CN110296946A - Transparent paper-based ultraviolet-visible light absorption measuring device and paper-based analysis method - Google Patents
Transparent paper-based ultraviolet-visible light absorption measuring device and paper-based analysis method Download PDFInfo
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- CN110296946A CN110296946A CN201910632652.XA CN201910632652A CN110296946A CN 110296946 A CN110296946 A CN 110296946A CN 201910632652 A CN201910632652 A CN 201910632652A CN 110296946 A CN110296946 A CN 110296946A
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- paper base
- cholesterol
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- protein
- detection
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- 238000004458 analytical method Methods 0.000 title claims abstract description 21
- 230000031700 light absorption Effects 0.000 title abstract 3
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 90
- 238000001514 detection method Methods 0.000 claims abstract description 53
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 45
- 238000002331 protein detection Methods 0.000 claims abstract description 24
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 23
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 23
- 238000000870 ultraviolet spectroscopy Methods 0.000 claims abstract description 19
- 239000000243 solution Substances 0.000 claims description 35
- 238000010521 absorption reaction Methods 0.000 claims description 26
- 238000005259 measurement Methods 0.000 claims description 21
- 229920002678 cellulose Polymers 0.000 claims description 13
- 239000001913 cellulose Substances 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 11
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- QPMIVFWZGPTDPN-UHFFFAOYSA-N Tetrabromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C(C(Br)=C(Br)C(Br)=C2Br)=C2S(=O)(=O)O1 QPMIVFWZGPTDPN-UHFFFAOYSA-N 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- 238000012360 testing method Methods 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 7
- 238000004090 dissolution Methods 0.000 claims description 7
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 claims description 6
- 101100366707 Arabidopsis thaliana SSL11 gene Proteins 0.000 claims description 6
- 101100366710 Arabidopsis thaliana SSL12 gene Proteins 0.000 claims description 6
- 108010055297 Sterol Esterase Proteins 0.000 claims description 5
- 102000000019 Sterol Esterase Human genes 0.000 claims description 5
- 239000011521 glass Substances 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- LHCPRYRLDOSKHK-UHFFFAOYSA-N 7-deaza-8-aza-adenine Chemical compound NC1=NC=NC2=C1C=NN2 LHCPRYRLDOSKHK-UHFFFAOYSA-N 0.000 claims description 4
- 108010089254 Cholesterol oxidase Proteins 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 4
- 208000022379 autosomal dominant Opitz G/BBB syndrome Diseases 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 235000019441 ethanol Nutrition 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- CJUDSKIRZCSXJA-UHFFFAOYSA-M sodium;3-(n-ethyl-3-methoxyanilino)-2-hydroxypropane-1-sulfonate Chemical compound [Na+].[O-]S(=O)(=O)CC(O)CN(CC)C1=CC=CC(OC)=C1 CJUDSKIRZCSXJA-UHFFFAOYSA-M 0.000 claims description 4
- 101100366711 Arabidopsis thaliana SSL13 gene Proteins 0.000 claims description 3
- 229920000742 Cotton Polymers 0.000 claims description 3
- 101100366561 Panax ginseng SS11 gene Proteins 0.000 claims description 3
- 101100366562 Panax ginseng SS12 gene Proteins 0.000 claims description 3
- 101100366563 Panax ginseng SS13 gene Proteins 0.000 claims description 3
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 3
- 239000002390 adhesive tape Substances 0.000 claims description 3
- 238000007605 air drying Methods 0.000 claims description 3
- 239000004202 carbamide Substances 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 3
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 210000000232 gallbladder Anatomy 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000010907 mechanical stirring Methods 0.000 claims description 3
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims description 3
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims description 3
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 3
- 229920002379 silicone rubber Polymers 0.000 claims description 3
- 238000003860 storage Methods 0.000 claims description 3
- 229930182558 Sterol Natural products 0.000 claims description 2
- 239000007979 citrate buffer Substances 0.000 claims description 2
- 230000002255 enzymatic effect Effects 0.000 claims description 2
- 150000003432 sterols Chemical class 0.000 claims description 2
- 235000003702 sterols Nutrition 0.000 claims description 2
