CN117007788B - Nitric oxide determination kit and preparation method and application thereof - Google Patents

Nitric oxide determination kit and preparation method and application thereof Download PDF

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CN117007788B
CN117007788B CN202310971214.2A CN202310971214A CN117007788B CN 117007788 B CN117007788 B CN 117007788B CN 202310971214 A CN202310971214 A CN 202310971214A CN 117007788 B CN117007788 B CN 117007788B
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nitric oxide
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following components
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CN117007788A (en
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王飞
刘明霞
隗勇
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China Kemeo Shandong Medical Laboratory Co ltd
Cicc Import & Export Co ltd
Zhongtuo Medical Laboratory Co ltd
Zhongtuo Biotechnology Co ltd
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China Kemeo Shandong Medical Laboratory Co ltd
Cicc Import & Export Co ltd
Zhongtuo Medical Laboratory Co ltd
Zhongtuo Biotechnology Co ltd
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    • 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
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • 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
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—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
    • 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
    • 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
    • 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
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—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
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Abstract

The invention provides a nitric oxide determination kit, which comprises a reagent R1 and a reagent R2; reagent R1 comprises the following components: citric acid buffer solution, dopamine hydrochloride, stabilizer, anti-interference agent, surfactant and sodium azide; reagent R2 comprises the following components: 4-hydroxyethyl piperazine ethane sulfonic acid buffer solution, 3-methyl-2-benzothiazolinone hydrazone hydrochloride, a stabilizer, a reaction enhancer, a surfactant and sodium azide. The kit is a liquid kit with strong stability, high accuracy, wide linear range, high sensitivity and interference resistance.

