WO2022120561A1 - 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 - Google Patents
一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 Download PDFInfo
- Publication number
- WO2022120561A1 WO2022120561A1 PCT/CN2020/134495 CN2020134495W WO2022120561A1 WO 2022120561 A1 WO2022120561 A1 WO 2022120561A1 CN 2020134495 W CN2020134495 W CN 2020134495W WO 2022120561 A1 WO2022120561 A1 WO 2022120561A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- chitosan
- water
- film
- additive
- preparation
- Prior art date
Links
- 239000012528 membrane Substances 0.000 title claims abstract description 39
- 241000700605 Viruses Species 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 19
- 239000004520 water soluble gel Substances 0.000 title abstract 3
- 229920001661 Chitosan Polymers 0.000 claims abstract description 41
- 239000000654 additive Substances 0.000 claims abstract description 34
- 230000000996 additive effect Effects 0.000 claims abstract description 34
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000000243 solution Substances 0.000 claims abstract description 26
- 239000007788 liquid Substances 0.000 claims abstract description 21
- 238000005266 casting Methods 0.000 claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 15
- 238000000034 method Methods 0.000 claims abstract description 14
- 230000015271 coagulation Effects 0.000 claims abstract description 8
- 238000005345 coagulation Methods 0.000 claims abstract description 8
- 150000003839 salts Chemical class 0.000 claims abstract description 8
- 239000000843 powder Substances 0.000 claims abstract description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 6
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000004202 carbamide Substances 0.000 claims abstract description 5
- 239000000706 filtrate Substances 0.000 claims abstract description 5
- 238000001914 filtration Methods 0.000 claims abstract description 5
- 238000004108 freeze drying Methods 0.000 claims abstract description 3
- 239000004014 plasticizer Substances 0.000 claims abstract description 3
- 239000012266 salt solution Substances 0.000 claims abstract description 3
- 239000004094 surface-active agent Substances 0.000 claims abstract description 3
- 239000002253 acid Substances 0.000 claims abstract 3
- 238000002156 mixing Methods 0.000 claims abstract 2
- 244000005700 microbiome Species 0.000 claims description 20
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 18
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 17
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 9
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 6
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 claims description 6
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 claims description 6
- 239000011148 porous material Substances 0.000 claims description 6
- 239000003929 acidic solution Substances 0.000 claims description 5
- HNJXPTMEWIVQQM-UHFFFAOYSA-M triethyl(hexadecyl)azanium;bromide Chemical group [Br-].CCCCCCCCCCCCCCCC[N+](CC)(CC)CC HNJXPTMEWIVQQM-UHFFFAOYSA-M 0.000 claims description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- 150000007522 mineralic acids Chemical class 0.000 claims description 4
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 claims description 4
- 241000894006 Bacteria Species 0.000 claims description 3
- 241000233866 Fungi Species 0.000 claims description 3
- 239000002202 Polyethylene glycol Substances 0.000 claims description 3
- 235000021355 Stearic acid Nutrition 0.000 claims description 3
- 229960003237 betaine Drugs 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 claims description 3
- 229920001223 polyethylene glycol Polymers 0.000 claims description 3
- 239000008117 stearic acid Substances 0.000 claims description 3
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 2
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 2
- 239000011248 coating agent Substances 0.000 claims description 2
- 238000000576 coating method Methods 0.000 claims description 2
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 2
- GVGUFUZHNYFZLC-UHFFFAOYSA-N dodecyl benzenesulfonate;sodium Chemical compound [Na].CCCCCCCCCCCCOS(=O)(=O)C1=CC=CC=C1 GVGUFUZHNYFZLC-UHFFFAOYSA-N 0.000 claims description 2
- 239000000194 fatty acid Substances 0.000 claims description 2
- 229930195729 fatty acid Natural products 0.000 claims description 2
- 239000000787 lecithin Substances 0.000 claims description 2
- 229940067606 lecithin Drugs 0.000 claims description 2
- 235000010445 lecithin Nutrition 0.000 claims description 2
- 229910017604 nitric acid Inorganic materials 0.000 claims description 2
- 229940080264 sodium dodecylbenzenesulfonate Drugs 0.000 claims description 2
- 238000009718 spray deposition Methods 0.000 claims description 2
- 238000005507 spraying Methods 0.000 claims description 2
- 229960004274 stearic acid Drugs 0.000 claims description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 claims 2
