CN114965644B - Electrochemiluminescence body, electrochemiluminescence aptamer sensor, and preparation methods and applications thereof - Google Patents
Electrochemiluminescence body, electrochemiluminescence aptamer sensor, and preparation methods and applications thereof Download PDFInfo
- Publication number
- CN114965644B CN114965644B CN202210640465.8A CN202210640465A CN114965644B CN 114965644 B CN114965644 B CN 114965644B CN 202210640465 A CN202210640465 A CN 202210640465A CN 114965644 B CN114965644 B CN 114965644B
- Authority
- CN
- China
- Prior art keywords
- lincomycin
- ucnps
- patp
- ptca
- electrode
- 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
- 108091023037 Aptamer Proteins 0.000 title claims abstract description 43
- 238000002360 preparation method Methods 0.000 title claims description 21
- OJMMVQQUTAEWLP-UHFFFAOYSA-N Lincomycin Natural products CN1CC(CCC)CC1C(=O)NC(C(C)O)C1C(O)C(O)C(O)C(SC)O1 OJMMVQQUTAEWLP-UHFFFAOYSA-N 0.000 claims abstract description 80
- OJMMVQQUTAEWLP-KIDUDLJLSA-N lincomycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@@H](C)O)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 OJMMVQQUTAEWLP-KIDUDLJLSA-N 0.000 claims abstract description 80
- 229960005287 lincomycin Drugs 0.000 claims abstract description 80
- 238000001514 detection method Methods 0.000 claims abstract description 23
- 239000002105 nanoparticle Substances 0.000 claims abstract description 23
- 229910021397 glassy carbon Inorganic materials 0.000 claims abstract description 22
- 229920001469 poly(aryloxy)thionylphosphazene Polymers 0.000 claims abstract description 15
- 238000006243 chemical reaction Methods 0.000 claims abstract description 13
- 239000010931 gold Substances 0.000 claims abstract description 13
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 9
- 229910052737 gold Inorganic materials 0.000 claims abstract description 9
- 239000002114 nanocomposite Substances 0.000 claims abstract description 7
- 101000804764 Homo sapiens Lymphotactin Proteins 0.000 claims abstract description 5
- 102100035304 Lymphotactin Human genes 0.000 claims abstract description 5
- 238000011068 loading method Methods 0.000 claims abstract description 5
- FVDOBFPYBSDRKH-UHFFFAOYSA-N perylene-3,4,9,10-tetracarboxylic acid Chemical compound C=12C3=CC=C(C(O)=O)C2=C(C(O)=O)C=CC=1C1=CC=C(C(O)=O)C2=C1C3=CC=C2C(=O)O FVDOBFPYBSDRKH-UHFFFAOYSA-N 0.000 claims abstract description 3
- 239000002773 nucleotide Substances 0.000 claims abstract 2
- 125000003729 nucleotide group Chemical group 0.000 claims abstract 2
- 239000000243 solution Substances 0.000 claims description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 238000003756 stirring Methods 0.000 claims description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 21
- 238000001035 drying Methods 0.000 claims description 19
- 238000012417 linear regression Methods 0.000 claims description 18
- 239000007853 buffer solution Substances 0.000 claims description 17
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 16
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 16
- 238000004020 luminiscence type Methods 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 14
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- 229910021607 Silver chloride Inorganic materials 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 229910052786 argon Inorganic materials 0.000 claims description 8
- 239000000872 buffer Substances 0.000 claims description 8
- 238000002484 cyclic voltammetry Methods 0.000 claims description 8
- 229910052697 platinum Inorganic materials 0.000 claims description 8
- 238000005498 polishing Methods 0.000 claims description 8
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 7
- 239000012086 standard solution Substances 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 239000012490 blank solution Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- CCCMONHAUSKTEQ-UHFFFAOYSA-N octadec-1-ene Chemical compound CCCCCCCCCCCCCCCCC=C CCCMONHAUSKTEQ-UHFFFAOYSA-N 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 4
- CLYVDMAATCIVBF-UHFFFAOYSA-N pigment red 224 Chemical compound C=12C3=CC=C(C(OC4=O)=O)C2=C4C=CC=1C1=CC=C2C(=O)OC(=O)C4=CC=C3C1=C42 CLYVDMAATCIVBF-UHFFFAOYSA-N 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims description 3
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims description 3
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims description 3
- 239000005642 Oleic acid Substances 0.000 claims description 3
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 239000006185 dispersion Substances 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 3
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- WCDSVWRUXWCYFN-UHFFFAOYSA-N 4-aminobenzenethiol Chemical compound NC1=CC=C(S)C=C1 WCDSVWRUXWCYFN-UHFFFAOYSA-N 0.000 claims description 2
- 239000005457 ice water Substances 0.000 claims description 2
- 239000002244 precipitate Substances 0.000 claims description 2
- 238000011895 specific detection Methods 0.000 claims 3
- 229910019142 PO4 Inorganic materials 0.000 claims 1
- LDDQLRUQCUTJBB-UHFFFAOYSA-N ammonium fluoride Chemical compound [NH4+].[F-] LDDQLRUQCUTJBB-UHFFFAOYSA-N 0.000 claims 1
- 238000004140 cleaning Methods 0.000 claims 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims 1
- 239000010452 phosphate Substances 0.000 claims 1
- 238000002203 pretreatment Methods 0.000 claims 1
- 238000001378 electrochemiluminescence detection Methods 0.000 abstract description 3
- 238000011896 sensitive detection Methods 0.000 abstract description 2
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 7
- 239000008367 deionised water Substances 0.000 description 7
- 229910021641 deionized water Inorganic materials 0.000 description 7
- 229910017604 nitric acid Inorganic materials 0.000 description 7
- 238000009210 therapy by ultrasound Methods 0.000 description 7
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 230000035945 sensitivity Effects 0.000 description 5
- 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 4
- 230000000694 effects Effects 0.000 description 4
- 235000019441 ethanol Nutrition 0.000 description 4
- 238000000504 luminescence detection Methods 0.000 description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound 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](C)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 UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 3
- 230000005611 electricity Effects 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 125000002080 perylenyl group Chemical group C1(=CC=C2C=CC=C3C4=CC=CC5=CC=CC(C1=C23)=C45)* 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 230000003068 static effect Effects 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- -1 Lincomycin amide Chemical class 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 229960005091 chloramphenicol Drugs 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
- 229960003405 ciprofloxacin Drugs 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 229960000318 kanamycin Drugs 0.000 description 2
- 229930027917 kanamycin Natural products 0.000 description 2
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 2
- 229930182823 kanamycin A Natural products 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- CSHWQDPOILHKBI-UHFFFAOYSA-N peryrene Natural products C1=CC(C2=CC=CC=3C2=C2C=CC=3)=C3C2=CC=CC3=C1 CSHWQDPOILHKBI-UHFFFAOYSA-N 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- XXMFJKNOJSDQBM-UHFFFAOYSA-N 2,2,2-trifluoroacetic acid;hydrate Chemical compound [OH3+].[O-]C(=O)C(F)(F)F XXMFJKNOJSDQBM-UHFFFAOYSA-N 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- CYDMQBQPVICBEU-UHFFFAOYSA-N chlorotetracycline Natural products C1=CC(Cl)=C2C(O)(C)C3CC4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-UHFFFAOYSA-N 0.000 description 1
- 229960004475 chlortetracycline Drugs 0.000 description 1