Description

Nitric oxide determination kit and preparation method and application thereof
Technical Field
The invention relates to the technical field of biochemical detection, in particular to a nitric oxide determination kit and application.
Background
Nitric Oxide (NO) is a colorless, odorless, poorly water-soluble, toxic gas. Since nitric oxide carries free radicals, this makes its chemistry very active. When it reacts with oxygen, corrosive gas nitrogen dioxide (NO 2 ) Nitrogen dioxide may react with water to form nitric acid. NO is produced in vivo under the catalysis of Nitric Oxide Synthase (NOS)And (3) forming the finished product. NOS is composed of 1025 amino acid residues, has a molecular weight of 13300 daltons, and is widely distributed in the body, and its isoforms are of three subtypes, namely neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) expressed in a normal state and Inducible Nitric Oxide Synthase (iNOS) induced to be expressed after injury. NO has an important role in the treatment of cardiovascular diseases and many other important chronic diseases.
In cardiovascular diseases, the synthesis and release of nitric oxide changes, as does the concentration of nitric oxide in the blood. Several studies have shown that the concentration of nitric oxide in serum is closely related to the occurrence and progression of cardiovascular disease. In patients with hypertension, the concentration of nitric oxide in serum is low, and the supplementation of nitric oxide can effectively reduce blood pressure; in patients with coronary heart disease, a decrease in serum nitric oxide concentration is associated with the severity and prognosis of the coronary heart disease condition. Pulmonary hypertension is a rare but serious cardiovascular disease characterized by elevated pressure in the pulmonary arteries, resulting in increased cardiac burden. Nitric oxide plays a key role in regulating pressure within the pulmonary artery. Thus, detecting nitric oxide levels in the blood can help assess the severity of pulmonary hypertension and monitor the effect of the treatment, and the use of nitric oxide therapy can improve the symptoms and prognosis of pulmonary hypertension.
Pulmonary infections are typically caused by bacterial or viral infections. During the early stages of pulmonary infection, nitric oxide levels may rise. Therefore, by detecting the level of nitric oxide in blood or exhaled air, early detection of lung infection and timely treatment can be facilitated. At the same time, the level of nitric oxide may also be used to monitor the therapeutic effect of pulmonary infection, as well as predict the extent of recovery from infection.
Nitric oxide can regulate the function of the immune system, promote the activity of macrophages and T lymphocytes, and thereby enhance the immunocompetence of the body. When nitric oxide levels are reduced, it may affect the immune function of the body, leading to the development and progression of immune system disorders.
Nitric oxide acts as a neurotransmitter that regulates the function of the nervous system. When nitric oxide levels are reduced, neurological disorders such as parkinson's disease, stroke, etc. may result.
The determination of serum NO mainly comprises nitrate reductase, griess reagent color development method and alpha-naphthylamine diazo method. The nitrate reductase determination method is stable, has higher accuracy and precision, and is convenient to operate, but the kit is more expensive and is a limitation to clinical application. The Griess reagent color development method and the alpha-naphthylamine diazo method are relatively low in precision and accuracy.
Patent CN112051259a discloses a method for measuring serum NO, which converts the in vivo metabolism into nitrate (NO 3 - ) And Nitrite (NO) 2 - ) Sample nitrate (NO 3 - ) And Nitrite (NO) 2 - ) The sum of the concentrations can accurately represent the NO level, with nitrate (NO 3 - ) Nitrite (NO) 2 - ) And dopamine hydrochloride and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) can test the NO content. However, dopamine hydrochloride is sensitive to air and light, and the reaction is easily disturbed by other substances in serum when clinical samples are detected.
Disclosure of Invention
In order to solve the problems, the invention provides a serum nitric oxide determination kit which is a liquid kit with strong stability, high accuracy, wide linear range, high sensitivity and interference resistance.
The invention is realized by the following technical scheme:
a serum nitric oxide assay kit consists of the following components, wherein the kit contains a reagent R1 and a reagent R2;
reagent R1 comprises the following components:
reagent R2 comprises the following components:
preferably, the reagent R1 has a pH of 4.0-6.0; the pH of the reagent R2 is 7.0-8.0.
Preferably, the anti-interference agent in the reagent R1 is trans-1, 2-cyclohexanediamine tetraacetic acid (CDTA).
Preferably, the stabilizer in the reagent R1 is selected from one or two of hydroxypropyl-beta-cyclodextrin and methyl-beta-cyclodextrin.
Preferably, the surfactant in the reagent R1 is selected from one or both of tetradecyltrimethylammonium bromide and tetradecyltrimethylammonium chloride.
Preferably, the stabilizer in the reagent R2 is acid hydrolyzed casein.
Preferably, the reaction enhancer in the reagent R2 is sodium polymethacrylate.
Preferably, the surfactant in the reagent R2 is selected from one or two of polyvinylpyrrolidone k30 and nonylphenol polyoxyethylene ether NP-40.
The application of the nitric oxide determination kit is used for determining the concentration of nitric oxide for the diagnosis and treatment purposes of non-diseases.
The invention has the beneficial effects that:
1. the surfactant in the reagent R1 of the system is tetradecyltrimethylammonium bromide or tetradecyltrimethylammonium chloride, has strong emulsifying and dispersing capabilities, effectively removes the interference of blood fat and chyle, ensures that the reagent R1 system is uniformly dispersed, greatly enhances the anti-interference capability of the reagent, and avoids dopamine and serum Fe by adding trans-1, 2-cyclohexanediamine tetraacetic acid (CDTA) 3+ A complexation reaction occurs to form a purple complex.
2. The protective agent hydroxypropyl-beta-cyclodextrin is added into the reagent R1 to protect dopamine hydrochloride and improve the stability of the reagent R1.
3. According to the reagent R2, the stabilizer is added to hydrolyze casein, and the stability of the 3-methyl-2-benzothiazolinone hydrazone hydrochloride aqueous solution is improved in a synergistic way with the surfactant, so that the reagent R2 has excellent stability.
