CN115975110A - Molecularly imprinted polymer and electrochemical sensor for specifically recognizing protocatechuic acid - Google Patents
Molecularly imprinted polymer and electrochemical sensor for specifically recognizing protocatechuic acid Download PDFInfo
- Publication number
- CN115975110A CN115975110A CN202211634175.9A CN202211634175A CN115975110A CN 115975110 A CN115975110 A CN 115975110A CN 202211634175 A CN202211634175 A CN 202211634175A CN 115975110 A CN115975110 A CN 115975110A
- Authority
- CN
- China
- Prior art keywords
- molecularly imprinted
- lafeo
- imprinted polymer
- protocatechuic acid
- enzyme cascade
- 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.)
- Granted
Links
- YQUVCSBJEUQKSH-UHFFFAOYSA-N 3,4-dihydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 title claims abstract description 104
- 229920000344 molecularly imprinted polymer Polymers 0.000 title claims abstract description 45
- 238000006116 polymerization reaction Methods 0.000 claims abstract description 60
- 229910052746 lanthanum Inorganic materials 0.000 claims abstract description 56
- FZLIPJUXYLNCLC-UHFFFAOYSA-N lanthanum atom Chemical compound [La] FZLIPJUXYLNCLC-UHFFFAOYSA-N 0.000 claims abstract description 55
- 229910000859 α-Fe Inorganic materials 0.000 claims abstract description 53
- 102000004190 Enzymes Human genes 0.000 claims abstract description 52
- 108090000790 Enzymes Proteins 0.000 claims abstract description 52
- 239000010931 gold Substances 0.000 claims abstract description 37
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 29
- 229910052737 gold Inorganic materials 0.000 claims abstract description 23
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 22
- 239000002105 nanoparticle Substances 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 17
- 239000000178 monomer Substances 0.000 claims abstract description 15
- 238000000137 annealing Methods 0.000 claims abstract description 12
- 102000003992 Peroxidases Human genes 0.000 claims abstract description 10
- 108040007629 peroxidase activity proteins Proteins 0.000 claims abstract description 9
- 230000006698 induction Effects 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims description 24
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 16
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 14
- 239000006185 dispersion Substances 0.000 claims description 13
- 229910052747 lanthanoid Inorganic materials 0.000 claims description 9
- 150000002602 lanthanoids Chemical class 0.000 claims description 9
- DBCAQXHNJOFNGC-UHFFFAOYSA-N 4-bromo-1,1,1-trifluorobutane Chemical compound FC(F)(F)CCCBr DBCAQXHNJOFNGC-UHFFFAOYSA-N 0.000 claims description 8
- 239000002253 acid Substances 0.000 claims description 8
- 239000011246 composite particle Substances 0.000 claims description 8
- STVZJERGLQHEKB-UHFFFAOYSA-N ethylene glycol dimethacrylate Substances CC(=C)C(=O)OCCOC(=O)C(C)=C STVZJERGLQHEKB-UHFFFAOYSA-N 0.000 claims description 8
- 238000004729 solvothermal method Methods 0.000 claims description 8
- 239000003638 chemical reducing agent Substances 0.000 claims description 7
- 230000000379 polymerizing effect Effects 0.000 claims description 7
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 229910052742 iron Inorganic materials 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 239000002131 composite material Substances 0.000 claims description 5
- 239000013110 organic ligand Substances 0.000 claims description 5
- KFDVPJUYSDEJTH-UHFFFAOYSA-N 4-ethenylpyridine Chemical compound C=CC1=CC=NC=C1 KFDVPJUYSDEJTH-UHFFFAOYSA-N 0.000 claims description 4
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 claims description 4
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 claims description 4
- VCJMYUPGQJHHFU-UHFFFAOYSA-N iron(3+);trinitrate Chemical compound [Fe+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O VCJMYUPGQJHHFU-UHFFFAOYSA-N 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- 230000009471 action Effects 0.000 claims description 3
- 230000009881 electrostatic interaction Effects 0.000 claims description 3
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 claims description 2
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 2
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 2
- RUTXIHLAWFEWGM-UHFFFAOYSA-H iron(3+) sulfate Chemical compound [Fe+3].[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O RUTXIHLAWFEWGM-UHFFFAOYSA-H 0.000 claims description 2
- 229910000360 iron(III) sulfate Inorganic materials 0.000 claims description 2
- FYDKNKUEBJQCCN-UHFFFAOYSA-N lanthanum(3+);trinitrate Chemical compound [La+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O FYDKNKUEBJQCCN-UHFFFAOYSA-N 0.000 claims description 2
- XKUYOJZZLGFZTC-UHFFFAOYSA-K lanthanum(iii) bromide Chemical compound Br[La](Br)Br XKUYOJZZLGFZTC-UHFFFAOYSA-K 0.000 claims description 2
- ICAKDTKJOYSXGC-UHFFFAOYSA-K lanthanum(iii) chloride Chemical compound Cl[La](Cl)Cl ICAKDTKJOYSXGC-UHFFFAOYSA-K 0.000 claims description 2
- FQPSGWSUVKBHSU-UHFFFAOYSA-N methacrylamide Chemical compound CC(=C)C(N)=O FQPSGWSUVKBHSU-UHFFFAOYSA-N 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- 229910017771 LaFeO Inorganic materials 0.000 claims 3
- 239000003480 eluent Substances 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 25
- 241000283690 Bos taurus Species 0.000 abstract description 15
- 102000001554 Hemoglobins Human genes 0.000 abstract description 14
- 108010054147 Hemoglobins Proteins 0.000 abstract description 14
- 238000001514 detection method Methods 0.000 abstract description 11
- 230000015572 biosynthetic process Effects 0.000 abstract description 9
- 229920000642 polymer Polymers 0.000 abstract description 9
- 238000003786 synthesis reaction Methods 0.000 abstract description 9
- 238000002848 electrochemical method Methods 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 4
- 239000004480 active ingredient Substances 0.000 abstract description 2
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 58
- 239000011805 ball Substances 0.000 description 26
- 230000000052 comparative effect Effects 0.000 description 18
- 238000012360 testing method Methods 0.000 description 15
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 12
- 239000000843 powder Substances 0.000 description 12
- 239000008367 deionised water Substances 0.000 description 9
- 229910021641 deionized water Inorganic materials 0.000 description 9
- 238000003760 magnetic stirring Methods 0.000 description 9
- 238000002156 mixing Methods 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 229910052760 oxygen Inorganic materials 0.000 description 8
- 238000003756 stirring Methods 0.000 description 8
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 7
- 238000004108 freeze drying Methods 0.000 description 7
- 239000000725 suspension Substances 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000001816 cooling Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000004833 X-ray photoelectron spectroscopy Methods 0.000 description 5
- 239000011259 mixed solution Substances 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 239000002243 precursor Substances 0.000 description 4
- 239000001509 sodium citrate Substances 0.000 description 4
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 4
- 240000003291 Armoracia rusticana Species 0.000 description 3
- 235000011330 Armoracia rusticana Nutrition 0.000 description 3
- 102000016938 Catalase Human genes 0.000 description 3
- 108010053835 Catalase Proteins 0.000 description 3
- 229960004543 anhydrous citric acid Drugs 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000005485 electric heating Methods 0.000 description 3
- SZQUEWJRBJDHSM-UHFFFAOYSA-N iron(3+);trinitrate;nonahydrate Chemical compound O.O.O.O.O.O.O.O.O.[Fe+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O SZQUEWJRBJDHSM-UHFFFAOYSA-N 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000002086 nanomaterial Substances 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- -1 polytetrafluoroethylene Polymers 0.000 description 3
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 3
- 239000004810 polytetrafluoroethylene Substances 0.000 description 3