- CYDMQBQPVICBEU-XRNKAMNCSA-N chlortetracycline Chemical compound C1=CC(Cl)=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O CYDMQBQPVICBEU-XRNKAMNCSA-N 0.000 description 1
- 235000019365 chlortetracycline Nutrition 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000000835 electrochemical detection Methods 0.000 description 1
- 239000007772 electrode material Substances 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000003547 immunosorbent Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 108091008104 nucleic acid aptamers Proteins 0.000 description 1
- 229940049964 oleate Drugs 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000010408 sweeping Methods 0.000 description 1
- 238000004454 trace mineral analysis Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
-
- 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
- B82Y40/00—Manufacture or treatment of nanostructures
-
- 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
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
- C09K11/07—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials having chemically interreactive components, e.g. reactive chemiluminescent compositions
-
- 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
- C09K11/77—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals
- C09K11/7783—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing rare earth metals containing two or more rare earth metals one of which being europium
- C09K11/7784—Chalcogenides
- C09K11/7787—Oxides
-
- 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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- 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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
- G01N21/763—Bioluminescence
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/70—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in livestock or poultry
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Physics & Mathematics (AREA)
- Nanotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plasma & Fusion (AREA)
- Materials Engineering (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Manufacturing & Machinery (AREA)
- Electrochemistry (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Optics & Photonics (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention belongs to the field of electrochemiluminescence detection, and relates to an electrochemiluminescence body and an electrochemiluminescenceThe luminescent aptamer sensor is formed by loading lincomycin aptamer on the surface of a glassy carbon electrode modified by UCNPs@Au/PTCA-PATP nanocomposite, wherein the UCNPs@Au/PTCA-PATP nanocomposite is a product formed by combining conversion nanoparticles modified by gold nanoparticles and thiolated perylene tetracarboxylic acid by gold sulfide bonds; the lincomycin aptamer nucleotide sequence is apt:5'-CGCG TGAT GTGG TCGA TGCG ATAC GGTG AGTC GCGC CACG GCTA CACA CGTC TCAG CGA-3'. The sensor can realize sensitive detection of lincomycin, and the minimum detection limit is 2.4 multiplied by 10 ‑16 mol/L。
Description
Technical Field
The invention belongs to the field of electrochemiluminescence detection, and relates to an electrochemiluminescence body, an electrochemiluminescence aptamer sensor, a preparation method and application thereof.
Background
Lincomycin (Lin for short) which is a Lincomycin amide antibiotic produced by fermentation of streptomyces has strong antibacterial activity and wide antibacterial spectrum, is widely applied to livestock and poultry cultivation, is often added into feed or drinking water, and most of the veterinary drug which is not absorbed by animal bodies enters the environment in a body or metabolite form, so that the antibiotic accumulation in the natural environment is caused, the balance of microorganisms is destroyed, and then the microorganism can enter human bodies through food chains, so that bacterial drug resistance is generated in the human bodies, and the human health is finally endangered. The problem of lincomycin residue in water environments has become a public health problem. The national Ministry of agriculture bulletin prescribes that the maximum residual limit of lincomycin in animal food is 100ng/mL.
The methods for detecting lincomycin are commonly used at present: photoelectric method, enzyme-linked immunosorbent method, molecular imprinting, high performance liquid chromatography, fluorescence method and the like, and the methods have the defects of expensive instrument and equipment, complex pretreatment of samples, low sensitivity, poor stability and the like. Therefore, developing a convenient and efficient analysis method for detecting the lincomycin content in the water body has very important significance.
Electrochemiluminescence (ECL) is also called electrochemiluminescence, and combines the advantages of wide electrochemical detection range, simple instrument, high sensitivity of chemiluminescence, good reproducibility, stable reagent and easy control. And the anti-interference capability of the sensor can be enhanced by introducing a nucleic acid aptamer chain with low price and strong specificity. ECL is a method for acquiring a luminous intensity spectrum with the aid of optical instruments such as photomultiplier tubes and the like, and establishing a relation between the luminous intensity spectrum and an object to be detected so as to realize trace analysis.
Disclosure of Invention
The invention aims at overcoming the defect of lincomycin detection in the prior art and provides an electrochemiluminescence body, an electrochemiluminescence aptamer sensor, a preparation method and application thereof. The electrochemical luminophor is UCNPs@Au/PTCA-PATP nanocomposite material, and is a substance which is modified by gold nano particles, converts nano particles UCNPs and is combined with mercapto perylene tetracarboxylic acid PTCA-PATP by gold-sulfur bonds. UCNPs@Au and PTCA-PATP are tightly combined through gold sulfide bonds, perylene substances have good electrochemical activity and good film forming property, gold nanoparticles have excellent conductivity, a relay barrier between a luminescent material and an electrode can be reduced, up-conversion nanoparticles (UCNPs) have stable cathode signals and excellent electrical and optical characteristics, and the combination of the three has good synergistic effect, so that the sensitivity and the strong anti-interference capability can be effectively improved.
When the sensor is used as an electrochemiluminescence aptamer of the electrochemiluminescence aptamer sensor, the sensitivity and stability of electrochemiluminescence are obviously improved, and then the electrochemiluminescence aptamer sensor (abbreviated as apt/UCNPs@Au/PTCA-PATP/GCE sensor) is obtained by loading an aptamer which is suitable for a target molecule to be detected under the action of static electricity, so that the target molecule can be specifically identified, and the selectivity of the target molecule (such as detection of antibiotics such as lincomycin) is improved.
Further, gold nanoparticle modified up-conversion nanoparticles UCNPs (ucnps@au) were obtained by the following method: uniformly dispersed with up-conversion nanoparticles UCNPs and HAuCl 4 Stirring and reacting for at least 30min under the protection of argon at 160-180 ℃, and then stirringGradually cooling to room temperature during stirring.
More specifically, Y 2 O 3 、Yb 2 O 3 、Tm 2 O 3 Respectively, in trifluoroacetic acid, and stirring under sealed conditions, filtering, and drying to give a white solid (Y (CF) 3 COO) 3 、Yb(CF 3 COO) 3 、Tm(CF 3 COO) 3 ). Dispersing the above solid in oleic acid and 1-octadecene at a certain mass ratio, stirring under vacuum, and adding NH 4 And (3) continuously stirring the methanol solution of F and NaOH, and centrifuging, washing and drying to obtain a white solid, namely UCNPs. Finally UCNPs and HAuCl 4 Mixing and stirring, centrifuging, washing and drying to obtain dark purple solid, namely UCNPs@Au.
