CN105928892B - Application of the cubic boron nitride as Mimetic Peroxidase - Google Patents
Application of the cubic boron nitride as Mimetic Peroxidase Download PDFInfo
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- CN105928892B CN105928892B CN201610231750.9A CN201610231750A CN105928892B CN 105928892 B CN105928892 B CN 105928892B CN 201610231750 A CN201610231750 A CN 201610231750A CN 105928892 B CN105928892 B CN 105928892B
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- boron nitride
- cubic boron
- peroxidase
- mimetic
- mimetic peroxidase
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- PZNSFCLAULLKQX-UHFFFAOYSA-N Boron nitride Chemical compound N#B PZNSFCLAULLKQX-UHFFFAOYSA-N 0.000 title claims abstract description 136
- 229910052582 BN Inorganic materials 0.000 title claims abstract description 134
- 102000003992 Peroxidases Human genes 0.000 title claims abstract description 72
- 108040007629 peroxidase activity proteins Proteins 0.000 title claims abstract description 72
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 70
- 102000004190 Enzymes Human genes 0.000 claims abstract description 19
- 108090000790 Enzymes Proteins 0.000 claims abstract description 19
- 230000003197 catalytic effect Effects 0.000 claims abstract description 13
- 230000000694 effects Effects 0.000 claims abstract description 13
- 108010001336 Horseradish Peroxidase Proteins 0.000 claims abstract description 11
- 238000002360 preparation method Methods 0.000 claims abstract description 11
- 230000002255 enzymatic effect Effects 0.000 claims abstract description 9
- 238000005259 measurement Methods 0.000 claims abstract description 8
- 238000001514 detection method Methods 0.000 claims abstract description 7
- 239000002253 acid Substances 0.000 claims abstract description 5
- 230000015556 catabolic process Effects 0.000 claims abstract description 5
- 238000006731 degradation reaction Methods 0.000 claims abstract description 5
- 229960002163 hydrogen peroxide Drugs 0.000 claims description 33
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 20
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 16
- 239000001257 hydrogen Substances 0.000 claims description 16
- 229910052739 hydrogen Inorganic materials 0.000 claims description 16
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 16
- 239000012498 ultrapure water Substances 0.000 claims description 16
- 238000006243 chemical reaction Methods 0.000 claims description 14
- 239000000758 substrate Substances 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 13
- 229910052757 nitrogen Inorganic materials 0.000 claims description 11
- 239000000725 suspension Substances 0.000 claims description 11
- 239000002994 raw material Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000000852 hydrogen donor Substances 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 238000001914 filtration Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 238000005303 weighing Methods 0.000 claims description 8
- 238000010521 absorption reaction Methods 0.000 claims description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 6
- ZOXJGFHDIHLPTG-UHFFFAOYSA-N Boron Chemical compound [B] ZOXJGFHDIHLPTG-UHFFFAOYSA-N 0.000 claims description 6
- 229910052796 boron Inorganic materials 0.000 claims description 6
- 239000007853 buffer solution Substances 0.000 claims description 6
- CBMPTFJVXNIWHP-UHFFFAOYSA-L disodium;hydrogen phosphate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].OP([O-])([O-])=O.OC(=O)CC(O)(C(O)=O)CC(O)=O CBMPTFJVXNIWHP-UHFFFAOYSA-L 0.000 claims description 6
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 claims description 6
- 229940043267 rhodamine b Drugs 0.000 claims description 6
- 238000002835 absorbance Methods 0.000 claims description 5
- 238000012545 processing Methods 0.000 claims description 5
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical group CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 claims description 4
- LRFVTYWOQMYALW-UHFFFAOYSA-N 9H-xanthine Chemical compound O=C1NC(=O)NC2=C1NC=N2 LRFVTYWOQMYALW-UHFFFAOYSA-N 0.000 claims description 4
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 4
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 claims description 3