- 229910052573 porcelain Inorganic materials 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- OZAIFHULBGXAKX-UHFFFAOYSA-N 2-(2-cyanopropan-2-yldiazenyl)-2-methylpropanenitrile Chemical compound N#CC(C)(C)N=NC(C)(C)C#N OZAIFHULBGXAKX-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- YASYEJJMZJALEJ-UHFFFAOYSA-N Citric acid monohydrate Chemical compound O.OC(=O)CC(O)(C(O)=O)CC(O)=O YASYEJJMZJALEJ-UHFFFAOYSA-N 0.000 description 2
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 2
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 2
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 2
- 229920006125 amorphous polymer Polymers 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 2
- 230000003197 catalytic effect Effects 0.000 description 2
- YCIMNLLNPGFGHC-UHFFFAOYSA-N catechol Chemical compound OC1=CC=CC=C1O YCIMNLLNPGFGHC-UHFFFAOYSA-N 0.000 description 2
- 229960004106 citric acid Drugs 0.000 description 2
- 229960002303 citric acid monohydrate Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000001903 differential pulse voltammetry Methods 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical group P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000000840 electrochemical analysis Methods 0.000 description 2
- 238000000445 field-emission scanning electron microscopy Methods 0.000 description 2
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 239000003999 initiator Substances 0.000 description 2
- HVMFKXBHFRRAAD-UHFFFAOYSA-N lanthanum(3+);trinitrate;hydrate Chemical compound O.[La+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O HVMFKXBHFRRAAD-UHFFFAOYSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- GHMLBKRAJCXXBS-UHFFFAOYSA-N resorcinol Chemical compound OC1=CC=CC(O)=C1 GHMLBKRAJCXXBS-UHFFFAOYSA-N 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- XYYVDQWGDNRQDA-UHFFFAOYSA-K trichlorogold;trihydrate;hydrochloride Chemical compound O.O.O.Cl.Cl[Au](Cl)Cl XYYVDQWGDNRQDA-UHFFFAOYSA-K 0.000 description 2
- NUXZAAJDCYMILL-UHFFFAOYSA-K trichlorolanthanum;hydrate Chemical compound O.Cl[La](Cl)Cl NUXZAAJDCYMILL-UHFFFAOYSA-K 0.000 description 2
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 2
- IHCCLXNEEPMSIO-UHFFFAOYSA-N 2-[4-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]piperidin-1-yl]-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethanone Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C1CCN(CC1)CC(=O)N1CC2=C(CC1)NN=N2 IHCCLXNEEPMSIO-UHFFFAOYSA-N 0.000 description 1
- ZBQCCTCQUCOXBO-UHFFFAOYSA-N 4-(4-aminophenyl)-2,2,6,6-tetramethylcyclohex-3-en-1-amine Chemical compound CC1(C)C(N)C(C)(C)CC(C=2C=CC(N)=CC=2)=C1 ZBQCCTCQUCOXBO-UHFFFAOYSA-N 0.000 description 1
- OMPJBNCRMGITSC-UHFFFAOYSA-N Benzoylperoxide Chemical compound C=1C=CC=CC=1C(=O)OOC(=O)C1=CC=CC=C1 OMPJBNCRMGITSC-UHFFFAOYSA-N 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 240000001238 Gaultheria procumbens Species 0.000 description 1
- 235000007297 Gaultheria procumbens Nutrition 0.000 description 1
- 235000005206 Hibiscus Nutrition 0.000 description 1
- 235000007185 Hibiscus lunariifolius Nutrition 0.000 description 1
- 244000284380 Hibiscus rosa sinensis Species 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 1
- 108700020962 Peroxidase Proteins 0.000 description 1
- 101000833014 Petromyzon marinus Globin-5 Proteins 0.000 description 1
- 240000007164 Salvia officinalis Species 0.000 description 1
- 229910021607 Silver chloride Inorganic materials 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 229940121363 anti-inflammatory agent Drugs 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019400 benzoyl peroxide Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- CREMABGTGYGIQB-UHFFFAOYSA-N carbon carbon Chemical compound C.C CREMABGTGYGIQB-UHFFFAOYSA-N 0.000 description 1
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- UQSQSQZYBQSBJZ-UHFFFAOYSA-N fluorosulfonic acid Chemical compound OS(F)(=O)=O UQSQSQZYBQSBJZ-UHFFFAOYSA-N 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 229910021397 glassy carbon Inorganic materials 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 150000002505 iron Chemical class 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- CSJDCSCTVDEHRN-UHFFFAOYSA-N methane;molecular oxygen Chemical compound C.O=O CSJDCSCTVDEHRN-UHFFFAOYSA-N 0.000 description 1
- OSWPMRLSEDHDFF-UHFFFAOYSA-N methyl salicylate Chemical compound COC(=O)C1=CC=CC=C1O OSWPMRLSEDHDFF-UHFFFAOYSA-N 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000011807 nanoball Substances 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000000634 powder X-ray diffraction Methods 0.000 description 1
- 238000001144 powder X-ray diffraction data Methods 0.000 description 1
- 235000005412 red sage Nutrition 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000027756 respiratory electron transport chain Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 208000013220 shortness of breath Diseases 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 description 1
- 239000012279 sodium borohydride Substances 0.000 description 1
- 229910000033 sodium borohydride Inorganic materials 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 239000012798 spherical particle Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The application discloses a molecularly imprinted polymer and an electrochemical sensor for specifically recognizing protocatechuic acid. The electrochemical sensor utilizes a complex enzyme cascade polymerization platform consisting of lanthanum ferrite porous spheres, gold nanoparticles and bovine hemoglobin to induce functional monomers to perform polymerization reaction in the presence of hydrogen peroxide to form a molecularly imprinted polymer, and an induction probe taking the polymer as a core realizes the specific recognition of protocatechuic acid as an active ingredient of traditional Chinese medicines in a solution under an electrochemical method. The synthesis of the lanthanum ferrite porous ball adopts a solvothermal and post-annealing method, the molecularly imprinted polymer adopts an enzyme-induced polymerization technology, and the polymer is synthesized by catalyzing hydrogen peroxide to generate hydroxyl radicals by using peroxidase and gold nanoparticles together. The enzyme-induced polymerization technology is adopted, so that secondary pollution to target molecules cannot be caused; compared with the traditional detection technology, the molecular imprinting electrochemical probe method is simple to operate and has a specific recognition function.
Description
Technical Field
The application relates to the technical field of high molecular material synthesis technology and electrochemical specificity identification, in particular to a molecularly imprinted polymer and an electrochemical sensor for specifically identifying protocatechuic acid.
Background
Over the past time, the problem of pharmaceutical safety around the world, and in particular the standardized use of traditional Chinese medicines, has been impressive and has attracted considerable attention. Among them, protocatechuic Acid (PA) is a natural active ingredient of Chinese herbs, widely distributed in stems and leaves of red sage root, hibiscus, acanthopanax, wintergreen, etc., and also distributed in small amounts in some edible vegetables, and is considered as an effective antioxidant, antibacterial agent and anti-inflammatory agent. However, studies have reported that medical use of residual protocatechuic acid or improper use (excessive ingestion) of protocatechuic acid can cause shortness of breath and convulsions in the limbs of humans and animals, and even death. Therefore, the method has important significance for monitoring the concentration of protocatechuic acid, particularly for the actual measurement of traditional Chinese medicine. Among many analytical methods, electrochemical analytical methods have been widely accepted by the academia because of their advantages such as simple operation, high sensitivity, economical efficiency, and high output efficiency. Therefore, it is valuable to construct a new electrochemical sensor to identify protocatechuic acid in the detection solution to fill the research gap.
