CN113637468B - Difunctional fluorescent film for detecting mercury ions and glutathione, preparation and application - Google Patents
Difunctional fluorescent film for detecting mercury ions and glutathione, preparation and application Download PDFInfo
- Publication number
- CN113637468B CN113637468B CN202110921350.1A CN202110921350A CN113637468B CN 113637468 B CN113637468 B CN 113637468B CN 202110921350 A CN202110921350 A CN 202110921350A CN 113637468 B CN113637468 B CN 113637468B
- Authority
- CN
- China
- Prior art keywords
- fluorescent film
- esm
- glutathione
- mmol
- fluorescence
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 title claims abstract description 94
- 229960003180 glutathione Drugs 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 108010024636 Glutathione Proteins 0.000 title claims abstract description 21
- 229910052753 mercury Inorganic materials 0.000 title claims abstract description 13
- -1 mercury ions Chemical class 0.000 title claims abstract description 12
- 229940012466 egg shell membrane Drugs 0.000 claims abstract description 107
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims abstract description 70
- 239000004201 L-cysteine Substances 0.000 claims abstract description 33
- 235000013878 L-cysteine Nutrition 0.000 claims abstract description 33
- 238000001514 detection method Methods 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 22
- 238000006243 chemical reaction Methods 0.000 claims abstract description 18
- 238000012360 testing method Methods 0.000 claims abstract description 11
- 239000000243 solution Substances 0.000 claims description 32
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 26
- 239000012498 ultrapure water Substances 0.000 claims description 26
- 230000000007 visual effect Effects 0.000 claims description 16
- 238000005406 washing Methods 0.000 claims description 15
- 239000012528 membrane Substances 0.000 claims description 12
- 239000007864 aqueous solution Substances 0.000 claims description 11
- 238000010791 quenching Methods 0.000 claims description 11
- 230000000171 quenching effect Effects 0.000 claims description 11
- 238000002791 soaking Methods 0.000 claims description 9
- 102000002322 Egg Proteins Human genes 0.000 claims description 7
- 108010000912 Egg Proteins Proteins 0.000 claims description 7
- 230000001588 bifunctional effect Effects 0.000 claims description 7
- 238000005520 cutting process Methods 0.000 claims description 7
- 210000003278 egg shell Anatomy 0.000 claims description 7
- 230000035484 reaction time Effects 0.000 claims description 5
- 230000005284 excitation Effects 0.000 claims description 4
- 150000002500 ions Chemical class 0.000 claims description 4
- 229910021645 metal ion Inorganic materials 0.000 claims description 4
- 230000008859 change Effects 0.000 claims description 3
- 230000001419 dependent effect Effects 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000007605 air drying Methods 0.000 claims description 2
- 238000012545 processing Methods 0.000 claims description 2
- 238000012800 visualization Methods 0.000 claims 1
- 125000000415 L-cysteinyl group Chemical group O=C([*])[C@@](N([H])[H])([H])C([H])([H])S[H] 0.000 abstract description 34
- 239000010949 copper Substances 0.000 abstract description 29
- 238000011065 in-situ storage Methods 0.000 abstract description 20
- 239000000463 material Substances 0.000 abstract description 19
- 229910052802 copper Inorganic materials 0.000 abstract description 14
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 abstract description 14
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 abstract description 13
- 229910000365 copper sulfate Inorganic materials 0.000 abstract description 12
- 239000003638 chemical reducing agent Substances 0.000 abstract description 9
- 239000003223 protective agent Substances 0.000 abstract description 6
- 238000011156 evaluation Methods 0.000 abstract description 4
- 238000012544 monitoring process Methods 0.000 abstract description 4
- 239000002243 precursor Substances 0.000 abstract description 4
- 230000000694 effects Effects 0.000 description 11
- 238000006722 reduction reaction Methods 0.000 description 10
- 230000009467 reduction Effects 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 8
- 230000003287 optical effect Effects 0.000 description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000004833 X-ray photoelectron spectroscopy Methods 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 239000000835 fiber Substances 0.000 description 5
- 238000002189 fluorescence spectrum Methods 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- VVNCNSJFMMFHPL-VKHMYHEASA-N D-penicillamine Chemical compound CC(C)(S)[C@@H](N)C(O)=O VVNCNSJFMMFHPL-VKHMYHEASA-N 0.000 description 4
- CTENFNNZBMHDDG-UHFFFAOYSA-N Dopamine hydrochloride Chemical compound Cl.NCCC1=CC=C(O)C(O)=C1 CTENFNNZBMHDDG-UHFFFAOYSA-N 0.000 description 4
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 4
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 4
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 4
- GSDSWSVVBLHKDQ-JTQLQIEISA-N Levofloxacin Chemical compound C([C@@H](N1C2=C(C(C(C(O)=O)=C1)=O)C=C1F)C)OC2=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-JTQLQIEISA-N 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 4
- 229960003405 ciprofloxacin Drugs 0.000 description 4
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 4
- 235000018417 cysteine Nutrition 0.000 description 4
- 229960001149 dopamine hydrochloride Drugs 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000015784 hyperosmotic salinity response Effects 0.000 description 4
- 229960003376 levofloxacin Drugs 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 229960001639 penicillamine Drugs 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000011068 loading method Methods 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 239000004475 Arginine Substances 0.000 description 2
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 2
- YTGJWQPHMWSCST-UHFFFAOYSA-N Tiopronin Chemical compound CC(S)C(=O)NCC(O)=O YTGJWQPHMWSCST-UHFFFAOYSA-N 0.000 description 2
- 108010058907 Tiopronin Proteins 0.000 description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000011066 ex-situ storage Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000012447 hatching Effects 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 238000002354 inductively-coupled plasma atomic emission spectroscopy Methods 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 229930182817 methionine Natural products 0.000 description 2
- 239000002086 nanomaterial Substances 0.000 description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 2
- 238000006862 quantum yield reaction Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000001878 scanning electron micrograph Methods 0.000 description 2
- 229960004402 tiopronin Drugs 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- OFNXOACBUMGOPC-HZYVHMACSA-N 5'-hydroxystreptomycin Chemical group CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](CO)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O OFNXOACBUMGOPC-HZYVHMACSA-N 0.000 description 1
- 239000012691 Cu precursor Substances 0.000 description 1
- 229920001410 Microfiber Polymers 0.000 description 1
- 238000000026 X-ray photoelectron spectrum Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000003636 chemical group Chemical group 0.000 description 1
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 1
- 239000013256 coordination polymer Substances 0.000 description 1