Further, gold nanoparticle modified up-conversion nanoparticles UCNPs and PTCA-PATP are respectively dispersed in DMF, and then the mass ratio is 1:2, mixing, stirring at room temperature until the mixture is fully combined after ultrasonic full mixing of ice water bath, collecting precipitate and drying in vacuum to obtain UCNPs@Au/PTCA-PATP.
More specifically, UCNPs@Au and PTCA-PATP are respectively dispersed in DMF to form a UCNPs@Au-DMF solution and a PTCA-PATP-DMF solution with the concentration of 1mg/mL, the two solutions are ultrasonically mixed (m (UCNPs@Au)/m (PTCA-PATP) =1:2), stirred (time is generally 16 h), and after the UCNPs@Au-PTCA-PATP is fully reacted, the UCNPs@Au/PTCA-PATP is obtained by centrifugation, washing and drying.
Further, the preparation method of the PTCA-PATP comprises the following steps: mixing perylene-3, 4,9, 10-tetracarboxylic dianhydride, imidazole and 4-amino thiophenol, heating and stirring (preferably at 100 ℃ for 3 hours) under the protection of argon, dispersing in ethanol after cooling, adding a certain amount of hydrochloric acid, continuously stirring (preferably for 16 hours), and filtering, washing and drying to obtain PTCA-PATP.
The invention also provides an electrochemiluminescence aptamer sensor for specifically detecting lincomycin, which is formed by loading an aptamer containing 5'-CGCG TGAT GTGG TCGA TGCG ATAC GGTG AGTC GCGC CACG GCTA CACA CGTC TCAG CGA-3' base sequence on the surface of a glassy carbon electrode modified by UCNPs@Au/PTCA-PATP nanocomposite.
The invention also provides a preparation method of the electrochemiluminescence aptamer sensor for specifically detecting lincomycin, which comprises the following steps:
preparation of apt/UCNPs@Au/PTCA-PATP/GCE sensor:
polishing a glassy carbon electrode, sequentially respectively carrying out ultrasonic treatment by using nitric acid, absolute ethyl alcohol and deionized water, naturally airing for standby, removing ultrapure aqueous dispersion liquid (1 mg/mL) of UCNPs@Au/PTCA-PATP, dripping the ultrapure aqueous dispersion liquid on the surface of the clean glassy carbon electrode, and drying at room temperature to obtain the UCNPs@Au/PTCA-PATP modified glassy carbon electrode; and then, dropwise adding an aptamer to the surface of UCNPs@Au/PTCA-PATP/GCE, combining the aptamer and UCNPs@Au/PTCA-PATP by static electricity, and placing the aptamer and UCNPs@Au/PTCA-PATP/GCE in a refrigerator with modified electrodes for 6-8h to obtain the ECL aptamer sensor.
Finally, the invention provides an application method of an electrochemiluminescence aptamer sensor for specifically detecting lincomycin, comprising the following steps:
firstly preparing a series of lincomycin standard solutions with different concentrations, preparing the lincomycin into a solution by using water, and then adding K-containing solution 2 S 2 O 8 In PBS buffer solution of (2) to obtain a series of lincomycin standard solutions with different concentrations, the concentration range is 1.0X10% -15 mol/L~1.0×10 -7 mol/L。
Then establishing a linear regression equation, taking an apt/UCNPs@Au/PTCA-PATP/GCE sensor as a working electrode, taking a platinum electrode as an auxiliary electrode and taking Ag/AgCl as a reference electrode to form a three-electrode system, and placing the three-electrode system in a series of lincomycin standard solutions with different concentrations and the K-containing solution 2 S 2 O 8 Soaking in PBS buffer solution, and carrying out cyclic voltammetry scanning at high pressure of 700V and scanning speed of 0.1V/s in an electrochemical window range of-1.8-0V, recording an electric potential-luminous intensity curve (E-ECL), and establishing a linear relation between a luminous intensity difference value before and after lincomycin is added and a lincomycin concentration logarithmic value to obtain a corresponding linear regression equation.
Finally, sample detection is carried out: the sample to be tested is subjected to a preliminary treatment (the preliminary treatment is mainly to filter impurities, e.g. taking a cell200mL of pond water sample, naturally standing for a period of time, centrifuging at 6000rpm for 10min, sucking supernatant, filtering with 0.22 μm filter membrane, and testing under the same electrochemiluminescence test conditions (the filtrate is formulated to contain 0.05mol/L K) 2 S 2 O 8 25mL of the obtained solution is taken in a detection pool, the luminous intensity is recorded, the obtained luminous intensity is used for calculating the concentration of lincomycin in the sample to be detected by using a linear regression equation corresponding to the obtained standard curve.
Further, contain K 2 S 2 O 8 The preparation method of the PBS buffer solution comprises the following steps: preparation of 0.05mol/L K in 0.1mol/L PBS buffer at pH7.4 2 S 2 O 8 Is a solution of PBS buffer.
Further, in the electrochemiluminescence test, the photomultiplier is high-pressure 700V, the sweeping speed is 0.1V/s, and the soaking time is 35min (the detection value is stable in 35 min).
Compared with the prior art, the invention has the following beneficial effects: the invention designs an electrochemiluminescence aptamer sensor based on perylene derivatives of up-conversion nano particles, UCNPs@Au and PTCA-PATP are combined by gold-sulfur bonds, and an electrochemiluminescence body with high intensity and stable performance can be obtained -15 ~1.0×10 -7 mol/L, the lowest detection limit is 2.4X10 -16 mol/L. The method for detecting lincomycin has the advantages of simple operation, good selectivity, low detection cost, high sensitivity and good stability.
Drawings
FIG. 1 is a scanning electron microscope (EDS) diagram of PTCA-PATP (A), UCNPs@Au (B), UCNPs@Au/PTCA-PATP (C), UCNPs@Au/PTCA-PATP (D).
FIG. 2 is a graph of ECL-potential curves for different concentrations of lincomycin.
Wherein the concentration of lincomycin is as follows from front to back according to the peak value of the curve: 1.0X10 -15 mol/L、1.0×10 -14 mol/L、1.0×10 -13 mol/L、1.0×10 -12 mol/L、1.0×10 -11 mol/L、1.0×10 -10 mol/L、1.0×10 -9 mol/L、1.0×10 -8 mol/L、1.0×10 -7 mol/L。
FIG. 3 is a standard curve of the difference in luminescence intensity and log concentration of lincomycin before and after addition of lincomycin.
FIG. 4 is a specific assay for lincomycin using the sensor of example 1.
FIG. 5 is an ECL-voltage diagram of modified electrodes of GCE, UCNPs@Au/GCE, PTCA-PATP/GCE, au/PTCA-PATP/GCE, UCNPs/PTCA-PATP/GCE, UCNPs@Au/PTCA-PATP/GCE in the invention.
Detailed Description
The present invention is not limited to the following embodiments, and those skilled in the art can implement the present invention in various other embodiments according to the present invention, or simply change or modify the design structure and thought of the present invention, which fall within the protection scope of the present invention. It should be noted that, without conflict, the embodiments of the present invention and features of the embodiments may be combined with each other.
