CN110003272B - Functional quaternary phosphonium salt self-assembly material and biosensor thereof - Google Patents
Functional quaternary phosphonium salt self-assembly material and biosensor thereof Download PDFInfo
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- 239000000463 material Substances 0.000 title claims abstract description 24
- 238000001338 self-assembly Methods 0.000 title claims abstract description 20
- 150000004714 phosphonium salts Chemical group 0.000 title claims abstract description 19
- 238000006243 chemical reaction Methods 0.000 claims abstract description 14
- 238000010560 atom transfer radical polymerization reaction Methods 0.000 claims abstract description 13
- -1 alkyl tributyl phosphonium bromide Chemical compound 0.000 claims abstract description 10
- KRLHYNPADOCLAJ-UHFFFAOYSA-N undecyl 2-methylprop-2-enoate Chemical compound CCCCCCCCCCCOC(=O)C(C)=C KRLHYNPADOCLAJ-UHFFFAOYSA-N 0.000 claims abstract description 7
- WAKFRZBXTKUFIW-UHFFFAOYSA-N 2-bromo-2-phenylacetic acid Chemical compound OC(=O)C(Br)C1=CC=CC=C1 WAKFRZBXTKUFIW-UHFFFAOYSA-N 0.000 claims abstract description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 50
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 26
- 239000012498 ultrapure water Substances 0.000 claims description 26
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 24
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 17
- 239000010931 gold Substances 0.000 claims description 17
- 229910052737 gold Inorganic materials 0.000 claims description 17
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 16
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 16
- 238000002791 soaking Methods 0.000 claims description 14
- UGZAJZLUKVKCBM-UHFFFAOYSA-N 6-sulfanylhexan-1-ol Chemical compound OCCCCCCS UGZAJZLUKVKCBM-UHFFFAOYSA-N 0.000 claims description 13
- 238000005406 washing Methods 0.000 claims description 13
- GGSUCNLOZRCGPQ-UHFFFAOYSA-N diethylaniline Chemical compound CCN(CC)C1=CC=CC=C1 GGSUCNLOZRCGPQ-UHFFFAOYSA-N 0.000 claims description 12
- 238000010992 reflux Methods 0.000 claims description 11
- 238000010438 heat treatment Methods 0.000 claims description 10
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 8
- 238000002390 rotary evaporation Methods 0.000 claims description 8
- 229910006213 ZrOCl2 Inorganic materials 0.000 claims description 7
- 238000011534 incubation Methods 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- VHRYZQNGTZXDNX-UHFFFAOYSA-N methacryloyl chloride Chemical compound CC(=C)C(Cl)=O VHRYZQNGTZXDNX-UHFFFAOYSA-N 0.000 claims description 6
- 238000001953 recrystallisation Methods 0.000 claims description 6
- 238000005498 polishing Methods 0.000 claims description 5
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 4
- XFGANBYCJWQYBI-UHFFFAOYSA-N 11-bromoundecan-1-ol Chemical compound OCCCCCCCCCCCBr XFGANBYCJWQYBI-UHFFFAOYSA-N 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 239000012074 organic phase Substances 0.000 claims description 4
- 229920006395 saturated elastomer Polymers 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- TUQOTMZNTHZOKS-UHFFFAOYSA-N tributylphosphine Chemical compound CCCCP(CCCC)CCCC TUQOTMZNTHZOKS-UHFFFAOYSA-N 0.000 claims description 4
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 claims description 3
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 3
- 239000012279 sodium borohydride Substances 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 240000000411 Sansevieria trifasciata Species 0.000 claims description 2
- 150000002343 gold Chemical class 0.000 claims description 2
- 238000007789 sealing Methods 0.000 claims description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 2
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- 238000000034 method Methods 0.000 claims 3
- 102000053602 DNA Human genes 0.000 claims 1
- 108020004414 DNA Proteins 0.000 claims 1
- QRZAKQDHEVVFRX-UHFFFAOYSA-N biphenyl-4-ylacetic acid Chemical compound C1=CC(CC(=O)O)=CC=C1C1=CC=CC=C1 QRZAKQDHEVVFRX-UHFFFAOYSA-N 0.000 abstract description 10
- 238000006116 polymerization reaction Methods 0.000 abstract description 6