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001857 fluorescence decay curve Methods 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 239000010806 kitchen waste Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000003658 microfiber Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000005693 optoelectronics Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000005424 photoluminescence Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 238000001161 time-correlated single photon counting Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 238000009827 uniform distribution Methods 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
- C09K11/025—Use of particular materials as binders, particle coatings or suspension media therefor non-luminescent particle coatings or suspension media
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y20/00—Nanooptics, e.g. quantum optics or photonic crystals
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/58—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing copper, silver or gold
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
-
- G—PHYSICS
- G09—EDUCATION; CRYPTOGRAPHY; DISPLAY; ADVERTISING; SEALS
- G09F—DISPLAYING; ADVERTISING; SIGNS; LABELS OR NAME-PLATES; SEALS
- G09F3/00—Labels, tag tickets, or similar identification or indication means; Seals; Postage or like stamps
- G09F3/02—Forms or constructions
- G09F3/0291—Labels or tickets undergoing a change under particular conditions, e.g. heat, radiation, passage of time
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
Abstract
The invention discloses a difunctional fluorescent film for detecting mercury ions and glutathione and preparation and application thereof, wherein the method takes eggshell membrane (ESM) as a reaction carrier at room temperature, and copper sulfate (CuSO) 4 ) As a precursor, L-cysteine (L-Cys) is used as a protective agent and a reducing agent, and fluorescent copper nanoclusters are loaded on the surface and inside of ESM in situ, so that a two-dimensional film material (Cys/CuNCs@ESM) capable of emitting intense red fluorescence under an ultraviolet lamp is prepared. The fluorescent film is specific to Hg 2+ And Glutathione (GSH) has higher 'On-Off-On' fluorescence sensing performance. The Hg can be realized respectively by combining the color recognition software of the modern smart phone 2+ And GSH, the detection limits are 1.49 respectivelymu.M, and 2.6. Mu.M. The preparation method of the fluorescent film is green, environment-friendly, economical and easy to operate, can be developed into a portable sensing test strip, and can realize on-site timely monitoring and evaluation of a target object.
Description
Technical Field
The invention belongs to the field of nano biological analysis chemistry, and relates to preparation of a difunctional fluorescent film, which is applied to visual detection of mercury ions and glutathione and high-grade fluorescence optical anti-counterfeiting.
Background
In the current society, sudden events occur in time, so that a real-time, rapid and portable on-site detection means is also receiving more and more attention. As a novel two-dimensional integral membrane material, the fluorescent membrane has good portability and stability, and is more widely applied to field visual detection than a solution. Meanwhile, the fluorescent light has the advantages of high selectivity, high sensitivity and the like. As a novel fluorescent nanomaterial, the metal Nanoclusters (NCs) have excellent light stability, larger Stokes shift, good biocompatibility and low toxicity. Particularly copper nanoclusters (CuNCs), are more suitable for various applications, such as sensing, optoelectronic devices, cell imaging, due to their mild synthesis conditions, abundant and inexpensive precursors. It is therefore interesting to use copper nanoclusters to build a fluorescent film.
The preparation method of the two-dimensional fluorescent film is mainly divided into two types: ex situ preparation and in situ preparation. Ex-situ preparation refers to the synthesis of the luminescent component prior to its loading onto a carrier, such as filter paper. The method has the following defects: (i) cumbersome steps; (ii) The luminous components are unevenly distributed in the carrier and are easy to fall off, so that a large-area uniform luminous film is difficult to prepare; (iii) The photoluminescence quantum yield of the luminescent component is reduced during the preparation of the fluorescent film. Whereas in situ preparation refers to the direct formation of the luminescent component in the matrix by chemical reaction. Because the luminescent component is synthesized in situ, the fluorescent material has the advantages of uniform distribution, high fluorescence quantum yield, good stability and the like. The novel fluorescent CuNCs are loaded/embedded on the biological film by adopting an in-situ reduction method, and the preparation of the novel fluorescent film functional material convenient for storage and transportation is a significant study.
Eggshell membrane (ESM) is a kitchen waste in daily life, and its unique fiber reticulin structure can be used as a good natural reaction carrier. However, until now, it was very rare to prepare functional film materials embedding fluorescent CuNCs in situ using ESM as a reaction carrier. Here we use ESM as the reaction carrier, copper sulfate (CuSO 4 ) The precursor is L-cysteine (L-Cys) as a protective agent and a reducing agent, and the two-dimensional film material (Cys/CuNCs@ESM) embedded with copper nanoclusters and having strong red fluorescence is prepared by in-situ reduction. In the field of fluorescent metal nanoclusters, many scholars point out that the fluorescence properties and applications of metal nanoclusters have a great relationship with the structure of the protective ligand used in the preparation process, the kind of the reducing agent and the synthesis conditions. Different protecting ligands and reducing agents can be used to synthesize metal nanoclusters with very different fluorescence properties, which is also attractive in this research area. Compared with the prior work of preparing a fluorescent film by adopting dithiothreitol organic small molecules as a reducing agent in our laboratory, the fluorescent film has the advantages that more stable, more green and environment-friendly amino acid molecules are adopted: l-cysteine is used as a protective agent and a reducing agent, and compared with dithiothreitol, the L-cysteine has better molecular biocompatibility, unique structure and richer functional groups (SH-, NH- 2 -, COOH-), which plays an important role in protecting the CuNCs during the in-situ reduction and formation process, and correspondingly, cuNCs with novel properties can be obtained. Experiments show that the fluorescent film prepared in situ by using L-cysteine as a protective agent and a reducing agent is used for Hg 2+ And Glutathione (GSH) has higher fluorescence sensing properties. Can respectively realize Hg by combining with color recognition software in a modern smart phone 2+ And visual detection of GSH. The preparation method of the fluorescent film is green, environment-friendly, economical and easy to operate, can be developed into portable sensing test paper, and can realize on-site timely monitoring and evaluation of a target object. Meanwhile, the application of the material in the aspect of advanced optical anti-counterfeiting is innovative to a certain extent.