- 239000011707 mineral Substances 0.000 claims 2
- -1 sban Chemical compound 0.000 claims 1
- 238000001035 drying Methods 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 239000007864 aqueous solution Substances 0.000 description 6
- 108010025899 gelatin film Proteins 0.000 description 6
- 108020004707 nucleic acids Proteins 0.000 description 5
- 150000007523 nucleic acids Chemical class 0.000 description 5
- 102000039446 nucleic acids Human genes 0.000 description 5
- 238000010828 elution Methods 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 239000005714 Chitosan hydrochloride Substances 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 238000000889 atomisation Methods 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000013618 particulate matter Substances 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000000967 suction filtration Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 241001678559 COVID-19 virus Species 0.000 description 1
- 241000711573 Coronaviridae Species 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 241001112090 Pseudovirus Species 0.000 description 1
- 108020000999 Viral RNA Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000003595 mist Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000002688 persistence Effects 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 206010041232 sneezing Diseases 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D65/00—Accessories or auxiliary operations, in general, for separation processes or apparatus using semi-permeable membranes
- B01D65/10—Testing of membranes or membrane apparatus; Detecting or repairing leaks
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
- B01D71/06—Organic material
- B01D71/74—Natural macromolecular material or derivatives thereof
-
- 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
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J5/00—Manufacture of articles or shaped materials containing macromolecular substances
- C08J5/18—Manufacture of films or sheets
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L5/00—Compositions of polysaccharides or of their derivatives not provided for in groups C08L1/00 or C08L3/00
- C08L5/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
Definitions
- the present invention relates to a hydrosol membrane with virus enrichment function.
- the pandemic of the virus has had a huge impact on people's physical health and psychology.
- Some viruses have a long incubation period.
- the new coronavirus is also highly contagious during the incubation period. It can be transmitted through contact and air. The virus can survive outside for several days without drugs. Treatment has a long course of disease, many severe cases, runs on medical resources and can bring down the medical system of a region.
- Viral RNA can exist on particulate matter in the outdoor air, the virus can cluster with outdoor fine particulate matter, and by reducing the diffusion coefficient, the persistence of the virus in the atmosphere is enhanced.
- the virus can be transmitted through fine particles in the air, and the infected person will spread the virus when coughing, sneezing, talking or even breathing. Therefore, the detection of the virus in the air is particularly important for the prevention and treatment of the virus.
- Airborne microbial samplers can be summarized into: 1 Anderson solid impact sampler; 2 centrifugal sampler; 3 liquid impact sampler; 4 cyclone sampler; 5 large-capacity electrostatic sedimentation sampler; 6 filter sampling device.
- the solid impact and filter methods are mostly used and are superior to other collection methods.
- the filter collection combined with the polymerase chain reaction (PCR) detection method has more advantages than the traditional impact sampler combined with the cell culture method.
- the filter sampler can collect samples for a long time (>30h) and can sample multiple times on site, while the impact sampler is not suitable for low temperature and long time collection, nor is it suitable for multiple sampling on site.
- the purpose of the present invention is to provide a hydrosol gel membrane with a virus enrichment function
- the gel membrane has a 3D network structure, the membrane pores are dense and transparent, and has an efficient virus enrichment capability, and can be used in an aqueous solution. dissolves rapidly.
- One aspect of the present invention provides a method for preparing a water-soluble chitosan film, comprising the following steps:
- chitosan salt solution chitosan is reacted with an acidic solution, and a filtrate is obtained, which is dried to obtain chitosan salt powder;
- the additive A is a plasticizer, and the additive B is a surfactant;
- the coagulation liquid is selected from one or both of lower alcohol or acetone, preferably, the lower alcohol is selected from one or both of methanol, glycerol, ethanol, ethylene glycol, isopropanol, and tert-butanol. one or more combinations.