- 125000000217 alkyl group Chemical group 0.000 abstract description 5
- 239000003999 initiator Substances 0.000 abstract description 2
- 238000011895 specific detection Methods 0.000 abstract description 2
- 101710141544 Allatotropin-related peptide Proteins 0.000 abstract 2
- 239000000178 monomer Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 25
- 235000019441 ethanol Nutrition 0.000 description 12
- VMGSQCIDWAUGLQ-UHFFFAOYSA-N n',n'-bis[2-(dimethylamino)ethyl]-n,n-dimethylethane-1,2-diamine Chemical compound CN(C)CCN(CCN(C)C)CCN(C)C VMGSQCIDWAUGLQ-UHFFFAOYSA-N 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical group OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 238000009210 therapy by ultrasound Methods 0.000 description 3
- 229910021590 Copper(II) bromide Inorganic materials 0.000 description 2
- 229910021135 KPF6 Inorganic materials 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- QTMDXZNDVAMKGV-UHFFFAOYSA-L copper(II) bromide Substances [Cu+2].[Br-].[Br-] QTMDXZNDVAMKGV-UHFFFAOYSA-L 0.000 description 2
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- 241001481789 Rupicapra Species 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
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- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
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- 238000013461 design Methods 0.000 description 1
- 230000005518 electrochemistry Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000005669 field effect Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000001453 impedance spectrum Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 239000002608 ionic liquid Substances 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000004377 microelectronic Methods 0.000 description 1
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- 108020004707 nucleic acids Proteins 0.000 description 1
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- 102000039446 nucleic acids Human genes 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- PMOIAJVKYNVHQE-UHFFFAOYSA-N phosphanium;bromide Chemical compound [PH4+].[Br-] PMOIAJVKYNVHQE-UHFFFAOYSA-N 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- XYFCBTPGUUZFHI-UHFFFAOYSA-O phosphonium Chemical compound [PH4+] XYFCBTPGUUZFHI-UHFFFAOYSA-O 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
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- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/54—Quaternary phosphonium compounds
- C07F9/5407—Acyclic saturated phosphonium compounds
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/001—Enzyme electrodes
- C12Q1/005—Enzyme electrodes involving specific analytes or enzymes
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Abstract
The invention discloses a functionalized quaternary phosphonium salt self-assembly material and a biosensor thereof. The (1-undecyl methacrylate) alkyl tributyl phosphonium bromide of the invention consists of an ionic head and a long chain alkyl tail, the long chain alkyl tail is functionalized, methacrylate ester bonds are introduced to lead the long chain alkyl tail to become a monomer for ATRP polymerization reaction, and Zr is added to react4+With initiator alpha-bromophenylacetic acid (BPAA)+An ATRP reaction system is introduced on a tDNA framework to construct a biosensor and realize the specific detection of the tDNA.
Description
Technical Field
The invention relates to a (1-undecyl methacrylate) alkyl tributyl phosphonium bromide self-assembly material and a biosensor constructed by the self-assembly material, belonging to the field of biosensing materials.
Background
As a novel functionalized self-assembly material, the quaternary phosphonium salt ionic liquid has the advantages of low steam pressure, nonflammability, good thermal and chemical stability, low corrosivity, high conductivity, large electrochemical window, certain specific functional groups and the like, and is applied to a plurality of fields of chemistry and chemical industry, electrochemistry, new materials, electronics and information technology, biology, medicine and the like.