4. The invention adopts a methodology as a speed method, and sodium polymethacrylate is added as a reaction enhancer to promote Nitrate (NO) 3 - ) Nitrite (NO) 2 - ) And the reaction of dopamine hydrochloride and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH), so as to improve the analysis sensitivity of the reagent.
5. The nitric oxide determination reagent prepared by the method has the advantages of good stability, high accuracy, high sensitivity and wide linear range, and is obviously superior to the commercial products of the same type.
Drawings
FIG. 1 is a correlation curve of the reagents of example 1 and comparative example 1;
fig. 2 and 3 show the concentration changes of the nitric oxide measuring reagents provided in example 1 and comparative examples 1,2, 3, 7 and 9 for stability tests.
Detailed Description
The following describes in detail the examples of the present invention, which are implemented on the premise of the technical solution of the present invention, and detailed embodiments and specific operation procedures are given, but the scope of protection of the present invention is not limited to the following examples.
The kit provided by the invention is used for measuring nitric oxide in serum, and the test conditions are as follows:
the method comprises the following steps: a rate method; primary/secondary wavelength: 545nm; temperature: 37 ℃; correction type: linearity; the calibration method comprises the following steps: two-point calibration; the reaction direction is as follows: upward.
The specific operation is shown in table 1.
TABLE 1 procedure for nitric oxide assay reagents
Calculation results:
sample requirements:
1. serum is not hemolyzed.
2. Sample stability: the specimen can be stored stably for 3 days at 2-8 ℃ and for 2 weeks at-20 ℃. The invention is further illustrated by the following examples:
example 1
Reagent R2 comprises the following components:
the preparation method of the nitric oxide assay kit comprises the following steps:
a) Preparation of reagent R1: and (3) weighing purified water with the same volume as the preparation volume, calculating and weighing a proper amount of citric acid, dopamine hydrochloride, hydroxypropyl-beta-cyclodextrin, trans-1.2-cyclohexanediamine tetraacetic acid (CDTA), tetradecyl trimethyl ammonium bromide and sodium azide according to the preparation volume, regulating the pH value to 4.5, and uniformly stirring to obtain the reagent R1.
b) Preparation of reagent R2: and (3) measuring the purified water with the same volume as the preparation volume, calculating according to the preparation volume, weighing a proper amount of 4-hydroxyethyl piperazine ethane sulfonic acid, 3-methyl-2-benzothiazolinone hydrazone hydrochloride, acid hydrolyzed casein, sodium polymethacrylate, polyvinylpyrrolidone k30 and sodium azide, regulating the pH value to 7.2, and uniformly stirring to obtain the reagent R2.
Example 2
Reagent R2 comprises the following components:
the preparation method of the nitric oxide assay kit is the same as that of example 1.
Example 3
Reagent R2 comprises the following components:
the preparation method of the nitric oxide assay kit is the same as that of example 1.
Comparative example 1
And importing a sigma nitric oxide assay kit.
Comparative example 2
The difference from the nitric oxide assay kit of example 1 is only that the reagent R1 does not contain hydroxypropyl-beta-cyclodextrin, otherwise the same as in example 1.
Comparative example 3
The only difference from the nitric oxide assay kit of example 1 is that the reagent R1 replaces hydroxypropyl-beta-cyclodextrin with an equivalent amount of beta-cyclodextrin, otherwise identical to example 1.
Comparative example 4
The difference from the nitric oxide assay kit of example 1 is that the reagent R1 does not contain trans-1.2-cyclohexanediamine tetraacetic acid (CDTA), and the other is the same as in example 1.
Comparative example 5
The only difference from the nitric oxide assay kit of example 1 is that reagent R1 replaces trans-1, 2-cyclohexanediamine tetraacetic acid (CDTA) with an equivalent amount of ethylenediamine tetraacetic acid (EDTA), otherwise identical to example 1.
Comparative example 6
The difference from the nitric oxide assay kit of example 1 is that the reagent R1 does not contain tetradecyltrimethylammonium bromide, and the other is the same as in example 1.
Comparative example 7
The difference from the nitric oxide assay kit of example 1 is only that the reagent R2 does not contain acid hydrolyzed casein, and otherwise is the same as in example 1.
Comparative example 8
The difference from the nitric oxide assay kit of example 1 is that the reagent R2 does not contain sodium polymethacrylate, and the other is the same as in example 1.
Comparative example 9
The difference from the nitric oxide assay kit of example 1 is that the reagent R2 does not contain polyvinylpyrrolidone k30, and the other is the same as in example 1.
Performance verification
1. Correlation experiments
And carrying out a correlation test, wherein the test scheme is as follows: example 1, comparative example 1, 40 clinical serum samples were simultaneously tested, correlation analysis was performed on two sets of test results, and a correlation coefficient r was calculated; the relative deviation (Bias%) of 40 pairs of data was calculated using the detection result of comparative example 1 as a target value. It is required that r is not less than 0.990 and the relative deviation is not more than + -10%.
TABLE 2 correlation comparison experiment results
TABLE 3 correlation coefficients for comparative example 1 and example 1
Correlation coefficient r
Example 1 and comparative example 1 0.9974
As can be seen from tables 2, 3 and 1, the maximum deviation of the serum samples measured by the reagents of the example 1 and the comparative example 1 is 1.96%, the correlation coefficient of the two reagents is 0.9974, and the detection results of the example 1 and the comparative example 1 are very close, so that the accuracy of the clinical sample measured by the detection reagent of the example 1 provided by the invention is higher.
Test two stability test
Stability tests were performed on the nitric oxide assay reagents provided in example 1 and comparative examples 1,2, 3, 7, 9, with the following test protocols: the reagents provided in example 1 and comparative examples 1,2, 3, 7 and 9 were put together in a 37 ℃ water bath, and a quality control product with a target value of 82 + -3.2 mu mol/L was detected every day, and the change in the measured value of the quality control product was monitored; the reagents provided in example 1 and comparative examples 1,2, 3, 7, and 9 were put into an instrument to conduct an open bottle stability test at 2-8℃and a quality control having a target value of 82.+ -. 3.2. Mu. Mol/L was detected weekly, and the change in the measured value of the quality control was monitored.