The invention is further described in detail below in connection with the examples:
in the embodiment, an electrochemiluminescence aptamer sensor for specifically detecting lincomycin is provided, wherein the electrochemiluminescence aptamer sensor is formed by loading an aptamer containing 5'-CGCG TGAT GTGG TCGA TGCG ATAC GGTG AGTC GCGC CACG GCTA CACA CGTC TCAG CGA-3' base sequences on the surface of a glassy carbon electrode modified by UCNPs@Au/PTCA-PATP nanocomposite, and the UCNPs@Au/PTCA-PATP nanocomposite is a product bonded by gold sulfide bonds between up-conversion nanoparticles (UCNPs@Au) and thiolated perylene tetracarboxylic acid (PTCA-PATP), and the electrochemiluminescence aptamer sensor can be obtained by a method described in the embodiment 1, and is specifically as follows:
example 1:
(1) Preparation of UCNPs@Au:
first, 0.3388g Y is weighed 2 O 3 ,0.2758g Yb 2 O 3 ,0.1158g Tm 2 O 3 Respectively dissolvingSealing and stirring in 14mL/8mL/4mL 50% trifluoroacetic acid water solution at 85deg.C until the solution becomes clear, filtering to remove insoluble substances, and slowly evaporating the obtained clear solution at 100-120deg.C to obtain white solid, namely Y (CF) 3 COO) 3 、Yb(CF 3 COO) 3 、Tm(CF 3 COO) 3 . Subsequently, 0.5950g Y (CF) was added to a flask containing 12mL of oleic acid and 15mL of 1-octadecene 3 COO) 3 、0.3072g Yb(CF 3 COO) 3 、0.0052g Tm(CF 3 COO) 3 The mixture was heated at 120 ℃ with vigorous stirring under vacuum to remove water and oxygen until no bubbles were generated, and thereby a clear solution containing the lanthanide-oleate complex was formed. The solution was then cooled naturally to room temperature under argon. Thereafter, 12mL of a catalyst containing NH was added 4 A solution of F (0.2963 g) and NaOH (0.2100 g) in methanol was stirred at 50℃for 20min, the methanol was evaporated, the solution was then heated to 300℃under argon for 1h, and the solution was then cooled rapidly to room temperature. The up-converted nanoparticles obtained by precipitation with ethanol were added to the reaction solution, and finally centrifuged through 9000rmp for 10min, washed three times with ethanol/cyclohexane (v: v=1:1), and dried at 60 ℃ under vacuum for 12h to obtain a white solid, UCPNs. 50mg UCNPs were dispersed in 10mL ethylene glycol and then further containing 20. Mu.M HAuCl 4 Mixing 10mL of ethylene glycol (mixed with 300mg of PVP) with the solution, heating to 180 ℃ under the protection of argon, preserving heat and stirring for 30min, continuously stirring for 16h, gradually cooling to room temperature, washing with acetone, centrifuging, and drying in vacuum to obtain dark purple solid, namely UCNPs@Au.
(2) Preparation of PTCA-PATP:
0.3209g of perylene-3, 4,9, 10-tetracarboxylic dianhydride (PTCDA), 6.1566g of imidazole and 0.7850g of 4-aminophenylsulfol were heated to 100℃under argon and stirred for 3h. After the reaction was cooled to room temperature, 100mL of ethanol and 300mL of hydrochloric acid (2M) were added, stirring was continued for 16h, followed by vacuum filtration through a 0.45 μm membrane filter, washing with ultrapure water to neutral pH, placing the solid in a vacuum oven, and drying at 60 ℃ for 12h to obtain a reddish-white solid, namely PTCA-PATP.
(3) Preparation of UCNPs@Au/PTCA-PATP:
and (3) dispersing UCNPs@Au and PTCA-PATP prepared in the step (1) and the step (2) in DMF respectively to form a UCNPs@Au-DMF solution and a PTCA-PATP-DMF solution of 1mg/mL, uniformly mixing the two solutions (m (UCNPs@Au)/m (PTCA-PATP) =1:2) by ultrasound, stirring for 16h, and obtaining a product, namely UCNPs@Au/PTCA-PATP, through centrifugation, washing and drying after the two solutions are fully reacted.
(4) Preparation of modified electrode apt/UCNPs@Au/PTCA-PATP/GCE:
and polishing the glassy carbon electrode, sequentially respectively carrying out ultrasonic treatment by using nitric acid, absolute ethyl alcohol and deionized water, naturally airing for standby, transferring 5 mu L of UCNPs@Au/PTCA-PATP solution to the surface of the clean glassy carbon electrode by using a pipetting gun, and drying at room temperature to obtain the UCNPs@Au/PTCA-PATP modified glassy carbon electrode. Then, 5 mu L of 3 mu mol/L of aptamer is dripped on the surface of UCNPs@Au/PTCA-PATP/GCE, and the aptamer and UCNPs@Au/PTCA-PATP are combined through static electricity to obtain the apt/UCNPs@Au/PTCA-PATP/GCE. Finally, placing the apt/UCNPs@Au/PTCA-PATP/GCE modified electrode in a refrigerator at the temperature of 4 ℃ for 6-8 hours to obtain the ECL aptamer sensor.
The recognition molecule apt sequence is: apt 5'-CGCG TGAT GTGG TCGA TGCG ATAC GGTG AGTC GCGC CACG GCTA CACA CGTC TCAG CGA-3' (manufacturer is a division of bioengineering (Shanghai))
(5) Drawing of a Standard Curve
The modified electrode apt/UCNPs@Au/PTCA-PATP/GCE is used as a working electrode, a platinum electrode is used as an auxiliary electrode, ag/AgCl is used as a reference electrode to form a three-electrode system, and the three-electrode system is placed in a series of lincomycin concentrations (1.0X10) -15 mol/L、1.0×10 -14 mol/L、1.0×10 -13 mol/L、1.0×10 -12 mol/L、1.0×10 -11 mol/L、1.0×10 -10 mol/L、1.0×10 -9 mol/L、1.0×10 -8 mol/L and 1.0X10 -7 mol/L) K containing 0.05mol/L 2 S 2 O 8 Soaking in 0.1mol/L PBS buffer solution with pH of 7.4 for 35min, and in electrochemical window range of-1.8-0V, performing cyclic voltammetry scanning at high pressure 700V and scanning speed of 0.1V/s, recording potential-luminous intensity curve (E-ECL), and establishingThe linear relation between the light-emitting intensity difference before and after adding lincomycin and the lincomycin concentration logarithmic value is obtained, and a corresponding linear regression equation is obtained as follows: deltaI ECL =6240.1245+1243.3891 lgc (mol/L), and the correlation coefficient (R) is 0.9993. The detection range of the linear regression equation is 1.0X10 -15 ~1.0×10 -7 mol/L, the lowest detection limit is 2.4X10 -16 mol/L。
(6) Detection of samples
Taking water sample of pond, naturally standing for a period of time, centrifuging to collect upper layer solution, filtering with 0.22 μm filter membrane to collect filtrate, adding water sample containing 0.05mol/LK 2 S 2 O 8 The pH of the solution was adjusted to 7.4 by using 0.1mol/L PBS buffer solution, 25mL of the obtained solution was used for electrochemiluminescence analysis, the electrochemiluminescence analysis was performed under the same electrochemiluminescence test conditions as in the step (4), the luminescence intensity was recorded, and the concentration of lincomycin in the sample to be tested was calculated according to the linear regression equation obtained in the step (5), and the results are shown in Table 1.