A biosensor is an analytical tool consisting of an identification element, a transducer and a signal amplifier. The recognition element, also called a sensitive element, comprises bioactive substances such as antigens, antibodies, cells, tissues, enzymes, nucleic acids, microorganisms and the like, which are used as targets to be detected and must ensure that the bioactive substances can cause certain physical or chemical changes; commonly used transducers include electrodes, field effect transistors, photodiodes, piezoelectric crystals, etc., which convert physical or chemical changes expressed by the biological activity of the identification element into electrical signals; then, the obtained biological signals are amplified by using the modern microelectronic and automatic instrument technology, and the detection of the target is realized. The biosensor has the characteristics of good selectivity, high sensitivity, high analysis speed, low cost, high automation, miniaturization and integration, and capability of carrying out online continuous monitoring in a complex system, and is widely applied to the fields of food, environment, fermentation, medicine and the like.
Disclosure of Invention
The invention aims to provide a functional quaternary phosphonium salt self-assembly material, and an ATRP polymerization reaction can be carried out by utilizing a methacrylic ester bond at the tail part of a long-chain alkyl of the self-assembly material to construct a biosensor. By performing an ATRP reaction on a phosphate backbone, an impedance signal is amplified, and the sensitivity of the biosensor is improved.
The technical scheme of the invention is as follows:
a functional quaternary phosphonium salt self-assembly material is (1-undecyl methacrylate) alkyl tributyl phosphonium bromide, and the molecular structural formula of the material is shown as a formula I:
the preparation method of the functional quaternary phosphonium salt self-assembly material comprises the following specific steps:
To (1-undecamet) alkyltributylphosphonium bromide.
Further, in the step 1, the recrystallization is carried out for 8-12 hours.
Further, in the step 2, the molar ratio of the methacryloyl chloride to the N, N-diethylaniline is 1:1, and the recrystallization time is 8-12 h.
The biosensor constructed by the functional quaternary phosphonium salt self-assembly material comprises the following specific preparation steps: sequentially modifying PNA, 6-mercapto-1-hexanol (MCH) and tDNA on the surface of the pretreated gold electrode; sequentially soaking the modified gold electrode in ZrOCl2Taking out the solution and an alpha-bromophenyl acetic acid (BPAA) solution, and then carrying out Atom Transfer Radical Polymerization (ATRP) polymerization reaction on the surface of a gold electrode with a modified tDNA framework in an eATRP solution prepared from a quaternary phosphonium salt self-assembly material. The biosensor is used for detecting the change of the impedance signal of the biosensor, so that the specificity detection of tDNA is realized.
Further, the pretreatment steps of the gold electrode are as follows: and (3) polishing by adopting conventional alumina turbid liquid, after soaking by using the sansevieria trifasciata acid and the sodium borohydride, polishing by using ultrapure water for more than 20min, and realizing that the impedance signal is small enough by using the pretreated gold electrode.
Further, the steps of sequentially modifying PNA, 6-mercapto-1-hexanol (MCH) and tDNA on the surface of the pretreated gold electrode are as follows: dripping 5.0 mu L of 0.5 mu M PNA on the surface of the treated gold electrode, then placing the electrode in a 37 ℃ incubator for incubation for 1-1.5 h, and washing the electrode with ultrapure water; preparing 2.0mM MCH solution by using 60% ethanol, soaking the surface of the electrode in the MCH solution, incubating for 15-30 min at 37 ℃, and then washing the electrode clean by using 60% ethanol and ultrapure water; 10 mu L of 1 fM-0.1 nM oligonucleotide (tDNA) is dripped on the surface of the electrode, then the electrode is placed in an incubator at 37 ℃ for incubation for 0.5-1.5 h, and the electrode is washed clean by ultrapure water.
Further, 5.0mM ZrOCl was prepared with 10% ethanol and 15% ethanol, respectively2Solution and 1.0mM BPAA solution, soaking the electrode in ZrOCl2Incubating the solution for 5-15 min at 37 ℃, and then washing the electrode with 10% ethanol and ultrapure water; and then soaking the electrode in a BPAA solution, incubating for 10-30 min at 37 ℃, and then washing the electrode clean by 15% ethanol and ultrapure water.
