CN110923128A - Soil catalase activity determination kit and detection method thereof - Google Patents
Soil catalase activity determination kit and detection method thereof Download PDFInfo
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- CN110923128A CN110923128A CN201910339881.2A CN201910339881A CN110923128A CN 110923128 A CN110923128 A CN 110923128A CN 201910339881 A CN201910339881 A CN 201910339881A CN 110923128 A CN110923128 A CN 110923128A
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- 239000002689 soil Substances 0.000 title claims abstract description 71
- 238000001514 detection method Methods 0.000 title claims abstract description 35
- 102000016938 Catalase Human genes 0.000 title claims abstract description 34
- 108010053835 Catalase Proteins 0.000 title claims abstract description 34
- 230000000694 effects Effects 0.000 title claims abstract description 32
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 36
- 238000003149 assay kit Methods 0.000 claims abstract description 12
- 102000004190 Enzymes Human genes 0.000 claims abstract description 11
- 108090000790 Enzymes Proteins 0.000 claims abstract description 11
- 238000006243 chemical reaction Methods 0.000 claims abstract description 6
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 35
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 239000012153 distilled water Substances 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 11
- 239000003795 chemical substances by application Substances 0.000 claims description 9
- 238000010521 absorption reaction Methods 0.000 claims description 7
- 238000000227 grinding Methods 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 230000015556 catabolic process Effects 0.000 claims description 3
- 238000006731 degradation reaction Methods 0.000 claims description 3
- 239000006261 foam material Substances 0.000 claims description 3
- 238000005259 measurement Methods 0.000 claims description 3
- 238000005070 sampling Methods 0.000 claims description 3
- 238000007873 sieving Methods 0.000 claims description 3
- 238000002835 absorbance Methods 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 238000002474 experimental method Methods 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 7
- 238000000034 method Methods 0.000 description 3
- 239000010453 quartz Substances 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 210000002824 peroxisome Anatomy 0.000 description 2
- 229910001868 water Inorganic materials 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
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- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention relates to the technical field of activity determination of soil catalase, and discloses a soil catalase activity determination kit and a detection method thereof. This soil catalase activity assay kit and detection method thereof, comprises a box bod, the inside left end movable mounting of box body has the cell, the inside fixed mounting of cell has a plurality of colour comparison tube, and the color comparison recess has all been seted up to the inside of every colour comparison tube, on the inner wall of cell and be located fixed mounting between the colour comparison tube have a plurality of baffle, a plurality of reagent groove has been seted up to the inside of box body and the right side that is located the cell. The soil catalase activity determination kit and the detection method thereof provide an intuitive and rapid detection method, the enzyme activity of a soil sample to be detected can be intuitively judged according to the color depth of an experimental result, and a terminator is adopted to uniformly stop the experimental reaction, so that the experimental operation difficulty is reduced, and the accuracy and the repeatability of detection data are improved.
Description
Technical Field
The invention relates to the technical field of activity determination of soil catalase, in particular to a soil catalase activity determination kit and a detection method thereof.
Background
Catalase (CAT), an enzyme that catalyzes the decomposition of hydrogen peroxide into oxygen and water, is present in the peroxide body of cells. Catalase is a marker enzyme for peroxisomes and accounts for about 40% of the total amount of peroxisome enzymes. Soil catalase can enzymatically decompose hydrogen peroxide to generate water and oxygen, and hydrogen peroxide itself has strong absorption at the ultraviolet wavelength of 240 nm. After a certain amount of hydrogen peroxide is added to act on soil for a period of time, the amount of the hydrogen peroxide remaining is measured by an ultraviolet spectrophotometry, the difference between the added amount and the remaining amount is the amount of the hydrogen peroxide reacting with enzyme, and the hydrogen peroxide consumed in a certain time is used for representing the activity of soil catalase.
Soil catalase is detected by the conventional method of titrating the residual hydrogen peroxide with potassium permanganate in the presence of sulfuric acid to determine the enzyme activity. The method has extremely high requirements on the detected acidic environment, and the key point of titration is difficult to grasp. The common method on the market at present is to measure the change of the absorbance value of the substrate hydrogen peroxide at 240nm by using an ultraviolet spectrophotometer. However, hydrogen peroxide is decomposed to generate bubbles easily, which affects the reading of a detection instrument, causes the problems of unstable reading, difficult reproduction of an experiment and the like, and in addition, components such as protein and the like contained in a detected sample have stronger absorption near 240nm, which can cause serious interference to the measurement. In addition, expensive quartz cuvettes and ultraviolet spectrophotometers with ultraviolet band detection are used in the operation process, so that the detection cost is overhigh.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a soil catalase activity determination kit and a detection method thereof, which have the advantages of convenience in operation, good implementation repeatability, high data preparation and the like, and solve the problems of high cost, low data accuracy and difficulty in experiment reproduction of the existing soil catalase detection kit.