Disclosure of Invention
The invention aims to provide a method for preparing a stable and uniform bi-functional fluorescent film with strong red fluorescence in situ. The method is green, environment-friendly, economical and easy to operate. And according to fluorescence quenching (Off) and fluorescence recovery (On), the double intelligent sensor strip is developed, and the visualized detection of mercury ions and glutathione is realized. Meanwhile, the application of the optical anti-counterfeiting film in the aspect of advanced optical anti-counterfeiting is researched. The aim of the invention can be achieved by the following technical scheme:
the preparation method of the difunctional fluorescent film for detecting the mercury ions and the glutathione comprises the following steps:
(1) Manually removing eggshell membrane (ESM) from fresh eggshell, washing ESM with ultrapure water, cutting into rectangular shape (about 1.0cm×1.5 cm), soaking in ultrapure water, and refrigerating at 4deg.C;
(2) One piece of ESM was soaked in 1.0mL of 100 mmol.L -1 CuSO of (C) 4 Incubating in water solution for 15min, taking out, carefully washing the surface of the membrane with ultrapure water, and removing superfluous CuSO 4 ；
(3) Will adsorb Cu 2+ Is soaked in 1.0mL of ESM with the concentration of 150 mmol.L -1 In the L-cysteine aqueous solution of (2), reacting for 15min to 3h at room temperature, and preparing the double-function membrane material (Cys/CuNCs@ESM) embedded with copper nanoclusters and having strong red fluorescence.
Furthermore, in the process of preparing the dual-function fluorescent film in situ, no toxic or harmful reagent is used, and the whole preparation method is more green and environment-friendly.
Further, cu is adsorbed 2+ The reaction time of ESM with L-cysteine solution was 1h.
Further, the copper nanoclusters are generated by in-situ reduction and embedded on the surface and inside of the ESM carrier, and the characteristic interweaved fiber structure of the ESM carrier is almost unchanged after the copper nanoclusters are generated.
Further, the prepared dual-function fluorescent film is milky white under sunlight, and emits bright red fluorescence under 365nm ultraviolet lamp, the maximum excitation wavelength and emission wavelength are 365nm and 615nm respectively, and Stokes shift is up to 250nm.
Further, the average fluorescence lifetime of the prepared red bifunctional fluorescent film is as high as 7.3 μs.
Further, the prepared dual-function fluorescent film has good salt tolerance, solvent stability and storage stability.
Further, the fluorescent film (L-Cys/CuNCs@ESM) prepared by the in-situ reduction method can be used as a dual-function test paper and applied to the visual detection of mercury ions and Glutathione (GSH), and is characterized in that: placing a plurality of fluorescent films with the same shape and luminous intensity in 1.0mL Hg with different concentrations 2+ Solution (0. Mu. Mol.L) -1 、5μmol·L -1 、10μmol·L -1 、50μmol·L -1 、100μmol·L -1 、200μmol·L -1 、300μmol·L -1 、400μmol·L -1 ) In the above, the reaction was carried out at room temperature for 30 minutes. The Hg-dependent performance was observed under 365nm UV light 2+ The more severe the red fluorescence quenching of the membrane the higher the concentration. While other metal ions (Ag + 、Mn 2+ 、PPi、CO 3 2- 、SCN - 、S 2- 、I - 、Mg 2+ 、Br - 、Cl - 、Ca 2+ 、Ba 2+ 、Cu 2+ 、Na + 、K + 、Cd 2+ ) Has little effect on its fluorescence. Based on the image shot by the camera of the smart phone, uploading the captured image to color recognition software in the smart phone and outputting a corresponding Lab-b value, thereby realizing Hg 2+ Visual quantitative detection of (2). Fluorescence in the membrane is Hg 2+ On the basis of quenching, 1.0mL of GSH solution (0 mmol.L) -1 、0.05mmol·L -1 、0.1mmol·L -1 、0.2mmol·L -1 、0.4mmol·L -1 、0.6mmol·L -1 、0.8mmol·L -1 、1mmol·L -1 ) In the reaction, the fluorescent light disappeared was recovered to various degrees by reacting for 1 hour at room temperature. The corresponding fluorescent film is taken out, washed by ultrapure water carefully, then placed on filter paper for natural air drying, and a more visual and bright fluorescent color change gradient can be observed under a 365nm ultraviolet lamp. Quantitative detection of Hg according to the sum 2+ The same analysis program collects images and data, and quantitative visual detection of GSH can be realized. Meanwhile, other amino acid molecules such as phenylalanine (Phe), aspartic acid (Asp), tryptophan (Trp), ciprofloxacin (CIP), dopamine hydrochloride (DA), levofloxacin (LEV), D-penicillamine (D-Pen), tiopronin (MPG), histidine (His), leucine (Leu), methionine (Met), glycine (Gly), arginine (Arg), lysine (Lys) and the like have almost no influence on fluorescence detection of the drug molecules.
Further, the red fluorescent film (L-Cys/CuNCs@ESM) disclosed by the invention can be used for the aspect of advanced optical anti-counterfeiting, and the specific technical scheme is as follows: first, a piece of washed eggshell membrane was immersed in 1.0 mLL-cysteine solution (150 mmol.L -1 ) After 30 minutes of incubation, the cells were removed. The surface was rinsed with ultra pure water to remove residual L-Cys and then placed on a glass plate. Then at 50 mmol.L -1 CuSO 4 The aqueous solution is used as 'ink', and the corresponding luminous pattern can be obtained under 365nm ultraviolet lamp by directly writing or drawing on the carrier. Further, 300. Mu. Mol.L was added dropwise to the pattern surface -1 Hg 2+ The pattern will disappear with an aqueous solution. Finally, 100 mmol.L of the mixture is added dropwise -1 GSH aqueous solution, the disappeared fluorescent pattern can reappear again, and the purpose of high-grade anti-counterfeiting is achieved.
