the molar ratio of the adsorbent to the activator or cross-linking agent is 1:0.05-0.5, stirring and reacting for 0.5-6h at room temperature, and then flushing with deionized water until no residual activator or cross-linking agent exists, thus obtaining the treated adsorbent;
(2) Mixing the treated adsorbent with a protease solution according to a solid-to-liquid ratio of 1:1-10, stirring at room temperature for reaction for 2-12h, centrifuging, washing with a detergent, and drying at room temperature to obtain the adsorbent.
Wherein the acetonitrile waste liquid is the waste liquid produced in any link of polypeptide synthesis, fermentation, purification and modification; the adsorbent is hydrophilic adsorbent such as mesoporous resin, macroporous resin, silica gel, molecular sieve, etc.; suitable functional groups are amino, carboxyl, alcohol groups; the activator or cross-linking agent is N-hydroxysuccinimide, carboxyl activator, disulfide cross-linking agent, dichloro isophorone and glutaraldehyde; the concentration of the protease solution is 1-10mg/mL. The detergent can be Tris-HCl, tween-20 or other salt solution or surfactant.
In the process of using the prepared adsorbent for recovering and preprocessing acetonitrile waste liquid, the content of residual polypeptide in the acetonitrile waste liquid to be processed is 0.001-0.1mg/mL, the adsorption temperature is 4-30 ℃, the pH of the acetonitrile waste liquid is 5.5-8.5, the salt concentration in the acetonitrile waste liquid is less than or equal to 500mM, the solid-liquid ratio is 0.01-10mg/mL, the adsorption time in the processing is 30-120min, the adsorption capacity of residual polypeptide is 1-10mg/g, the residual peptide removal rate is more than 99%, and the adsorbed residual polypeptide is degraded into micromolecular amino acid.
The following experimental examples and examples serve to further illustrate the invention but are not limited thereto.
Example 1
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 1 comprises the following steps:
mixing amino silica gel and 2% glutaraldehyde solution according to a solid-to-liquid ratio of 1:1, stirring at 20 ℃ for 0.5h, filtering and washing with water to obtain a silica gel matrix containing glutaraldehyde crosslinking agent, mixing the obtained matrix with 1mg/mL protease solution according to a solid-to-liquid ratio of 1:1, stirring at 20 ℃ for reacting for 2h, and naturally drying at room temperature to obtain the adsorbent. Activity measurement of protease Fu Lin Fenfa in Pronase preparation GB/T23527-2009, the enzyme activity was measured and 1. Mu.g of casein was hydrolyzed per minute by 1g of enzyme solution at 20℃to give 1. Mu.g of tyrosine, which was defined as an enzyme activity unit (U). The recovery of the enzyme activity was calculated to be 14%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 6.5, the salt concentration is 90mM, and the content of residual polypeptide is 0.01 mg/mL) according to the solid-to-liquid ratio of 1mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for adsorption for 30min. After the adsorption is completed, collecting supernatant, measuring absorbance at 280nm, and determining adsorption quantity according to the value of absorbance measured before and after the adsorption, wherein the adsorption quantity reaches 10mg/g, and the removal rate of residual polypeptide in acetonitrile waste liquid reaches 99.8%.
Example 2
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 2 comprises the following steps:
mixing carboxyl silica gel and 0.01% glutaraldehyde solution according to a solid-to-liquid ratio of 1:1, stirring for 3 hours at 25 ℃, filtering and washing with water to obtain a silica gel matrix containing glutaraldehyde crosslinking agent, mixing the obtained matrix with 1mg/mL protease solution according to a solid-to-liquid ratio of 1:1, stirring for reacting for 2 hours at 20 ℃, and naturally drying at room temperature to obtain the adsorbent. The recovery rate of the enzyme activity was measured in the same manner as in example 1 and was 26%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 6.8, the salt concentration is 100mM, and the content of residual polypeptide is 0.1 mg/mL) according to the solid-to-liquid ratio of 10mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for adsorption for 30min. After the adsorption was completed, the adsorption amount and the removal rate of the residual polypeptide in the acetonitrile waste liquid were measured in the same manner as in example 1, the adsorption amount was 9.2mg/g, and the removal rate of the residual polypeptide in the acetonitrile waste liquid was 99.6%.
Example 3
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 3 comprises the following steps:
mixing alcohol-based silica gel and 0.5% glutaraldehyde solution according to a solid-to-liquid ratio of 1:1, stirring for 6 hours at 30 ℃, filtering and washing with water to obtain a silica gel matrix containing glutaraldehyde crosslinking agent, mixing the obtained matrix with a protease solution with a solid-to-liquid ratio of 1:10, stirring for reaction for 12 hours at 20 ℃, and naturally drying at room temperature to obtain the adsorbent. The recovery rate of the enzyme activity was measured in the same manner as in example 1 and found to be 46%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 7.2, the salt concentration is 40mM, the content of residual polypeptide is 0.001 mg/mL) according to the solid-to-liquid ratio of 1mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for adsorption for 120min. After the adsorption was completed, the adsorption amount and the removal rate of the residual polypeptide in the acetonitrile waste liquid were measured in the same manner as in example 1, the adsorption amount was 1mg/g, and the removal rate of the residual polypeptide in the acetonitrile waste liquid was 99.9%.