- CWPKTBMRVATCBL-UHFFFAOYSA-N 3-[1-[1-[(2-methylphenyl)methyl]piperidin-4-yl]piperidin-4-yl]-1h-benzimidazol-2-one Chemical compound CC1=CC=CC=C1CN1CCC(N2CCC(CC2)N2C(NC3=CC=CC=C32)=O)CC1 CWPKTBMRVATCBL-UHFFFAOYSA-N 0.000 claims 1
- 235000011330 Armoracia rusticana Nutrition 0.000 claims 1
- 240000003291 Armoracia rusticana Species 0.000 claims 1
- 238000000862 absorption spectrum Methods 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract 1
- 239000000126 substance Substances 0.000 description 7
- 230000007613 environmental effect Effects 0.000 description 5
- 229920000298 Cellophane Polymers 0.000 description 4
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000004737 colorimetric analysis Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000012827 research and development Methods 0.000 description 3
- 238000004847 absorption spectroscopy Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 150000001860 citric acid derivatives Chemical class 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 150000004683 dihydrates Chemical class 0.000 description 2
- 239000010408 film Substances 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 238000005375 photometry Methods 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 238000012805 post-processing Methods 0.000 description 2
- 238000007639 printing Methods 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- CGNOCUSLPSCMLL-UHFFFAOYSA-N 3-o-benzyl 1-o-ethyl propanedioate Chemical compound CCOC(=O)CC(=O)OCC1=CC=CC=C1 CGNOCUSLPSCMLL-UHFFFAOYSA-N 0.000 description 1
- 108010025188 Alcohol oxidase Proteins 0.000 description 1
- 235000001258 Cinchona calisaya Nutrition 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- LOUPRKONTZGTKE-WZBLMQSHSA-N Quinine Natural products C([C@H]([C@H](C1)C=C)C2)C[N@@]1[C@@H]2[C@H](O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-WZBLMQSHSA-N 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- LOUPRKONTZGTKE-UHFFFAOYSA-N cinchonine Natural products C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-UHFFFAOYSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000007641 inkjet printing Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- UZKWTJUDCOPSNM-UHFFFAOYSA-N methoxybenzene Substances CCCCOC=C UZKWTJUDCOPSNM-UHFFFAOYSA-N 0.000 description 1
- RDOXTESZEPMUJZ-UHFFFAOYSA-N methyl phenyl ether Natural products COC1=CC=CC=C1 RDOXTESZEPMUJZ-UHFFFAOYSA-N 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229960000948 quinine Drugs 0.000 description 1
- -1 quinine imines Chemical class 0.000 description 1
- 150000004060 quinone imines Chemical class 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 238000007650 screen-printing Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- PNGLEYLFMHGIQO-UHFFFAOYSA-M sodium;3-(n-ethyl-3-methoxyanilino)-2-hydroxypropane-1-sulfonate;dihydrate Chemical compound O.O.[Na+].[O-]S(=O)(=O)CC(O)CN(CC)C1=CC=CC(OC)=C1 PNGLEYLFMHGIQO-UHFFFAOYSA-M 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 239000010409 thin film Substances 0.000 description 1
- 229940116269 uric acid Drugs 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
-
- 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
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- General Physics & Mathematics (AREA)
- Hematology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Endocrinology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a transparent paper-based ultraviolet-visible light absorption determination device, which comprises a protein detection layer A, a cholesterol detection layer B, a sample and an ultraviolet-visible spectrophotometer, wherein the protein detection layer A and the cholesterol detection layer B are respectively arranged at the output end of the ultraviolet-visible spectrophotometer, the sample is respectively added into the protein detection layer A and the cholesterol detection layer B to be stacked after being dried, and then the ultraviolet-visible spectrophotometer emits light beams to the sample to realize the simultaneous detection of the protein and the cholesterol. The invention also discloses an analysis method of the transparent paper-based ultraviolet-visible light absorption determination device, which can realize simultaneous detection of protein and cholesterol, has high sensitivity and good selectivity for the detection of the protein and the cholesterol, and has detection limits of 0.09 mM and 0.075 mu M respectively.
Description
Technical field
The present invention relates to transparent paper base UV-Visible absorption measurement device and paper base analysis methods, and in particular to saturating
On bright, after being reacted using protein with cholesterol with substance absorbance Bu Tong and meanwhile detect the paper of protein and cholesterol
Base analytical equipment and method belong to paper base technical field of analysis and detection.
Background technique
Paper base analytical equipment is because it is cheap, consumption reagent is few, environmental-friendly, good biocompatibility obtains extensively due to the advantages that
Using, the colorimetric detection that carries out on paper base for not needing any instrument is liked by researcher deeply, but due to chromatographic paper,
When carrying out colorimetric detection on the opaque paper base such as filter paper, office paper, the quantitative of the colorimetric analysis determinand of paper base is usually required
The intensity of the light reflected from papery equipment surface is measured using smart phone or flat bed scanner, this detection method realizes
Real-time response and high portability.However, being quantitatively not always to be easy to, and need the coloured of high concentration using colorimetric method
Product, detection limit are usually very high.Therefore, using the detection of the colorimetric method of transparent paper base, detection uv-visible absorption spectroscopy
Quantitative detection and prepare multi-layer transparent paper base analytical equipment realize many kinds of substance quantitative detection it is particularly important.
The transparent nano paper being made of nano-cellulose is a kind of up-and-coming detection device substrate, has transparency
High, at low cost, good biocompatibility, the smooth feature of degradability and surface.A series of flexible electronic devices containing touch screen
It is successfully assembled on cellophane paper including Organic Light Emitting Diode (OLED), thin film transistor (TFT) (TFT) etc..Cellophane paper based device can
To pass through ink jet printing, wax printing, flexographic printing and silk-screen printing production.Detected with the method for colorimetric detection lactic acid, uric acid,
The example of protein and cholesterol is successfully reported in paper base equipment.But pass through absorption spectrometry pair on multi-layer transparent paper
The detection of Cucumber progress simultaneous quantitative is also fewer in biofluid.
UV-Vis Spectrophotometry refers under the action of light, the transmitting light that is generated by measurement substance, absorb light or
The method for scattering the wavelength of light to be analyzed.In spectrum analysis, the selective absorbing of light is set up according to substance
Absorption process be known as absorption photometry.Absorption photometry is widely used in the qualitative and quantitative survey of substance because its is easy to operate
In examination.
Horizontal increase in human body of protein and cholesterol may cause some diseases, including liver or kidney trouble,
And heart disease.
Summary of the invention
It is an object of the present invention to overcome defect of the existing technology, above-mentioned technical problem is solved, proposes transparent paper base
UV-Visible absorption measurement device and paper base analysis method, transparent paper base detection method of the invention are Proteins in Serum
A kind of convenient, fast, effective method is provided with the quantitative detection of cholesterol, can be diagnosed in vitro, the side such as environmental monitoring
Find more applications in face.
The technical problem to be solved in the present invention includes: first, and it is cheap to research and develop transparent paper base analytical equipment, solves inspection
The problem of surveying somewhat expensive；Second, the transparent paper base analysis method of research and development can detect two kinds of substances simultaneously.
The present invention adopts the following technical scheme: transparent paper base UV-Visible absorption measurement device, which is characterized in that packet
Include protein detection layer A, cholesterol detection layer B, sample, ultraviolet-visible spectrophotometer, the protein detection layer A and institute
The output end that cholesterol detection layer B is respectively arranged at the ultraviolet-visible spectrophotometer is stated, the sample is added into respectively
It is stacked after the protein detection layer A and cholesterol detection layer B is dry, then passes through the spectrophotometry
It is detected while realization on meter outgoing beam to the sample to protein and cholesterol.
The present invention also proposes the paper base analysis method of transparent paper base UV-Visible absorption measurement device, which is characterized in that
Include the following steps:
Step SS1: transparent paper base is prepared；
Step SS2: testing device for transparent paper is prepared；
Step SS3: using UV-Visible absorption method measurement protein and cholesterol.
As a kind of preferred embodiment, the step SS1 is specifically included:
Step SS11: cellulose dissolution liquid is prepared；
Step SS12: cellulose solution is prepared；
Step SS13: cellulose solution prepared by the step SS12 is poured on glass plate, is formed one layer uniform thin
Then film places it in the sulfuric acid solution of 5w% and is formed by curing gel, thoroughly impregnated using distilled water, then solid using adhesive tape
It is fixed, transparent paper base is made after drying at room temperature.
As a kind of preferred embodiment, the step SS11 is specifically included: by sodium hydroxide, urea and water according to quality
Than the mass ratio wiring solution-forming for 7:12:81, it is put into spare in -12 DEG C of refrigerator.
As a kind of preferred embodiment, the step SS12 is specifically included: the step SS11 institute is added in cotton linter
In the cellulose dissolution liquor of preparation, using mechanical stirring, mixing time is 15 min, 3000 rpm of mixing speed, later
It is under 8000 rpm mixing speeds that solution centrifugation bubble removing is spare.
As a kind of preferred embodiment, the testing device for transparent paper for preparing in the step SS2 is specifically included:
Step SS21: with mass ratio being that 10:1 is mixed by PDMS and silicone elastomer curing agent, and room temperature vacuumizes half an hour and removes
Go bubble spare；
Step SS22: solution described in step SS21 is printed on the transparent paper base by layout using screen process press,
Using vacuum oven, toasted 12 hours at 80 DEG C, to be formed by curing transparent paper base analytical equipment, the final Kong Zhi of detection zone
Diameter is 5 mm, and normal temperature storage is spare.
As a kind of preferred embodiment, the step SS3 is specifically included:
Step SS31: the preparation of protein detection solution；
Step SS32: the preparation of cholesterol detection solution；
Step SS33: it is detected using ultraviolet-visible spectrophotometer.
As a kind of preferred embodiment, the step SS31 is specifically included: the pH 125 mM citrates for being 1.8 are delayed
The tetrabromophenol blue TBPB solution of fliud flushing and 9 mM dissolved with 95% ethyl alcohol and 5% water in the ratio of volume ratio 10:1 to mix
It closes, 4 DEG C store for future use.
As a kind of preferred embodiment, the step SS32 is specifically included: cholesterol esterase CE(14.1 U), gallbladder is solid
Alcohol oxidase CO(10 mg/mL), 4-AA 4-APP(10 mM), horseradish peroxidase HRP(1 mg/mL) and
N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline sodium salt dihydrate ADOS(1mM) with volume ratio 1:1:1:1:1
Mixing, is put into 4 DEG C and stores for future use.
As a kind of preferred embodiment, the step SS33 is specifically included: by the protein detection of the step SS31
The cholesterol detection solution of solution and the step SS32 are added separately to protein detection layer A and cholesterol detection layer B, later
It is separately added into sample, air drying stacks, and uses the absorption light of ultraviolet-visible spectrophotometer detection protein and cholesterol
Spectrum.
Advantageous effects of the invention: first, transparent paper base analytical equipment of the invention and paper base analysis method can
To be detected while realization to protein and cholesterol, and all have to the detection of protein and cholesterol highly sensitive and good
Good selectivity, detection limit are respectively 0.09 mM and 0.075 μM；Second, the cellophane paper that the present invention uses is that one kind can drop
The ideal material of solution replaces the materials such as traditional plastics, glass to alleviate environmental pressure with it, cheap, environmental protection, post-processing
Simply, it thoroughly solves that the transparent paper base analytical equipment of research and development is cheap, solves the problems, such as testing cost valuableness and research and development
Transparent paper base analysis method can detect the technical need of two kinds of substances simultaneously.
Detailed description of the invention
Fig. 1 is the schematic diagram of the invention measured using UV-Visible absorption.
Fig. 2 is schematic diagram of the present invention in transparent paper base analytical equipment detection protein.
Fig. 3 is schematic diagram of the present invention in transparent paper base analytical equipment cholesterol detection, and meaning marked in the figure: BSA is
Citrate buffer；Tetrabromophenol blue is Tetrabromophenol Blue (TBPB)；4-APP = 4-
Aminoantipyrine, i.e. 4-AA；CE=Cholesterol esterase, i.e. cholesterol esterase；CO =
Cholesterol oxidase, i.e. cholesterol oxidase；Cho=Colesterol, i.e. sterol；HRP = Horseradish
Peroxidase, i.e. horseradish peroxidase；ADOS=N-ethyl-N-(2-hydroxy-3-sulfopropyl) -3-
Methoxyaniline, sodium salt, dihydrate, i.e. N- ethyl-N-(2- hydroxyl -3- sulfopropyl) -3- methoxybenzene
Amine, sodium salt, dihydrate；Quinoneimine is quinine imines.
Fig. 4 is the effect picture of present invention detection protein and cholesterol.
Meaning marked in the figure: 1- protein detection layer A, 2- cholesterol detection layer B, 3- ultraviolet-visible spectrophotometer,
4- sample, 5- light beam, 6- hanging piece one, 7- hanging piece two.
Specific embodiment
The invention will be further described below in conjunction with the accompanying drawings.Following embodiment is only used for clearly illustrating the present invention
Technical solution, and not intended to limit the protection scope of the present invention.
Explanation of nouns: the sulfuric acid solution of 5w% refers to that mass ratio is 5% sulfuric acid solution；Rpm representative rev/min；MM is represented
Mmol/L, i.e., mM every liter；U is unit of enzyme activity, i.e. the enzyme amount that 1 μm of ol substrate can be converted in 1min is known as 1 enzyme unit
(U)。
The present invention proposes transparent paper base UV-Visible absorption measurement device, including the inspection of protein detection layer A1, cholesterol
Layer B2, sample 4, ultraviolet-visible spectrophotometer 3 are surveyed, the protein detection layer A is set respectively with the cholesterol detection layer B
It is placed in the output end of the ultraviolet-visible spectrophotometer, the sample is added into the protein detection layer A and institute respectively
It is stacked after stating cholesterol detection layer B drying, the sample is then arrived by the ultraviolet-visible spectrophotometer outgoing beam 5
It is detected while upper realization is to protein and cholesterol.
As a kind of preferred embodiment, hanging piece 1 is set on protein detection layer A as needed, is examined in cholesterol
It surveys and hanging piece 27 is set on layer 2, for adjusting light beam 5 at a distance from protein detection layer A, cholesterol detection layer 2 and inclination angle
Degree.
The present invention also proposes the paper base analysis method of transparent paper base UV-Visible absorption measurement device, including walks as follows
It is rapid:
Step SS1: transparent paper base is prepared；
Step SS2: testing device for transparent paper is prepared；
Step SS3: using UV-Visible absorption method measurement protein and cholesterol.
As a kind of preferred embodiment, the step SS1 is specifically included:
Step SS11: cellulose dissolution liquid is prepared；
Step SS12: cellulose solution is prepared；
Step SS13: cellulose solution prepared by the step SS12 is poured on glass plate, is formed one layer uniform thin
Then film places it in the sulfuric acid solution of 5w% and is formed by curing gel, thoroughly impregnated using distilled water, then solid using adhesive tape
It is fixed, transparent paper base is made after drying at room temperature.
As a kind of preferred embodiment, the step SS11 is specifically included: by sodium hydroxide, urea and water according to quality
Than the mass ratio wiring solution-forming for 7:12:81, it is put into spare in -12 DEG C of refrigerator.
As a kind of preferred embodiment, the step SS12 is specifically included: the step SS11 institute is added in cotton linter
In the cellulose dissolution liquor of preparation, using mechanical stirring, mixing time is 15 min, 3000 rpm of mixing speed, later
It is under 8000 rpm mixing speeds that solution centrifugation bubble removing is spare.
As a kind of preferred embodiment, the testing device for transparent paper for preparing in the step SS2 is specifically included:
Step SS21: with mass ratio being that 10:1 is mixed by PDMS and silicone elastomer curing agent, and room temperature vacuumizes half an hour and removes
Go bubble spare；
Step SS22: solution described in step SS21 is printed on the transparent paper base by layout using screen process press,
Using vacuum oven, toasted 12 hours at 80 DEG C, to be formed by curing transparent paper base analytical equipment, the final Kong Zhi of detection zone
Diameter is 5 mm, and normal temperature storage is spare.
As a kind of preferred embodiment, the step SS3 is specifically included:
Step SS31: the preparation of protein detection solution；
Step SS32: the preparation of cholesterol detection solution；
Step SS33: it is detected using ultraviolet-visible spectrophotometer.
As a kind of preferred embodiment, the step SS31 is specifically included: the pH 125 mM citrates for being 1.8 are delayed
The tetrabromophenol blue TBPB solution of fliud flushing and 9 mM dissolved with 95% ethyl alcohol and 5% water in the ratio of volume ratio 10:1 to mix
It closes, 4 DEG C store for future use, as shown in Figure 2.
As a kind of preferred embodiment, the step SS32 is specifically included: being 14.1 U's by enzymatic activity unit
Cholesterol oxidase CO that cholesterol esterase, concentration are 10 mg/mL, the 4-AA 4-APP that concentration is 10 mM,
N- ethyl-N-3- aminoanisole the sodium salt two that the horseradish peroxidase HRP and concentration that concentration is 1 mg/mL are 1mM is hydrated
Object ADOS is put into 4 DEG C and stores for future use, as shown in Figure 3 with volume ratio 1:1:1:1:1 mixing.
As a kind of preferred embodiment, the step SS33 is specifically included: by the protein detection of the step SS31
The cholesterol detection solution of solution and the step SS32 are added separately to protein detection layer A and cholesterol detection layer B, later
It is separately added into sample, air drying stacks, and uses the absorption light of ultraviolet-visible spectrophotometer detection protein and cholesterol
Spectrum, test results are shown in figure 4.