Disclosure of Invention
In view of the above, the present application provides a molecularly imprinted polymer for specifically recognizing protocatechuic acid and an electrochemical sensor using the same as a probe, which can realize specific detection of protocatechuic acid while ensuring high sensitivity.
<Creation process>
In the related art, there is a lack of an effective electrochemical sensor for specifically recognizing protocatechuic acid in a detection solution.
In order to overcome the obstacles of certain biomolecules that may coexist in practical assays, molecular Imprinting (MIT) has emerged from various methods of modified preparation of electrochemical sensors. It is noteworthy that in typical molecular imprinting techniques, the target molecule is specifically recognized only if the template molecule specifically binds to the functional monomer through directed interaction. More importantly, although the molecularly imprinted polymer has a specific recognition function, the contamination of toxic traditional initiators (azobisisobutyronitrile, dibenzoyl peroxide, etc.) is a problem to be solved in the polymerization and subsequent detection processes, especially in the case of selective detection of protocatechuic acid in real samples. Fortunately, hydroxyl radical (. OH) has been extensively studied and considered as a green efficient initiator for polymer synthesis, avoiding secondary contamination during polymerization and detection. At present, researchers find that bovine hemoglobin (BHb) as a natural peroxidase not only can effectively decompose hydrogen peroxide to generate OH, but also has low price and high stability among a plurality of peroxidases, and has practical application value. In addition, bovine hemoglobin, as an important metalloporphyrin, can be modified and fixed on an electrochemical active carrier, so that the sensitivity of the electrochemical sensor is improved.
In terms of the selection of the carrier for fixing the enzyme, the perovskite nano material has more and more attention due to the incomparable effect of excellent electron transfer rate, remarkable biocompatibility and satisfactory stability in the aspect of electrochemical sensing. In particular, the perovskite nano material not only can be used as a carrier for immobilizing biological enzyme or bionic enzyme, but also can be selected as a photo-assisted biosensing probe, so that the sensor has higher electrochemical activity. Therefore, we can expect that the combination of the perovskite nano material and the molecularly imprinted polymer is really a potential candidate material for realizing the specific recognition and detection of protocatechuic acid in the traditional Chinese medicine.
Based on the above search and development of the related art, the present inventors have hard found the following:
firstly, lanthanide series inorganic salt, iron series inorganic salt and citric acid are subjected to solvothermal reaction at a fixed molar ratio to obtain a brownish red precursor, and residual organic ligands are removed in a high-temperature annealing process to obtain the covalent bond-bonded lanthanum ferrite porous ball. The lanthanum ferrite porous ball generally has the porous ball shape with uneven surface, can load more electrochemical active components, and has high working efficiency, the average diameter of the ball body is 800-900 nanometers, and the stability is high. The method comprises the following steps of (1) taking a lanthanum ferrite porous ball as a carrier, fixing gold nanoparticles on the surface of the lanthanum ferrite porous ball through electrostatic action, wherein the average diameter of the gold nanoparticles is 10 nanometers; then peroxidase is fixed on the surface of the gold nanoparticles through Jin Qiu covalent bonds to obtain Au @ LaFeO 3 @ BHb composite particles (also known as a multienzyme cascade polymerization platform).
Compared with the original peroxidase, the compound enzyme cascade polymerization platform has higher hydroxyl radical productivity, can spontaneously initiate polymerization reaction to synthesize the molecularly imprinted polymer after introducing hydrogen peroxide, has higher synthesis efficiency, and is green, environment-friendly and pollution-free. According to the invention, the lanthanum ferrite porous ball presents a complex cross-linked structure under the wrapping of the molecularly imprinted polymer, and the polymer is used as a basis to construct an induction probe, so that the specific recognition and detection of protocatechuic acid in a solution are realized under an electrochemical condition.
Based on the discovery of the innovation, the invention is created.
<Complex enzyme cascade polymerization platform>
The complex enzyme cascade polymerization platform is Au @ LaFeO 3 @ BHb composite particles;
wherein, the Au @ LaFeO 3 In the @ BHb composite particle, a lanthanum ferrite porous ball is used as a carrier, gold nanoparticles are loaded on the surface of the lanthanum ferrite porous ball through electrostatic interaction, and peroxidase is bonded on the gold nanoparticles through Jin Qiu covalent bonds.
Suitably but not limitatively, the average particle size of the lanthanum ferrite porous spheres is 800-900nm, the average particle size of the gold nanoparticles is 10nm, and the topographic surface of the lanthanum ferrite porous spheres is uneven spherical particles.
<Preparation of complex enzyme cascade polymerization platform>
The preparation method of the complex enzyme cascade polymerization platform comprises the following steps:
providing a lanthanum ferrite porous ball;
allowing the lanthanum ferrite porous spheres and peroxidase to form a mixed dispersion;
reacting the mixed dispersion with chloroauric acid solution and reducing agent solution, and centrifuging and drying to obtain Au @ LaFeO 3 @ BHb composite particles.
As an exemplary specific example of the reducing agent, one or more of citric acid, sodium citrate, sodium borohydride, and ascorbic acid; preferably, the preparation method of the gold nanoparticles adopts a one-step reduction method with sodium citrate as a reducing agent.
Suitably but not limitatively, the peroxidase is selected from one of bovine hemoglobin or horseradish catalase, suitably but not limitatively, the reaction temperature is normal temperature, and the reaction time is 3-5h;
preferably, the concentration of the mixed dispersion is 1-2mg mL -1 ；
Preferably, the concentration of the chloroauric acid solution is 0.015-0.025M;
preferably, the concentration of the reducing agent solution is 0.015 to 0.025M.
Preferably, the volume ratio of the mixed dispersion liquid to the chloroauric acid solution to the organic source solution is 25:1:1.
the lanthanum ferrite porous ball is suitably prepared by a method of carrying out a solvothermal reaction on a lanthanide inorganic salt and an iron inorganic salt with the addition of an organic ligand.
Here, the organic ligand includes one or more of anhydrous citric acid or citric acid monohydrate.
Suitably but not limitatively, the concentration of said lanthanide inorganic salt is the same as the molar amount of the iron-based inorganic salt, the molar amount of organic source being 2-3 times the concentration of the lanthanide inorganic salt;
preferably, the lanthanide inorganic salt is selected from one or more of lanthanum nitrate, lanthanum chloride or lanthanum bromide;
preferably, the iron-based inorganic salt is selected from one or more of ferric nitrate, ferric chloride or ferric sulfate;
preferably, the temperature of the solvothermal reaction is 175-185 ℃;
preferably, the preparation method of the lanthanum ferrite porous ball further comprises annealing the target product after the solvothermal reaction;
preferably, the annealing temperature is 790-810 ℃, and the annealing time is 1-3h.
<Molecularly imprinted polymers>
The molecular imprinting polymer is generated by polymerizing a polymerization functional monomer under the action of the complex enzyme cascade polymerization platform by adopting enzyme induction polymerization.
Suitable but non-limiting specific examples of the polymeric functional monomer are derived from one or more of acrylic acid, methacrylic acid, acrylamide, methacrylamide or 4-vinylpyridine.
<Preparation of molecularly imprinted polymers>
The process of enzyme-induced polymerization comprises the following steps:
dispersing a catalyst in a hydrogen peroxide solution to form a composite enzyme cascade polymerization platform dispersion liquid;
polymerizing the complex enzyme cascade polymerization platform dispersion liquid and the polymerization functional monomer under the condition of adding protocatechuic acid and ethylene glycol dimethacrylate, and separating the molecularly imprinted polymer from a polymerization mixed product;
preferably, the protocatechuic acid is a solution dissolved in anhydrous acetonitrile to obtain a concentration of 0.5-2M;
preferably, the molar amount of the polymerization functional monomer is 3 to 5 times of the molar amount of the protocatechuic acid;
preferably, the molar amount of the ethylene glycol dimethacrylate is 10 to 20 times that of protocatechuic acid.
