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Detection method of nitrate and nitrite in blood, urine and tissue

A technology of nitrite and detection method, which is applied to the content of nitrate and nitrite in urine and tissue, and the field of accurate determination of human or animal blood. It can solve the problems of difficult to meet the detection requirements, interference with experimental results, and low content. , to achieve the effect of improving detection sensitivity, eliminating mutual interference, and eliminating interference

Inactive Publication Date: 2013-08-07
云南健牛环境监测有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, NO in blood or urine 2 - Low content, due to individual differences, complex components in blood or urine often interfere with experimental results, making it difficult to meet detection requirements
In the prior art, there is no method for simultaneously and accurately determining the content of nitrate and nitrite in urine, blood and tissue by HPLC

Method used

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  • Detection method of nitrate and nitrite in blood, urine and tissue
  • Detection method of nitrate and nitrite in blood, urine and tissue
  • Detection method of nitrate and nitrite in blood, urine and tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The invention is used to detect the content of nitrate and nitrite in human urine.

[0026] 1. Production of nitrate and nitrite working curve: take 0.20, 0.4, 0.6, 0.80, 1.00mL NO respectively 3 - (200μg / mL) and NO 2 - (20μg / mL) The standard stock solution is diluted to 5mL in five 10mL centrifuge tubes, and the concentrations are respectively 8.0, 16.0, 24.0, 32.0, 40.0μg / mL and 0.8, 1.6, 2.4, 3.2, 4.0μg / mL. Add 150μL GriessA reagent and mix well. Add 150μL GriessB reagent after 1min, mix well, leave it for 5min, inject 10μL under the mobile phase conditions described in Table 1, and measure at 220nm and 510nm wavelength respectively; regression equation, correlation coefficient, relative standard deviation, recovery rate, etc. See Table 2.

[0027] Table 2 Linear equation of nitrate and nitrite

[0028]

[0029]

[0030] 2. Sample processing and measurement results

[0031] Take 10mL urine in a 10mL centrifuge tube, add 1.0g activated carbon, mix well, centrifuge at 4000r / ...

Embodiment 2

[0033] The invention is used to detect the content of nitrate and nitrite in rabbit urine.

[0034] 1. According to Example 1, make the working curve of nitrate and nitrite.

[0035] 2. Sample processing and measurement results

[0036] Take 2.0mL urine in a 10mL centrifuge tube, add 0.2g sepiolite, mix well, centrifuge at 5000r / min for 15min, filter paper to remove sepiolite. Take 1.0 mL of decolorized urine in a 10 mL centrifuge tube, add 30 μL GriessA reagent, and mix well. Add 30μL GriessB reagent after 1min, mix well, leave it for 5min, use Table 1 as the mobile phase conditions, inject 10μL, measure at 220nm and 510nm wavelengths respectively, and substitute the peak areas obtained into the working curve of Table 2 to obtain The nitrate content was 11.43μg / mL, and nitrite was not detected.

Embodiment 3

[0038] The invention is used to detect the content of nitrate and nitrite in human blood.

[0039] 1. According to Example 1, make the working curve of nitrate and nitrite.

[0040] 2. Sample processing and measurement results

[0041] Take 5mL of fresh blood and centrifuge at 5000r / min for 25min. Take the supernatant in a centrifuge tube, add 0.1 mL of 10% (w / v) sodium hydroxide solution and 1 mL of 5% (w / v) zinc sulfate solution, mix well, centrifuge at 10000r / min for 10 minutes, and take the supernatant 1.0 Add 50μL GriesssA reagent to 10mL centrifuge tube and mix well. After 1 min, add 50μL GriesssB reagent, mix well, leave it for 5min, use Table 1 as the mobile phase conditions, inject 10μL, measure at 220nm and 510nm wavelengths respectively, and substitute the peak areas obtained into the working curve of Table 2 to obtain The nitrate content is 9.53μg / mL, and the nitrite content is 0.08μg / mL.

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Abstract

The invention discloses a detection method of nitrate and nitrite in blood, urine and tissue. In the weak acidic condition, nitrate ions and an ion pair reagent form an ion pair, and nitrite and a Griess reagent are reacted for a diazotization coupling reaction for forming an azo compound, thereby nitrate and nitrite which can not be retained on the chromatographic column without UV absorption can be analyzed by liquid chromatogram at different wavelengths, and mutual interferences are eliminated. The nitrate and nitrite detection in blood, urine and tissue which have large matrix interferences and low contents have larger applicability, and the detection limit of nitrate and nitrite are 0.2 mu g / mL and 0.05 mu g / mL respectively. The invention has the characteristics of simple operation, rapid determination, efficiency, accuracy, etc.

Description

Technical field [0001] The invention belongs to the technical field of drug residue detection, and specifically relates to a method for accurately determining the content of nitrate and nitrite in human or animal blood, urine and tissues. Background technique [0002] Nitric oxide (NO) is an important vasodilator. Low concentration of NO relaxes bronchial smooth muscle and pulmonary blood vessels. Its effective release plays an important role in maintaining normal vascular pressure in humans or animals. At the same time, NO is also an important neurotransmitter. It plays an important and mysterious role in different central nervous systems. It participates in the regulation of physiological processes and the development of pathophysiological processes in almost all systems, while inhibiting platelet aggregation and atherosclerosis. The role of pulmonary sclerosis, muscle relaxation, inflammation, blood pressure, immune response and cardiovascular regulation is particularly import...

Claims

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Application Information

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IPC IPC(8): G01N33/02G01N33/06
Inventor 杨亚玲赵娇吕云辉
Owner 云南健牛环境监测有限公司
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