TABLE 4 results of thermal stability validation of reagents
Time (Tian) Example 1 Comparative example 1 Comparative example 2 Comparative example 3 Comparative example 7 Comparative example 9
1 82.47 82.92 80.89 79.29 82.97 82.59
2 82.47 82.79 79.19 74.13 80.14 79.34
3 82.46 82.37 77.46 71.11 78.21 78.86
4 81.93 82.15 73.48 66.84 76.51 77.12
5 81.5 81.85 70.13 64.09 75.42 74.39
6 81.37 81.79 69.69 63.68 70.94 71.25
7 81.35 81.38 69.19 59.73 70.06 70.65
8 80.55 80.87 66.37 57.87 68.73 67.72
9 80.12 80.51 66.18 54.73 61.46 65.19
10 80.06 80.48 65.63 53.72 60.40 64.32
TABLE 5 results of reagent bottle opening stability verification
Time (week) Example 1 Comparative example 1 Comparative example 2 Comparative example 3 Comparative example 7 Comparative example 9
1 82.82 80.02 82.53 81.74 83.09 80.59
2 81.8 81.51 74.6 72.15 70.41 68.54
3 81.23 82.77 62.5 69.11 69.86 65.18
4 80.36 81.47 53.12 51.26 53.64 59.23
As can be seen from tables 4, 5, 2 and 3, the reagent of example 1 provided by the invention has better stability in 10 days under the water bath condition of 37 ℃ similar to that of comparative example 1, and the quality control values of the reagents of comparative examples 2, 3, 7 and 9 are obviously reduced in 10 days under the water bath condition of 37 ℃; the reagent of the embodiment 1 provided by the invention has better stability for one month under the condition of 2-8 ℃ bottle opening, and the quality control values of the reagents of the comparative examples 2, 3, 7 and 9 are obviously reduced from week 2 under the condition of 2-8 ℃ bottle opening, so that the acceleration and the bottle opening stability of the reagent of the embodiment 1 of the invention are superior to those of the reagent kit of the comparative examples 2, 3, 7 and 9, and the addition of the hydroxypropyl-beta-cyclodextrin in the reagent R1 is favorable for improving the stability of the reagent R1; the acid hydrolysis casein and polyvinylpyrrolidone k30 are added into the reagent R2, so that the stability of the reagent R2 is improved synergistically.
Test three anti-interference test
The nitric oxide assay kits provided in example 1 and comparative examples 1, 4, 5, 6 were subjected to an anti-interference test, with the following test protocol: and (3) taking a nitric oxide quality control product with a target value of 38+/-2.6 mu mol/L, preparing a triglyceride sample, a chyle interference sample and a ferric chloride sample with different concentration levels, measuring and adding the samples with different concentrations of the interference substances by using the reagents provided in the embodiment 1 and the comparative examples 1, 4, 5 and 6, and calculating the deviation between the measured value of the nitric oxide in the interference sample and the base value by taking the measurement result of the nitric oxide in the level 1 sample as the base value, wherein the deviation is more than or equal to +/-10 percent, and the deviation is interference. The results are shown in tables 6, 7 and 8.
TABLE 6 Triglycerides interference experimental determination results
TABLE 7 chylomicron interference experimental determination results
TABLE 8 determination of iron chloride interference experiments
The results show that: example 1 similarly to comparative example 1, when the concentration of triglyceride in the sample to be measured was 14.3mmol/L and chyle was 1000mg/dL,has no interference to the measurement result of nitric oxide. In comparative example 6, when the concentration of triglyceride in the sample to be measured is 4.5mmol/L and the chyle is 400mg/dL, obvious interference is generated to the measurement result of nitric oxide. Comparative example 4, when the ferric chloride concentration of the sample to be measured is 8 mug/mL, interference to the measuring result of nitric oxide is generated; comparative example 5 although ethylenediamine tetraacetic acid was added in an equivalent amount, 8. Mu.g/mL Fe could be eliminated 3+ When the concentration of ferric chloride reaches 13 mug/mL, the interference effect is obvious. Therefore, the system of the invention has important function on eliminating interference of chyle and blood fat by adding tetradecyl trimethyl ammonium bromide, and the addition of trans-1, 2-cyclohexanediamine tetraacetic acid (CDTA) is beneficial to eliminating Fe 3+ Is a function of (a) and (b).
Test four sensitivity test
Samples of known concentrations at 150. Mu. Mol/L were measured with the reagents provided in example 1, comparative examples 1, 8, respectively, and the absorbance change rate (. DELTA.A/min) was recorded. The detection results are shown in Table 9:
table 9 analytical sensitivity vs. experimental results
As can be seen from the detection data, the absorbance change rate of the assay kit of example 1 is similar to that of the kit of comparative example 1, which shows that the kit of the invention has better analysis sensitivity. Comparative example 8 has poor sensitivity, demonstrating that the present invention promotes nitrate (NO 3 -), nitrite (NO) 2 - ) And the reaction of dopamine hydrochloride and 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH), so as to improve the analysis sensitivity of the reagent.
Test five-linearity test
Taking high-value nitric oxide samples of 450 mu mol/L, carrying out gradient dilution, preparing 6 samples with different concentrations, and sequentially detecting 0 mu mol/L, 25 mu mol/L, 75 mu mol/L, 150 mu mol/L, 225 mu mol/L and 450 mu mol/L by using the reagents provided in example 1 and comparative example 1, wherein each sample with each concentration level is measured for 3 times, and the average value is obtained. The test results are shown in Table 10:
table 10 results of reagent Linear validation
Linear sample theory concentration (mu mol/L) Example 1 Comparative example 1
0 0.45 0.35
25 23.6 21.5
75 73.4 72.6
150 151 148.4
225 234 228.2
450 456 441
Correlation coefficient r 0.9997 0.9996
As can be seen from Table 10, similar to comparative example 1, inventive example 1 varied linearly with dilution concentration, and the linear correlation coefficient reached 0.9997, indicating that the linear range of example 1 satisfies the clinical examination requirements.
The specific embodiments described herein are offered by way of example only to illustrate the spirit of the invention. Various modifications or additions to the described embodiments may be made by those skilled in the art to which the invention pertains or may be substituted in a similar manner without departing from the spirit of the invention or beyond the scope of the appended claims.
While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.