The principle of the invention is that
Eggshell membrane (ESM) as a protein-rich biomaterial contains a microscopic interwoven fiber structure with a large surface area, in which a large number of multifunctional chemical groups are present to provide it with a strong ability to coordinate metal ions, so that its staggered microfiber structure can serve as a good reaction carrier. In addition, ESM has good flexibility, and the operation is simple and convenient. Placing ESM in copper sulfate water solutionIn the liquid, after incubation for several minutes, cu 2+ I.e. adsorbed on ESM carrier to form Cu 2+ ESM complex. Further, cu is as follows 2+ And (3) placing the ESM in an L-Cys aqueous solution, and embedding the copper nanoclusters with red fluorescence generated by in-situ reduction on the surface and the inside of the ESM due to the reduction and protection effects of the L-Cys, so as to obtain a uniform and stable red fluorescent film material (L-Cys/CuNCs@ESM). Meanwhile, hg is added according to the prepared fluorescent film 2+ And GSH fluorescence quenching and recovery sensing characteristics, and can respectively realize Hg by combining with the color recognition software of a modern smart phone 2+ And GSH, the detection limits were 1.49. Mu.M and 2.6. Mu.M, respectively. The preparation method of the fluorescent film is green, environment-friendly, economical and easy to operate, can be developed into portable sensing test paper, and can realize on-site timely monitoring and evaluation of a target object. In addition, the ESM is sequentially immersed in an aqueous L-Cys solution and CuSO 4 And obtaining a corresponding light-emitting mode in the solution. The surface luminous pattern is further processed by Hg 2+ And GSH can disappear and reappear after being processed respectively, thereby successfully constructing the advanced optical anti-counterfeiting mode.
The invention has the beneficial effects that:
(1) The invention uses cheap and easily available eggshell membrane (ESM) as a reaction carrier, copper sulfate as a metallic copper precursor, L-cysteine as a protective agent and a reducing agent, and adopts an in-situ reduction method to prepare a uniform and stable red fluorescent membrane (Cys/CuNCs@ESM) at room temperature. All materials/reagents related by the invention are nontoxic and harmless, and the characteristic interweaved fiber structure of the ESM carrier is almost unchanged in the preparation process of the product. The method is green, environment-friendly, economical and strong in operability, and completely accords with the development concept of green chemistry;
(2) The fluorescence lifetime of the prepared fluorescent film is longer, and the average fluorescence lifetime is as long as 7.3 mu s;
(3) The prepared fluorescent film product emits bright red fluorescence under a 365nm ultraviolet lamp, has better salt tolerance, solvent stability and storage stability, and lays a good foundation for various applications of the product under other complex conditions;
(4)Hg 2+ the ions will make the fluorescent film effectiveThe phenomenon of fluorescence quenching occurs, and the membrane sample after fluorescence quenching is placed in a Glutathione (GSH) solution, so that the fluorescence of the membrane can be quickly recovered due to the competition effect. The Hg can be realized respectively by combining the color recognition software of the modern smart phone 2+ And GSH, "On-Off-On" visual detection limited to 1.49. Mu.M and 2.6. Mu.M, respectively. The preparation method of the fluorescent film is green, environment-friendly, economical and easy to operate, can be developed into a portable sensing test strip, and can realize on-site timely monitoring and evaluation of a target object, and similar reports are not yet seen at present.
(5) Hg based on the fluorescent film 2+ The invention also proves the application of the fluorescent film in the aspect of advanced optical anti-counterfeiting by the unique response of 'On-Off-On' of ions and GSH. Compared with a single fluorescence quenching (off) anti-counterfeiting mode developed before a laboratory, the technology has higher anti-counterfeiting value, is simple to operate and has high cost efficiency.
(6) The invention opens up a new way for preparing the two-dimensional functional material in situ with green and low cost, and combines with a modern smart phone sensing platform, so that the invention has wide application prospect in the fields of strip sensing chemistry, photoelectric devices, advanced anti-counterfeiting and the like.
Of course, it is not necessary for any one product to practice the invention to achieve all of the advantages set forth above at the same time.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is: wherein A is the pictures (b, c, d and b ', c ', d ' of Cys/CuNCs@ESM product (a) and control group under 365nm ultraviolet light (up) and under sunlight (down) corresponding to ESM, cu in sequence 2+ /ESM、L-Cys/ESM)；
B is ESM, cu 2+ Fluorescence emission spectra of ESM, L-Cys/ESM control and Cys/CuNCs@ESM product(excitation wavelength is 365 nm);
panel C shows the fluorescence decay curve of Cys/CuNCs@ESM product at 615 nm;
fig. 2 is: panel A is an SEM image of an ESM carrier; panel B is an SEM image of a Cys/CuNCs@ESM sample; panel C is a fluorescence microscope image of a Cys/CuNCs@ESM sample;
fig. 3 is: panel A shows XPS (XPS) full spectrum of Cys/CuNCs@ESM fluorescent film; panel B shows XPS high-resolution spectrum of Cu2 p;
FIG. 4 is a CuSO 4 Effect of concentration on Cys/cuncs@esm fluorescent film fluorescence intensity plot;
FIG. 5 is a graph of the effect fluorescence intensity of L-cysteine concentration on Cys/CuNCs@ESM fluorescent films;
FIG. 6 is a graph showing the effect fluorescence intensity of incubation time of ESM and CuSO4 on Cys/CuNCs@ESM fluorescent film;
FIG. 7 is a graph showing the effect of reaction time in cysteine on the fluorescence intensity of L-Cys/CuNCs@ESM;
FIG. 8 is a graph of the effect fluorescence intensity of NaCl concentration on Cys/CuNCs@ESM fluorescent film;
FIG. 9 is a graph showing the effect fluorescence intensity of different organic solvents on Cys/CuNCs@ESM fluorescent films;
in fig. 10: panel A shows Hg 2+ Visual detection of (2); panel B shows a selectivity experiment; panel C shows b and Hg 2+ A plot of linear relationship between logarithms of concentrations; FIG. D shows Cys/CuNCs@ESM fluorescent film and Hg at different concentrations 2+ A fluorescence spectrum after the action;
in fig. 11: panel A is visual detection of GSH; panel B shows a selectivity experiment; panel C is a linear plot of b value versus GSH concentration; FIG. D is a graph of fluorescence recovery spectra of different concentrations of GSH for a Hg2+ -fluorescent film system;
FIG. 12 is a diagram of an example of an advanced fluorescent anti-counterfeiting application.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
With ESM as a reaction carrier, copper sulfate (CuSO 4 ) As a precursor, L-cysteine (L-Cys) is used as a protective agent and a reducing agent, and the specific steps of preparing a film material (Cys/cuncs@esm) with intense red fluorescence at room temperature by an in-situ reduction method are as follows:
(1) Manually removing eggshell membrane (ESM) from fresh eggshell, washing ESM with ultrapure water, cutting into rectangular shape (about 1.0cm×1.5 cm), soaking in ultrapure water, and refrigerating at 4deg.C;
(2) One piece of ESM was soaked in 1.0mL of 100 mmol.L -1 CuSO of (C) 4 Incubating in the solution for 15min, taking out ESM, carefully washing with ultrapure water, and removing redundant CuSO on the surface of the carrier 4 ；
(3) Will adsorb Cu 2+ Is soaked in 1.0mL of ESM with the concentration of 150 mmol.L -1 In the L-cysteine solution of (2), reacting for 1h at room temperature to obtain the film material (Cys/CuNCs@ESM) with intense red fluorescence.