- the acidic solution is selected from the mixed solution of inorganic acid and lower alcohol, preferably, the inorganic acid is selected from hydrochloric acid, sulfuric acid or nitric acid; the lower alcohol is selected from methanol, glycerol, ethanol, ethyl alcohol A combination of one or more of glycol, isopropanol, tert-butanol.
- the volume ratio of the inorganic acid to the lower alcohol is 1-2:3-5.
- the acidic solution is selected from a mixture of hydrochloric acid and ethanol.
- the additive A is one or a combination of glycerol, ethylene glycol, polyethylene glycol, and propylene glycol.
- the additive B is cetyltriethylammonium bromide (CTAB), stearic acid, sodium dodecylbenzenesulfonate, lecithin, span, Tween, One or more of betaine and fatty acid glycerides.
- CTAB cetyltriethylammonium bromide
- stearic acid sodium dodecylbenzenesulfonate
- lecithin lecithin
- span span
- Tween One or more of betaine and fatty acid glycerides.
- the concentration of the urea solution is 0.1-2wt%, preferably 0.8-1.2wt%.
- the concentration of the additive A in the casting liquid is 0.1-10 wt%.
- the concentration of additive B in the casting liquid is 0.1-8 wt%.
- the concentration of the chitosan salt in the casting liquid is 0.1-10 wt%.
- the method for forming the film in step 3) is selected from blade coating, spray coating or casting.
- Another aspect of the present invention provides the water-soluble chitosan film obtained by the above preparation method.
- the water-soluble chitosan film has a structure of layer-by-layer stacking of membrane pores.
- Another aspect of the present invention provides an air filter device or component, which comprises the water-soluble chitosan membrane of the present invention as a filter material.
- Another aspect of the present invention provides the use of the water-soluble chitosan membrane as a filter material, preferably as a filter material for microorganisms in the air.
- Another aspect of the present invention provides a method for filtering and separating microorganisms, comprising the steps of:
- a method for detecting microorganisms in the air comprising the steps of:
- step iii) using the microorganism solution in step ii) for qualitative or quantitative detection.
- the microorganisms are selected from viruses, bacteria or fungi.
- the qualitative or quantitative detection includes culturing the microorganism, or directly detecting the nucleic acid of the microorganism.
- the traditional filter membrane uses a cellulose filter membrane, which is insoluble in water, because the virus particles are small and easy to be caught in the membrane gap, resulting in difficult elution, resulting in low actual detection sensitivity and difficult operation.
- the hydrosol membrane is directly dissolved in water after intercepting and enriching the virus, which can subvert the traditional filtration and elution mode, and use all the retained virus for detection, which greatly improves the detection sensitivity.
- this is also the first application of chitosan film in the field of virus (including but not limited to 2019-nCoV) enrichment and detection, which has good application prospects.
- the preparation process of the chitosan filter membrane of the present invention is simple, it has a 3D network structure, the membrane pores are dense and transparent, and it has efficient virus enrichment ability, and can be rapidly dissolved in an aqueous solution at the same time.
- Example 1 is a photo of the hydrosol membrane with virus enrichment function in Example 1 of the present invention; it can be seen from the figure that the surface of the hydrosol membrane is relatively flat.
- Example 2 is an electron microscope photo of the hydrosol membrane with virus enrichment function in Example 1 of the present invention; it can be seen from the figure that both sides of the hydrosol membrane have pores with a three-dimensional network structure, and the membrane pores are transparent.
- Example 3 is a comparison diagram of the hydrosol gel membrane with virus enrichment function in deionized water for two minutes before and after water solubility in Example 1 of the present invention; it can be seen from the figure that the hydrosol gel membrane is in deionized water It can be completely dissolved in water.
- the glass plate containing the gel film into the coagulation liquid for replacement (>24h) to obtain a chitosan wet film;
- the coagulation liquid is an alcohol solvent;
- Example 2 The same method as in Example 1 was adopted, except that the additive A was polyethylene glycol, the concentration of the additive A was 2 wt %, the additive B was stearic acid, and the concentration of the additive B was 6 wt %. Hydrosol gel films can also be prepared.