Claims (2)

1. A nitric oxide assay kit is characterized by comprising the following components;
reagent R1 comprises the following components:
citrate buffer 50-100mM
Dopamine hydrochloride 5-8g/L
Hydroxypropyl-beta-cyclodextrin 2-5g/L
1-2g/L of trans-1, 2-cyclohexanediamine tetraacetic acid
Tetradecyl trimethyl ammonium bromide 1-5g/L
Sodium azide 1g/L
Reagent R2 comprises the following components:
50-100mM 4-hydroxyethyl piperazine ethane sulfonic acid buffer
5-10 g/L3-methyl-2-benzothiazolinone hydrazone hydrochloride
Acid hydrolyzed casein 1-10 g/L
2-5g/L of sodium polymethacrylate
Polyvinylpyrrolidone k 30-2 g/L
Sodium azide 1g/L
The pH of the reagent R1 is 4.0-6.0; the pH of the reagent R2 is 7.0-8.0.
2. Use of the nitric oxide assay kit according to claim 1 for determining the concentration of nitric oxide for non-disease diagnostic and therapeutic purposes.
CN202310971214.2A 2023-08-03 2023-08-03 Nitric oxide determination kit and preparation method and application thereof Active CN117007788B (en)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101204649B1 (en) * 2011-07-14 2012-11-23 서울시립대학교 산학협력단 nitrite concentration measuring agents and the kit uising thereof
CN111289501A (en) * 2020-02-21 2020-06-16 安徽大千生物工程有限公司 Kit for detecting NO based on indirect colorimetric method and preparation and use methods thereof
CN112051259A (en) * 2020-08-31 2020-12-08 武汉生之源生物科技股份有限公司 Nitric oxide detection kit and detection method
CN114965453A (en) * 2022-05-23 2022-08-30 苏州络森生物科技有限公司 Nitrate radical detection reagent containing 3-methyl-2-benzothiazolinone hydrazone hydrochloride, using method and vegetable nitrate nitrogen detection kit

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060121620A1 (en) * 2004-12-06 2006-06-08 Basant Bhandari Nitrate/nitrite assay reagents, kit, and method of use

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101204649B1 (en) * 2011-07-14 2012-11-23 서울시립대학교 산학협력단 nitrite concentration measuring agents and the kit uising thereof
CN111289501A (en) * 2020-02-21 2020-06-16 安徽大千生物工程有限公司 Kit for detecting NO based on indirect colorimetric method and preparation and use methods thereof
CN112051259A (en) * 2020-08-31 2020-12-08 武汉生之源生物科技股份有限公司 Nitric oxide detection kit and detection method
CN114965453A (en) * 2022-05-23 2022-08-30 苏州络森生物科技有限公司 Nitrate radical detection reagent containing 3-methyl-2-benzothiazolinone hydrazone hydrochloride, using method and vegetable nitrate nitrogen detection kit

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