The pictures of the synthesized product and the control group are shown in FIG. 1-A, and the material is bright red (a) in the whole under 365nm ultraviolet lamp and is milky yellow (a') under fluorescent lamp. The fluorescence spectra of the synthesized product and the control group at 365nm excitation wavelength are shown in FIG. 1-B, and the Cys/CuNCs@ESM product has a distinct strong emission peak at 615nm, which corresponds to the red fluorescence emitted by the film in FIG. 1-A (a), while the control group has no fluorescence peak in the wave band, which shows that the two-dimensional fluorescent film material embedded with CuNCs is successfully prepared. The time-resolved FL decay pattern of the fluorescent film (FIG. 1-C) was further measured by transient fluorescence TCSPC technique, the decay curve of which conforms to a three-exponential fit function, and the average fluorescence lifetime of the product was calculated to be as high as 7.3 μs.
FIGS. 2-A and 2-B show Scanning Electron Microscope (SEM) images of blank ESM and Cys/CuNCs@ESM products, with a number of uniformly dispersed CuNCs aggregates on the surface of the Cys/CuNCs@ESM fluorescent film material compared to the ESM itself, indicating successful loading of the CuNCs onto and into the ESM support. The laser confocal fluorescence microscopy pictures of Cys/CuNCs@ESM samples are shown in FIG. 2-C, which shows that after copper nanoclusters are loaded in situ, the characteristic interwoven fiber structure of the ESM carrier remains almost unchanged, which benefits from the unique composition and good flexibility of the ESM carrier. The Cu loading in the fluorescent film sample was quantitatively determined to be 1.5% by inductively coupled plasma-atomic emission spectrometry (ICP-AES).
FIG. 3-A is an X-ray photoelectron spectroscopy (XPS) chart of Cys/CuNCs@ESM product confirming the presence of all the expected elements C, O, N, S and Cu. To investigate the chemical valence of the Cu element in the product, high resolution XPS spectra of Cu2p were recorded (fig. 3-B). It can be seen that the two peaks of 931.34eV and 951.26eV correspond to Cu2P in the Cu (0) state, respectively 3/2 And Cu2P 1/2 The presence of copper nanoclusters in the fluorescent film was confirmed. About 942eV does not have satellite peak, and the prepared fluorescent film does not contain Cu (II), so that the in-situ reduction reaction is more complete. It is noted that the binding energy between Cu (0) and Cu (I) differs by only 0.1eV, and the valence state of copper in the fluorescent film may be between 0 to +1 according to the formation process of fluorescent copper nanoclusters.
Example 2
Influence of copper sulfate concentration on the preparation
(1) Manually removing eggshell membrane (ESM) from fresh eggshell, washing ESM with ultrapure water, cutting into rectangular shape (about 1.0cm×1.5 cm), soaking in ultrapure water, and refrigerating at 4deg.C;
(2) 7 pieces of ESM were respectively immersed in 1.0mL of copper sulfate aqueous solutions of different concentrations (concentration: 50 mmol.L in order) -1 、100mmol·L -1 、150mmol·L -1 、200mmol·L -1 、300mmol·L -1 、400mmol·L -1 、50mmol·L -1 ). Incubating for 15min, taking out, carefully washing with ultrapure water, and removing redundant CuSO on ESM carrier surface 4 ；
(3) Will adsorb Cu 2+ Is placed in a concentration of 150 mmol.L at 1.0mL -1 Is reacted in the cysteine solution for 1h. Placing the sample under 365nm ultraviolet lampAs shown in FIG. 4, when CuSO 4 The concentration reaches 50 mmol.L -1 The prepared product can emit red fluorescence. When the concentration of the copper sulfate solution is too high (more than or equal to 150 mmol.L) -1 ) In this case, the generated CuNCs are partially exfoliated. Therefore, cuSO is selected 4 The concentration of the solution was 100 mmol.L -1 In the process, the copper nanoclusters in the prepared sample are distributed most uniformly, and the fluorescence intensity is high.
Example 3
Effect of L-cysteine concentration on preparation
(1) Manually removing eggshell membrane (ESM) from fresh eggshell, washing ESM with ultrapure water, cutting into rectangular shape (about 1.0cm×1.5 cm), soaking in ultrapure water, and refrigerating at 4deg.C;
(2) 6 pieces of ESM were respectively immersed in 1.0mL of 100 mmol.L -1 After incubating in copper sulfate solution for 15min, taking out, carefully washing with ultrapure water, and removing redundant CuSO on the surface of ESM carrier 4 ；
(3) Will adsorb Cu 2+ The ESM vectors of (a) were placed in 1.0mL of L-cysteine solutions of different concentrations (concentration: 50 mmol.L in order) -1 、100mmol·L -1 、150mmol·L -1 、200mmol·L -1 、300mmol·L -1 、400mmol·L -1 ) The reaction was carried out for 1h. The sample was observed under a 365nm ultraviolet lamp, as shown in FIG. 5, the L-cysteine concentration was 50 mmol.L -1 And then preparing the film material which emits red fluorescence. However, it was observed in the experiment that the L-cysteine concentration was low (. Ltoreq.100 mmol.L) -1 ) At the same time, cuNCs may undergo partial shedding and sedimentation. When the concentration of cysteine is too high (more than or equal to 300 mmol.L) -1 ) When the fluorescence of the resulting product is reduced. Therefore, the concentration of L-cysteine is selected to be 150-200 mmol.L -1 The fluorescent light distribution of the obtained product is most uniform and the intensity is strongest.
Example 4
Influence of incubation time in copper sulphate on preparation
(1) Manually removing eggshell membrane (ESM) from fresh eggshell, washing ESM with ultrapure water, cutting into rectangular shape (about 1.0cm×1.5 cm), soaking in ultrapure water, and refrigerating at 4deg.C;
(2) 6 pieces of ESM were respectively immersed in 1.0mL of 100 mmol.L -1 After hatching in copper sulfate solution for different time (time: 5min, 10min, 15min, 20min, 25min in order), taking out, carefully washing with ultrapure water, and removing redundant CuSO on the surface of the carrier 4 ；
(3) Will adsorb Cu 2+ Respectively soaking in 1.0mL of 150 mmol.L -1 Is reacted in the cysteine solution for 1h. When the sample is observed under an ultraviolet lamp with the wavelength of 365nm, as shown in fig. 6, when the ESM carrier is incubated in the copper sulfate aqueous solution for more than 5min, the membrane material capable of emitting red fluorescence can be prepared.
