CN114184584A - Enzyme-determination-based micelle liquid and method for evaluating antioxidant activity of marine natural product - Google Patents
Enzyme-determination-based micelle liquid and method for evaluating antioxidant activity of marine natural product Download PDFInfo
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- 239000000693 micelle Substances 0.000 title claims abstract description 50
- 229930014626 natural product Natural products 0.000 title claims abstract description 37
- 239000007788 liquid Substances 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 24
- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 20
- 239000011521 glass Substances 0.000 claims abstract description 125
- 239000000758 substrate Substances 0.000 claims abstract description 104
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- POKOASTYJWUQJG-UHFFFAOYSA-M 1-butylpyridin-1-ium;chloride Chemical compound [Cl-].CCCC[N+]1=CC=CC=C1 POKOASTYJWUQJG-UHFFFAOYSA-M 0.000 claims abstract description 39
- 102000004190 Enzymes Human genes 0.000 claims abstract description 30
- 108090000790 Enzymes Proteins 0.000 claims abstract description 30
- 238000001514 detection method Methods 0.000 claims abstract description 26
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000001768 carboxy methyl cellulose Substances 0.000 claims abstract description 14
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 claims abstract description 14
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 claims abstract description 14
- 238000001952 enzyme assay Methods 0.000 claims abstract description 12
- 239000000084 colloidal system Substances 0.000 claims abstract description 10
- 238000002156 mixing Methods 0.000 claims abstract description 10
- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229920000604 Polyethylene Glycol 200 Polymers 0.000 claims abstract description 6
- 159000000011 group IA salts Chemical class 0.000 claims abstract description 6
- 229940079877 pyrogallol Drugs 0.000 claims abstract description 4
- 238000003756 stirring Methods 0.000 claims abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 70
- 239000000243 solution Substances 0.000 claims description 52
- 239000011248 coating agent Substances 0.000 claims description 35
- 238000000576 coating method Methods 0.000 claims description 35
- 239000012153 distilled water Substances 0.000 claims description 35
- 239000002245 particle Substances 0.000 claims description 22
- 238000005406 washing Methods 0.000 claims description 19
- 238000007824 enzymatic assay Methods 0.000 claims description 5
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 4
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 claims description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 2
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 2
- 235000011181 potassium carbonates Nutrition 0.000 claims description 2
- 239000001632 sodium acetate Substances 0.000 claims description 2
- 235000017281 sodium acetate Nutrition 0.000 claims description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 2
- 235000017550 sodium carbonate Nutrition 0.000 claims description 2
- 235000010265 sodium sulphite Nutrition 0.000 claims description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims 1
- 238000007664 blowing Methods 0.000 claims 1
- 238000012360 testing method Methods 0.000 description 9
- 238000005259 measurement Methods 0.000 description 8
- 229930182970 xyloketal Natural products 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000007760 free radical scavenging Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000002000 scavenging effect Effects 0.000 description 2
- ZKZYJAUOKOKEOC-UHFFFAOYSA-N 2H-chromene furan Chemical class O1CC=CC2=C1C=CC=C2.O2C=CC=C2 ZKZYJAUOKOKEOC-UHFFFAOYSA-N 0.000 description 1
- 206010024769 Local reaction Diseases 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
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- 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
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
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- 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)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The application discloses enzyme assay micelle liquid and a method for evaluating the antioxidant activity of marine natural products, wherein the enzyme assay micelle liquid comprises the following steps: dissolving sodium carboxymethylcellulose in water to form 1% sodium carboxymethylcellulose hydrosol, adding 3-5g of PEG-200 into 100mol of the quantitative 1% sodium carboxymethylcellulose hydrosol, stirring to form colloid solution, treating pyrogallol with alkaline salt water solution under a light shading condition at a constant temperature to obtain a determination solution, and mixing the determination solution and the colloid solution according to a molar ratio of 6: 10: 0.5: 1 mixing to form a micelle solution for enzyme assay. The method obtains the enzyme detection micelle liquid, uniformly coats the enzyme detection micelle liquid on the substrate, uniformly coats the natural product solution to be detected on the glass sheet, then presses the glass sheet and the substrate, obtains a light-emitting result by using the BPCL tester, and analyzes the light-emitting result to obtain the antioxidant activity detection. The method is simple, and the conditions required by the determination are low.
Description
Technical Field
The application relates to an enzyme determination micelle testing solution and a method for evaluating the antioxidant activity of a marine natural product based on the enzyme determination micelle testing solution, belonging to the technical field of natural product determination.
Background
In the literature of the current research, Xyloketals compounds exist in marine natural products, and the Xyloketals compounds are benzopyran furan compounds; there are 9 similar structures, and specific reference is made to the following figure, which schematically depicts 3 species of Xyloketals, which have good activity and can be used to synthesize pharmaceutically acceptable carriers. Therefore, the Xyloketals compounds have extensive research at home and abroad.