- 238000000862 absorption spectrum Methods 0.000 claims description 3
- 230000009471 action Effects 0.000 claims description 3
- 238000000137 annealing Methods 0.000 claims description 3
- 238000006555 catalytic reaction Methods 0.000 claims description 3
- 150000002978 peroxides Chemical class 0.000 claims description 3
- HSTOKWSFWGCZMH-UHFFFAOYSA-N 3,3'-diaminobenzidine Chemical compound C1=C(N)C(N)=CC=C1C1=CC=C(N)C(N)=C1 HSTOKWSFWGCZMH-UHFFFAOYSA-N 0.000 claims description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 2
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 235000019441 ethanol Nutrition 0.000 claims description 2
- 238000002795 fluorescence method Methods 0.000 claims description 2
- MEUKEBNAABNAEX-UHFFFAOYSA-N hydroperoxymethane Chemical compound COO MEUKEBNAABNAEX-UHFFFAOYSA-N 0.000 claims description 2
- MCPLVIGCWWTHFH-UHFFFAOYSA-L methyl blue Chemical compound [Na+].[Na+].C1=CC(S(=O)(=O)[O-])=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[NH+]C=2C=CC(=CC=2)S([O-])(=O)=O)C=2C=CC(NC=3C=CC(=CC=3)S([O-])(=O)=O)=CC=2)C=C1 MCPLVIGCWWTHFH-UHFFFAOYSA-L 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 239000002957 persistent organic pollutant Substances 0.000 claims description 2
- 230000008569 process Effects 0.000 claims description 2
- 229940075420 xanthine Drugs 0.000 claims description 2
- ORZHVTYKPFFVMG-UHFFFAOYSA-N xylenol orange Chemical compound OC(=O)CN(CC(O)=O)CC1=C(O)C(C)=CC(C2(C3=CC=CC=C3S(=O)(=O)O2)C=2C=C(CN(CC(O)=O)CC(O)=O)C(O)=C(C)C=2)=C1 ORZHVTYKPFFVMG-UHFFFAOYSA-N 0.000 claims description 2
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims 1
- 238000005468 ion implantation Methods 0.000 claims 1
- 230000004048 modification Effects 0.000 claims 1
- 238000012986 modification Methods 0.000 claims 1
- 239000007974 sodium acetate buffer Substances 0.000 claims 1
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 claims 1
- 230000003796 beauty Effects 0.000 abstract description 2
- 238000003759 clinical diagnosis Methods 0.000 abstract description 2
- 239000000356 contaminant Substances 0.000 abstract description 2
- 238000003018 immunoassay Methods 0.000 abstract description 2
- 238000010586 diagram Methods 0.000 description 6
- 239000000872 buffer Substances 0.000 description 5
- 229910052734 helium Inorganic materials 0.000 description 5
- 239000001307 helium Substances 0.000 description 5
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 description 5
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 238000002441 X-ray diffraction Methods 0.000 description 3
- XKRFYHLGVUSROY-UHFFFAOYSA-N argon Substances [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 3
- -1 peroxide Compound Chemical class 0.000 description 3
- 229910052786 argon Inorganic materials 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- HFACYLZERDEVSX-UHFFFAOYSA-N benzidine Chemical compound C1=CC(N)=CC=C1C1=CC=C(N)C=C1 HFACYLZERDEVSX-UHFFFAOYSA-N 0.000 description 2
- 239000003575 carbonaceous material Substances 0.000 description 2
- UBEWDCMIDFGDOO-UHFFFAOYSA-N cobalt(2+);cobalt(3+);oxygen(2-) Chemical compound [O-2].[O-2].[O-2].[O-2].[Co+2].[Co+3].[Co+3] UBEWDCMIDFGDOO-UHFFFAOYSA-N 0.000 description 2
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 229910044991 metal oxide Inorganic materials 0.000 description 2
- 150000004706 metal oxides Chemical class 0.000 description 2
- 239000002086 nanomaterial Substances 0.000 description 2
- 239000005416 organic matter Substances 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- OCUIPDGJXYIRKY-UHFFFAOYSA-N C(CC(O)(C(=O)O)CC(=O)O)(=O)O.[P] Chemical compound C(CC(O)(C(=O)O)CC(=O)O)(=O)O.[P] OCUIPDGJXYIRKY-UHFFFAOYSA-N 0.000 description 1
- QPLDLSVMHZLSFG-UHFFFAOYSA-N Copper oxide Chemical compound [Cu]=O QPLDLSVMHZLSFG-UHFFFAOYSA-N 0.000 description 1
- 239000005751 Copper oxide Substances 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical compound [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- ZSEMWHCVIJETNE-UHFFFAOYSA-N N1=CC=CC2=CC(=C3C=CC=NC3=C12)S(=O)(=O)O.C(C)N1CSC2=C1C=CC=C2 Chemical compound N1=CC=CC2=CC(=C3C=CC=NC3=C12)S(=O)(=O)O.C(C)N1CSC2=C1C=CC=C2 ZSEMWHCVIJETNE-UHFFFAOYSA-N 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- SXEHKFHPFVVDIR-UHFFFAOYSA-N [4-(4-hydrazinylphenyl)phenyl]hydrazine Chemical compound C1=CC(NN)=CC=C1C1=CC=C(NN)C=C1 SXEHKFHPFVVDIR-UHFFFAOYSA-N 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- YBCVMFKXIKNREZ-UHFFFAOYSA-N acoh acetic acid Chemical compound CC(O)=O.CC(O)=O YBCVMFKXIKNREZ-UHFFFAOYSA-N 0.000 description 1
