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Rapid detection method for nitrites in blood and urine

A nitrite and detection method technology, which is applied in the direction of material analysis by observing the influence of chemical indicators, and analysis by making materials undergo chemical reactions, can solve the problems of difficult detection requirements, interference with experimental results, complex matrix, etc. problems, to achieve the effects of increased sensitivity, elimination of interference, and mild operating conditions

Inactive Publication Date: 2015-01-14
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

NO in blood or urine 2 - The detection of Griess detection kit is usually used, however, NO in blood or urine 2 - The content is low, the matrix is ​​complex, often interferes with the experimental results, and it is difficult to meet the detection requirements

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The invention is used to detect the content of nitrite in human urine.

[0024] 1. Preparation of nitrite standard color scale: take NO with a concentration of 10μg / mL 2 - Standard solution 0.01, 0.05, 0.10, 0.30, 0.70, 1.0mL in a graduated centrifuge tube, dilute to 5mL with distilled water, add 250 μL of GriessA reagent, mix well; add 250 μL of GriessB reagent after 1 min, mix well, stand for 5 min, add Non-ionic surfactant Triton X-114 0.5mL, ammonium sulfate 0.2g, vortexed for 1min, centrifuged to separate the phases, a red droplet color scale solution from light to dark appeared at the bottom of the centrifuge tube;

[0025] 2. Sample processing and measurement results

[0026] Take 10mL of urine into a 10mL centrifuge tube, add 1.0g of activated carbon, mix well, centrifuge at 4000r / min for 10min, filter through filter paper to remove the activated carbon. Take 5.0 mL of decolorized urine into a 10 mL centrifuge tube, add 250 μL of Griess A reagent, and ...

Embodiment 2

[0028] The invention is used to detect the content of nitrite in human blood.

[0029] 1. make the standard color scale of nitrite by embodiment 1, nonionic surfactant is Triton X-100, and salt is sodium chloride 0.5g.

[0030] 2. Sample processing and measurement results

[0031] Take 5 mL of fresh blood and centrifuge at 4000r / min for 15min. Take the supernatant in a centrifuge tube, add 0.2mL of 0.1mol / L sodium hydroxide solution and 0.2g of zinc sulfate, mix well, add 1mL of acetonitrile, centrifuge, take 2mL of the supernatant in a 10mL centrifuge tube and dilute to volume with distilled water To 5mL, add 250μL of GriessA reagent, mix well, add 250μL of GriesssB reagent after 1min, mix well, add non-ionic surfactant Triton X-100 0.5mL, sodium chloride 0.5g, vortex mix for 1min, centrifuge to separate phases, centrifuge The red drop at the bottom of the tube is between 0.30 μg / mL and 0.70 μg / mL of nitrite.

Embodiment 3

[0033] The invention is used to detect the content of nitrite in human blood.

[0034] 1. make the standard color scale of nitrite by embodiment 1, nonionic surfactant is NP-7, and salt is ammonium chloride 0.3g.

[0035] 2. Sample processing and measurement results

[0036] Take 5mL of fresh blood in a centrifuge tube, centrifuge at 6000r / min for 8min, take the supernatant in a centrifuge tube, add 0.1mol / L sodium hydroxide solution 0.2mL and zinc sulfate 0.2g, mix well, add acetonitrile 1mL, and centrifuge , Take 2 mL of the supernatant and dilute to 5 mL with distilled water in a 10 mL centrifuge tube, add 250 μL of GriessA reagent, and mix well. After 1 min, add 250 μL of GriessB reagent, mix evenly, and after standing for 5 min, add 0.5 mL of nonionic surfactant NP-7, 0.3 g of ammonium chloride, vortex and mix for 1 min, and centrifuge to separate the phases. Nitrate was not detected.

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PUM

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Abstract

The invention discloses a rapid detection method for nitrites in blood and urine. The method comprises: utilizing a diazotization coupling reaction of nitrites and Griess reagent to form an azo compound, performing cloud point extraction, precipitating the colored azo compound at the bottom of a centrifuge tube, comparing the red color at the tube bottom with standard color gradation, and determining the content of the nitrite according to the color depth. Because the enriching multiple of cloud point extraction is large and blood and urine are subjected to decoloring and protein-removal processing, the interference is substantially eliminated, and the detection limit of nitrites in the system can reach 0.01 mu g / mL. The method has the characteristics of simpleness, rapidness, high sensitivity, on-site detection and the like.

Description

technical field [0001] The invention belongs to the technical field of drug detection in vivo, and in particular relates to a rapid detection method for nitrite in blood and urine. Background technique [0002] The biochemical, physiological and pathological research of nitric oxide (NO) is widely carried out in the biological and medical circles in the world. As a relatively stable metabolite of NO, nitrite (NO 2 - ) has become a hot spot in the world's medical circles in the last 10 years. Nitrite is a common inorganic salt in daily diet and drinking water. It is generally believed that excessive intake of such substances may lead to methemoglobinemia and transform into carcinogenic nitrosamines in the body. By measuring NO in urine and blood 2 - The content of NO in the body reflects the NO content in the body and serves as an important indicator of many body functions. [0003] At present, the most common method for nitrite detection is the national food safety s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78
Inventor 杨亚玲赵娇王军
Owner 云南健牛环境监测有限公司
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