The most accepted method for measuring the antioxidant activity of natural products is free radical scavenging, wherein the free radical scavenging comprises DPPH and ABTS scavenging, hydroxyl radical scavenging and superoxide anion scavenging, the measurement requirements of the methods are high, and factors needing to be considered are many, including the colloidal property of a system, the oxidation conditions of antioxidants in different phases, different steps and the like all cause local reactions. Therefore, the measurement results have great variability.
Disclosure of Invention
According to the application, an enzyme assay test micelle liquid and a marine natural product antioxidant activity evaluation method are provided.
The technical scheme of the application is as follows:
an enzymatic assay micelle solution comprising:
dissolving sodium carboxymethylcellulose in water to form 1% sodium carboxymethylcellulose hydrosol,
adding 3-5g of PEG-200 into 100mol of quantitative 1% sodium carboxymethylcellulose hydrosol, stirring to form colloid solution,
treating pyrogallol with an alkaline salt water solution under a shading condition at a constant temperature to obtain a determination solution, and mixing the determination solution and a colloid solution according to a molar ratio of 6: 10: 0.5: 1 mixing to form a micelle solution for enzyme assay.
In the above, the measurement solution and the solution for colloid use are mixed in a molar ratio of 10: 1 mixing to form a micelle solution for enzyme assay.
In the above, the temperature of the constant temperature condition is 20 to 25 ℃.
In the above, the alkaline salt comprises one of sodium carbonate, potassium carbonate, sodium sulfite and sodium acetate in an aqueous solution.
In the above, the pH of the alkaline aqueous salt solution is 8.5 to 10.5.
The method for evaluating the antioxidant activity of the marine natural product comprises the enzyme detection micelle liquid, and is used for detecting the antioxidant activity of the marine natural product based on the enzyme detection micelle liquid;
the method comprises the following steps:
step 1: uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 300-450 nm;
step 2: uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a vessel to be treated for 10-20min by a water bath method;
and step 3: and obtaining a light emitting result by using a BPCL tester.
The substrate is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing.
In the step 2), the water bath method adopts the temperature of 35-45 ℃.
In step 2), the vessel is a glass vessel.
In the step 2), the glass sheet is firstly cleaned by ethanol, and then is washed by a large amount of distilled water and dried after washing.
In the step 3), the wavelength range of the BPCL tester is selected from 180-500 nm; the temperature is 15-40 deg.C, and one group of data is read at intervals of 2-5 seconds for 3-5 min.
The beneficial effects that this application can produce include:
the method obtains the enzyme detection micelle liquid, uniformly coats the enzyme detection micelle liquid on the substrate, uniformly coats the natural product solution to be detected on the glass sheet, then presses the glass sheet and the substrate, obtains a light-emitting result by using the BPCL tester, and analyzes the light-emitting result to obtain the antioxidant activity detection. The method is simple, and the conditions required by the determination are low.
In the process, no other reagent is needed to be added, no pollutant is introduced, no complex post-treatment is needed, and the measurement result can be obtained by combining the two through a BPCL tester to obtain a luminescence result.
Drawings
FIG. 1 shows the results of the measurement of the average particle size of the test micelle solution of the present invention at different wavelength ranges and different enzymes;
FIGS. 2 to 4 are measurement maps of antioxidant activity of Xyloketals compounds with average particle size of 400nm in enzyme measurement test micelle liquid.
Detailed Description
The present application will be described in detail with reference to examples, but the present application is not limited to these examples.
An enzymatic test micelle solution comprising
Dissolving sodium carboxymethylcellulose in water to form 1% sodium carboxymethylcellulose hydrosol,
adding 3-5g of PEG-200 into 100mol of quantitative 1% sodium carboxymethylcellulose hydrosol, stirring to form colloid solution,
treating pyrogallol with an alkaline salt water solution under a shading condition at a constant temperature to obtain a determination solution, and mixing the determination solution and a colloid solution according to a molar ratio of 6: 10: 0.5: 1 mixing to form a micelle solution for enzyme assay.
In the above, the measurement solution and the solution for colloid use are mixed in a molar ratio of 10: 1 mixing to form a micelle solution for enzyme assay.
In the above, sodium carboxymethylcellulose is dissolved in water as the main part of the sol, PEG-200 is used as the regulator of the degree of adhesion, and the sodium carboxymethylcellulose and PEG-200 do not affect the determination of the antioxidant activity of the enzyme.
