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Method for regulating nitration substitution degree of nitrocellulose

A technology of nitrocellulose and nitrification, applied in instruments, analytical materials, measuring devices, etc., can solve problems such as uneven fluorescence background, reduce background noise, overcome high green fluorescence background and uneven fluorescence background, background noise low effect

Inactive Publication Date: 2015-04-29
LONGYAN NINEHEALTH BIOCHIP INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to accurately adjust and control the degree of nitration of nitrocellulose to obtain a nitrocellulose substrate with stable and superior performance, and to overcome the technical defects of high green fluorescence background and uneven fluorescence background, the present invention provides a method for accurately adjusting the degree of nitration substitution of nitrocellulose Methods

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  • Method for regulating nitration substitution degree of nitrocellulose
  • Method for regulating nitration substitution degree of nitrocellulose
  • Method for regulating nitration substitution degree of nitrocellulose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Preparation of nitrocellulose with precisely adjusted degree of nitration:

[0033] (1) Preparation of nitrocellulose with a high degree of substitution:

[0034] A. Accurately measure 30-45 mL of fuming nitric acid and 50-80 mL of concentrated sulfuric acid, mix them and place them on a magnetic stirrer;

[0035] B. Accurately weigh 4 g of cellulose and add it to the mixture of fuming nitric acid and concentrated sulfuric acid, and react for 1 to 4 hours to make the nitration reaction as complete as possible and the degree of substitution as high as possible;

[0036] C. Inject the reaction solution into 2 L of ultrapure water, and add sodium carbonate while stirring until no carbon dioxide bubbles are produced;

[0037] D. Precipitate overnight, discard the supernatant, boil the precipitate with 2% sodium carbonate solution for 20 min, adjust the pH to neutral with hydrochloric acid, and centrifuge in a 50 mL centrifuge tube for 5 times, centrifuge the precipitate a...

Embodiment 2

[0050] Comparison of background intensity and uniformity for red and green fluorescent channels:

[0051] (1) Use nitrocellulose obtained by accurately adjusting the degree of nitration to make a nitrocellulose substrate: weigh 0.2-1 g of nitrocellulose and dissolve it in 4-25 mL of acetone, then add 1-3 mL of isopropanol , 1 to 5 mL of methanol, and 0.1 to 0.5 mL of water were used to form a film-forming liquid, and the film-forming liquid was evenly coated on a glass slide with a film applicator to prepare the nitrocellulose substrate of the present invention.

[0052] (2) Take each of the substrate made by the present invention and the Whatman substrate, scan with an Axon GenePix 4200AL fluorescence scanner, and compare the substrate made by the present invention with the Whatman substrate to check the difference in background uniformity and background intensity . When scanning, the signal amplification uses the same setting. When displaying the visual effect of the backgro...

Embodiment 3

[0060] Comparison of background noise for red and green fluorescent channels:

[0061] (1) Use nitrocellulose obtained by accurately adjusting the degree of nitration to make a nitrocellulose substrate: weigh 0.2-1 g of nitrocellulose and dissolve it in 4-25 mL of acetone, then add 1-3 mL of isopropanol , 1 to 5 mL of methanol, and 0.1 to 0.5 mL of water were used to form a film-forming liquid, and the film-forming liquid was evenly coated on a glass slide with a film applicator to prepare the nitrocellulose substrate of the present invention.

[0062] (2) According to the method disclosed in the literature of Diagnostic markers of ovarian cancer by high-throughput antigen cloning and detection on arrays published by Chatterjee et al. in 2006, the antigen protein clone library was prepared, and two 384-well plates were prepared with this clone library, with a total of 768 antigenic protein marker samples. Use the ArrayJet Marathon (Arrayjet Ltd., Edinbough, Scottland) spottin...

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Abstract

The invention relates to a method for regulating the nitration substitution degree of nitrocellulose, belonging to the technical field of biochemical analysis product development. The method for regulating the nitration substitution degree of the nitrocellulose mainly comprises the steps of: preparing high substitution nitrocellulose and sodium ethoxide and re-regulating nitration degree to the target nitration degree. The actual nitration degree of cellulose exceeds the target nitration degree, and then is accurately re-regulated by adopting the sodium ethoxide. Because the adding quantity of the sodium ethoxide can be accurately controlled, the nitration degree of the cellulose can be accurately controlled. The nitrocellulose for accurately regulating the nitration degree is made into a nitrocellulose base sheet made of a film forming agent, backgrounds of a red and green fluorescence channels are more uniform, the background noise of the green fluorescence channel is greatly reduced while the lower background noise of the red fluorescence channel is ensured, and the technical defects of non-uniform fluorescence background and high green fluorescence background are overcome successfully.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis product development, in particular to a method for adjusting the degree of nitration substitution of nitrocellulose. Background technique [0002] Nitrocellulose substrate is currently the most widely used solid phase support, that is, nitrocellulose forms a thin film about 10 microns thick on a solid plane, relying on its good hydrophobicity, it can be combined with most negatively charged DNA, RNA and protein, etc. produce high affinity adsorption. After adsorption, it can be fixed by baking at 80°C or cross-linked by ultraviolet irradiation. Different biomolecules are arranged in a microarray, attached to the surface of the biochip substrate, and react with the corresponding molecules in the sample to complete the analysis on the chip. Experimental process, test results are given. Because of this property, nitrocellulose substrates have been widely used in the fields of biochemi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B5/02G01N5/00
Inventor 翁长仁庄明仙
Owner LONGYAN NINEHEALTH BIOCHIP INST
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