Further, the preparation steps of the eATRP solution are as follows: reacting CuBr2And Me6TREN was dissolved in DMF at a ratio of 1:1.1 to make 10mM CuⅡBr/Me6TREN, 1.0M KBr and 0.1M KPF in ultrapure water respectively6Preparing 10 fM-100 mM quaternary phosphonium salt self-assembly material by using DMF, and then respectively taking 1.8mL of DMF and 0.1mL of CuⅡBr/Me6TREN, 0.1mL of quaternary phosphonium salt self-assembly material, 1.0mL of KBr, and 7.0mL of KPF6Preparing an eATRP solution.
Further, the electrode is soaked in an eATRP solution to generate an ATRP reaction through electropolymerization, and the ATRP reaction time is 5 min-2 h.
Compared with the prior art, the (1-undecyl methacrylate) alkyl tributyl phosphonium bromide has the advantages that the tail part of the long alkyl chain contains the methacrylic ester bond, ATRP polymerization reaction can be carried out on a phosphoric acid skeleton by using the alkyl tributyl phosphonium bromide, impedance signals are amplified, a biosensor is constructed, the specific detection of DNA is realized, and the alkyl tributyl phosphonium bromide can be used in the high-sensitivity biosensing field.
Drawings
FIG. 1 is a schematic representation of (1-undecamrylate) alkyltributylphosphonium bromide1H NMR spectrum.
FIG. 2 is an impedance profile of various stages of a biosensor constructed in accordance with the present invention.
Detailed Description
The present invention will be described in more detail with reference to the following examples and the accompanying drawings.
The synthesis route of the functional quaternary phosphonium salt self-assembly material is as follows.
The design principle of the biosensor is as follows: in the scheme, the gold electrode is sequentially soaked in ZrOCl2Solution and BPAA solution by Zr4+Respectively connecting with a phosphoric acid framework and an initiator BPAA to realize the connection of the BPAA to the tDNA framework, and then realizing ATRP polymerization in an eATRP solution prepared by the quaternary phosphonium salt self-assembly material.
Example 1
2.0g (8.0mmol) of 11-bromoundecanol, 1.9g (9.6mmol) of tributylphosphine and 40mL of chloroform were mixed at room temperature, sealed and the reaction was heated under stirring and refluxed for 48h under a nitrogen blanket. Stopping the reaction, removing chloroform by rotary evaporation, adding 50mL of n-hexane and 0.5mL of ethyl acetate, heating and refluxing for 10min, naturally cooling to room temperature, standing for 12h for recrystallization to obtain 3.3g of 11-alkoxy tributyl phosphonium bromide yellow liquid with the yield of 91%.
3.3g (7.2mmol) of 11-alkoxytributylphosphonium bromide, 1.8g (17.3mmol) of methacryloyl chloride, 2.6g (17.3mmol) of N, N-diethylaniline and 50mL of 1, 4-dioxane were mixed under ice-bath, the reaction was transferred to 55 ℃ for 3h and excess saturated NaHCO was added3Adding 50mL of dichloromethane and 50mL of ultrapure water for extraction and liquid separation, performing rotary evaporation on an organic phase to remove dichloromethane, adding 40mL of n-hexane and 0.5mL of ethyl acetate, heating and refluxing for 10min, naturally cooling to room temperature, standing for 8h, and recrystallizing to obtain 2.9g of (1-undecyl methacrylate) alkyl tributyl phosphonium bromide with the yield of 91%.
FIG. 1 is a drawing of phosphonium (3,4, 5-tris (dodecyloxy) benzyl) triphenyltetrafluoroborate1As can be seen from FIG. 1, the synthesized quaternary phosphonium salt has a correct structure and no impurities, and is very clean.
Example 2
2.0g (8.0mmol) of 11-bromoundecanol, 2.4g (12mmol) of tributylphosphine and 40mL of chloroform were mixed at room temperature, sealed and the reaction was heated under reflux for 72h under nitrogen protection. Stopping the reaction, removing chloroform by rotary evaporation, adding 50mL of n-hexane and 0.5mL of ethyl acetate, heating and refluxing for 10min, naturally cooling to room temperature, standing for 12h for recrystallization to obtain 3.3g of 11-alkoxy tributyl phosphonium bromide yellow liquid with the yield of 91%.