- 150000001448 anilines Chemical class 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000002041 carbon nanotube Substances 0.000 description 1
- 229910021393 carbon nanotube Inorganic materials 0.000 description 1
- 239000011545 carbonate/bicarbonate buffer Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- RIVZIMVWRDTIOQ-UHFFFAOYSA-N cobalt iron Chemical compound [Fe].[Co].[Co].[Co] RIVZIMVWRDTIOQ-UHFFFAOYSA-N 0.000 description 1
- 229910000431 copper oxide Inorganic materials 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229940056319 ferrosoferric oxide Drugs 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 229910021389 graphene Inorganic materials 0.000 description 1
- ILHIHKRJJMKBEE-UHFFFAOYSA-N hydroperoxyethane Chemical compound CCOO ILHIHKRJJMKBEE-UHFFFAOYSA-N 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 1
- 150000004767 nitrides Chemical class 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- POECFFCNUXZPJT-UHFFFAOYSA-M sodium;carbonic acid;hydrogen carbonate Chemical compound [Na+].OC(O)=O.OC([O-])=O POECFFCNUXZPJT-UHFFFAOYSA-M 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
- G01N2021/3155—Measuring in two spectral ranges, e.g. UV and visible
Landscapes
- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A kind of application the invention discloses cubic boron nitride as Mimetic Peroxidase, specially application of the cubic boron nitride as Mimetic Peroxidase measurement concentration of hydrogen peroxide, the wherein cubic boron nitride Mimetic enzyme, partial size is unlimited, all has Mimetic Peroxidase activity.Present invention firstly provides cubic boron nitrides to have the function of that Mimetic enzyme, enzymatic activity can match in excellence or beauty natural horseradish peroxidase (HRP), can be used for detecting the content of the hydrogen peroxide in solution example.Relative to native enzyme, cubic boron nitride analogue enztme is higher in the extreme environments stability inferior such as high temperature and strong acid and strong base, reusable and be able to maintain efficient catalytic activity.Preparation process of the present invention is simple, reproducible, and stability is high, at low cost.Cubic boron nitride can replace peroxidase to be applied to the fields such as immunoassay, biological detection, clinical diagnosis and contaminant degradation as a kind of new Mimetic enzyme of peroxidase.
Description
Technical field
The present invention relates to biomaterial field of chemical detection, refers in particular to a kind of cubic boron nitride and simulated as peroxide
The application of enzyme.
Background technique
Native enzyme is because of its highly selective and efficient catalytic activity, in fields such as biology, chemistry, agricultural and food processings
It has important application.However, native enzyme vulnerable to the experiment conditions such as temperature, pH value influence and can deactivation cause it steady
Qualitative reduction, expensive, purification difficult, storage and use cost are high, these disadvantages significantly limit the application range of enzyme.
Therefore, it is necessary to develop the catalysis characteristics with enzyme and the more stable analogue enztme of structure.
Compared with native enzyme, there is nano material at low cost, catalytic activity to be adjusted, stability is high, is easily handled and stores up
The advantages that depositing.In recent years, the various inorganic material analogue enztmes with peroxidase activity are developed successively.Current receives
Rice material Mimetic Peroxidase mainly includes three categories: metal, such as platinum, gold and ferrocobalt etc.；Metal oxide, such as
Ferroso-ferric oxide, cobaltosic oxide and copper oxide etc.；Carbon-based material, such as graphene oxide, carbon quantum dot and helical structure
Carbon nanotube etc..These the disadvantages of generally existing catalytic efficiency of nano material analogue enztme is low, tolerance is poor and non-renewable.