Example 4
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 4 comprises the following steps:
mixing amino resin and 0.05% glutaraldehyde solution according to a solid-to-liquid ratio of 1:5, stirring for 3h at 30 ℃, filtering and washing with water to obtain a resin matrix containing glutaraldehyde crosslinking agent, mixing the obtained matrix with 1mg/mL protease solution according to a solid-to-liquid ratio of 1:5, stirring for reaction for 3h at 25 ℃, and naturally drying at room temperature to obtain the adsorbent. The recovery rate of the enzyme activity was measured in the same manner as in example 1 and found to be 34%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 8, the salt concentration is 60mM, and the content of residual polypeptide is 0.01 mg/mL) according to the solid-to-liquid ratio of 1mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for 100min for adsorption. After the adsorption was completed, the adsorption amount and the removal rate of the residual polypeptide in the acetonitrile waste liquid were measured in the same manner as in example 1, the adsorption amount was 8.4mg/g, and the removal rate of the residual polypeptide in the acetonitrile waste liquid was 99.2%.
Example 5
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 5 comprises the following steps:
mixing an amino molecular sieve and 0.1% glutaraldehyde solution according to a solid-to-liquid ratio of 1:5, stirring for 3 hours at 30 ℃, filtering and washing with water to obtain a molecular sieve matrix containing glutaraldehyde crosslinking agent, mixing the obtained matrix with 2mg/mL protease solution according to a solid-to-liquid ratio of 1:3, stirring and reacting for 6 hours at 25 ℃, and naturally drying at room temperature to obtain the adsorbent. The recovery rate of the enzyme activity was measured in the same manner as in example 1 and was 40%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 7.5, the salt concentration is 100mM, the content of residual polypeptide is 0.01 mg/mL) according to the solid-to-liquid ratio of 1mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for adsorption for 100min. After the adsorption was completed, the adsorption amount and the removal rate of the residual polypeptide in the acetonitrile waste liquid were measured in the same manner as in example 1, the adsorption amount was 9.6mg/g, and the removal rate of the residual polypeptide in the acetonitrile waste liquid was 99.8%.
Example 6
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 6 comprises the following steps:
mixing amino silica gel and 0.1% isophorone solution according to a solid-to-liquid ratio of 1:1, stirring at 30 ℃ for 2 hours, filtering and washing with water to obtain a silica gel matrix containing isophorone, mixing the obtained matrix with 1mg/mL protease solution according to a solid-to-liquid ratio of 1:1, stirring at 25 ℃ for reacting for 10 hours, and naturally drying at room temperature to obtain the adsorbent. The recovery rate of the enzyme activity was measured in the same manner as in example 1 and found to be 37%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 8.5, the salt concentration is 500mM, the content of residual polypeptide is 0.01 mg/mL) according to the solid-to-liquid ratio of 1mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for adsorption for 120min. After the adsorption was completed, the adsorption amount and the removal rate of the residual polypeptide in the acetonitrile waste liquid were measured in the same manner as in example 1, the adsorption amount was 8.6mg/g, and the removal rate of the residual polypeptide in the acetonitrile waste liquid was 99.1%.
Example 7
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 7 comprises the following steps:
mixing amino silica gel and 1% polyethylene glycol solution according to a solid-to-liquid ratio of 1:3, stirring for 4 hours at 30 ℃, filtering and washing with water to obtain a silica gel matrix containing polyethylene glycol, mixing the obtained matrix with 2mg/mL protease solution according to a solid-to-liquid ratio of 1:1, stirring and reacting for 6 hours at 25 ℃, and naturally drying at room temperature to obtain the adsorbent. The recovery rate of the enzyme activity was measured in the same manner as in example 1 and found to be 34%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 7, the salt concentration is 20mM, and the content of residual polypeptide is 0.001 mg/mL) according to the solid-to-liquid ratio of 0.01mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for adsorption for 120min. After the adsorption was completed, the adsorption amount and the removal rate of the residual polypeptide in the acetonitrile waste liquid were measured in the same manner as in example 1, the adsorption amount was 5.8mg/g, and the removal rate of the residual polypeptide in the acetonitrile waste liquid was 99.4%.
Example 8
The preparation method of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid in the embodiment 8 comprises the following steps:
mixing alcohol-based resin and 0.5% dimeric aldol solution according to a solid-to-liquid ratio of 1:2, stirring for 3 hours at 20 ℃, filtering and washing with water to obtain a dimeric aldol-containing resin matrix, mixing the obtained matrix with 1mg/mL protease solution according to a solid-to-liquid ratio of 1:1, stirring and reacting for 10 hours at 25 ℃, and naturally drying at room temperature to obtain the adsorbent. The recovery rate of the enzyme activity was measured in the same manner as in example 1 and found to be 33%.