Example 5
Influence of the reaction time in the L-cysteine solution on the preparation
(1) Manually removing eggshell membrane (ESM) from fresh eggshell, washing ESM with ultrapure water, cutting into rectangular shape (about 1.0cm×1.5 cm), soaking in ultrapure water, and refrigerating at 4deg.C;
(2) 7 pieces of ESM were respectively immersed in 1.0mL of 100 mmol.L -1 Taking out after hatching in copper sulfate solution for 10min, carefully washing with ultrapure water, and removing redundant CuSO on the surface of the carrier 4 ；
(3) Will adsorb Cu 2+ The ESM vectors of (C) were placed at a concentration of 150 mmol.L, respectively -1 After reaction in the L-cysteine solution for different times (time: 0.25h, 0.5h, 1h, 1.5h, 2h, 2.5h, 3h in order). The sample is placed under an ultraviolet lamp of 365nm for observation, as shown in fig. 7, the reaction of preparing the fluorescent film in situ is faster, and the film material which can emit obvious red fluorescence can be prepared when the reaction time in the L-cysteine solution is 15min at room temperature.
Example 6
Salt tolerance of Cys/CuNCs@ESM fluorescent film
Respectively placing Cys/CuNCs@ESM fluorescent films into 1.0mL NaCl aqueous solutions with different concentrations, and standing at room temperature for 1hA picture of the sample under a 365nm uv lamp was recorded (figure 8). As can be seen, when the NaCl concentration in the solution is lower than 2mol.L -1 When the fluorescence intensity of Cys/CuNCs@ESM is basically not affected, the NaCl concentration reaches 5 mol.L -1 When the fluorescent film is used, the fluorescence intensity of Cys/CuNCs@ESM is obviously quenched, so that the prepared fluorescent film of Cys/CuNCs@ESM has better salt tolerance.
Example 7
Solvent stability
Cys/CuNCs@ESM are respectively put into 1.0mL of different organic solvents, and the solvents are as follows in sequence: absolute ethanol (EtOH), isopropyl alcohol (IPA), absolute Methanol (MT), acetone (CP), ethyl Acetate (EAC), cyclohexane (CYH), lactic acid (HL), N-Dimethylformamide (DMF). After standing at room temperature for 1h, a picture of the sample under 365nm ultraviolet light was recorded (FIG. 9). The graph shows that the fluorescence intensity of the Cys/CuNCs@ESM fluorescent film in different organic solvents is basically unchanged, and the product has better solvent stability.
Example 8
The red fluorescent film of the invention relates to Hg 2+ And Glutathione (GSH) has higher 'On-Off-On' fluorescence sensing performance. The Hg can be realized respectively by combining the color recognition software of the modern smart phone 2+ And GSH visual detection, the specific technical scheme is as follows:
the prepared fluorescent films are respectively placed in 1.0mL Hg with different concentrations 2+ Solution (0. Mu. Mol.L) -1 、5μmol·L -1 、10μmol·L -1 、50μmol·L -1 、100μmol·L -1 、200μmol·L -1 、300μmol·L -1 、400μmol·L -1 ) In the above, the reaction was carried out at room temperature for 30 minutes. The Hg-dependent performance was observed under 365nm UV light 2+ The more pronounced the red fluorescence quenching of the membrane with increasing concentration (fig. 10-a). While other metal ions (Ag + 、Mn 2+ 、PPi、CO 3 2- 、SCN - 、S 2- 、I - 、Mg 2+ 、Br - 、Cl - 、Ca 2+ 、Ba 2+ 、Cu 2+ 、Na + 、K + 、Cd 2+ ) Has little effect on fluorescence(FIG. 10-B). Shooting an image by a camera of the smart phone, analyzing and processing the captured image by color recognition software in the smart phone, and outputting a corresponding Lab-b value to realize Hg 2+ Is a quantitative detection of (a). As shown in FIG. 10-C, the b value of the image is compared with Hg 2+ The concentration logarithm has two sections of good linear relation. In the range of 5-100. Mu.M, the linear regression equation is b=36.23-17.26Log [ Hg ] 2+ ](R 2 = 0.9848), the limit of detection (LOD) was 1.49 μm. At the same time, we also tested the fluorescence spectrum of each sample with a fluorescence spectrometer (FIG. 10-D), which can be seen as Hg 2+ The fluorescence intensity of the fluorescent film gradually decreases with increasing concentration, but the position of the emission peak is substantially unchanged. The experimental result shows that the prepared fluorescent film can be used as an effective 'On-Off' intelligent sensing test strip for Hg 2+ Visual inspection of (c).
Fluorescence at Cys/CuNCs@ESM is blocked by Hg 2+ On the basis of quenching, the mixture was put into 1.0mL of Glutathione (GSH) solution (0 mmol.L) with different concentrations -1 、0.05mmol·L -1 、0.1mmol·L -1 、0.2mmol·L -1 、0.4mmol·L -1 、0.6mmol·L -1 、0.8mmol·L -1 、1mmol·L -1 ) In the above reaction, the reaction was carried out at room temperature for 1 hour, and the vanishing fluorescence was found to gradually recover. The treated fluorescent film was taken out, carefully rinsed with ultra-pure water, and then naturally air-dried on a filter paper, and a more visual and bright gradient of fluorescent color change was obtained under a 365nm ultraviolet lamp (FIG. 11-A). Meanwhile, other amino acid molecules such as phenylalanine (Phe), aspartic acid (Asp), tryptophan (Trp), ciprofloxacin (CIP), dopamine hydrochloride (DA), levofloxacin (LEV), D-penicillamine (D-Pen), tiopronin (MPG), histidine (His), leucine (Leu), methionine (Met), glycine (Gly), arginine (Arg), lysine (Lys) and the like have little influence on fluorescence recovery of the drug molecules (FIG. 11-B). Quantitative detection of Hg according to the sum 2+ The same analysis program collects images and data. As shown in fig. 11-C, the b value of the image has a good linear relationship with the log GSH concentration: b=109.007 [ gsh ]]+43.57(R 2 = 0.9883), LOD is 2.6 μm. At the same time, we also tested each with a fluorescence spectrometerThe fluorescence spectra of the individual samples (FIG. 11-D) shows that with increasing GSH concentration, the fluorescence intensity of the film recovered at progressively higher increases, with the position of the emission peak being substantially unchanged. The experimental results show that Hg is prepared 2+ The fluorescent film can be used as an effective Off-On intelligent sensing test strip for the visual detection of GSH.