Cubic boron nitride is a kind of superhard material, has very strong chemical inertness, is resistant to the poles such as high temperature and strong acid and strong base
End ring border, and can be separated by the straightforward procedures such as being centrifuged and filtering with liquid phase medium realization.Cubic boron nitride has simulation peroxide
Compound enzyme viability.Cubic boron nitride is typical nitride, therefore is different from metal, metal oxide and carbon-based material, is
4th class inorganic peroxide analogue enztme.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art and disadvantage, and the characteristic based on cubic boron nitride is (as simulated
Peroxidase activity), a kind of application of cubic boron nitride as Mimetic Peroxidase is provided.
To achieve the above object, technical solution provided by the present invention are as follows: cubic boron nitride is as Mimetic Peroxidase
Application, specially application of the cubic boron nitride as Mimetic Peroxidase measurement concentration of hydrogen peroxide, wherein described cube
Boron nitride Mimetic enzyme, partial size is unlimited, all has Mimetic Peroxidase activity.
Application of the cubic boron nitride as Mimetic Peroxidase in measurement concentration of hydrogen peroxide, comprising the following steps:
1) preparation of cubic boron nitride analogue enztme suspension
The cubic boron nitride powder for weighing set amount is placed in container, is added after ultrapure water ultrasonic disperse 15 minutes, is prepared
Obtain cubic boron nitride suspension；
2) cubic boron nitride Mimetic Peroxidase characteristic
It takes buffer solution to be placed in warm bath 10 minutes in water bath with thermostatic control, sequentially adds cubic boron nitride Mimetic Peroxidase
Hydrogen donor substrate, hydrogenperoxide steam generator, cubic boron nitride solution, shake up, 45 DEG C after reaction 4 minutes, are taken out, after filtering
To sample to be tested；Using ultrapure water as blank, is measured in 400-800nm wave-length coverage with ultraviolet-uisible spectrophotometer and absorb light
Spectrum；
3) cubic boron nitride measures hydrogen peroxide as Mimetic Peroxidase
It takes buffer solution to be placed in warm bath 10 minutes in water bath with thermostatic control, sequentially adds cubic boron nitride Mimetic Peroxidase
Hydrogen donor substrate, with hydrogenperoxide steam generator, the cubic boron nitride solution of step 2) various concentration, shake up, 45 DEG C are reacted 20 points
Zhong Hou takes out, obtains sample to be tested after syringe filter filters；Using ultrapure water as blank, with UV, visible light spectrophotometric
The absorbance of meter measurement maximum absorption wave strong point, obtains absorbance-concentration of hydrogen peroxide relation curve.
The concentration of hydrogen peroxide is detected, the range of linearity is 10 μm of ol/L to 100 μm of ol/L, and detection is limited to 5 μm of ol/L.
The hydrogen donor of horseradish peroxidase (HRP) can be used as the hydrogen donor bottom of cubic boron nitride Mimetic Peroxidase
Object, preferably 3,3', 5,5'- tetramethyl benzidines (TMB), 3,3'- diaminobenzidine (DAB), o-phenylenediamine (OPD) or 2,
2'- joins bis- (3- ethyl benzo thiazole phenanthroline -6- sulfonic acid) di-ammonium salts of nitrogen, and chromogenic reaction occurs under the action of hydrogen peroxide.
The hydrogen acceptor substrate of cubic boron nitride Mimetic Peroxidase is the hydrogen acceptor bottom of horseradish peroxidase (HRP)
Object is not limited to hydrogen peroxide, further includes methyl hydroperoxide and ethyl hydrogen peroxide, can be in the effect of HRP hydrogen donor substrate
Lower generation chromogenic reaction.
Its buffer solution of cubic boron nitride is unlimited, and preferably citrate-phosphate disodium hydrogen or acetic acid-acetate buffer is molten
Liquid, the pH range with catalytic activity are 2 to 10.
For cubic boron nitride as Mimetic Peroxidase, the temperature range with catalytic activity is 4 DEG C to 90 DEG C.
It further include through excessively high in addition to cubic boron nitride raw material as the cubic boron nitride material of Mimetic Peroxidase
Temperature, the processed cubic boron nitride of strong acid and strong base.Temperature and pH value when processing is unlimited；The time of processing is unlimited, and preferably 2 is small
When, cubic boron nitride can keep mimetic enzyme catalysis active.