3.3g (7.2mmol) of 11-alkoxytributylphosphonium bromide, 2.7g (26.0mmol) of methacryloyl chloride, 3.9g (26.0mmol) of N, N-diethylaniline and 50mL of 1, 4-dioxane were mixed under ice-bath, the reaction was transferred to 55 ℃ for 5h, and excess saturated NaHCO was added3Adding 50mL of dichloromethane and 50mL of ultrapure water for extraction and liquid separation, performing rotary evaporation on an organic phase to remove dichloromethane, adding 40mL of n-hexane and 0.5mL of ethyl acetate, heating and refluxing for 10min, naturally cooling to room temperature, standing for 8h, and recrystallizing to obtain 2.9g of (1-undecyl methacrylate) alkyl tributyl phosphonium bromide with the yield of 91%.
Example 3
Polishing the gold electrode on chamois leather for 3min by using alumina suspension with the particle size of 0.3 mu m, and then performing ultrasonic treatment on the electrode for 2-3 min by using absolute ethyl alcohol and ultrapure water respectively; soaking the electrode in newly-prepared goby acid for 30min, and then respectively carrying out ultrasonic treatment on the goby acid and ultrapure water for 2-3 min; soaking the electrode in sodium borohydride for 15min, and then performing ultrasonic treatment for 2-3 min by using absolute ethyl alcohol and ultrapure water respectively; the electrodes were polished with ultrapure water for 20 min.
Dripping 5.0 mu L of 0.5 mu M PNA on the surface of the treated gold electrode, then placing the electrode in an incubator at 37 ℃ for incubation for 1.5h, and washing the electrode with ultrapure water to remove the PNA of which the surface is not reacted; preparing 2.0mM MCH solution by using 60% ethanol, soaking the surface of the electrode in the MCH solution, incubating for 30min at 37 ℃, and then washing the electrode clean by using 60% ethanol and ultrapure water; mu.L of 0.1nM oligonucleotide (tDNA) was added dropwise to the electrode surface, incubated in a 37 ℃ incubator for 1.5h, and the electrode was rinsed clean with ultra pure water.
5.0mM ZrOCl was prepared with 10% ethanol and 15% ethanol, respectively2Solution and 1.0mM BPAA solution, soaking the electrode in ZrOCl2Incubating in the solution at 37 deg.C for 15min, and washing the electrode with 10% ethanol and ultrapure water; then the electrode is soaked in the BPAA solution,incubating at 37 deg.C for 30min, and washing the electrode with 15% ethanol and ultrapure water; reacting CuBr2And Me6TREN was dissolved in DMF at a ratio of 1:1.1 to make 10mM CuⅡBr/Me6TREN; 1.8mL of DMF, 0.1mL of 10mM quaternary phosphonium salt ((1-undecamet-yl) alkyltributyl phosphonium bromide), and 0.1mL of 10mM Cu were each usedⅡBr/Me6TREN, 1.0mL of 1.0M KBr and 7.0mL of 0.1M KPF6Preparing an eATRP solution; the electrode is immersed in an etatr solution to undergo an ATRP reaction by electropolymerization.
FIG. 2 is an impedance spectrum of each stage of the experiment, and curve 1 is the impedance of the bare electrode after pretreatment; curves 2 and 3 are superposed to show the impedance of PNA modified on the surface of gold electrode and the impedance of MCH modified on the surface of gold electrode; curves 4 and 5 are the impedance after tDNA is modified on the surface of the gold electrode and the electrode is soaked in ZrOCl2Impedance after incubation in solution; curve 6 is the impedance after soaking the electrode in BPAA solution for incubation; curve 7 is the impedance after ATRP polymerization has occurred. It can be seen from FIG. 2 that the signal is still very strong at a detection sensitivity of 1 nM.