In summary, considering the increasing demands of low cost, fast response and field detection techniques of the developed solutions, the prepared Cys/CuNCs@ESM two-dimensional fluorescent film can be used as portable Hg 2+ And GSH 'On-Off-On' type dual-function test paper, and can quantitatively analyze the visual result through a smart phone, thereby improving the accuracy and sensitivity of analysis. Meanwhile, in view of the fact that the Cys/CuNCs@ESM test strip has better specificity and higher sensitivity to a target object, the test strip can be directly applied to Hg in an actual sample 2+ And rapid detection of GSH in the tablets.
Example 9
The invention can be applied to the aspects of high-grade fluorescence anti-counterfeiting and the like, and the specific technical scheme is as follows:
first, a piece of the ESM after washing was immersed in 1.0 mLL-cysteine solution (150 mmol.L -1 ) After incubation for 30 minutes, the mixture was taken out. The residual L-Cys on the surface of the carrier was removed by rinsing with ultrapure water, and then placed on a clean glass sheet. Then, at 50 mmol.L -1 CuSO 4 The aqueous solution was used as an "ink" to write or draw directly on the film, and the corresponding fluorescent pattern was observed under a 365nm ultraviolet lamp (FIG. 12-A). Next, 300. Mu. Mol.L was added dropwise to the pattern surface -1 Hg 2+ The fluorescent pattern disappeared from the solution (fig. 12-B). Finally, 100 mmol.L of the mixture is added dropwise -1 GSH solution, the vanishing fluorescent pattern reappears again (FIG. 12-C). Therefore, the secondary high-grade fluorescent anti-counterfeiting mode is successfully constructed, and a way is provided for developing the biological matrix luminescent nano material for high-grade optical anti-counterfeiting, hidden writing, information encryption and security paper.
The foregoing is merely illustrative and explanatory of the principles of the invention, as various modifications and additions may be made to the specific embodiments described, or similar thereto, by those skilled in the art, without departing from the principles of the invention or beyond the scope of the appended claims.
Claims (6)
1. A preparation method of a difunctional fluorescent film for detecting mercury ions and glutathione is characterized by comprising the following steps: the method comprises the following steps:
(1) Manually removing eggshell membrane from eggshell, washing with ultrapure water, cutting into rectangular shape with the same size of 1.0cm ×1.5cm, soaking in ultrapure water, and refrigerating at 4deg.C;
(2) Soaking eggshell membrane in a concentration of 1.0mL to 100 mmol.L -1 CuSO of (C) 4 Incubating in water solution for 15min, and taking out the Cu adsorbed 2+ Washing eggshell membrane with ultrapure water to remove redundant CuSO on the surface of the carrier 4 ；
(3) Will adsorb Cu 2+ Is soaked in 1.0mL with concentration of 150mmol.L -1 In the L-cysteine aqueous solution, the reaction is carried out for 15min to 3h at room temperature, thus obtaining the difunctional fluorescent film.
2. The method for preparing the bifunctional fluorescent film for detecting mercury ions and glutathione according to claim 1, wherein the method comprises the following steps: the reaction time in step (3) was 1h.
3. The method for preparing the bifunctional fluorescent film for detecting mercury ions and glutathione according to claim 1, wherein the method comprises the following steps: the dual-function fluorescent film is milky yellow under sunlight, and emits bright red fluorescence under 365nm ultraviolet lamp, the maximum excitation and emission wavelength are 365nm and 615nm respectively, and Stokes shift is 250nm.
4. The method for preparing the bifunctional fluorescent film for detecting mercury ions and glutathione according to claim 1, wherein the method comprises the following steps: the average fluorescence life of the dual-function fluorescent film is as long as 7.3 mu s.
5. A bifunctional fluorescent film for detecting mercury ions and glutathione prepared by the method of any one of claims 1 to 4.
6. The use of the bifunctional fluorescent film for detecting mercuric ions and glutathione as a bifunctional test strip of claim 5, which is applied to the visualized detection of mercuric ions and glutathione, characterized in that: (1) Respectively placing the difunctional fluorescent films in 1.0mL with the concentration of 0 mu mol.L -1 、5 µmol·L -1 、10 µmol·L -1 、50 µmol·L -1 、100 µmol·L -1 、200 µmol·L -1 、300 µmol·L -1 、400 µmol·L -1 Is (1) Hg of 2+ In the solution, reacting for 30 minutes at room temperature; the Hg-dependent performance was observed under 365nm UV light 2+ The more severe the red fluorescence quenching of the membrane the concentration increases; while other metal ions Ag + 、Mn 2+ 、PPi、CO 3 2- 、SCN - 、S 2- 、I - 、Mg 2+ 、Br - 、Cl - 、Ca 2+ 、Ba 2+ 、Cu 2+ 、Na + 、K + 、Cd 2+ Has no influence on fluorescence; based on the image shot by the camera of the smart phone, analyzing and processing the captured image by adopting color recognition software, and outputting a corresponding Lab-b value, namely Hg 2+ Visual quantitative detection of (2);
(2) Fluorescence of the film at step (1) is Hg 2+ Quenching, and then placing it into a reactor with a concentration of 1.0mL and 0 mmol.L -1 、0.05 mmol·L -1 、0.1 mmol·L -1 、0.2 mmol·L -1 、0.4 mmol·L -1 、0.6 mmol·L -1 、0.8 mmol·L -1 、1 mmol·L -1 In the glutathione aqueous solution of (2), the reaction is carried out at room temperature for 1h, and the disappeared fluorescence can be found to be recovered to different degrees; the corresponding fluorescent film was taken out, rinsed with ultrapure water, then placed on a filter paper for natural air-drying, and a bright fluorescent color change was observed under a 365nm ultraviolet lampA gradient; quantitative detection of Hg according to visualization in step (1) 2+ The same analysis program collects images and data, and can realize quantitative visual detection of glutathione.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110921350.1A CN113637468B (en) | 2021-08-11 | 2021-08-11 | Difunctional fluorescent film for detecting mercury ions and glutathione, preparation and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110921350.1A CN113637468B (en) | 2021-08-11 | 2021-08-11 | Difunctional fluorescent film for detecting mercury ions and glutathione, preparation and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113637468A CN113637468A (en) | 2021-11-12 |