As the cubic boron nitride material of Mimetic Peroxidase, in addition to cubic boron nitride raw material, further include by from
The modified cubic boron nitride of son injection.Ionic species is unlimited, preferably nitrogen, boron, carbon, oxygen, helium and argon ion, and modified cube
Boron nitride all has catalytic activity.
As the cubic boron nitride material of Mimetic Peroxidase, in addition to cubic boron nitride raw material, further include by
It anneals under atmosphere or vacuum environment modified cubic boron nitride.Annealing atmosphere is unlimited, preferably nitrogen, oxygen, helium, argon gas and
Air；Annealing time is unlimited, and preferably 2-8 hours, modified cubic boron nitride all had activity.
Its reaction dissolvent of cubic boron nitride is water, ethyl alcohol, isopropanol, the mixing of one or more of acetonitrile.It is molten
Agent is preferably water.
Cubic boron nitride can keep 50% or more enzymatic activity within reusing 10 times.
It, can be with other than it can detect concentration of hydrogen peroxide as the cubic boron nitride material of Mimetic Peroxidase
The content of detecting glucose, amino acid and xanthine.
As the cubic boron nitride material of Mimetic Peroxidase, concentration of hydrogen peroxide is characterized in addition to using ultraviolet-visible
Outside optical absorption method, fluorescence method, chemoluminescence method and electrochemical process can also be used.
Cubic boron nitride in addition to for being catalytically decomposed and detect hydrogen peroxide, being except excellent Mimetic Peroxidase,
Still the good catalyst of degradation of phenol, rhodamine B, aniline, methyl blue and xylenol orange organic pollutant.
Compared with prior art, the present invention have the following advantages that with the utility model has the advantages that
1, present invention firstly provides cubic boron nitrides to have the function of that Mimetic enzyme, enzymatic activity can match in excellence or beauty
Natural horseradish peroxidase (HRP), can be used for detecting the content of the hydrogen peroxide in solution example.
2, relative to native enzyme, cubic boron nitride analogue enztme the extreme environments such as high temperature and strong acid and strong base stability inferior more
Height, it is reusable and be able to maintain efficient catalytic activity.
3, preparation process of the present invention is simple, reproducible, and stability is high, at low cost.Cubic boron nitride is as a kind of new
Mimetic enzyme of peroxidase can replace peroxidase to be applied to immunoassay, biological detection, clinical diagnosis and contaminant degradation
Equal fields.
Detailed description of the invention
Fig. 1 is the X-ray diffraction spectrogram of cubic boron nitride Mimetic Peroxidase of the invention.
Fig. 2 is the scanning electron microscope diagram of cubic boron nitride Mimetic Peroxidase of the invention.
Fig. 3 is cubic boron nitride Mimetic Peroxidase of the invention in peroxidase substrate 3,3', 5,5'- tetramethyl
Chromogenic reaction schematic diagram under benzidine (TMB) effect.
Fig. 4 a is the relative activity figure of cubic boron nitride Mimetic Peroxidase of the invention at various ph values.
Fig. 4 b is the relative activity figure of cubic boron nitride Mimetic Peroxidase of the invention at different temperatures.
Fig. 5 a be cubic boron nitride Mimetic Peroxidase raw material of the invention and respectively through injection helium, boron, nitrogen from
The enzymatic activity figure of the cubic boron nitride of son.
Fig. 5 b is for cubic boron nitride Mimetic Peroxidase raw material of the invention and respectively through nitrogen and air anneal
Cubic boron nitride enzymatic activity figure.
Fig. 6 is the application signal that cubic boron nitride of the invention measures concentration of hydrogen peroxide as Mimetic Peroxidase
Figure.
Fig. 7 is the application signal that cubic boron nitride of the invention decomposes rhodamine B organic matter as Mimetic Peroxidase
Figure.
Specific embodiment
Below with reference to multiple specific embodiments, the invention will be further described.
Embodiment 1
The cubic boron nitride powder for weighing 50mg is placed in centrifuge tube, is added after 10ml ultrapure water ultrasonic disperse 10 minutes,
Preparation obtains the cubic boron nitride suspension of 5mg/ml.Taking 3.7mlpH is that 4 citrate-phosphate disodium hydrogen buffers are placed in constant temperature
45 DEG C warm bath 10 minutes in water-bath sequentially add 3,3', 5, the 5'- tetramethyl benzidines that 0.1ml concentration is 0.1mmol/L
(TMB) solution, the hydrogenperoxide steam generator of the 10mmol/L of 0.1ml and the cubic boron nitride solution of 0.1ml, shake up, 45 DEG C of reactions
After ten minutes, it takes out, obtains sample to be tested after filtering.Using ultrapure water as blank, with ultraviolet-uisible spectrophotometer in 400-
Absorption spectrum is measured in 800nm wave-length coverage.