- Example 2 The same method as in Example 1 was adopted, except that the additive A was ethylene glycol, the concentration of additive A was 7 wt%, and the concentration of additive B was betaine, and the concentration of additive B was 1 wt%. Hydrosol gel films can also be prepared.
- the aqueous solution of dissolving the water-soluble membrane is extracted using nano-magnetic beads, and the nucleic acid in the aqueous solution dissolved in the water-soluble membrane is detected by automatic amplification and detection of the viral nucleic acid extraction product based on RT-PCR technology.
Abstract
一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用，具体公开了所述壳聚糖水溶性膜的制备方法，其包括以下步骤：1)壳聚糖盐溶液的制备：将壳聚糖与酸性溶液反应，并获得滤液，将干燥得到壳聚糖盐粉末；2)铸膜液的制备：将壳聚糖盐粉末与尿素溶液混合，加入添加剂A和添加剂B，得到铸膜液；3)将铸膜液制成膜；4)以凝固液固定所述膜；5)将固定后的膜冻干，得到壳聚糖水溶性膜；其中，所述添加剂A为增塑剂，添加剂B为表面活性剂；所述凝固液选自低级醇或丙酮中的一种或两种。所述水溶凝胶膜颠覆了现有技术中对于空气中富集检测领域的方法，无需过滤。
Description
本发明涉及一种具有病毒富集功能的水溶凝胶膜。
病毒的大流行对人们的身体健康以及心理造成了巨大影响，有些病毒潜伏期长，例如新冠病毒在潜伏期也极具传染性，接触和空气都能传播，病毒能在外界存活数天，没有药物能治疗，病程长、重症多、挤兑医疗资源并能拖垮一个地区的医疗系统。
病毒RNA可以存在于室外空气中的颗粒物上，病毒可以与室外细颗粒物成簇，并且通过降低扩散系数，增强病毒在大气中的持久性。病毒可通过空气中的细小颗粒物传播，感染者在咳嗽、打喷嚏、说话甚至呼吸状态下都会传播病毒，因此对空气中病毒的检测对病毒的防治尤为重要。
空气中微生物采样器归纳起来可分为：①安德森固体撞击式采样器；②离心式采样器；③液体冲击式采样器；④气旋式采样器；⑤大容量静电沉降采样器；⑥过滤式采样器。目前以固体撞击式和过滤式居多并优于其它采集方式，然而，过滤器收集结合聚合酶链式反应(PCR)的检测方法相对于传统的撞击式采样器结合细胞培养的方法更具优势，过滤式采样器可以进行长时间(>30h)的样品收集，可现场多次采样，而撞击式采样器不适于低温和长时间收集，也不适用现场多次采样。
普通的过滤式滤膜采用纤维素类滤膜不可水溶，因为病毒颗粒较小容易夹在膜缝隙中，造成洗脱困难，导致实际检测灵敏度不高且操作难度大。因此，若能采用水溶性膜，在截留富集病毒之后直接溶于水，则可以颠覆传统过滤洗脱模式，将截留下的病毒全部用于检测，极大地提高检测灵敏度。
发明内容
有鉴于此，本发明目的在于提供一种具有病毒富集功能的水溶凝胶膜，该凝胶膜具有3D网络结构，膜孔致密且通透，具有高效的病毒富集能力，同时可以在水溶液中迅速溶解。
本发明一个方面提供了一种壳聚糖水溶性膜的制备方法，其包括以下步骤：
1)壳聚糖盐溶液的制备：将壳聚糖与酸性溶液反应，并获得滤液，将干燥得到壳聚糖盐粉末；
2)铸膜液的制备：将壳聚糖盐粉末与尿素溶液混合，加入添加剂A和添加剂B，得到铸 膜液；
3)将铸膜液制成膜；
4)以凝固液固定所述膜；
5)将固定后的膜冻干，得到壳聚糖水溶性膜；
其中，所述添加剂A为增塑剂，添加剂B为表面活性剂；
所述凝固液选自低级醇或丙酮中的一种或两种，优选地，所述的低级醇选自甲醇、丙三醇、乙醇、乙二醇、异丙醇、叔丁醇中的一种或多种的组合。