<Electrochemical sensor for specific recognition of protocatechuic acid>
The chemical sensor comprises a molecularly imprinted polymer modified on the surface of a working electrode and provided with the molecularly imprinted polymer.
It is understood that, here, the modification of the molecularly imprinted polymer on the working electrode can be performed by conventional electrode surface modification, such as conventional dispersion impregnation.
Compared with the related art, the method has the following advantages and beneficial effects: (1) The composite enzyme cascade polymerization platform synthesized by the method can generate hydroxyl radicals to initiate polymerization under the condition of introducing hydrogen peroxide, and is economical and practical.
(2) The molecularly imprinted polymer provided by the application has the advantages of stable structure, simple and efficient synthesis method, mild reaction conditions and environmental friendliness.
(3) The method for detecting the protocatechuic acid specificity in the solution by using the electrochemical method is simple to operate and high in working efficiency.
(4) In the application, the molecularly imprinted polymer can specifically identify protocatechuic acid in a solution, and can effectively avoid interference of other organic molecules with similar structures in actual detection.
Drawings
The technical solution and other advantages of the present application will become apparent from the detailed description of the embodiments of the present application with reference to the accompanying drawings.
FIG. 1 shows a porous lanthanum ferrite sphere (LaFeO) obtained in example 1-1 of the present application 3 ) And three platforms (Au @ LaFeO) prefabricated by the same 3 、Au@LaFeO 3 @BHb、Au@LaFeO 3 @ BHb-MIPs).
FIGS. 2 (a) -2 (d) are porous lanthanum ferrite spheres (LaFeO) obtained in example 1-1 of the present application 3 ) And three platforms (Au @ LaFeO) prefabricated by the same 3 、Au@LaFeO 3 @BHb、Au@LaFeO 3 @ BHb-MIPs).
FIG. 3 shows a molecularly imprinted electrochemical probe (Au @ LaFeO) obtained in example 3 of the present application 3 @ BHb-MIPs).
FIGS. 4 (a) -4 (f) are the molecular imprinting electrochemical probe (Au @ LaFeO) obtained in example 3 of the present application 3 @ BHb-MIPs).
FIG. 5 shows lanthanum ferrite porous spheres (LaFeO) obtained in examples 1-2 of the present application 3 ) Field emission scanning electron microscopy images of (a).
FIG. 6 shows the porous lanthanum ferrite spheres (LaFeO) obtained in comparative example 1 of the present application 3 ) Field emission scanning electron microscopy images of (a).
FIG. 7 shows the complex enzyme cascade polymerization platform (Au @ LaFeO) obtained in comparative example 2 of the present application 3 @ BHb-2).
FIG. 8 is a view showing a molecularly imprinted electrochemical probe obtained in comparative example 3 of the present application.
(Au@LaFeO 3 @ BHb-MIPs-2).
FIG. 9 shows a molecularly imprinted electrochemical probe (Au @ LaFeO) obtained in comparative example 4 of the present application 3 @ BHb-MIPs-3).
FIGS. 10 (a) -10 (b) are views showing the Complex enzyme Cascade polymerization platform (Au @ LaFeO) obtained in example 3 of the present application 3 @ BHb) with original bovine hemoglobin.
FIGS. 11 (a) -11 (b) are the complex enzyme cascade polymerization platform (Au @ LaFeO) obtained in example 3 of the present application 3 @ BHb) was fitted to the uv kinetics of the original bovine hemoglobin.
FIGS. 12 (a) -12 (c) are views showing a molecularly imprinted electrochemical probe (Au @ LaFeO) obtained in example 3 of the present application 3 @ BHb-MIPs) in solution by electrochemical methodsTest pattern for protocatechuic acid.
FIG. 13 shows a molecularly imprinted electrochemical probe (Au @ LaFeO) obtained in example 3 of the present application 3 @ BHb-MIPs) test charts specifically recognizing protocatechuic acid in a solution by an electrochemical method.
Detailed Description
The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the embodiments of the present application and the accompanying drawings. It should be apparent that the described embodiments are only a few embodiments of the present application, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
Example 1-1 lanthanum ferrite porous sphere (LaFeO) 3 ) Synthesis of (2)
The embodiment provides a preparation method of a lanthanum ferrite porous ball, which comprises the following main steps of:
(1) Dissolving 0.4g of lanthanum nitrate hydrate in 50mL of deionized water, and keeping magnetic stirring;
(2) Adding 0.4g of ferric nitrate nonahydrate into the solution in the step (1), and keeping magnetic stirring;
(3) Adding 0.4g of anhydrous citric acid into the solution obtained in the step (2), and keeping magnetic stirring;
(4) Transferring the solution in the step (3) into a polytetrafluoroethylene reaction kettle, reacting for 10 hours at 180 ℃ in an electric heating constant temperature air blast drying oven, and naturally cooling;
(5) Centrifuging the solution obtained in the step (4), and freeze-drying to obtain organic precursor powder;
(6) Transferring the powder in the step (5) into a porcelain boat, annealing for 2 hours in a tube furnace at 800 ℃, naturally cooling to obtain the lanthanum ferrite porous ball named LaFeO 3 。
Example 1-2 lanthanum ferrite porous spheres (LaFeO) 3 Synthesis of (2)
The embodiment provides a preparation method of a lanthanum ferrite porous ball, which comprises the following main steps:
(1) Dissolving 0.4g of lanthanum nitrate hydrate in 50mL of deionized water, and keeping magnetic stirring;
(2) Adding 0.4g of ferric nitrate nonahydrate into the solution in the step (1), and keeping magnetic stirring;
(3) Adding 0.5g of citric acid monohydrate into the solution obtained in the step (2), and keeping magnetic stirring;
(4) Transferring the solution in the step (3) into a polytetrafluoroethylene reaction kettle, reacting for 10 hours at 180 ℃ in an electric heating constant temperature air blast drying oven, and naturally cooling;
(5) Centrifuging the solution obtained in the step (4), and freeze-drying to obtain organic precursor powder;
(6) Transferring the powder in the step (5) into a porcelain boat, annealing for 2 hours in a tube furnace at 800 ℃, and naturally cooling to obtain the lanthanum ferrite porous ball named LaFeO 3 -2。
Example 2 Synthesis of Complex enzyme Cascade polymerization platform (Au @ LaFeO) 3 @BHb)
The embodiment provides a preparation method of a complex enzyme cascade polymerization platform, which comprises the following main steps:
(1) 50mg of lanthanum ferrite porous ball and 50mg of bovine hemoglobin are dispersed in 50mL of deionized water to obtain the solution with the concentration of 1.0mg mL -1 The brownish red suspension of (a);
(2) Dissolving 0.4g of tetrachloroauric acid trihydrate in 50mL of deionized water to obtain a chloroauric acid solution with the concentration of 0.02M;
(3) Dissolving 0.2g of anhydrous sodium citrate in 50mL of deionized water to obtain a sodium citrate solution with the concentration of 0.02M;
(4) Respectively dropwise adding 2mL of the solution in the step (2) and 2mL of the solution in the step (3) into the suspension in the step (1), and keeping stirring;
(5) Reacting the mixed solution obtained in the step (4) for 4 hours at normal temperature, centrifuging and freeze-drying to obtain a complex enzyme cascade polymerization platform named as Au @ LaFeO 3 @BHb。
Example 2 is a complex enzyme cascade polymerization platform formed by combining three active components through electrostatic interaction and covalent bond, gold nanoparticles and bovine hemoglobin can be observed to be successfully loaded on a lanthanum ferrite porous ball through a field emission scanning electron microscope, and the existence of lanthanum ferrite and gold is proved through powder X-ray diffraction, which respectively correspond to JCPDS card Nos. 75-0541 and 04-0784.