In addition, also carrying out X-ray diffraction, scanning electron microscope and photoelectron to above-mentioned cubic boron nitride in this embodiment
Energy spectrum analysis characterization.
The X-ray diffraction spectrum of cubic boron nitride is as shown in Figure 1, all diffraction maximums of material can all be demarcated as cubic system
Boron nitride matches with standard diagram JCPDS79-0623.
The scanning electron microscope diagram of cubic boron nitride is as shown in Fig. 2, the average grain diameter that cubic boron nitride can be obtained is
410nm。
Cubic boron nitride Mimetic Peroxidase is made in peroxidase substrate 3,3', 5,5'- tetramethyl benzidine (TMB)
Chromogenic reaction under is as shown in Figure 3.
Embodiment 2
The cubic boron nitride powder for weighing 50mg is placed in centrifuge tube, is added after 10ml ultrapure water ultrasonic disperse 10 minutes,
Preparation obtains the cubic boron nitride suspension of 5mg/ml.Taking 3.7mlpH respectively is respectively 2,3,4,5,6,7 and 8 citric acid-phosphorus
Sour disodium hydrogen buffer and pH are respectively that 9,10 and 11 sodium carbonate-bicarbonate buffer is placed in 45 DEG C of temperature in water bath with thermostatic control
Bath 10 minutes sequentially adds 3,3', 5,5'- tetramethyl benzidine (TMB) solution that 0.1ml concentration is 1mmol/L, 0.1ml
50mmol/L hydrogenperoxide steam generator and 0.1ml cubic boron nitride solution, shake up, 45 DEG C reaction after ten minutes, take out,
Sample to be tested is obtained after filtering.Using ultrapure water as blank, with ultraviolet-uisible spectrophotometer measurement maximum absorption wave strong point
Absorbance.
The opposite peroxidase activity of cubic boron nitride Mimetic Peroxidase at various ph values is as shown in fig. 4 a.
Embodiment 3
The cubic boron nitride powder for weighing 50mg is placed in centrifuge tube, is added after 10ml ultrapure water ultrasonic disperse 10 minutes,
Preparation obtains the cubic boron nitride suspension of 5mg/ml.Taking 3.7mlpH is that 4 citrate-phosphate disodium hydrogen buffer is placed in perseverance
Warm bath 10 minutes in tepidarium, temperature are set to 25,30,35,40,45,50,55 and 60 DEG C, sequentially add 0.1ml concentration
It is the 3,3' of 1mmol/L, 5,5'- tetramethyl benzidines (TMB) solution, the hydrogenperoxide steam generator of the 50mmol/L of 0.1ml, and
The cubic boron nitride solution of 0.1ml, shakes up, and reacts after ten minutes respectively in corresponding temperature, takes out, obtained after filtering to test sample
Product.Using ultrapure water as blank, with the absorbance of ultraviolet-uisible spectrophotometer measurement maximum absorption wave strong point.
The opposite peroxidase activity of cubic boron nitride Mimetic Peroxidase at different temperatures is as shown in Figure 4 b.
Embodiment 4
Cubic boron nitride powder is carried out to helium, boron and N~+ implantation processing respectively or is handled in nitrogen and air anneal.
The above-mentioned cubic boron nitride powder for weighing 50mg respectively is placed in centrifuge tube, is added after 10ml ultrapure water ultrasonic disperse 10 minutes,
Preparation obtains the cubic boron nitride suspension of 5mg/ml.Taking 3.7mlpH is that 4 citrate-phosphate disodium hydrogen buffers are placed in constant temperature
45 DEG C warm bath 10 minutes in water-bath sequentially add 3,3', 5, the 5'- tetramethyl benzidines that 0.1ml concentration is 0.1mmol/L
(TMB) solution, the hydrogenperoxide steam generator of the 10mmol/L of 0.1ml and the cubic boron nitride solution of 0.1ml, shake up, 45 DEG C of reactions
After ten minutes, it takes out, obtains sample to be tested after filtering.Using ultrapure water as blank, with ultraviolet-uisible spectrophotometer in 400-
Absorption spectrum is measured in 800nm wave-length coverage.