The technical scheme adopted by the invention for solving the technical problems is as follows: a soil catalase activity determination kit and a detection method thereof comprise a box body, wherein a cuvette is movably arranged at the left end inside the box body, a plurality of colorimetric tubes are fixedly arranged inside the cuvette, a colorimetric groove is arranged inside each colorimetric tube, a plurality of partition plates are fixedly arranged on the inner wall of the cuvette and between the colorimetric tubes, a plurality of reagent grooves are arranged in the box body and at the right side of the cuvette, reagent bottles are movably arranged in each reagent groove, a dropper groove is arranged at the right side of the reagent groove, a dropper is movably arranged in the dropper groove, the utility model discloses a test tube box, including box body, equal fixed mounting has the connecting block on the outer wall of the box body left and right sides, and a plurality of test tube groove has all been seted up to the inside of every connecting block, and the equal movable mounting in inside of every test tube groove has the EP pipe, the top movable mounting of box body has the lid.
Further, all be provided with on the top of cell left and right sides outer wall and carry and take the handle, and the inner wall swing joint of cell and box body.
Furthermore, the number of the reagent tanks and the number of the reagent bottles are five, and the five reagent bottles are respectively filled with a soil sample, distilled water, a hydrogen peroxide solution, a hydrochloric acid solution and a color developing agent.
Furthermore, the left side and the right side in the middle of the burette groove are both provided with rectangular grooves.
Furthermore, the inner walls of the reagent tank, the dropper tank and the test tube tank are all made of foam materials.
The detection method comprises the following steps:
1) and preparing a soil sample:
and (3) taking a fresh soil sample to be air-dried or an oven with 37 degrees to be air-dried, firstly carrying out coarse grinding, sieving by a 40-mesh sieve, then grinding by a 60-mesh sieve again, and placing the sieved soil sample in a reagent bottle for experimental detection.
2) Sampling and detecting a soil sample:
s1, taking three 1.5mL EP tubes, adding 0.1g of sieved soil samples into two of the EP tubes, opening the box cover, taking out a reagent bottle filled with distilled water, adding 800 mu L of distilled water into the three EP tubes by using a dropper, and culturing for 10 minutes at 25-37 ℃ in a shaking incubator (30 rpm/min).
S2, adding 100 mu L (300- & lt 400 & gtmM) of hydrogen peroxide solution into one of the soil tube and the soil-free tube, and adding equal volume of 100 mu L of distilled water into the other soil tube.
S3, culturing at 25-37 deg.C for 15min (precisely controlling time), and adding 5M hydrochloric acid solution to terminate the reaction.
S4, putting the three EP tubes into a centrifuge, centrifuging at 12000rpm and 4 ℃ for 10min, and taking out the supernatant for later use.
S5, dripping 25 mu L of supernatant obtained in the previous step and 975 mu L of color developing agent into a colorimetric groove in a cuvette, standing for 5min at room temperature (25 ℃), enabling the color developing agent to react with residual hydrogen peroxide specifically to generate a red (powder) color complex, wherein the color substance has a characteristic absorption peak at 510nm, reading the absorption values A of a soil-free tube, a determination tube and a matrix-free tube, and △ A is A soil-free tube- (A determination tube-A matrix-free tube).
3) And calculating a result:
the kit also provides a standard curve, is convenient for a client to directly bring into a calculation formula for calculation,
and S1, wherein the standard curve equation is that y is 0.1677x-0.0005, x is the molar mass of the standard substance, and y is △ A.
S2, soil catalase (S-CAT) Unit definition: one unit of enzyme activity was defined as the 1. mu. mol H2O2 degradation catalyzed by each gram of air-dried soil sample per hour.
S-CAT (μmol/h/g dry soil) [ (△ a +0.0005) ÷ 0.1677 ]/(W ÷ T)/(23.9 × (△ a + 0.0005)/(W ÷ 23.9 × (△ a +0.0005) /)
The invention has the beneficial effects that:
1. the soil catalase activity determination kit and the detection method thereof provide an intuitive and rapid detection method, the enzyme activity of a soil sample to be detected can be intuitively judged according to the color depth of an experimental result, and a terminator is adopted to uniformly stop the experimental reaction, so that the experimental operation difficulty is reduced, and the accuracy and the repeatability of detection data are improved.