| CN113637468B true CN113637468B (en) | 2023-12-26 |
Family
ID=78420947
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110921350.1A Active CN113637468B (en) | 2021-08-11 | 2021-08-11 | Difunctional fluorescent film for detecting mercury ions and glutathione, preparation and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113637468B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103336039A (en) * | 2013-07-15 | 2013-10-02 | 长沙理工大学 | Sensing chip method for detecting mercury ions based on L-cysteine |
| WO2014088512A1 (en) * | 2012-12-03 | 2014-06-12 | National University Of Singapore | Ratiometric fluorescent dye for the detection of glutathione in cell and tissue |
| CN105527267A (en) * | 2016-01-30 | 2016-04-27 | 山西大学 | Red fluorescence gold nanocluster as well as preparation method and application thereof |
| CN108535230A (en) * | 2018-04-12 | 2018-09-14 | 淮北师范大学 | Based on egg shell membrane platform red fluorescence copper nanocluster in-situ synthetic method and application |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9139550B2 (en) * | 2011-08-09 | 2015-09-22 | Los Gatos Research | Fluorescence dye tagging scheme for mercury quantification and speciation |
-
2021
- 2021-08-11 CN CN202110921350.1A patent/CN113637468B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014088512A1 (en) * | 2012-12-03 | 2014-06-12 | National University Of Singapore | Ratiometric fluorescent dye for the detection of glutathione in cell and tissue |
| CN103336039A (en) * | 2013-07-15 | 2013-10-02 | 长沙理工大学 | Sensing chip method for detecting mercury ions based on L-cysteine |
| CN105527267A (en) * | 2016-01-30 | 2016-04-27 | 山西大学 | Red fluorescence gold nanocluster as well as preparation method and application thereof |
| CN108535230A (en) * | 2018-04-12 | 2018-09-14 | 淮北师范大学 | Based on egg shell membrane platform red fluorescence copper nanocluster in-situ synthetic method and application |
Non-Patent Citations (5)
| Title |
|---|
| In situ fabrication of a luminescent copper nanocluster/eggshell membrane composite and its application in visual detection of Ag+ ions, light-emitting diodes and surface patterning;Chunbo Li, et al.;《Photochemical & Photobiological Sciences》;第18卷(第12期);第2942-2951页 * |
| In Situ Generation of Fluorescent Copper Nanoclusters Embedded in Monolithic Eggshell Membrane Properties and Applications;Lu Li, et al.;《Materials》;第11卷(第10期);第1913页 * |
| 新型荧光铜和荧光碳纳米材料的制备及其性能研究;李露;《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》(第8期);第B014-42页 * |
| 红色荧光铜纳米材料的制备及其性能研究;李春波;《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》(第12期);第B014-350页 * |
| 荧光金纳米团簇的制备及其在分析化学中的应用;肖敏;《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》(第9期);第B014-41页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113637468A (en) | 2021-11-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kieber et al. | Photooxidation of triglycerides and fatty acids in seawater: Implication toward the formation of marine humic substances | |
| Li et al. | A luminescent Ln-MOF thin film for highly selective detection of nitroimidazoles in aqueous solutions based on inner filter effect | |
| CN108840879A (en) | A kind of double ligand MOF complexs and its synthesis and the application in fluorescence identifying iron ion | |
| CN110484233A (en) | Zinc oxide nano-crystal, zinc oxide nano-crystal composition, preparation method and electroluminescent device | |
| Wang et al. | Tb (iii) postsynthetic functional coordination polymer coatings on ZnO micronanoarrays and their application in small molecule sensing | |
| CN109179377B (en) | Preparation method of hydrophobic fluorescent carbon dots | |
| CN113637468B (en) | Difunctional fluorescent film for detecting mercury ions and glutathione, preparation and application | |
| CN108587607B (en) | Luminescent metal organic framework material for detecting thiocyanate ions in water and preparation method and application thereof | |
| CN107312527A (en) | One kind is without matrix phosphorescence carbon quantum dot and its preparation method and application | |
| Xu et al. | Metal–organic framework-5 as a novel phosphorescent probe for the highly selective and sensitive detection of Pb (II) in mussels | |
| Xue et al. | Preparation of AgInS2 quantum dots and their application for Pb2+ detection based on fluorescence quenching effect | |
| Tong et al. | Polymer ligand-sensitized lanthanide metal–organic frameworks for an on-site analysis of a radionuclide | |
| CN110270365B (en) | Preparation and application of carbon nitride/lanthanum oxychloride composite material | |
| EP3282043A1 (en) | Functionalized silk fibroin security marker | |
| Ye et al. | A smartphone-assisted paper-based analytical device for fluorescence assay of Hg2+ | |
| CN107011892A (en) | A kind of preparation method and application of Cu Mn codopes ZnS quantum dot solution | |
| CN114045166B (en) | Multi-stimulus-response functional anti-counterfeiting material and preparation method and application thereof | |
| Li et al. | A turn-on fluorescence sensing strategy for rapid detection of flumequine in water environments using covalent-coordination functionalized MOFs | |
| CN101697873A (en) | Method for developing latent fingerprints by adopting nano zinc oxide | |
| CN114544599A (en) | Synthesis of rare Eu (II) -MOF, method for constructing near-infrared sensor by using Eu (II) -MOF as electroluminescent probe and application of Eu (II) -MOF | |
| CN114774112A (en) | Hydrogel of agarose-coated Eu-MOF luminescent material, and preparation method and application thereof | |
| CN107954891A (en) | A kind of rare earth Tetracyclines complex and its preparation method and application | |
| Zhang et al. | Nitrogen-doped carbon dots from Hutai-8 grape skin and their application in Hg2+ detection | |
| CN108982471B (en) | AgNR/O-g-C3N4Substrate, preparation method thereof and application thereof in recyclable SERS (surface enhanced Raman scattering) sensitive detection | |
| CN113502155B (en) | Fluorescent probe for visually detecting doxycycline and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |