A kind of detection method of nitrite in blood

A technology of nitrite and detection method, which is applied in the preparation of test samples and the measurement of color/spectral properties, etc., can solve the problems of reduced detection sensitivity, large matrix interference, and multi-sample volume, etc., and achieves improved detection sensitivity and detection efficiency. High, the effect of overcoming detection errors

Active Publication Date: 2017-07-28
云南健牛环境监测有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Further studies have found that a quantitative, accurate and convenient method for detecting nitrite in blood is more needed in clinical practice. Since clinical blood nitrite detection has the characteristics of small sample size, large matrix interference, and large sample size, if using The current photometric method requires a large amount of sample, otherwise it will be further diluted to reduce the detection sensitivity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. Preparation of nitrite standard working curve

[0022] ①Preparation of matrix blank solution: Take 10 mL of fresh blood without nitrite plus anticoagulant, put it into a centrifuge, centrifuge at 6000r / min for 8min, take 5 mL of the supernatant into a clean centrifuge tube, add 0.2 A protein precipitation and decolorization agent consisting of 1 mL of mol / L sodium hydroxide solution and 1 g of zinc sulfate was mixed evenly, centrifuged at 5000 r / min for 10 min, and the supernatant was taken out to obtain an almost colorless and transparent matrix blank solution.

[0023] ②Take NO with a concentration of 1.0µg / mL 2 - 20, 40, 60, 80, 100 µL standard solutions in micro test tubes, dilute to 500 µL with the matrix blank solution prepared in step ①, add 25 µL of GriessA reagent, mix well; add 25 µL of GriessB reagent after 1 min, and mix well. Take 200µL of the mixed chromogenic solution in the microplate plate, measure the absorbance at the wavelength of 490nm of the m...

Embodiment 2

[0027] 1. make nitrite standard working curve by embodiment 1.

[0028] 2. Sample processing and measurement results

[0029] Blood test: Take 5 mL of fresh blood with anticoagulant, put it in a centrifuge and centrifuge at 8000r / min for 5 minutes, take 2 mL of supernatant, add 0.4 mL of 0.2 mol / L sodium hydroxide solution and 0.4 g of zinc sulfate The composed protein precipitation and decolorizing agent, mix well, centrifuge at 6000r / min for 6min to separate, take out the supernatant, add 1mL of acetonitrile, centrifuge at 6000r / min for 5min and separate, take the supernatant into 500µL micro test tube, add 25µL of GriessA reagent, mix well ; After 1 min, add 25 µL of GriessB reagent and mix well. Take 200µL of the mixed chromogenic solution in the microplate plate, measure the absorbance at the wavelength of 490nm of the microplate reader to be 0.011, and compare the linear regression equation obtained in step (1), calculate the content of nitrite in the sample to be 0.062...

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Abstract

The invention discloses a rapid detection method for nitrite in blood. Using a multi-well plate and a microplate reader to measure the absorbance of the colored complex formed by nitrite and Griess, the blood without nitrite is used as the matrix solution for making the working curve, which eliminates the interference of the blood matrix and improves the sensitivity and accuracy of detection . The method has the characteristics of high accuracy, large detection sample volume, small detection sample volume, and high detection efficiency.

Description

technical field [0001] The invention belongs to the technical field of drug detection in vivo, and in particular relates to a method for detecting blood nitrite. Background technique [0002] Nitric oxide (NO) is a biological signal molecule that regulates the cardiovascular, endocrine, nervous and immune systems. The development of a simple and accurate detection method for NO in blood and body fluids is important for its clinical and pharmaceutical applications. very important. NO has the characteristics of simple structure, low molecular weight, high lipophilicity and free radicals. It is extremely unstable in the body, has a very short half-life of only a few seconds, and easily forms nitrite and nitrate. Direct NO detection is difficult, as a relatively stable NO metabolite, nitrite (NO 2 - ) has become a hot topic in the world medical field in the last 10 years, by measuring NO in blood and body fluids 2 - The content of NO can be used to reflect the content of NO...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N1/28G01N1/38
Inventor 杨亚玲赵娇宁金艳王军吕云辉
Owner 云南健牛环境监测有限公司
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