In the adsorption process, the adsorbent is added into acetonitrile waste liquid (the pH of the acetonitrile waste liquid is 6.8, the salt concentration is 50mM, and the content of residual polypeptide is 0.001 mg/mL) according to the solid-to-liquid ratio of 1mg/mL, and the mixture is placed in a shaking table at the constant temperature of 30 ℃ for adsorption for 80min. After the adsorption was completed, the adsorption amount and the removal rate of the residual polypeptide in the acetonitrile waste liquid were measured in the same manner as in example 1, the adsorption amount was 1.6mg/g, and the removal rate of the residual polypeptide in the acetonitrile waste liquid was 99.1%.
The removal rate of the residual polypeptide in the acetonitrile waste liquid is known by referring to examples 1-8, the use of the adsorbent for the recovery pretreatment of the acetonitrile waste liquid prepared by the invention overcomes the problems of complicated steps, high cost, introduction of new impurities and the like in the existing acetonitrile waste liquid purification process, expands the research of using the enzyme active substance modified adsorbent for the pretreatment of the acetonitrile waste liquid, and provides a novel scheme for using adsorption materials such as resin, silica gel, molecular sieve and the like as adsorbents, using covalent coupling or crosslinking enzyme active substances for the pretreatment of the acetonitrile waste liquid, and adsorbing and degrading the residual polypeptide in the acetonitrile waste liquid. The adsorbent prepared by the invention has high adsorption rate and fast adsorption rate on the polypeptide, the adsorption process can degrade the polypeptide, the adsorbent can be recycled, the regeneration process is simple, the residual polypeptide can be removed greatly by using the adsorbent prepared by the invention to pretreat the acetonitrile waste liquid, and the difficulty of purification recovery of the subsequent acetonitrile waste liquid and the residual polypeptide adsorption post-treatment is reduced.
Finally, it is noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered by the scope of the claims of the present invention.
Claims (10)
1. The adsorbent for recovering and preprocessing the acetonitrile waste liquid is characterized by comprising the following components: an adsorbent containing a functional group, an activator or a cross-linking agent solution, and a protease solution.
2. The adsorbent for pretreatment of acetonitrile waste liquid recovery according to claim 1, wherein the adsorbent containing a functional group is an adsorbent containing any one of an amino group, a carboxyl group and an alcohol group.
3. The adsorbent for pretreatment of acetonitrile waste liquid recovery according to claim 2, wherein the adsorbent containing functional groups comprises a mesoporous resin, a macroporous resin, silica gel and a molecular sieve.
4. The adsorbent for pretreatment of acetonitrile waste liquid recovery according to claim 1, wherein the activator or cross-linker solution comprises any one or two of N-hydroxysuccinimide, carboxyl activator, disulfide cross-linker, dichloro isophorone and glutaraldehyde.
5. The adsorbent for pretreatment of acetonitrile waste liquid recovery according to claim 1, wherein the concentration of the protease solution is 1-10mg/mL.
6. A method for producing the adsorbent for pretreatment of acetonitrile waste liquid recovery according to any one of claims 1 to 5, comprising the steps of:
(1) Adding an activating agent or a cross-linking agent solution into the adsorbent containing the functional groups, stirring at room temperature for reaction, and flushing with deionized water until no residual activating agent or cross-linking agent exists after the reaction is completed to obtain a treated adsorbent;
(2) Mixing the treated adsorbent with protease solution, stirring at room temperature for reaction, centrifuging, washing with detergent, and drying at room temperature to obtain adsorbent.
7. The method for producing an adsorbent for pretreatment of acetonitrile waste liquid recovery according to claim 6, wherein in the step (1), a molar ratio of the adsorbent containing a functional group to the activator or crosslinker solution is 1:0.05-0.5, and the stirring reaction time is 0.5-6 hours.
8. The method according to claim 6, wherein in the step (2), the solid-to-liquid ratio of the treated adsorbent to the protease solution is 1:1-10, the stirring reaction time is 2-12 hours, and the detergent is a salt solution or a surfactant including Tris-HCl and Tween-20.
9. Use of an adsorbent according to any one of claims 1 to 5 in the recovery pretreatment of acetonitrile waste liquid.
10. The use according to claim 9, wherein the acetonitrile waste liquid is a waste liquid produced at any stage of polypeptide synthesis, fermentation, purification and modification.
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| CN118022701A (en) * | 2024-04-11 | 2024-05-14 | 齐鲁工业大学(山东省科学院) | Small-pore silica gel clarifying agent and preparation method and application thereof |
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| CN118022701A (en) * | 2024-04-11 | 2024-05-14 | 齐鲁工业大学(山东省科学院) | Small-pore silica gel clarifying agent and preparation method and application thereof |
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