EXAMPLE 3 preparation of molecularly imprinted polymer (Au @ LaFeO) 3 @BHb-MIPs)
The embodiment provides a preparation method of a molecularly imprinted polymer for specifically recognizing protocatechuic acid, which comprises the following main steps:
(1) Mixing 0.1g protocatechuic acid, 0.2mL methacrylic acid and 50mL acetonitrile at room temperature for more than 8 hours;
(2) Dispersing 50mg of the complex enzyme cascade polymerization platform obtained in the embodiment 2 into 5mL of hydrogen peroxide (30%) solution, and sealing and stirring for more than 2 hours;
(3) Adding 0.8mL of ethylene glycol dimethacrylate into the solution obtained in the step (1), and uniformly stirring;
(4) Introducing nitrogen into the solution obtained in the steps (2) and (3) for more than 30 minutes to remove oxygen, and mixing;
(5) Polymerizing the mixed solution obtained in the step (4) for 8 hours at room temperature under a closed condition, and centrifuging to obtain white powder;
(6) And (3) mixing the powder obtained in the step (5) with 100mL of a mixture with the volume ratio of 9:1 for more than 3 times, centrifuging, and freeze drying to obtain molecularly imprinted polymer named as Au @ LaFeO 3 @BHb-MIPs。
Example 3 is a molecularly imprinted polymer that can specifically recognize protocatechuic acid in solution by an electrochemical method, the cross-linked structure of the polymer can be observed by a field emission scanning electron microscope, the vibrational peaks of carbonyl groups in the cross-linking agent and the functional monomer can be observed by infrared spectroscopy, and the existence of carbon-carbon single bond, carbon-oxygen single bond and carbon-oxygen double bond in the polymer is proved by X-ray photoelectron spectroscopy, which proves the successful synthesis of the molecularly imprinted polymer.
Comparative example 1
The comparative example provides a lanthanum ferrite porous ball (LaFeO) 3 -3) a process for the preparation comprising the main steps of:
(1) Dissolving 0.3g of lanthanum chloride hydrate in 50mL of deionized water, and keeping magnetic stirring;
(2) Adding 0.4g of ferric nitrate nonahydrate into the solution obtained in the step (1), and keeping magnetic stirring;
(3) Adding 0.4g of anhydrous citric acid into the solution obtained in the step (2), and keeping magnetic stirring;
(4) Transferring the solution in the step (3) into a polytetrafluoroethylene reaction kettle, and reacting for 10 hours at 180 ℃ in an electric heating constant-temperature air blast drying oven;
(5) Naturally cooling the solution obtained in the step (4), centrifuging and drying to obtain precursor powder;
(6) Transferring the powder in the step (5) into a porcelain boat, annealing for 2 hours in a tube furnace at 800 ℃, naturally cooling to obtain the lanthanum ferrite porous ball named LaFeO 3 -3。
Comparative example 2
The comparative example provides a complex enzyme cascade polymerization platform (Au @ LaFeO) 3 The preparation method of @ BHb-2) comprises the following main steps:
(1) 50mg of lanthanum ferrite porous ball and 25mg of horseradish catalase are dispersed in 50mL of deionized water to obtain the solution with the concentration of 1.0mg mL -1 The red-brown suspension of (a);
(2) Dissolving 0.4g of tetrachloroauric acid trihydrate in 50mL of deionized water to obtain a chloroauric acid solution with the concentration of 0.02M;
(3) Dissolving 0.2g of anhydrous sodium citrate in 50mL of deionized water to obtain a sodium citrate solution with the concentration of 0.02M;
(4) Respectively dropwise adding 2mL of the solution in the step (2) and 2mL of the solution in the step (3) to the suspension in the step (1), and keeping stirring;
(5) Reacting the mixed solution obtained in the step (4) for 4 hours at normal temperature, centrifuging, and freeze-drying to obtain a compound enzyme cascade polymerization platform named as Au @ LaFeO 3 @BHb-2。
Comparative example 3
This comparative example provides a molecularly imprinted polymer (Au @ LaFeO) that specifically recognizes protocatechuic acid 3 The preparation method of @ BHb-MIPs-2) comprises the following main steps:
(1) Dissolving 0.1g of protocatechuic acid and 0.2g of acrylamide in 50mL of anhydrous acetonitrile, and mixing for more than 8 hours at normal temperature;
(2) Adding 0.8mL of ethylene glycol dimethacrylate into the solution obtained in the step (1), and uniformly stirring;
(3) Dispersing 50mg of the complex enzyme cascade polymerization platform obtained in the embodiment 2 in 5mL of hydrogen peroxide (30%) solution, and stirring for more than 2 hours in a closed manner;
(4) Introducing nitrogen into the solution in the step (2) and the suspension in the step (3) for more than 30 minutes to remove oxygen, and mixing;
(5) Polymerizing the mixed solution obtained in the step (4) for 8 hours at room temperature under a closed condition, and centrifuging to obtain white powder;
(6) And (3) mixing the powder obtained in the step (5) with 100mL of a mixture with the volume ratio of 9:1 for more than 3 times, centrifuging, and freeze drying to obtain molecularly imprinted polymer named as Au @ LaFeO 3 @BHb-MIPs-2。
Comparative example 4
This comparative example provides a molecularly imprinted polymer (Au @ LaFeO) that specifically recognizes protocatechuic acid 3 The preparation method of @ BHb-MIPs-3) comprises the following main steps:
(1) Dissolving 0.1g of protocatechuic acid and 0.3mL of 4-vinylpyridine in 50mL of anhydrous acetonitrile, and mixing at normal temperature for more than 8 hours;
(2) Adding the solution obtained in the step (1) into 0.8mL of ethylene glycol dimethacrylate, and uniformly stirring;
(3) Dispersing 50mg of the complex enzyme cascade polymerization platform obtained in the embodiment 2 into 5mL of hydrogen peroxide (30%) solution, and sealing and stirring for more than 2 hours;
(4) Introducing nitrogen into the solution in the step (2) and the suspension in the step (3) for more than 30 minutes to remove oxygen, and mixing;
(5) Polymerizing the mixed solution obtained in the step (4) for 8 hours at room temperature under a closed condition, and centrifuging to obtain white powder;
(6) And (3) mixing the powder obtained in the step (5) with 100mL of a mixture with the volume ratio of 9:1 for more than 3 times, centrifuging, and freeze drying to obtain molecularly imprinted polymer named as Au @ LaFeO 3 @BHb-MIPs-3。
FIG. 1 shows a lanthanum ferrite porous pellet (LaFeO) prepared in example 1-1 of the present invention 3 ) And preparation thereofThree platforms of (2): lanthanum ferrite porous ball loaded with gold nanoparticles (Au @ LaFeO) 3 ) The complex enzyme cascade polymerization platform prepared in example 2 (Au @ LaFeO) 3 @ BHb), namely bovine hemoglobin modified by lanthanum ferrite porous spheres loaded with gold nanoparticles, and the sensing probe loaded with molecularly imprinted polymer prepared in example 3 (Au @ LaFeO) 3 @ BHb-MIPs) in a Field Emission Scanning Electron Microscope (FESEM) image.