The invention has the remarkable advantages that the electrochemical luminescence sensor for detecting lincomycin and the preparation method thereof are developed, and compared with the common electrochemical luminescence sensor, the electrochemical luminescence sensor has the remarkable advantages of: firstly, UCNPs@Au and PTCA-PATP are tightly combined through gold-sulfur bonds, and the synergistic effect of the UCNPs@Au and the PTCA-PATP is reflected in ECL; secondly, the perylene material has good electrochemical activity and film forming property, the gold nanoparticles have excellent conductivity, the relay barrier between the luminescent material and the electrode can be reduced, and the up-conversion nanoparticles (UCNPs) have stable cathode signals and excellent electrical and optical characteristics. The invention fully utilizes the advantages of the aptamer and the electrochemical luminescence sensor, successfully realizes the sensitive detection of the lincomycin by the enhancement mechanism of the lincomycin on the ECL signal intensity of the system, and the sensing platform can specifically identify and detect the lincomycin and has high selectivity. The invention has important significance for popularizing the practical application of the aptamer sensor in the aspects of water environment monitoring and the like.
And the electrochemical luminescence sensor for detecting lincomycin prepared in the embodiment 1 is used for improving anti-interference detection, wherein the working after the aptamer is incubatedThe electrodes are respectively at 10 -6 Ciprofloxacin (CIP), chloramphenicol (CAP), kanamycin (KAN), aureomycin (CTC), streptomycin (STP) interferents, and at 10 -8 M was tested in lincomycin (ENR) standard solution, and the working electrode was again tested in a mixture of the above materials, the test results are shown in FIG. 4.
As can be seen from fig. 4, the modified electrode with excellent electrochemical performance has a selective recognition effect on lincomycin after the aptamer is incubated thereon, and the detection effect of the 100-fold concentration of the interferent on lincomycin after mixing is also very small. Therefore, the working electrode can realize the anti-interference selective detection of lincomycin.
Comparative example 1:
(1) Preparation of UCNPs@Au/PTCA-PATP/GCE modified electrode
Polishing the glassy carbon electrode, sequentially using nitric acid, absolute ethyl alcohol and deionized water for respectively ultrasonic treatment, and naturally airing for later use. And (3) transferring 5.0 mu L of 1.0mg/mL UCNPs@Au/PTCA-PATP material aqueous solution to the surface of a clean glassy carbon electrode by using a pipetting gun, and drying at room temperature to obtain the UCNPs@Au/PTCA-PATP/GCE modified electrode serving as a working electrode for electrochemiluminescence test.
(2) Drawing of a Standard Curve
The UCNPs@Au/PTCA-PATP/GCE modified electrode is used as a working electrode, a platinum electrode is used as an auxiliary electrode, ag/AgCl is used as a reference electrode to form a three-electrode system, and the electrode system contains 0.05mol/L K 2 S 2 O 8 The luminescence intensity was measured using a 0.1mol/L PBS buffer at pH7.4 as a blank solution, and the three-electrode system was placed in a series of lincomycin concentrations (1.0X10) - 15 mol/L、1.0×10 -14 mol/L、1.0×10 -13 mol/L、1.0×10 -12 mol/L、1.0×10 -11 mol/L、1.0×10 - 10 mol/L、1.0×10 -9 mol/L、1.0×10 -8 mol/L and 1.0X10 -7 mol/L) K containing 0.05mol/L 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS with pH of 7.4, in an electrochemical window range of-1.8-0V, a photomultiplier tube is subjected to high pressure 700V and a scanning speed of 0.1V/s, cyclic voltammetry scanning is carried out, an E-ECL curve is recorded, and a luminous intensity difference before and after lincomycin is added is establishedAnd (3) obtaining a corresponding linear regression equation by the linear relation between the value and the lincomycin concentration logarithmic value.
(3) Detection of samples
25mL of treated pond water was added to a pond containing 0.05mol/L K 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS having a pH of 7.4, for electrochemical luminescence detection, the concentration of lincomycin in the sample to be detected was calculated according to the linear regression equation corresponding to the above step (2), and the results are shown in Table 1.
Comparative example 2:
(1) Preparation of apt/UCNPs@Au/GCE modified electrode
Polishing the glassy carbon electrode, sequentially using nitric acid, absolute ethyl alcohol and deionized water for respectively ultrasonic treatment, and naturally airing for later use. Transferring 5.0 mu L of 1.0mg/mL UCNPs@Au aqueous solution to the surface of a clean glassy carbon electrode by using a pipetting gun, and drying at room temperature to obtain a UCNPs@Au/GCE modified electrode; and (3) dripping 5.0 mu L of 3.0 mu M aptamer on the surface of the UCNPs@Au/GCE modified electrode (same as in example 1), and placing in a refrigerator at 4 ℃ for 6 hours to obtain an apt/UCNPs@Au/GCE sensor serving as a working electrode for electrochemiluminescence test.
(2) Drawing of a Standard Curve
The three-electrode system is formed by using an apt/UCNPs@Au/GCE modified electrode as a working electrode, a platinum electrode as an auxiliary electrode and Ag/AgCl as a reference electrode, and K containing 0.05mol/L 2 S 2 O 8 The luminescence intensity was measured using a 0.1mol/L PBS buffer with pH=7.4 as a blank solution, and the three-electrode system was placed in a series of lincomycin concentrations (1.0X10) -15 mol/L、1.0×10 -14 mol/L、1.0×10 -13 mol/L、1.0×10 -12 mol/L、1.0×10 -11 mol/L、1.0×10 -10 mol/L、1.0×10 -9 mol/L、1.0×10 -8 mol/L and 1.0X10 -7 mol/L) K containing 0.05mol/L 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS with pH of 7.4, carrying out cyclic voltammetry scanning at high pressure of 700V and scanning speed of 0.1V/s by a photomultiplier in an electrochemical window range of-1.8-0V, recording E-ECL curve, and establishing a luminous intensity difference value before and after adding lincomycin and lincomycin concentrationAnd obtaining a corresponding linear regression equation by the linear relation of the degree to the numerical value.
(3) Detection of samples
25mL of treated pond water was added to a pond containing 0.05mol/L K 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS having a pH of 7.4, for electrochemical luminescence detection, the concentration of lincomycin in the sample to be detected was calculated according to the linear regression equation corresponding to the above step (2), and the results are shown in Table 1.