在本发明的技术方案中，所述的酸性溶液选自无机酸与低级醇的混合溶液，优选地，无机酸选自盐酸、硫酸或硝酸；低级醇选自甲醇、丙三醇、乙醇、乙二醇、异丙醇、叔丁醇中的一种或多种的组合。在一些优选的实施方案中，所述无机酸与低级醇体积比为1～2∶3～5。在一个优选的实施例中，所述的酸性溶液选自盐酸与乙醇的混合物。
在本发明的技术方案中，所述的添加剂A是甘油、乙二醇、聚乙二醇、丙二醇中的一种或几种的组合。
在本发明的技术方案中，所述的添加剂B为十六烷基三乙基溴化铵(CTAB)、硬脂酸、十二烷基苯磺酸钠、卵磷脂、司班、吐温、甜菜碱、脂肪酸甘油酯中的一种或几种。
在本发明的技术方案中，尿素溶液的浓度为0.1-2wt％，优选为0.8-1.2wt％。
在本发明的技术方案中，铸膜液中添加剂A的浓度为0.1-10wt％。
在本发明的技术方案中，铸膜液中添加剂B的浓度为0.1-8wt％。
在本发明的技术方案中，铸膜液中壳聚糖盐的浓度为0.1-10wt％。
在本发明的技术方案中，步骤3)中制成膜的方法选自刮涂、喷涂或流延。
本发明另一个方面提供了上述制备方法获得的壳聚糖水溶性膜。
在本发明的技术方案中，所述的壳聚糖水溶性膜内部具有膜孔通透层层堆叠的结构。
本发明再一个方面提供了一种空气过滤装置或部件，其中包含本发明所述的壳聚糖水溶性膜作为过滤材料。
本发明再一个方面提供了壳聚糖水溶性膜作为过滤材料的用途，优选作为空气中微生物过滤材料的用途。
本发明再一个方面提供了一种过滤分离微生物的方法，其中包括以下步骤：
i)以本发明所述的壳聚糖水溶性膜作为过滤材料过滤空气中的微生物；
ii)将壳聚糖水溶性膜溶解，然后获得微生物的溶液。
一种检测空气中微生物的方法，其中包括以下步骤：
i)以权利要求6所述的壳聚糖水溶性膜作为过滤材料过滤空气中的微生物；
ii)将壳聚糖水溶性膜溶解，然后获得富集的微生物溶液；
iii)将步骤ii)中的微生物溶液用于定性或定量检测。
在本发明的技术方案中，所述的微生物选自病毒、细菌或真菌。
在本发明的技术方案中，所述定性或定量检测包括对于微生物的培养，或直接检测微生物的核酸。
1)传统的过滤式滤膜采用纤维素类滤膜不可水溶，因为病毒颗粒较小容易夹在膜缝隙中，造成洗脱困难，导致实际检测灵敏度不高且操作难度大。水溶凝胶膜在截留富集病毒之后直接溶于水，则可以颠覆传统过滤洗脱模式，将截留下的病毒全部用于检测，极大地提高检测灵敏度。而且，这也是壳聚糖膜首次应用于病毒(包括并不仅限于新冠病毒)富集检测领域，具有很好的应用前景。
2)本发明的壳聚糖滤膜制备工艺简单，其具有3D网络结构，膜孔致密且通透，具有高效的病毒富集能力，同时可以在水溶液中迅速溶解。
图1为本发明实施例一中具有病毒富集功能的水溶凝胶膜照片；从图中可以看出水溶凝胶膜表面较为平整。
图2为本发明实施例一中具有病毒富集功能的水溶凝胶膜电镜照片；从图中可以看出水溶凝胶膜两面都具有三维网络结构的孔，且膜孔通透。
图3为本发明实施例一中具有病毒富集功能的水溶凝胶膜在去离子水中两分钟前后情况，水溶前和水溶后的对比图；从图中可以看出水溶凝胶膜在去离子水中可以完全彻底溶解。
为了使本发明的上述目的、特征和优点能够更加明显易懂，下面对本发明的具体实施方式做详细的说明，但不能理解为对本发明的可实施范围的限定。
实施例1制备水溶凝胶膜
1)将196g壳聚糖粉末与784mL盐酸乙醇溶液(盐酸溶液与乙醇体积比1.5∶4)混合均匀，搅拌反应2h，过滤得滤液575mL和滤渣262.5g，所述滤液备用，用500mL乙醇洗涤所述滤渣，合并滤液后65℃真空干燥箱中烘干、粉碎至80目，得到壳聚糖盐酸盐219g。
2)取上述制备得到的壳聚糖盐酸盐溶于1％的尿素水溶液中形成壳聚糖盐酸盐浓度为5wt％的混合溶液，随后加入适量添加剂A、添加剂B，使添加剂A浓度为5wt％，添加剂B的浓度为4wt％，强力搅拌0.5～3h，得到铸膜液；所述的添加剂A是甘油；所述的添加剂B为CTAB。
3)将上述铸膜液倒在玻璃板上刮制成膜；
4)将含有凝胶膜的玻璃板放入凝固液中进行置换(>24h)，得到壳聚糖湿膜；所述凝固液是醇溶剂；
5)将得到的壳聚糖湿膜冻干(>24h)，即可得到具有病毒富集功能的水溶凝胶膜。
实施例2制备水溶凝胶膜
采用和实施例1相同的方法，区别仅在于盐酸溶液与乙醇体积比1∶5。也能够制备得到水溶凝胶膜。
实施例3制备水溶凝胶膜
采用和实施例1相同的方法，区别仅在添加剂A为聚乙二醇，添加剂A浓度为2wt％，添加剂B为硬脂酸，添加剂B的浓度为6wt％。也能够制备得到水溶凝胶膜。