Claims (8)
1. The functional quaternary phosphonium salt self-assembly material is characterized in that the molecular structural formula of the material is shown as a formula I:
2. the method for preparing the functionalized quaternary phosphonium salt self-assembly material according to claim 1, comprising the following steps:
step 1, dissolving 11-bromoundecanol and tributyl phosphine in a molar ratio of 1: 1.2-1.5 in chloroform, sealing under nitrogen protection, heating, stirring, refluxing for 48-72 h, stopping the reaction, removing chloroform by rotary evaporation, adding n-hexane and ethyl acetate, heating, refluxing for 10min, naturally cooling to room temperature, standing, recrystallizing to obtain 11-alkoxy tributyl phosphonium bromide II,
step 2, carrying out ice bath, dissolving 11-alkoxy tributyl phosphonium bromide II, methacryloyl chloride and N, N-diethylaniline with the molar ratio of 1: 2.4-3.6 in 1, 4-dioxane, transferring the reaction to 55 ℃, reacting for 3-5 h, adding excessive saturated NaHCO3Adding dichloromethane and ultrapure water for extraction and liquid separation, removing dichloromethane by organic phase rotary evaporation, adding n-hexane and ethyl acetate, heating and refluxing for 10min, naturally cooling to room temperature, standing for 8-12 h for recrystallization to obtain (1-undecyl methacrylate) alkyl tributyl phosphonium bromide I,
3. the method according to claim 2, wherein in step 1, the recrystallization time is 8 to 12 hours.
4. The process of claim 2, wherein in step 2, the molar ratio of methacryloyl chloride to N, N-diethylaniline is 1:1.
5. The biosensor constructed by using the functionalized quaternary phosphonium salt self-assembly material of claim 1, which comprises the following steps: sequentially modifying PNA, 6-mercapto-1-hexanol and tDNA on the surface of the pretreated gold electrode; sequentially soaking the modified gold electrode in ZrOCl2Taking out the solution and an alpha-bromophenyl acetic acid solution, and then carrying out atom transfer radical polymerization reaction on the surface of a gold electrode modified with a tDNA framework in an eATRP solution prepared from a quaternary phosphonium salt self-assembly material.
6. The biosensor of claim 5, wherein the gold electrode is pretreated by: and (3) polishing by adopting an alumina suspension, soaking by using the sansevieria trifasciata acid and sodium borohydride, and polishing for more than 20min by using ultrapure water.
7. The biosensor of claim 5, wherein the steps of sequentially modifying PNA, 6-mercapto-1-hexanol, and tDNA on the surface of the pre-treated gold electrode are as follows: dripping 5.0 mu L of 0.5 mu M PNA on the surface of the pretreated gold electrode, then placing the electrode in a 37 ℃ incubator for incubation for 1-1.5 h, and washing the electrode clean with ultrapure water; preparing 2.0mM 6-mercapto-1-hexanol solution by using 60% ethanol, soaking the surface of the electrode in the 6-mercapto-1-hexanol solution, incubating for 15-30 min at 37 ℃, and then washing the electrode clean by using 60% ethanol and ultrapure water; and (3) dropwise adding 10 mu L of tDNA (deoxyribonucleic acid) with the concentration of 1fM to 0.1nM to the surface of the electrode, then placing the electrode in an incubator at 37 ℃ for incubation for 0.5 to 1.5 hours, and washing the electrode with ultrapure water.
8. The biosensor of claim 5, wherein 5.0mM ZrOCl is formulated with 10% ethanol and 15% ethanol, respectively2The solution and 1.0mM alpha-bromophenylacetic acid solution were immersed in ZrOCl2Incubating the solution for 5-15 min at 37 ℃, and then washing the electrode with 10% ethanol and ultrapure water; and then soaking the electrode in an alpha-bromophenyl acetic acid solution, incubating for 10-30 min at 37 ℃, and then washing the electrode clean by 15% ethanol and ultrapure water.
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