FESEM was used to observe the external morphology and physical structure of each sample, as described below:
LaFeO 3 : the lanthanum ferrite porous ball prepared by the solvothermal method has a relatively regular form, the physical appearance is a nano ball structure which is locally and orderly arranged, and the average diameter is about 800-900 nanometers.
Au@LaFeO 3 : gold nanoparticles were successfully immobilized on the surface of lanthanum ferrite porous spheres with an average diameter of about 10 nm.
Au@LaFeO 3 @ BHb: the biological enzyme bovine hemoglobin is fixed on the surfaces of lanthanum ferrite and gold nanoparticles, and the shape of the sphere is uneven from surface to smooth.
Au@LaFeO 3 @ BHb-MIPs: the rugged lanthanum ferrite porous ball is wrapped by a molecularly imprinted polymer and shows a smooth shape with a complex three-dimensional cross-linked structure.
FIG. 2 shows a lanthanum ferrite porous pellet (LaFeO) prepared in example 1-1 of the present invention 3 ) And three platforms prepared therewith: lanthanum ferrite porous ball loaded with gold nanoparticles (Au @ LaFeO) 3 ) The complex enzyme cascade polymerization platform prepared in example 2 (Au @ LaFeO) 3 @ BHb), namely bovine hemoglobin modified by gold nanoparticles loaded on lanthanum ferrite porous spheres, and the electrochemical sensing probe loaded with molecularly imprinted polymer prepared in example 3 (Au @ LaFeO) 3 @ BHb-MIPs) powder X-ray diffraction (PXRD) pattern.
XRD was used to identify the crystal structure of each sample, and the results are described below:
LaFeO 3 : the XRD diffraction pattern exhibited typical peaks of crystalline character corresponding to JCPDS card No.75-0541.
Au@LaFeO 3 : XRD diffraction pattern in LaFeO 3 On the basis, the crystal face diffraction peak of the gold nanoparticle is displayed and corresponds to JCPDS card No.04-0784.
Au@LaFeO 3 @ BHb: the bovine hemoglobin is amorphous polymer, and has low content, XRD diffraction pattern and Au @ LaFeO 3 The spectra of (A) are substantially identical.
Au@LaFeO 3 @ BHb-MIPs: molecularly Imprinted Polymers (MIPs) are amorphous macromolecules with significant amorphous polymer bulges around 20 degrees, proving that they are supported on a polymerization platform.
FIG. 3 shows an electrochemical sensing probe (Au @ LaFeO) loaded with a molecularly imprinted polymer prepared in example 3 of the present invention 3 @ BHb-MIPs) in a fourier infrared (FT-IR) spectrum.
FT-IR is used for identifying characteristic groups of the molecularly imprinted polymer, and the results are described as follows:
Au@LaFeO 3 @BHb-MIPs：3444cm -1 belonging to the tensile mode of the hydroxyl group (O-H), 2956cm -1 (C-H) stretching mode due to carbon-hydrogen bond, 1728cm -1 The peak at (D) should be attributed to carbonyl (C = O) tensile vibration, 1480cm -1 The characteristic peak of (A) is 1225cm which is the expansion of the C-O single bond -1 The characteristic peaks of (A) correspond to the extension of a saturated C-C single bond, and the characteristic peaks are derived from functional monomer methacrylic acid and crosslinking agent ethylene glycol dimethacrylate, thus proving the successful synthesis of the molecularly imprinted polymer.
FIG. 4 is a view showing an inductive probe (Au @ LaFeO) loaded with a molecularly imprinted polymer prepared in example 3 of the present invention 3 @ BHb-MIPs) in X-ray photoelectron spectroscopy (XPS).
XPS was used to confirm the constituent elements of the sensing probe and their combination state, and the results are described below:
Au@LaFeO 3 @ BHb-MIPs: 5 obvious elements can be seen in the XPS total spectrum of the induction probe, from left to right are respectively La, fe, O, C and Au, specifically to a fitting graph of each element, the convolution of the lanthanum element is 4 characteristic peaks, from left to right are respectively a satellite peak and a 3d peak 3/2 Peak, satellite Peak and 3d 5/2 Peak, knotThe fruit shows La as the positive trivalent; the iron element is convoluted into 3 characteristic peaks which are respectively 2p from left to right 1/2 Peak, satellite Peak and 2p 3/2 Peak, results show Fe is trivalent positive; the gold element is 2 characteristic peaks which are 4f from left to right 5/2 Peak sum 4f 7/2 Peak, result shows Au is zero valence; the convolution of oxygen element is 3 characteristic peaks, namely C = O, C-O and the peak of metal oxide from left to right; carbon is 3 characteristic peaks, from left to right, of C = O, C-O and C-C, respectively. The above results confirmed Au @ LaFeO 3 Successful preparation of @ BHb-MIPs.
Lanthanum ferrite porous spheres (LaFeO) obtained in example 1-2 3 The physical morphology of-2) is shown in FIG. 5, and the degree of regularity of the morphology is similar to that of the lanthanum ferrite porous spheres (LaFeO) obtained in example 1-1 3 ) And (4) the equivalent.
Porous lanthanum ferrite spheres (LaFeO) obtained in comparative example 1 3 The physical morphology of-3) is shown in FIG. 6, the lanthanum source is selected from lanthanum chloride hydrate, other components are unchanged, and the material preparation is carried out under the same solvothermal and annealing conditions. Compared with the example 1-1, the lanthanum ferrite porous ball formed in the comparative example 1 is smoother, cannot load more active components, and has low electrochemical activity.
The Complex enzyme Cascade polymerization platform obtained in comparative example 2 (Au @ LaFeO) 3 @ BHb-2) is shown in FIG. 7, the bio-enzyme is selected from horseradish catalase, other components are unchanged, and the material preparation is carried out under the same solution conditions. Compared with example 2, the polymerization platform formed by comparative example 2 has no big difference, but the unit price of the enzyme purchased from the same manufacturer is increased by 2.2 times, and the cost performance is lower.
Molecularly imprinted polymer (Au @ LaFeO) obtained in comparative example 3 3 The physical morphology of @ BHb-MIPs-2) is shown in figure 8, the functional monomer is selected from acrylamide, other components are unchanged, and the material preparation is carried out under the same solution condition. Compared with example 3, the polymer has too high degree of crosslinking, poor dispersibility and low protocatechuic acid recognition efficiency.
Molecularly imprinted polymer (Au @ LaFeO) obtained in comparative example 4 3 The physical morphology of @ BHb-MIPs-3) is shown in FIG. 9, the functional monomer is selected from 4-vinylpyridine, other components are unchanged, and the functional monomer is in the same solution stripThe material preparation was carried out under the conditions. Compared with example 3, the polymer has an excessively high degree of crosslinking, poor dispersibility and low protocatechuic acid recognition efficiency.