实施例4制备水溶凝胶膜
采用和实施例1相同的方法，区别仅在添加剂A为乙二醇，添加剂A浓度为7wt％，，添加剂B为甜菜碱，添加剂B的浓度为1wt％。也能够制备得到水溶凝胶膜。
实施例5病毒富集实验
1)消毒灭菌，开始抽滤实验前，将实验仪器(抽吸泵表面、雾化器、烧杯)用75％酒精消毒烘干；
2)根据实验需求，选择不同浓度假病毒样(5-10mL)放入雾化器中，选择实施例1-4中的任意一种水溶凝胶膜固定在抽吸泵上；
3)将抽吸泵置于雾化系统中，关闭顶盖以形成密闭空间，开启雾化器，工作1～5min，以保证雾化系统中水雾分布均匀；
4)接通抽吸泵电源，根据不同需求，抽滤不同时长(1～60min)；
5)抽滤结束，将抽吸泵取出，卸下水溶膜溶解于去离子水中；
6)核酸检测溶解水溶膜的水溶液。使用纳米磁珠对水溶液中的病毒核酸进行提取，针对病毒核酸提取产物基于RT-PCR技术自动化扩增检测溶解水溶膜的水溶液中的核酸。
Claims (10)
- 一种壳聚糖水溶性膜的制备方法，其包括以下步骤：1)壳聚糖盐溶液的制备：将壳聚糖与酸性溶液反应，并获得滤液，将干燥得到壳聚糖盐粉末；2)铸膜液的制备：将壳聚糖盐粉末与尿素溶液混合，加入添加剂A和添加剂B，得到铸膜液；3)将铸膜液制成膜；4)以凝固液固定所述膜；5)将固定后的膜冻干，得到壳聚糖水溶性膜；其中，所述添加剂A为增塑剂，添加剂B为表面活性剂；所述凝固液选自低级醇或丙酮中的一种或两种；优选地，铸膜液中壳聚糖盐的浓度为0.1-10wt％。
- 根据权利要求1所述的制备方法，所述的添加剂A是甘油、乙二醇、聚乙二醇、丙二醇中的一种或几种的组合；优选地，铸膜液中添加剂A的浓度为0.1-10wt％。
- 根据权利要求1所述的制备方法，所述的添加剂B为十六烷基三乙基溴化铵(CTAB)、硬脂酸，十二烷基苯磺酸钠、卵磷脂、司班、吐温、甜菜碱、脂肪酸甘油酯中的一种或几种；优选地，铸膜液中添加剂B的浓度为0.1-8t％。
- 根据权利要求1所述的制备方法，所述的酸性溶液选自无机酸与低级醇的混合溶液，优选地，无机酸选自盐酸、硫酸或硝酸；低级醇选自甲醇、丙三醇、乙醇、乙二醇、异丙醇、叔丁醇中的一种或多种的组合；优选地，所述无机酸与低级醇体积比为1-2∶3-5。
- 根据权利要求1所述的制备方法，步骤3)中制成膜的方法选自刮涂、喷涂或流延。
- 根据权利要求1-5任一项所述的制备方法获得的壳聚糖水溶性膜，壳聚糖水溶性膜内部具有膜孔通透层层堆叠的结构。
- 一种空气过滤装置或部件，其中包含权利要求6所述的壳聚糖水溶性膜作为过滤材料。
- 权利要求6所述的壳聚糖水溶性膜作为空气过滤材料的用途。
- 一种过滤分离空气中微生物的方法，其中包括以下步骤：i)以权利要求6所述的壳聚糖水溶性膜作为过滤材料过滤空气中的微生物；ii)将壳聚糖水溶性膜溶解，然后获得富集的微生物的溶液；优选地；所述的微生物选自病毒、细菌或真菌。
- 一种检测空气中微生物的方法，其中包括以下步骤：i)以权利要求6所述的壳聚糖水溶性膜作为过滤材料过滤空气中的微生物；ii)将壳聚糖水溶性膜溶解，然后获得富集的微生物溶液；iii)将步骤ii)中的微生物溶液用于定性或定量检测；优选地；所述的微生物选自病毒、细菌或真菌。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2020/134495 WO2022120561A1 (zh) | 2020-12-08 | 2020-12-08 | 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/CN2020/134495 WO2022120561A1 (zh) | 2020-12-08 | 2020-12-08 | 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2022120561A1 true WO2022120561A1 (zh) | 2022-06-16 |
Family
ID=81973989