Comparative example 3:
(1) Preparation of apt/PTCA-PATP/GCE modified electrode
Polishing the glassy carbon electrode, sequentially using nitric acid, absolute ethyl alcohol and deionized water for respectively ultrasonic treatment, and naturally airing for later use. Transferring 5.0 mu L of PTCA-PATP aqueous solution to the surface of a clean glassy carbon electrode by using a liquid transferring gun, and drying at room temperature to obtain a PTCA-PATP/GCE modified electrode; 5.0 mu L of 3.0 mu M aptamer (same as in example 1) is dripped on the surface of the PTCA-PATP/GCE modified electrode, and the mixture is placed in a refrigerator at 4 ℃ for 6 hours, so that an apt/PTCA-PATP/GCE sensor serving as a working electrode for electrochemiluminescence testing is obtained.
(2) Drawing of a Standard Curve
The three-electrode system is formed by using an apt/PTCA-PATP/GCE modified electrode as a working electrode, a platinum electrode as an auxiliary electrode and Ag/AgCl as a reference electrode, and K containing 0.05mol/L 2 S 2 O 8 The luminescence intensity was measured using a 0.1mol/L PBS buffer with pH=7.4 as a blank solution, and the three-electrode system was placed in a series of lincomycin concentrations (1.0X10) -15 mol/L、1.0×10 -14 mol/L、1.0×10 -13 mol/L、1.0×10 -12 mol/L、1.0×10 -11 mol/L、1.0×10 -10 mol/L、1.0×10 -9 mol/L、1.0×10 -8 mol/L and 1.0X10 -7 Soaking in the standard solution of mol/L) for 35min, taking out, leaching, taking out as a working electrode, carrying out cyclic voltammetry scanning on a photomultiplier tube with high voltage of 700V and a scanning speed of 0.1V/s in an electrochemical window range of-1.8-0V, recording an E-ECL curve, and establishing a linear relation between a luminous intensity difference value before and after lincomycin is added and a lincomycin concentration logarithmic value to obtain a corresponding linear regression equation.
(3) Detection of samples
25mL of treated pond water was added to a pond containing 0.05mol/L K 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS having a pH of 7.4, for electrochemical luminescence detection, the concentration of lincomycin in the sample to be detected was calculated according to the linear regression equation corresponding to the above step (2), and the results are shown in Table 1.
Comparative example 4:
(1) Preparation of Au/PTCA-PATP/GCE modified electrode
Polishing the glassy carbon electrode, sequentially using nitric acid, absolute ethyl alcohol and deionized water for respectively ultrasonic treatment, and naturally airing for later use. And (3) transferring 5.0 mu L of 1.0mg/mL of the aqueous solution of the Au/PTCA-PATP material to the surface of the clean glassy carbon electrode by using a liquid transfer gun, and drying at room temperature to obtain the Au/PTCA-PATP/GCE modified electrode serving as a working electrode for electrochemiluminescence test.
(2) Drawing of a Standard Curve
The Au/PTCA-PATP/GCE modified electrode is used as a working electrode, a platinum electrode is used as an auxiliary electrode, ag/AgCl is used as a reference electrode to form a three-electrode system, and the three-electrode system contains 0.05mol/L K 2 S 2 O 8 The luminescence intensity was measured using a 0.1mol/L PBS buffer at pH7.4 as a blank solution, and the three-electrode system was placed in a series of lincomycin concentrations (1.0X10) -15 mol/L、1.0×10 -14 mol/L、1.0×10 -13 mol/L、1.0×10 -12 mol/L、1.0×10 -11 mol/L、1.0×10 -10 mol/L、1.0×10 -9 mol/L、1.0×10 -8 mol/L and 1.0X10 -7 mol/L) K containing 0.05mol/L 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS with the pH of 7.4, in an electrochemical window range of-1.8-0V, carrying out cyclic voltammetry scanning on a photomultiplier at high pressure of 700V and a scanning speed of 0.1V/s, recording an E-ECL curve, and establishing a linear relation between a luminous intensity difference value before and after lincomycin is added and a lincomycin concentration logarithmic value to obtain a corresponding linear regression equation.
(3) Detection of samples
25mL of treated pond water was added to a pond containing 0.05mol/L K 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS having a pH of 7.4, for electrochemical luminescence detection, the concentration of lincomycin in the sample to be detected was calculated according to the linear regression equation corresponding to the above step (2), and the results are shown in Table 1.
Comparative example 5:
(1) Preparation of UCNPs/PTCA-PATP/GCE modified electrode
Polishing the glassy carbon electrode, sequentially using nitric acid, absolute ethyl alcohol and deionized water for respectively ultrasonic treatment, and naturally airing for later use. And (3) transferring 5.0 mu L of 1.0mg/mL of the aqueous solution of UCNPs/PTCA-PATP material to the surface of a clean glassy carbon electrode by using a liquid transfer gun, and drying at room temperature to obtain the UCNPs/PTCA-PATP/GCE modified electrode serving as a working electrode for electrochemiluminescence test.
(2) Drawing of a Standard Curve
UCNPs/PTCA-PATP/GCE modified electrode is used as working electrode, platinum electrode is used as auxiliary electrode, ag/AgCl is used as reference electrode to form a three-electrode system, and K containing 0.05mol/L 2 S 2 O 8 The luminescence intensity was measured using a 0.1mol/L PBS buffer at pH7.4 as a blank solution, and the three-electrode system was placed in a series of lincomycin concentrations (1.0X10) -15 mol/L、1.0×10 -14 mol/L、1.0×10 -13 mol/L、1.0×10 -12 mol/L、1.0×10 -11 mol/L、1.0×10 -10 mol/L、1.0×10 -9 mol/L、1.0×10 -8 mol/L and 1.0X10 -7 mol/L) K containing 0.05mol/L 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS with the pH of 7.4, in an electrochemical window range of-1.8-0V, carrying out cyclic voltammetry scanning on a photomultiplier at high pressure of 700V and a scanning speed of 0.1V/s, recording an E-ECL curve, and establishing a linear relation between a luminous intensity difference value before and after lincomycin is added and a lincomycin concentration logarithmic value to obtain a corresponding linear regression equation.
(3) Detection of samples
25mL of treated pond water was added to a pond containing 0.05mol/L K 2 S 2 O 8 In a buffer solution of 0.1mol/L PBS (pH 7.4) for electrochemiluminescence detection, according to the linear regression equation corresponding to the step (2)The concentration of lincomycin in the sample to be tested was determined and the results are shown in Table 1.
TABLE 1 determination of lincomycin in pond water
Remarks: a is the average value of three determinations
As shown in Table 1, the samples were tested 3 times in parallel with a relative standard deviation of less than 5% and a labeled recovery ranging from 95% to 101%. The results show that the lincomycin is difficult to detect after the sensing element is further assembled after the UCNPs@Au/PTCA-PATP composite material is not used for modification and the UCNPs@Au or PTCA-PATP modified glassy carbon electrode is used for detection, and the composite electrode material is feasible for detecting the lincomycin in pond water.
The present invention is not limited to the above-mentioned embodiments, but is not limited to the above-mentioned embodiments, and any simple modification, equivalent changes and modification made to the above-mentioned embodiments according to the technical matters of the present invention can be made by those skilled in the art without departing from the scope of the present invention.