Cubic boron nitride Mimetic Peroxidase raw material and respectively through injection helium, boron, Nitrogen ion cubic boron nitride
Enzymatic activity it is as shown in Figure 5 a.
Cubic boron nitride Mimetic Peroxidase raw material and cubic boron nitride respectively through nitrogen and air anneal
Enzymatic activity is as shown in Figure 5 b.
Embodiment 5
The above-mentioned cubic boron nitride powder for weighing 50mg is placed in centrifuge tube, and ultrasonic disperse 10 divides after 10ml ultrapure water is added
Clock, preparation obtain the cubic boron nitride suspension of 5mg/ml.Taking 3.1mlpH respectively is 4 citrate-phosphate disodium hydrogen buffer
45 DEG C warm bath 10 minutes in water bath with thermostatic control are placed in, 3,3', 5, the 5'- tetramethyls that 0.4ml concentration is 10mmol/L are sequentially added
Benzidine (TMB) solution, the concentration of 0.4ml are respectively the hydrogenperoxide steam generator of 0.1,0.3,0.6,1,3,6 and 10mmol/L,
It with the cubic boron nitride solution of 0.1ml, shakes up, 45 DEG C of reactions after ten minutes, take out, obtain sample to be tested after filtering.With super
Pure water is blank, with the absorbance of ultraviolet-uisible spectrophotometer measurement maximum absorption wave strong point.
Fig. 6 is the application schematic diagram that cubic boron nitride measures concentration of hydrogen peroxide as Mimetic Peroxidase.It is detected
The concentration of hydrogen peroxide, the range of linearity are 10 μm of ol/L to 100 μm of ol/L, and detection is limited to 5 μm of ol/L.
Embodiment 6
The above-mentioned cubic boron nitride powder for weighing 50mg is placed in centrifuge tube, and ultrasonic disperse 10 divides after 10ml ultrapure water is added
Clock, preparation obtains the cubic boron nitride suspension of 5mg/ml, and suspension is injected into 3 end to end syringe filters
In.The rhodamine B solution that 20ml concentration is 10mmol/L is prepared, the hydrogenperoxide steam generator of 1mmol/L is added, is placed in 25ml injection
In device, and syringe and filter group are spliced, rhodamine B solution is allowed slowly to flow through the cubic boron nitride mistake in filter
Oxide analogue enztme simultaneously reacts.Picking up efflux is the rhodamine B solution through cubic boron nitride catalytic degradation.
Cubic boron nitride is as shown in Figure 7 as the application schematic diagram that Mimetic Peroxidase decomposes phenyl amines organic matter.
The examples of implementation of the above are only the preferred embodiments of the invention, and implementation model of the invention is not limited with this
It encloses, therefore all shapes according to the present invention, changes made by principle, should all be included within the scope of protection of the present invention.
Claims (10)
1. application of the cubic boron nitride as Mimetic Peroxidase, it is characterised in that: be cubic boron nitride as peroxide
The application of enzymatic determination concentration of hydrogen peroxide is simulated, wherein the cubic boron nitride Mimetic enzyme, partial size is unlimited, has
There is Mimetic Peroxidase active.
2. application of the cubic boron nitride according to claim 1 as Mimetic Peroxidase, which is characterized in that cube nitrogen
Change application of the boron as Mimetic Peroxidase in measurement concentration of hydrogen peroxide, comprising the following steps:
1) preparation of cubic boron nitride analogue enztme suspension
The cubic boron nitride powder for weighing set amount is placed in container, is added after ultrapure water ultrasonic disperse 15 minutes, preparation obtains
Cubic boron nitride suspension；
2) cubic boron nitride Mimetic Peroxidase characteristic
It takes buffer solution to be placed in warm bath 10 minutes in water bath with thermostatic control, sequentially adds the confession of cubic boron nitride Mimetic Peroxidase
Hydrogen body substrate, hydrogenperoxide steam generator, cubic boron nitride solution, shake up, 45 DEG C reaction 4 minutes after, take out, obtain after filtering to
Sample；Using ultrapure water as blank, absorption spectrum is measured in 400-800nm wave-length coverage with ultraviolet-uisible spectrophotometer；
3) cubic boron nitride measures hydrogen peroxide as Mimetic Peroxidase
It takes buffer solution to be placed in warm bath 10 minutes in water bath with thermostatic control, sequentially adds the confession of cubic boron nitride Mimetic Peroxidase
Hydrogen body substrate, hydrogenperoxide steam generator, cubic boron nitride solution with step 2) various concentration, shake up, and 45 DEG C are reacted after twenty minutes,
It takes out, obtains sample to be tested after syringe filter filters；Using ultrapure water as blank, measured with ultraviolet-uisible spectrophotometer
The absorbance of maximum absorption wave strong point obtains absorbance-concentration of hydrogen peroxide relation curve.