Test example 1 testing of working efficiency of Complex enzyme Cascade polymerization platform
This test example 1 provides a preformed complex enzyme cascade polymerization platform (Au @ LaFeO) 3 @ BHb) and the original biological enzyme bovine hemoglobin (BHb) catalytic efficiency for hydrogen peroxide, i.e. productivity of hydroxyl radicals, under the same test conditions. Wherein the concentration of the test body is kept at 0.1mg mL -1 The total volume of the experimental solution is kept at 3mL, and the experimental solution comprises 1mL of experimental body suspension, 1mL of hydrogen peroxide solution and 1mL of color developing agent (3,3,5,5-tetramethyl benzidine), and ultraviolet-visible spectrophotometry is used as a testing means to respectively carry out ultraviolet kinetic testing on hydrogen peroxide solutions with different concentrations, and the ultraviolet absorbance is recorded once every 10s, and the result is shown in fig. 10. Meanwhile, the average slope of the curve is calculated to calculate the working efficiency of each prefabricated platform, and the dynamic fitting is performed according to the Michaelis-Menten equation and the Lineweaver-Burk equation, and the result is shown in FIG. 11, and the related description is as follows:
the curves in fig. 10 represent the uv kinetics of hydrogen peroxide catalyzed by the experimental body at different concentrations, respectively, and the hydrogen peroxide concentrations represented from top to bottom by the curves are: 50mM;20mM;10mM;5mM;2mM;1mM;0.5mM, the slope of the curve being the catalytic rate of the test body at the corresponding hydrogen oxide concentration. The Michaelis-Menten equation fitting in FIG. 11 shows that the working efficiency of the complex enzyme cascade polymerization platform is far higher than that of the original bovine hemoglobin under the condition of the same concentration of hydrogen peroxide, and the intercept between the curve and the x axis is 1/V in the double reciprocal curve Lineweaverer-Burk equation fitting max The intercept between the curve and the y-axis is-1/K m The calculation result shows that the composite enzyme cascade polymerization platform is 11.11, and is the original bovine hemoglobin V max 2.26 times of (4.91).
Test example 2 specificity recognition test of a preformed electrochemical sensing Probe for protocatechuic acid
This test 2 provides a prefabricated electrochemical sensing probe (Au @ LaFeO) 3 @ BHb-MIPs) specifically recognize protocatechuic acid in solution,specifically, an induction probe is fixed on the surface of a central carbon layer of a glassy carbon electrode through a perfluorosulfonic acid resin solution (5 wt.%), another silver/silver chloride electrode is used as a reference electrode, a platinum electrode is used as a counter electrode, and a three-electrode test system is formed to identify and detect protocatechuic acid in the solution on an electrochemical workstation. Specifically, the test electrolyte is selected from Phosphate Buffered Saline (PBS) at a concentration of 0.1M (pH = 7.2), a concentration gradient of protocatechuic acid is set to 0.1 μ M, 0.2 μ M, 0.5 μ M, 1 μ M, 2 μ M, 5 μ M, 10 μ M, 20 μ M, 50 μ M, 100 μ M, 200 μ M, 500 μ M, 1000 μ M, and protocatechuic acid in the solution is identified and detected by Differential Pulse Voltammetry (DPV) at normal temperature and pressure, and electrochemical test results of the pre-formed sensing probe for protocatechuic acid are shown in fig. 12, in which corresponding current density values are linearly increased as the concentration of protocatechuic acid is increased, and curves are DPV test curves of protocatechuic acid at concentrations of 0.1 μ M, 0.2 μ M, 0.5 μ M, 1 μ M, 2 μ M, 5 μ M, 10 μ M, 20 μ M, 50 μ M, 100 μ M, 200 μ M, 500 μ M, 1000 μ M, respectively from bottom to top. The detection range of the prefabricated inductive probe to protocatechuic acid under line through an electrochemical method is 0.2 mu M-1000 mu M, the linear correlation coefficients are 0.9919 and 0.9838 respectively, and the detection limit is 50nM. In the results of specific recognition of protocatechuic acid shown in fig. 13, only protocatechuic acid molecules are captured by the preformed molecularly imprinted polymer, and an obvious signal is obtained in an electrochemical test, while other organic interfering molecules with similar structures, such as hydroquinone, catechol, and resorcinol, have no obvious electrochemical response, and the sensing probe has a good specific recognition effect and can be practically applied.
The above description is only for the preferred embodiment of the present application, but the scope of the present application is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present application should be covered within the scope of the present application.
Claims (10)
1. A complex enzyme cascade polymerization platform for catalyzing hydrogen peroxide to generate hydroxyl free radicals is characterized in that the complex enzyme cascade polymerization platform is Au @ LaFeO 3 @ BHb composite particles;
wherein, the Au @ LaFeO 3 In the @ BHb composite particles, a lanthanum ferrite porous ball is used as a carrier, gold nanoparticles are loaded on the surface of the lanthanum ferrite porous ball through electrostatic interaction, and peroxidase is bonded on the gold nanoparticles through Jin Qiu covalent bonds.
2. The complex enzyme cascade polymerization platform of claim 1, wherein the average particle size of the lanthanum ferrite porous spheres is 800-900nm, the average particle size of the gold nanoparticles is 10nm, and the surface of the lanthanum ferrite porous spheres is uneven.
3. The complex enzyme cascade polymerization platform of claim 1, wherein the au @ lafeo is 3 The preparation method of the @ BHb composite particle comprises the following steps of:
providing a lanthanum ferrite porous ball;
allowing the lanthanum ferrite porous spheres and peroxidase to form a mixed dispersion;
reacting the mixed dispersion with chloroauric acid solution and reducing agent solution, and centrifuging and drying to obtain Au @ LaFeO 3 @ BHb composite particles.
4. The complex enzyme cascade polymerization platform of claim 3, wherein the reaction temperature is normal temperature, and the reaction time is 3-5h;
preferably, the concentration of the mixed dispersion is 1-2mg mL -1 ；
Preferably, the concentration of the chloroauric acid solution is 0.015-0.025M;
preferably, the concentration of the reducing agent solution is 0.015 to 0.025M.
Preferably, the volume ratio of the mixed dispersion liquid to the chloroauric acid solution to the reducing agent solution is 25:1:1.
5. the complex enzyme cascade polymerization platform as claimed in claim 3, wherein the preparation method of the lanthanum ferrite porous ball is obtained by subjecting lanthanide inorganic salt and iron inorganic salt to solvothermal reaction with the addition of an organic ligand.
6. The complex enzyme cascade polymerization platform as claimed in claim 5, wherein the concentration of the lanthanide inorganic salt is the same as the molar amount of the iron-based inorganic salt, and the molar amount of the organic ligand is 2-3 times of the concentration of the lanthanide inorganic salt;
preferably, the lanthanide inorganic salt is selected from one or more of lanthanum nitrate, lanthanum chloride or lanthanum bromide;
preferably, the iron inorganic salt is selected from one or more of ferric nitrate, ferric chloride or ferric sulfate;
preferably, the temperature of the solvothermal reaction is 175-185 ℃;
preferably, the preparation method of the lanthanum ferrite porous ball further comprises annealing the target product after the solvothermal reaction;
preferably, the annealing temperature is 790-810 ℃, and the annealing time is 1-3h.
7. A molecularly imprinted polymer for specifically recognizing protocatechuic acid, which is generated by polymerizing a polymerization functional monomer under the action of the complex enzyme cascade polymerization platform of claim 1 by enzyme induction.
8. The molecularly imprinted polymer according to claim 7, wherein the polymeric functional monomer is derived from one or more of acrylic acid, methacrylic acid, acrylamide, methacrylamide or 4-vinylpyridine.
9. The molecularly imprinted polymer according to claim 7, wherein the process of enzyme-induced polymerization comprises the steps of:
dispersing the composite enzyme cascade polymerization platform in a hydrogen peroxide solution to form a composite enzyme cascade polymerization platform mixed dispersion liquid;
polymerizing the mixed dispersion liquid of the complex enzyme cascade polymerization platform and the polymerization functional monomer under the condition of adding protocatechuic acid and ethylene glycol dimethacrylate, and separating the molecularly imprinted polymer from the polymerization mixed product by eluent;
preferably, the protocatechuic acid is dissolved in anhydrous acetonitrile to obtain a solution with the concentration of 0.5-2M;
preferably, the molar amount of the polymerized functional monomer is 3 to 5 times of the molar amount of the protocatechuic acid;
preferably, the molar amount of the ethylene glycol dimethacrylate is 10 to 20 times that of protocatechuic acid.