Claims (9)
1. An electrochemical light emitter, characterized by: the nano composite material is UCNPs@Au/PTCA-PATP, which is a substance modified by gold nano particles to convert nano particles UCNPs and combined with thiolated perylene tetracarboxylic acid PTCA-PATP by gold-sulfur bonds;
gold nanoparticle modified up-conversion nanoparticle UCNPs are prepared by the following stepsThe method comprises the following steps: uniformly dispersed with up-conversion nanoparticles UCNPs and HAuCl 4 Stirring and reacting for at least 30min at 160-180 ℃ under the protection of argon, and then gradually cooling to room temperature under stirring;
the preparation method of UCNPs comprises the steps of preparing Y (CF 3 COO) 3 、Yb(CF 3 COO) 3 And Tm (CF) 3 COO) 3 Dispersing in oleic acid and 1-octadecene in a certain mass ratio, stirring in vacuum, then adding methanol solution containing NH4F and NaOH, continuing stirring, and obtaining white solid through centrifugation, washing and drying.
2. An electrochemical light emitter according to claim 1, wherein: UCNPs@Au/PTCA-PATP is obtained by the following method: the gold nanoparticle modified up-conversion nanoparticles UCNPs and PTCA-PATP are respectively dispersed in DMF, and the mass ratio is 1:2, mixing, stirring at room temperature until the mixture is fully combined after ultrasonic full mixing of ice water bath, collecting precipitate and drying in vacuum to obtain UCNPs@Au/PTCA-PATP.
3. An electrochemical light emitter according to claim 1, wherein: the preparation method of the PTCA-PATP comprises the following steps: mixing perylene-3, 4,9, 10-tetracarboxylic dianhydride, imidazole and 4-amino thiophenol, heating and stirring under the protection of argon, dispersing in ethanol after cooling, adding a certain amount of hydrochloric acid, continuously stirring, and filtering, washing and drying to obtain PTCA-PATP.
4. Use of an electrochemical light-emitting body according to any one of claims 1 to 3, characterized in that: an electrochemiluminescence body for use as an electrochemiluminescence aptamer sensor.
5. An electrochemiluminescent aptamer sensor for specifically detecting lincomycin, characterized in that: is formed by loading lincomycin aptamer on the surface of the electrochemical luminophor modified glassy carbon electrode in any one of claims 1 to 3;
the lincomycin aptamer nucleotide sequence is as follows:
apt: 5′-CGCG TGAT GTGG TCGA TGCG ATAC GGTG AGTC GCGC CACG GCTA CACA CGTC TCAG CGA-3′。
6. the method for preparing the electrochemiluminescence aptamer sensor for specifically detecting lincomycin according to claim 5, wherein the method comprises the following steps: the method comprises the following steps:
and (3) polishing and cleaning a glassy carbon electrode, dropwise adding DMF dispersion liquid of UCNPs@Au/PTCA-PATP on the surface of the GCE, naturally airing to obtain a UCNPs@Au/PTCA-PATP/GCE modified electrode, then, dropwise adding an aptamer to obtain an apt/UCNPs@Au/PTCA-PATP/GCE modified electrode, and storing at a low temperature for 6-8 hours for standby to obtain the ECL aptamer sensor.
7. Use of an electrochemiluminescent aptamer sensor for the specific detection of lincomycin according to claim 5, characterized in that it comprises the following steps:
(1) Preparing K-containing 2 S 2 O 8 Phosphate PBS buffer solution of (C);
(2) Contains different concentrations of lincomycin and 0.05mol/L K 2 S 2 O 8 PBS buffer solution is prepared;
preparing aqueous solution of lincomycin, and adding a certain amount of the aqueous solution of lincomycin into the aqueous solution containing 0.05mol/L K 2 S 2 O 8 In PBS buffer solution with pH of 0.1 mol/L=7.4 to obtain a series of lincomycin standard solutions with different concentrations, the concentration range is 1.0x10 -15 ~1.0×10 -7 mol/L;
(3) Drawing of a Standard Curve
The modified electrode apt/UCNPs@Au/PTCA-PATP/GCE is taken as a working electrode, a platinum electrode is taken as an auxiliary electrode, ag/AgCl is taken as a reference electrode to form a three-electrode system, the three-electrode system is placed in a series of lincomycin standard solutions with different concentrations to be soaked for a certain time, and the three-electrode system is soaked in a solution containing 0.05mol/L K 2 S 2 O 8 The luminescence intensity was measured as a blank solution with 0.1mol/L PBS buffer at ph=7.4; photomultiplier tube high voltage within electrochemical window range of-1.8-0V700V, scanning at 0.1V/s, performing cyclic voltammetry, recording potential-luminous intensity curve (E-ECL), and establishing a linear relation between luminous intensity difference values before and after lincomycin is added and lincomycin concentration logarithmic value to obtain a corresponding linear regression equation;
(4) Actual sample detection
And (3) detecting an actual sample, performing pretreatment, adjusting the pH value, performing test according to the same electrochemiluminescence test conditions in the step (3), recording the luminescence intensity, and calculating the concentration of lincomycin in the sample to be detected by using a linear regression equation corresponding to the obtained standard curve according to the obtained luminescence intensity.
8. Use of an electrochemiluminescent aptamer sensor for the specific detection of lincomycin according to claim 7, in the detection of lincomycin in water, characterized in that: and (3) soaking the modified electrode apt/UCNPs@Au/PTCA-PATP/GCE for 35min.