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2020/134495 WO2022120561A1 (zh) | 2020-12-08 | 2020-12-08 | 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2022120561A1 (zh) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1480039A (zh) * | 2003-06-12 | 2004-03-10 | �Ϻ���ͨ��ѧ | 一种病原微生物滤杀材料及其制法 |
| CN1593735A (zh) * | 2004-06-29 | 2005-03-16 | 浙江大学 | 用于空气中细菌和病毒采样检测滤膜的制备方法 |
| CN101177494A (zh) * | 2007-10-26 | 2008-05-14 | 天津商业大学 | 以甘油和硬脂酸为增塑剂的壳聚糖复合膜的制备方法 |
| CN106310978A (zh) * | 2016-10-17 | 2017-01-11 | 江苏新风环保科技有限公司 | 一种基于壳聚糖和氧化石墨烯的复合空气滤膜及其制备方法 |
| CN111518296A (zh) * | 2019-02-01 | 2020-08-11 | 武汉大学 | 一种壳聚糖膜材料的制备方法 |
| CN112675718A (zh) * | 2020-12-08 | 2021-04-20 | 深圳先进技术研究院 | 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 |
-
2020
- 2020-12-08 WO PCT/CN2020/134495 patent/WO2022120561A1/zh active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1480039A (zh) * | 2003-06-12 | 2004-03-10 | �Ϻ���ͨ��ѧ | 一种病原微生物滤杀材料及其制法 |
| CN1593735A (zh) * | 2004-06-29 | 2005-03-16 | 浙江大学 | 用于空气中细菌和病毒采样检测滤膜的制备方法 |
| CN101177494A (zh) * | 2007-10-26 | 2008-05-14 | 天津商业大学 | 以甘油和硬脂酸为增塑剂的壳聚糖复合膜的制备方法 |
| CN106310978A (zh) * | 2016-10-17 | 2017-01-11 | 江苏新风环保科技有限公司 | 一种基于壳聚糖和氧化石墨烯的复合空气滤膜及其制备方法 |
| CN111518296A (zh) * | 2019-02-01 | 2020-08-11 | 武汉大学 | 一种壳聚糖膜材料的制备方法 |
| CN112675718A (zh) * | 2020-12-08 | 2021-04-20 | 深圳先进技术研究院 | 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 |
Non-Patent Citations (3)
| Title |
|---|
| HUI LI, ZHE WANG, HAIYAN ZHANG, ZHIJUAN PAN : "Nanoporous PLA/(Chitosan Nanoparticle) Composite Fibrous Membranes with Excellent Air Filtration and Antibacterial Performance", POLYMERS, vol. 10, no. 10, 30 September 2018 (2018-09-30), pages 1085, XP055941009 * |
| LI, JIAZHE: "Development of Chitosan as Membrane Materials", ACTA AGRICULTURAE JIANGXI, JIANGXI SHENG NONGYE KEXUEYUAN, CN, vol. 22, no. 4, 1 April 2010 (2010-04-01), CN , pages 124 - 126, XP055941017, ISSN: 1001-8581, DOI: 10.19386/j.cnki.jxnyxb.2010.04.040 * |