10. An electrochemical sensor for specifically recognizing protocatechuic acid, which is modified on the surface of a working electrode and has a molecularly imprinted polymer according to claim 7.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211634175.9A CN115975110B (en) | 2022-12-19 | 2022-12-19 | Molecularly imprinted polymer for specifically recognizing protocatechuic acid and electrochemical sensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211634175.9A CN115975110B (en) | 2022-12-19 | 2022-12-19 | Molecularly imprinted polymer for specifically recognizing protocatechuic acid and electrochemical sensor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115975110A true CN115975110A (en) | 2023-04-18 |
| CN115975110B CN115975110B (en) | 2024-01-26 |
Family
ID=85957376
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211634175.9A Active CN115975110B (en) | 2022-12-19 | 2022-12-19 | Molecularly imprinted polymer for specifically recognizing protocatechuic acid and electrochemical sensor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115975110B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103913499A (en) * | 2014-03-26 | 2014-07-09 | 青岛大学 | Preparation method of molecular imprinting electrochemical sensor for detecting bovine hemoglobin |
| CN104181217A (en) * | 2014-04-29 | 2014-12-03 | 南昌大学 | Construction and application of impedance type electrochemical sensor based on molecularly imprinted polymer with magnetic surface |
| CN109019696A (en) * | 2018-08-01 | 2018-12-18 | 济南大学 | A kind of Au-LaFeO3The preparation method of nanocomposite |
| CN111346648A (en) * | 2018-12-24 | 2020-06-30 | 上海萃励电子科技有限公司 | Preparation method of Au-loaded lanthanum ferrite nanoparticles |
| CN111378961A (en) * | 2020-03-20 | 2020-07-07 | 中国科学院合肥物质科学研究院 | Gold nanoparticle-loaded lanthanum ferrite film and preparation method and application thereof |
| CN114560975A (en) * | 2022-01-05 | 2022-05-31 | 临沂大学 | Lanthanum metal organic framework, cascade nano polymerization platform and preparation method of molecularly imprinted polymer |
-
2022
- 2022-12-19 CN CN202211634175.9A patent/CN115975110B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103913499A (en) * | 2014-03-26 | 2014-07-09 | 青岛大学 | Preparation method of molecular imprinting electrochemical sensor for detecting bovine hemoglobin |
| CN104181217A (en) * | 2014-04-29 | 2014-12-03 | 南昌大学 | Construction and application of impedance type electrochemical sensor based on molecularly imprinted polymer with magnetic surface |
| CN109019696A (en) * | 2018-08-01 | 2018-12-18 | 济南大学 | A kind of Au-LaFeO3The preparation method of nanocomposite |
| CN111346648A (en) * | 2018-12-24 | 2020-06-30 | 上海萃励电子科技有限公司 | Preparation method of Au-loaded lanthanum ferrite nanoparticles |
| CN111378961A (en) * | 2020-03-20 | 2020-07-07 | 中国科学院合肥物质科学研究院 | Gold nanoparticle-loaded lanthanum ferrite film and preparation method and application thereof |
| CN114560975A (en) * | 2022-01-05 | 2022-05-31 | 临沂大学 | Lanthanum metal organic framework, cascade nano polymerization platform and preparation method of molecularly imprinted polymer |
Non-Patent Citations (5)
| Title |
|---|
| 张凌燕;袁若;柴雅琴;曹淑瑞;黎雪莲;王娜;: "基于辣根过氧化物酶/纳米金/辣根过氧化物酶/多壁纳米碳管修饰的过氧化氢生物传感器的研究", 化学学报, no. 16 * |
| 李增威;王娜;雷婵;曾月;付珍珍;何利;刘书亮;周康;敖晓琳;陈姝娟;: "磁性分子印迹聚合物的制备及对食品中氯霉素残留的检测", 浙江农业学报, no. 06 * |
| 李文娟;袁若;柴雅琴;王娜;陈时洪;高凤仙;杨淦;: "基于层层静电吸附天青Ⅰ和纳米金固定过氧化物酶生物传感器的研究", 分析试验室, no. 10 * |
| 李艳霞;陈毅挺;黄露;王秀平;: "识别牛血红蛋白的分子印迹电化学传感器的制备与研究", 理化检验(化学分册), no. 12 * |
| 黎雪莲;袁若;柴雅琴;张凌燕;王娜;朱强;: "基于静电吸附甲苯胺蓝和纳米金固定过氧化物酶生物传感器的研究", 分析化学, no. 03 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115975110B (en) | 2024-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Baldim et al. | Polymer-coated cerium oxide nanoparticles as oxidoreductase-like catalysts | |
| Chen et al. | A Facile synthesis of superparamagnetic Fe3O4 nanofibers with superior peroxidase-like catalytic activity for sensitive colorimetric detection of L-cysteine | |
| Díaz-Díaz et al. | New materials for analytical biomimetic assays based on affinity and catalytic receptors prepared by molecular imprinting | |
| Margel et al. | Acrolein polymerization: monodisperse, homo, and hybrido microspheres, synthesis, mechanism, and reactions | |
| Karami et al. | A catechol biosensor based on immobilizing laccase to Fe3O4@ Au core-shell nanoparticles | |
| Lahcen et al. | Fast route for the synthesis of decorated nanostructured magnetic molecularly imprinted polymers using an ultrasound probe | |
| Çorman et al. | Preparation of nanoparticles which contains histidine for immobilization of Trametes versicolor laccase | |
| Gan et al. | The chain-like Au/carbon dots nanocomposites with peroxidase-like activity and their application for glucose detection | |
| CN109019659A (en) | A kind of synthetic method of chiral oxidization copper nano-particle | |
| Xi et al. | Iron doped graphitic carbon nitride with peroxidase like activity for colorimetric detection of sarcosine and hydrogen peroxide | |
| Ju et al. | Modification and application of Fe3O4 nanozymes in analytical chemistry: A review | |
| Xian et al. | One-pot fabrication of Fe-doped carbon nitride nanoparticles as peroxidase mimetics for H2O2 and glucose detection | |
| CN106732232B (en) | A kind of preparation method of the yolk type Fenton microreactor of inner surface supported precious metal nano-particle | |
| Bithi et al. | Cationic polyelectrolyte grafted mesoporous magnetic silica composite particles for targeted drug delivery and thrombolysis | |
| Aghayan et al. | Micellar catalysis of an iron (III)-MOF: enhanced biosensing characteristics | |
| Mumtaz et al. | Facile method to synthesize dopamine-capped mixed ferrite nanoparticles and their peroxidase-like activity | |
| Kaang et al. | Polyelectrolyte brush-grafted polydopamine-based catalysts with enhanced catalytic activity and stability | |
| Zhang et al. | Structure-activity mechanism of iron oxide nanozymes | |
| Lan et al. | Superparamagnetic nanocomposites based on surface imprinting for biomacromolecular recognition | |
| Luo et al. | Water dispersible cobalt single-atom catalysts with efficient chemiluminescence enhancement for sensitive bioassay | |
| Fu et al. | An ultraviolet self-initiated polymerized platform for specific recognition and elimination of caffeic acid based on the molecular imprinting technology | |
| Hamed et al. | Single-atom nanozymes with peroxidase-like activity: A review | |
| Guo et al. | Research progress of metal–organic frameworks-molecularly imprinted polymers for specific recognition | |
| Chen et al. | Biocompatible BSA-AuNP@ ZnCo2O4 nanosheets with oxidase-like activity: Colorimetric biosensing and antitumor activity | |
| WO2011074742A1 (en) | Nanohybrid nitrogen monoxide detecting sensor and a production method therefor |
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 |