2. According to the soil catalase activity determination kit and the detection method thereof, the required instruments and materials are cheap, an expensive ultraviolet spectrophotometer with an ultraviolet band detection function and a matched quartz cuvette are not needed, the experiment cost is reduced, the experiment is easy to realize, the experiment can be completed in a common laboratory, and a simple, accurate and economic experiment means is provided for the majority of scientific research workers.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a side view of the reagent tank of the present invention;
fig. 3 is a front view at the cassette of the present invention.
Description of reference numerals: 1 box body, 2 cuvettes, 3 colour comparison tube, 4 colorimetric grooves, 5 baffles, 6 reagent grooves, 7 reagent bottles, 8 dropper grooves, 9 droppers, 10 connecting blocks, 11 test tube grooves, 12EP tubes, 13 box covers.
Detailed Description
The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention.
Referring to fig. 1-3, a soil catalase activity assay kit and a detection method thereof, comprising a box body 1, wherein a cuvette 2 is movably mounted at the left end inside the box body 1, the top ends of the outer walls at the left side and the right side of the cuvette 2 are respectively provided with a lifting handle, the cuvette 2 is movably connected with the inner wall of the box body 1, and the cuvette 2 can be taken out from the inside of the box body 1 through the lifting handles, so that the cuvette 2 can be conveniently cleaned. The inside fixed mounting of cell 2 has a plurality of colour comparison tube 3, and color comparison groove 4 has all been seted up to the inside of every colour comparison tube 3, and fixed mounting has a plurality of baffle 5 on the inner wall of cell 2 and between being located colour comparison tube 3, and several regions are cut apart into with cell 2 to baffle 5 to in distinguish and observe. The inside of box body 1 just is located the right side of cell 2 and has seted up a plurality of reagent groove 6, and the equal movable mounting in inside of every reagent groove 6 has reagent bottle 7, and the quantity of reagent groove 6 and reagent bottle 7 is five, and is equipped with soil sample, distilled water, hydrogen peroxide solution, hydrochloric acid solution, colour-developing agent in five reagent bottles 7 respectively. A burette groove 8 is formed in the right side of the reagent groove 6, a burette 9 is movably mounted inside the burette groove 8, rectangular grooves are formed in the left side and the right side of the middle of the burette groove 8, and the rectangular grooves are arranged so that the burette 9 can be taken out conveniently. Equal fixed mounting has connecting block 10 on the outer wall of the 1 left and right sides of box body, and a plurality of test tube groove 11 has all been seted up to the inside of every connecting block 10, and the equal movable mounting in inside of every test tube groove 11 has EP pipe 12, and the inner wall of reagent groove 6, burette groove 8 and test tube groove 11 is foam material and makes, makes reagent bottle 7, burette 9, EP pipe 12 place steadily, and can not damaged by the extrusion. The top of the box body 1 is movably provided with a box cover 13.
The detection method comprises the following steps:
1) and preparing a soil sample:
and (3) taking a fresh soil sample to be air-dried or air-dried in a 37-degree oven, coarsely grinding the soil sample, sieving the soil sample by a 40-mesh sieve, grinding the soil sample by a 60-mesh sieve again, and placing the sieved soil sample in a reagent bottle 7 for detection of an experiment.
2) Sampling and detecting a soil sample:
s1, three 1.5mL EP tubes 12 are taken, 0.1g of sieved soil sample is added to each of two of the EP tubes 12, a box cover 13 is opened, a reagent bottle 7 filled with distilled water is taken out, 800 mu L of distilled water is added to each of the three EP tubes 12 by using a dropper 9, and the mixture is cultured for 10 minutes at a temperature of 25-37 ℃ in a shaking incubator (30 rpm/min).
S2, adding 100 mu L (300- & lt 400 & gtmM) of hydrogen peroxide solution into one of the soil tube and the soil-free tube, and adding equal volume of 100 mu L of distilled water into the other soil tube.
S3, culturing at 25-37 deg.C for 15min (precisely controlling time), and adding 5M hydrochloric acid solution to terminate the reaction.
S4, putting the three EP tubes 12 into a centrifuge, centrifuging at 12000rpm and 4 ℃ for 10min, and taking out the supernatant for later use.
S5, dripping 25 μ L of supernatant and 975 μ L of color developing agent in the above steps into the colorimetric groove 4 in the cuvette 2, standing at room temperature (25 ℃) for 5min, wherein the color developing agent reacts specifically with the residual hydrogen peroxide to generate a red (pink) color complex, and the colored substance has a characteristic absorption peak at 510nm, reading the absorption values A of the soil-free tube, the determination tube and the matrix-free tube respectively, and △ A is A soil-free tube- (A determination tube-A matrix-free tube).
3) And calculating a result:
the kit also provides a standard curve, is convenient for a client to directly bring into a calculation formula for calculation,
and S1, wherein the standard curve equation is that y is 0.1677x-0.0005, x is the molar mass of the standard substance, and y is △ A.
S2, soil catalase (S-CAT) Unit definition: one unit of enzyme activity was defined as the 1. mu. mol H2O2 degradation catalyzed by each gram of air-dried soil sample per hour.
S-CAT (μmol/h/g dry soil) [ (△ a +0.0005) ÷ 0.1677 ]/(W ÷ T)/(23.9 × (△ a + 0.0005)/(W ÷ 23.9 × (△ a +0.0005) /)
When the kit is used, the kit provides an intuitive and quick detection method, the enzyme activity of a soil sample to be detected can be intuitively judged according to the color depth of an experimental result, and the experimental reaction is uniformly stopped by adopting a terminator, so that the experimental operation difficulty is reduced, and the accuracy and the repeatability of detection data are improved; in addition, the required instruments and materials are cheap, an expensive ultraviolet spectrophotometer with an ultraviolet band detection function and a matched quartz cuvette are not needed, the experiment cost is reduced, the experiment is easy to realize, a common laboratory can complete the experiment, and a simple, accurate and economic experiment means is provided for vast researchers.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (6)
1. A soil catalase activity determination kit and a detection method thereof comprise a kit body (1), and are characterized in that: the utility model discloses a reagent bottle box, including box body (1), cell (2), a plurality of colorimetric tube (3), colorimetric groove (4) have all been seted up to the inside left end movable mounting of box body (1), fixed mounting has a plurality of colorimetric tube (3) in the inside of cell (2), on the inner wall of cell (2) and lie in between colorimetric tube (3) a plurality of baffle (5), the inside of box body (1) just lies in the right side of cell (2) and has seted up a plurality of reagent groove (6), the inside of each reagent groove (6) is equal movable mounting has reagent bottle (7), the right side of reagent groove (6) has seted up burette groove (8), the inside movable mounting of burette groove (8) has burette (9), all fixed mounting has connecting block (10) on the outer wall of the box body (1) left and right sides, and the inside of each connecting block (10) has all seted up a plurality of test tube groove (11), an EP pipe (12) is movably mounted in each test tube groove (11), and a box cover (13) is movably mounted at the top of the box body (1).
2. The soil catalase activity assay kit and the detection method thereof according to claim 1, wherein the soil catalase activity assay kit comprises: all be provided with on the top of cell (2) left and right sides outer wall and carry and take the handle, and the inner wall swing joint of cell (2) and box body (1).
3. The soil catalase activity assay kit and the detection method thereof according to claim 1, wherein the soil catalase activity assay kit comprises: the number of the reagent tanks (6) and the number of the reagent bottles (7) are five, and the five reagent bottles (7) are respectively filled with a soil sample, distilled water, a hydrogen peroxide solution, a hydrochloric acid solution and a color developing agent.
4. The soil catalase activity assay kit and the detection method thereof according to claim 1, wherein the soil catalase activity assay kit comprises: the left side and the right side of the middle part of the dropper groove (8) are both provided with rectangular grooves.
5. The soil catalase activity assay kit and the detection method thereof according to claim 1, wherein the soil catalase activity assay kit comprises: the inner walls of the reagent tank (6), the dropper tank (8) and the test tube tank (11) are all made of foam materials.
6. The soil catalase activity assay kit and the detection method thereof according to claim 1, wherein the soil catalase activity assay kit comprises: the detection method comprises the following steps:
1) and preparing a soil sample:
and (3) taking a fresh soil sample to be air-dried or air-dried in a 37-degree oven, coarsely grinding the soil sample, sieving the soil sample by a 40-mesh sieve, grinding the soil sample by a 60-mesh sieve again, and placing the sieved soil sample in a reagent bottle (7) for experimental detection.
2) Sampling and detecting a soil sample:
s1, three 1.5mL EP tubes (12) are taken, 0.1g of sieved soil sample is added into two of the EP tubes, a box cover (13) is opened, a reagent bottle (7) filled with distilled water is taken out, 800 mu L of distilled water is added into the three EP tubes (12) by using a dropper (9), and the mixture is cultured for 10 minutes at the temperature of 25-37 ℃ in a shaking incubator (30 rpm/min).
S2, adding 100 mu L (300- & lt 400 & gtmM) of hydrogen peroxide solution into one of the soil tube and the soil-free tube, and adding equal volume of 100 mu L of distilled water into the other soil tube.
S3, culturing at 25-37 deg.C for 15min (precisely controlling time), and adding 5M hydrochloric acid solution to terminate the reaction.
S4, putting the three EP tubes (12) into a centrifuge, centrifuging at 12000rpm and 4 ℃ for 10min, and taking out the supernatant for later use.
S5, dropping 25 μ L of the supernatant from the above steps and 975 μ L of a color developing agent into the colorimetric groove (4) of the cuvette (2), standing at room temperature (25 ℃) for 5min, wherein the color developing agent reacts specifically with the remaining hydrogen peroxide to generate a red (powder) color complex, which has a characteristic absorption peak at 510nm, and reading the absorbance values a of the soil-free tube, the measurement tube and the matrix-free tube, △ a ═ a soil-free tube- (a measurement tube-a matrix-free tube), respectively.
3) And calculating a result:
the kit also provides a standard curve, is convenient for a client to directly bring into a calculation formula for calculation,
and S1, wherein the standard curve equation is that y is 0.1677x-0.0005, x is the molar mass of the standard substance, and y is △ A.
S2, soil catalase (S-CAT) Unit definition: one unit of enzyme activity was defined as the 1. mu. mol H2O2 degradation catalyzed by each gram of air-dried soil sample per hour.
S-CAT (μmol/h/g dry soil) [ (△ a +0.0005) ÷ 0.1677 ]/(W ÷ T) × (△ a +0.0005) ÷ W).
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111505244A (en) * | 2020-05-01 | 2020-08-07 | 杜大庆 | Soil detection case for environmental protection |
| CN113777064A (en) * | 2021-09-10 | 2021-12-10 | 合肥工业大学 | Kit and method for detecting in-vitro antioxidant activity of xylaria intracellular polysaccharide |
| RU2835507C1 (en) * | 2024-03-21 | 2025-02-26 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Кабардино-Балкарский государственный университет им. Х.М. Бербекова" (КБГУ) | Device for determining catalase activity of soil |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0186476A2 (en) * | 1984-12-21 | 1986-07-02 | Hemochek Corporation | Diagnostic kit and method for assaying blood components |
| JPH095330A (en) * | 1995-04-19 | 1997-01-10 | Konica Corp | Method and kit for detecting protein |
| CN205470422U (en) * | 2016-04-08 | 2016-08-17 | 环境保护部华南环境科学研究所 | Environment soil sample detection and analysis kit |
| CN205786627U (en) * | 2016-06-29 | 2016-12-07 | 浙江中通检测科技有限公司 | A kind of total nitrogen of soil detection kit |
| CN206431119U (en) * | 2017-01-22 | 2017-08-22 | 北京普赞生物技术有限公司 | A kind of self-retaining formula gutter oil detection reagent box |
-
2019
- 2019-04-25 CN CN201910339881.2A patent/CN110923128A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0186476A2 (en) * | 1984-12-21 | 1986-07-02 | Hemochek Corporation | Diagnostic kit and method for assaying blood components |
| JPH095330A (en) * | 1995-04-19 | 1997-01-10 | Konica Corp | Method and kit for detecting protein |
| CN205470422U (en) * | 2016-04-08 | 2016-08-17 | 环境保护部华南环境科学研究所 | Environment soil sample detection and analysis kit |
| CN205786627U (en) * | 2016-06-29 | 2016-12-07 | 浙江中通检测科技有限公司 | A kind of total nitrogen of soil detection kit |
| CN206431119U (en) * | 2017-01-22 | 2017-08-22 | 北京普赞生物技术有限公司 | A kind of self-retaining formula gutter oil detection reagent box |
Non-Patent Citations (1)
| Title |
|---|
| 郑婷: "间伐对重阳木细根特性及土壤酶活性的测定及影响", 《中国农业信息》 * |
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| CN113777064A (en) * | 2021-09-10 | 2021-12-10 | 合肥工业大学 | Kit and method for detecting in-vitro antioxidant activity of xylaria intracellular polysaccharide |
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