3. application of the cubic boron nitride according to claim 2 as Mimetic Peroxidase, it is characterised in that: as mistake
The cubic boron nitride material of oxide analogue enztme further includes by high temperature, strong acid and strong base processing in addition to cubic boron nitride raw material
The cubic boron nitride crossed, treated, and cubic boron nitride is able to maintain mimetic enzyme catalysis activity.
4. application of the cubic boron nitride according to claim 2 as Mimetic Peroxidase, it is characterised in that: as mistake
The cubic boron nitride material of oxide analogue enztme further includes by the vertical of ion implantation modification in addition to cubic boron nitride raw material
Square boron nitride, modified cubic boron nitride all have catalytic activity.
5. application of the cubic boron nitride according to claim 2 as Mimetic Peroxidase, it is characterised in that: as mistake
The cubic boron nitride material of oxide analogue enztme further includes by atmosphere or vacuum environment in addition to cubic boron nitride raw material
The modified cubic boron nitride of lower annealing, modified cubic boron nitride all have activity.
6. application of the cubic boron nitride according to claim 2 as Mimetic Peroxidase, it is characterised in that: peroxidating
The range of linearity of hydrogen concentration detection is 10 μm of ol/L to 100 μm of ol/L, and detection is limited to 5 μm of ol/L；The cubic boron nitride mistake
Oxide analogue enztme characteristic, the temperature range with catalytic activity are 25 DEG C to 60 DEG C；Cubic boron nitride is reusing 10
Within secondary, it is able to maintain 50% or more enzymatic activity；The hydrogen donor substrate of cubic boron nitride Mimetic Peroxidase is
The hydrogen donor of horseradish peroxidase HRP；The hydrogen acceptor substrate of cubic boron nitride Mimetic Peroxidase is horseradish peroxidase
The hydrogen acceptor substrate of enzyme HRP；The buffer solution is citrate-phosphate disodium hydrogen or acetic acid-sodium acetate buffer solution, is had
The pH range of catalytic activity is 2 to 7；The reaction dissolvent of cubic boron nitride be one of water, ethyl alcohol, isopropanol, acetonitrile or
Two or more mixing.
7. application of the cubic boron nitride according to claim 6 as Mimetic Peroxidase, it is characterised in that: cube nitrogen
The hydrogen donor substrate for changing boron Mimetic Peroxidase is 3,3', 5,5'- tetramethyl benzidine TMB, 3,3'-diaminobenzidine
DAB, o-phenylenediamine OPD or 2,2'- join one of double di-ammonium salts of nitrogen, chromogenic reaction occur under the action of hydrogen peroxide；It is vertical
The hydrogen acceptor substrate of square boron nitride Mimetic Peroxidase further includes methyl hydroperoxide and ethyl peroxidating in addition to hydrogen peroxide
Under the action of HRP hydrogen donor substrate chromogenic reaction can occur for hydrogen.
8. application of the cubic boron nitride according to any one of claims 1 to 6 as Mimetic Peroxidase, feature exist
In: as the cubic boron nitride material of Mimetic Peroxidase, other than it can detect concentration of hydrogen peroxide, additionally it is possible to detect Portugal
The content of grape sugar, amino acid and xanthine.
9. application of the cubic boron nitride according to any one of claims 1 to 6 as Mimetic Peroxidase, feature exist
In: as the cubic boron nitride material of Mimetic Peroxidase, concentration of hydrogen peroxide is characterized in addition to using UV-Visible absorption
Outside method, additionally it is possible to use fluorescence method, chemoluminescence method and electrochemical process.
10. application of the cubic boron nitride according to any one of claims 1 to 6 as Mimetic Peroxidase, feature
Be: cubic boron nitride is in addition to that can be catalytically decomposed and detect hydrogen peroxide, additionally it is possible to catalytic degradation phenol, rhodamine B, benzene
Amine, methyl blue and xylenol orange organic pollutant.
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