| WANG ZHEN; YAN FENG; PEI HONGCHANG; LI JIANXIN; CUI ZHENYU; HE BENQIAO: "Antibacterial and environmentally friendly chitosan/polyvinyl alcohol blend membranes for air filtration", CARBOHYDRATE POLYMERS, APPLIED SCIENCE PUBLISHERS , LTD BARKING, GB, vol. 198, 21 June 2018 (2018-06-21), GB , pages 241 - 248, XP085437675, ISSN: 0144-8617, DOI: 10.1016/j.carbpol.2018.06.090 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Milton | What was the primary mode of smallpox transmission? Implications for biodefense | |
| CN104722285B (zh) | 一种细菌纤维素膜/多孔碳吸附剂及其制备 | |
| CN105111711B (zh) | 分解型净化甲醛的组合物及制备方法与应用 | |
| CN101565190B (zh) | 一种微米级超细氯化钠的制备方法 | |
| CN103724635B (zh) | 一种醋酸纤维素多孔微球的制备方法及其产品 | |
| CN109959649B (zh) | 一种具有化学发光性质的卷烟焦油提取物及其化学发光体系 | |
| CN108054387A (zh) | 一种钯-介孔氧化硅空心多核纳米催化材料的制备方法 | |
| CN110449127A (zh) | 一种生物炭基载铁复合材料制备方法及其应用 | |
| WO2022120561A1 (zh) | 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 | |
| CN112675718B (zh) | 一种具有病毒富集功能的水溶凝胶膜及其制备方法和应用 | |
| US6562583B1 (en) | Method for detecting microorganisms in gases | |
| CN103992712A (zh) | 一种自清洁空气型外墙涂料及其制备方法 | |
| CN102233257A (zh) | 一种医院甲醛污染清除剂及其制备方法 | |
| CN107362697B (zh) | 一种空气过滤膜的制备方法 | |
| CN106727446A (zh) | 成膜组合物、保护膜及应用 | |
| CN110368795A (zh) | 一种天然木质素甲醛净化剂及其制备方法 | |
| CN102489182B (zh) | 非离子表面活性剂凝胶气体分离膜及其制备方法 | |
| CN108373715B (zh) | 一种具有甲醛净化功能的涂料及其制备方法 | |
| CN103894057B (zh) | 一种空气净化器用复合空气净化剂 | |
| CN108148477A (zh) | 环保吸附墙纸基膜及其加工方法 | |
| CN113583950A (zh) | 一种制备干细胞活性因子的方法及其应用 | |
| CN103611433B (zh) | 一种壳聚糖/氯化锂混合膜及其制备方法和应用 | |
| CN109110796B (zh) | 一种可水分散的氧化铜纳米管及其胶体溶液和制备方法 | |
| CN102440962B (zh) | 丹皮酚微小的储库型载体及提高丹皮酚抗氧化能力的方法 | |
| CN111500423A (zh) | 一种采样液、气溶胶中rna病毒的采样系统及采样方法 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 20964507 Country of ref document: EP Kind code of ref document: A1 |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 20964507 Country of ref document: EP Kind code of ref document: A1 |