Advantageous effects of the invention: first, transparent paper base analytical equipment of the invention and paper base analysis method can
To be detected while realization to protein and cholesterol, and all have to the detection of protein and cholesterol highly sensitive and good
Good selectivity, detection limit are respectively 0.09 mM and 0.075 μM；Second, the present invention is a kind of degradable using cellophane paper
Ideal material, replace the materials such as traditional plastics, glass to alleviate environmental pressure with it, it is cheap, environmental protection, post-processing letter
It is single.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the technical principles of the invention, several improvement and deformations can also be made, these improvement and deformations
Also it should be regarded as protection scope of the present invention.
Claims (10)
1. transparent paper base UV-Visible absorption measurement device, which is characterized in that including protein detection layer A, cholesterol detection
Layer B, sample, ultraviolet-visible spectrophotometer, the protein detection layer A and the cholesterol detection layer B are respectively arranged at institute
The output end of ultraviolet-visible spectrophotometer is stated, the sample is added into the protein detection layer A respectively and the gallbladder is solid
It is stacked after alcohol detection layers B is dry, then passes through realization pair on the ultraviolet-visible spectrophotometer outgoing beam to the sample
It is detected while protein and cholesterol.
2. based on the paper base analysis method of transparent paper base UV-Visible absorption measurement device described in claim 1, feature
It is, includes the following steps:
Step SS1: transparent paper base is prepared；
Step SS2: testing device for transparent paper is prepared；
Step SS3: using UV-Visible absorption method measurement protein and cholesterol.
3. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 2, feature
It is, the step SS1 is specifically included:
Step SS11: cellulose dissolution liquid is prepared；
Step SS12: cellulose solution is prepared；
Step SS13: cellulose solution prepared by the step SS12 is poured on glass plate, is formed one layer uniform thin
Then film places it in the sulfuric acid solution of 5w% and is formed by curing gel, thoroughly impregnated using distilled water, then solid using adhesive tape
It is fixed, transparent paper base is made after drying at room temperature.
4. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 3, feature
It is, the step SS11 is specifically included: sodium hydroxide, urea and water is matched according to the mass ratio that mass ratio is 7:12:81
At solution, it is put into spare in -12 DEG C of refrigerator.
5. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 3, feature
It is, the step SS12 is specifically included: cotton linter is added in cellulose dissolution liquor prepared by the step SS11,
Using mechanical stirring, mixing time is 15 min, 3000 rpm of mixing speed, later under 8000 rpm mixing speeds by solution from
Heart bubble removing is spare.
6. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 2, feature
It is, the testing device for transparent paper for preparing in the step SS2 specifically includes:
Step SS21: with mass ratio being that 10:1 is mixed by PDMS and silicone elastomer curing agent, and room temperature vacuumizes half an hour and removes
Go bubble spare；
Step SS22: solution described in step SS21 is printed on the transparent paper base by layout using screen process press,
Using vacuum oven, toasted 12 hours at 80 DEG C, to be formed by curing transparent paper base analytical equipment, the final Kong Zhi of detection zone
Diameter is 5 mm, and normal temperature storage is spare.
7. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 2, feature
It is, the step SS3 is specifically included:
Step SS31: the preparation of protein detection solution；
Step SS32: the preparation of cholesterol detection solution；
Step SS33: it is detected using ultraviolet-visible spectrophotometer.
8. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 7, feature
Be, the step SS31 is specifically included: by pH be 1.8 125 mM citrate buffers with 95% ethyl alcohol and 5% water
To mix in the ratio of volume ratio 10:1,4 DEG C store for future use the tetrabromophenol blue TBPB solution of 9 mM of dissolution.
9. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 7, feature
It is, the step SS32 is specifically included: is 10 mg/mL by cholesterol esterase that enzymatic activity unit is 14.1 U, concentration
Cholesterol oxidase CO, the 4-AA 4-APP that concentration is 10 mM, the horseradish peroxidating that concentration is 1 mg/mL
Object enzyme HRP and concentration are the N- ethyl-N-3- aminoanisole sodium salt dihydrate ADOS of 1mM mixed with volume ratio 1:1:1:1:1
It closes, is put into 4 DEG C and stores for future use.
10. the paper base analysis method of transparent paper base UV-Visible absorption measurement device according to claim 7, special
Sign is that the step SS33 is specifically included: by the gallbladder of the protein detection solution of the step SS31 and the step SS32
Sterol detection solution is added separately to protein detection layer A and cholesterol detection layer B, is separately added into sample later, air drying,
It stacks, uses the absorption spectrum of ultraviolet-visible spectrophotometer detection protein and cholesterol.
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