9. Use of an electrochemiluminescent aptamer sensor for the specific detection of lincomycin according to claim 7, in the detection of lincomycin in water, characterized in that: the pretreatment method in the step (4) comprises the following steps: and naturally standing the sample to be tested for a period of time, centrifuging to absorb the supernatant, and filtering the supernatant through a 0.22 mu m filter membrane.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210640465.8A CN114965644B (en) | 2022-06-08 | 2022-06-08 | Electrochemiluminescence body, electrochemiluminescence aptamer sensor, and preparation methods and applications thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210640465.8A CN114965644B (en) | 2022-06-08 | 2022-06-08 | Electrochemiluminescence body, electrochemiluminescence aptamer sensor, and preparation methods and applications thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114965644A CN114965644A (en) | 2022-08-30 |
CN114965644B true CN114965644B (en) | 2024-01-23 |
Family
ID=82960584
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210640465.8A Active CN114965644B (en) | 2022-06-08 | 2022-06-08 | Electrochemiluminescence body, electrochemiluminescence aptamer sensor, and preparation methods and applications thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114965644B (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115785462B (en) * | 2022-11-02 | 2023-08-22 | 常州大学 | Zr-PTC luminophor material, electrochemical luminescence sensor, and preparation method and application thereof |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2809647A1 (en) * | 2012-09-07 | 2014-03-07 | First Affiliated Hospital, Third Military Medical University, Chinese People's Liberation Army, P.R. China | Erythropoietin receptor modified electrode and its preparation method and application |
CN111337661A (en) * | 2020-03-25 | 2020-06-26 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Up-conversion bar code fluorescence sensor for detecting veterinary drug residues in food and preparation method thereof |
CN112098484A (en) * | 2020-09-11 | 2020-12-18 | 常州大学 | Sensor for detecting acetamiprid based on electrochemical luminescence method, preparation method and application |
CN112505116A (en) * | 2020-11-05 | 2021-03-16 | 常州大学 | Electrochemical luminescence aptamer sensor for specifically detecting kanamycin, and preparation method and application thereof |
CN113075269A (en) * | 2021-03-26 | 2021-07-06 | 常州大学 | Electrochemical luminescence aptamer sensor for specifically detecting chloramphenicol and preparation method and application thereof |
CN114527113A (en) * | 2022-02-18 | 2022-05-24 | 常州大学 | Electrochemical luminescence aptamer sensor for specifically detecting enrofloxacin, preparation method and application |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20190119731A1 (en) * | 2017-10-24 | 2019-04-25 | The Hong Kong Polytechnic University | Nanoprobe sandwich assay for nucleotide sequence detection |
-
2022
- 2022-06-08 CN CN202210640465.8A patent/CN114965644B/en active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2809647A1 (en) * | 2012-09-07 | 2014-03-07 | First Affiliated Hospital, Third Military Medical University, Chinese People's Liberation Army, P.R. China | Erythropoietin receptor modified electrode and its preparation method and application |
CN111337661A (en) * | 2020-03-25 | 2020-06-26 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Up-conversion bar code fluorescence sensor for detecting veterinary drug residues in food and preparation method thereof |
CN112098484A (en) * | 2020-09-11 | 2020-12-18 | 常州大学 | Sensor for detecting acetamiprid based on electrochemical luminescence method, preparation method and application |
CN112505116A (en) * | 2020-11-05 | 2021-03-16 | 常州大学 | Electrochemical luminescence aptamer sensor for specifically detecting kanamycin, and preparation method and application thereof |
CN113075269A (en) * | 2021-03-26 | 2021-07-06 | 常州大学 | Electrochemical luminescence aptamer sensor for specifically detecting chloramphenicol and preparation method and application thereof |
CN114527113A (en) * | 2022-02-18 | 2022-05-24 | 常州大学 | Electrochemical luminescence aptamer sensor for specifically detecting enrofloxacin, preparation method and application |
Non-Patent Citations (7)
Title |
---|
Au nanoparticle plasmon-enhanced electrochemiluminescence aptasensor based on the 1D/2D PTCA/CoP for diclofenac assay;Jingxian Li et al.;Microchimica Acta;第188卷;全文 * |
CuS as co-reaction accelerator in PTCA-K2S2O8 system for enhancing electrochemiluminescence behavior of PTCA and its application in detection of amyloid-β protein;Xianzhen Song et al.;Biosensors and Bioelectronics;第126卷;222-229 * |
Electrochemiluminescence sensor based on upconversion nanoparticles and oligoaniline-crosslinked gold nanoparticles imprinting recognition sites for the determination of dopamine;Ying Gu et al.;Biosensors and Bioelectronics;第128卷;129-136 * |
Hemin as electrochemically regenerable co-reaction accelerator for construction of an ultrasensitive PTCA-based electrochemiluminescent aptasensor;Wei-Jia Zeng et al.;Biosensors and Bioelectronics;第100卷;490-496 * |
Ni Gao et al..Near-infrared-emitting NaYF4:Yb,Tm/Mn upconverting nanoparticle/gold nanorod electrochemiluminescence resonance energy transfer system for sensitive prostate-specific antigen detection.Analytical and Bioanalytical Chemistry.2017,第409卷2675–2683. * |
基于CdS纳米颗粒和HRP-AuNPs-apt的卡那霉素电化学发光适配体传感器研究;宋海燕;康天放;鲁理平;程水源;;化学研究与应用(05);全文 * |
基于金纳米粒子/巯基-β-环糊精修饰的柔红霉素传感器;吴静;刘桂华;闫雅倩;黄杉生;;化学传感器(04);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN114965644A (en) | 2022-08-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN112505116B (en) | Electrochemical luminescence aptamer sensor for specifically detecting kanamycin, and preparation method and application thereof | |
CN113075269B (en) | Electrochemical luminescence aptamer sensor for specifically detecting chloramphenicol and preparation method and application thereof | |
CN110907511B (en) | Gold-curcumin nanoparticle quenched CdS hybrid TiO2Electrochemical luminescence sensor for detecting insulin by nanobelt | |
CN112098484B (en) | Sensor for detecting acetamiprid based on electrochemical luminescence method, and preparation method and application thereof | |
CN110308286B (en) | Enhanced thyroglobulin electrochemiluminescence immunosensor based on photothermal release signals | |
CN108226252B (en) | Preparation method and application of current type immunosensor for detecting breast cancer | |
CN114965644B (en) | Electrochemiluminescence body, electrochemiluminescence aptamer sensor, and preparation methods and applications thereof | |
Thiruppathiraja et al. | Development of electrochemical based sandwich enzyme linked immunosensor for Cryptosporidium parvum detection in drinking water | |
CN114527113A (en) | Electrochemical luminescence aptamer sensor for specifically detecting enrofloxacin, preparation method and application | |
CN103616423A (en) | Preparation method and application of competitive type aptamer sensor for detecting oxytetracycline | |
CN113406168B (en) | Electrochemical sensor for detecting chloramphenicol by molecular imprinting and preparation method and application thereof | |
He et al. | Electrochemical aptasensor based on aptamer-complimentary strand conjugate and thionine for sensitive detection of tetracycline with multi-walled carbon nanotubes and gold nanoparticles amplification | |
CN111198222B (en) | Preparation and use methods of sandwich type electrochemical immunosensor for detecting prostate specific antigen | |
CN114441616B (en) | Method for modifying new coronavirus biological probe on electrochemical biosensor | |
CN112630279B (en) | Gold nanoparticle-based plasma resonance enhanced electrochemical luminescence sensor for detecting dichlorophenolic acid and preparation method thereof | |
CN105866211B (en) | A kind of preparation method and application of ampicillin molecular engram sensor | |
CN104483481B (en) | A kind of preparation method of the carcinoma of urinary bladder mark-NMP22 immunosensor based on carbon nano-tube/PdPt nanocages structure and application | |
CN113702461B (en) | Preparation method of photoelectrochemistry self-energized sensor and application of photoelectrochemistry self-energized sensor in detection of lincomycin | |
CN116124848A (en) | Preparation method and application of molecularly imprinted electrochemical sensor | |
CN111398388B (en) | Method for detecting Listeria monocytogenes based on bifunctional sensor for detecting nucleic acid hly and acetoin | |
CN111272600B (en) | Quartz crystal microbalance sensor and modification method and application of gold electrode thereof | |
CN114235936A (en) | Time induction sensor and preparation method and application thereof | |
CN114441614A (en) | Electrochemical microorganism rapid detector and modification method of biological probe | |
CN110308142B (en) | Tetracycline flexible gold-like foil electrochemical luminescence sensor and preparation method and detection method thereof | |
CN115932000B (en) | Electrochemical biosensor for detecting EGFR-ctDNA and preparation method and application 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 |