Example 1:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 300 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 35 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 180 nm; the temperature was 15 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 2:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 300 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 35 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 180 nm; the temperature was 15 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 3:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 300 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 35 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 180 nm; the temperature was 15 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 4:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 330 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 38 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 240 nm; the temperature was 25 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 5:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 330 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 38 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 240 nm; the temperature was 25 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 6:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 330 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 38 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 240 nm; the temperature was 25 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 7:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 360 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 38 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 240 nm; the temperature was 25 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 8:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 360 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 38 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 240 nm; the temperature was 25 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 9:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 360 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 10min at 38 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 240 nm; the temperature was 25 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 10:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 420 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 15min at 40 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 320 nm; the temperature was 30 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 11:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 420 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 15min at 40 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 320 nm; the temperature was 30 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 12:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 420 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 15min at 40 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 320 nm; the temperature was 30 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 13:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 500 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 20min at 40 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 400 nm; the temperature was 30 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 14:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 500 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 20min at 40 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 400 nm; the temperature was 30 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Example 15:
uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 500 nm; uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a flat-bottomed glassware to treat for 20min at 40 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 400 nm; the temperature was 30 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the substrate is first cleaned with ethanol, and then rinsed with a large amount of distilled water and dried after rinsing. The glass sheet is firstly cleaned by ethanol, and then washed by a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Comparative example 16:
uniformly coating the natural product solution to be measured on a glass sheet, and then placing the glass sheet in a flat-bottomed glassware to treat for 20min at 40 ℃; and obtaining a light emitting result by using a BPCL tester. The wavelength range of the BPCL tester is 400 nm; the temperature was 30 ℃ and one set of data was read every 2 seconds for 3 min. In this embodiment, the glass sheet is firstly cleaned with ethanol, and then washed with a large amount of distilled water and dried after washing. The substrate and the glass sheet are ensured not to be polluted, a black substrate is basically adopted, and the glass sheet is a transparent glass sheet.
Referring to fig. 1, examples 1 to 15 and comparative example 16, the average particle size of the enzyme assay micelle liquid significantly varied at different wavelengths, and had a significant absorption peak at a wavelength of about 200nm and a secondary absorption peak at a wavelength of about 270 nm. The average particle size of the optimal enzyme detection micelle liquid is between 300 and 400nm, and other temperatures such as temperature and pH value are selected, and the effect of the temperature selected in the test on the result is not obvious.
Referring to fig. 2 and 4, fig. 2-4 are graphs showing the antioxidant activity of three Xyloketals compounds mentioned in the background art when the average particle size of the test micelle liquid is 400nm and the test micelle liquid is combined with the three Xyloketals compounds and is not measured by using enzyme. The results of fig. 2 and 4 also validate the above assay.
Although the present application has been described with reference to a few embodiments, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the application as defined by the appended claims.
Claims (10)
1. An enzymatic assay micelle solution, comprising:
dissolving sodium carboxymethylcellulose in water to form 1% sodium carboxymethylcellulose hydrosol,
adding 3-5g of PEG-200 into 100mol of quantitative 1% sodium carboxymethylcellulose hydrosol, stirring to form colloid solution,
treating pyrogallol with an alkaline salt water solution under a shading condition at a constant temperature to obtain a determination solution, and mixing the determination solution and a colloid solution according to a molar ratio of 6: 10: 0.5: 1 mixing to form a micelle solution for enzyme assay.
2. The enzymatic assay micelle solution according to claim 1, wherein the temperature under the constant temperature condition is 20 to 25 ℃.
3. The enzymatic assay micelle solution of claim 1, wherein the alkaline salt comprises one of sodium carbonate, potassium carbonate, sodium sulfite and sodium acetate in an aqueous solution.
4. The enzymatic assay micelle solution of claim 1, wherein the PH of the alkaline saline solution is 8.5 to 10.5.
5. The method for evaluating the antioxidant activity of a marine natural product, which comprises the enzyme assay micelle liquid according to any one of claims 1 to 5, and is characterized in that the antioxidant activity of the marine natural product is determined based on the enzyme assay micelle liquid;
the method comprises the following steps:
step 1: uniformly coating the enzyme detection micelle liquid on a substrate, wherein the average particle size is 300-450 nm;
step 2: uniformly coating a natural product solution to be measured on a glass sheet, then pressing the glass sheet and a substrate together, and then placing the glass sheet and the substrate in a vessel to be treated for 10-20min by a water bath method;
and step 3: and obtaining a light emitting result by using a BPCL tester.
6. The method for evaluating antioxidant activity of marine natural products according to claim 5, wherein the substrate is firstly cleaned by ethanol, and then washed by a large amount of distilled water and then dried by blowing after washing.
7. The method for evaluating antioxidant activity of marine natural products according to claim 1, wherein in the step 2), the water bath method is adopted at a temperature of 35-45 ℃.
8. The method for evaluating antioxidant activity of marine natural products according to claim 1, wherein in step 2), the vessel is a glass vessel.
9. The method for evaluating the antioxidant activity of marine natural products as claimed in claim 1, wherein in the step 2), the glass sheet is firstly cleaned by ethanol, and then is washed by a large amount of distilled water after being cleaned, and is dried after being washed.
10. The method for evaluating the antioxidant activity of marine natural products as claimed in claim 1, wherein in the step 3), the wavelength range of the BPCL tester is selected from 180-500 nm; the temperature is 15-40 deg.C, and one group of data is read at intervals of 2-5 seconds for 3-5 min.
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