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Method for detecting content of aldehyde groups contained in biological tissue

A technology for the content of biological tissue and aldehyde groups, applied in the detection field, can solve the problems of inability to detect products, affect the content of free aldehyde groups, and many reaction steps, and achieve the effects of rapid detection, high detection accuracy and simple operation

Active Publication Date: 2020-06-09
SHANGHAI NEWMED MEDICAL CO LTD
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, since the aldehyde group is connected to the biological tissue through a chemical bond, the reactant reacts with the aldehyde group and is also connected to the biological tissue through a chemical bond, because the product cannot be dissolved in the solution Therefore, the product cannot be directly detected, and the method of chemical digestion, because strong acid is usually used, may cause the release of reacted aldehyde groups in the tissue, thereby affecting the actual content of free aldehyde groups
Therefore, there has been no effective method to detect the content of aldehyde groups in biological tissues.
In an article published by Edwards Lifesciences in 2006 (Carpentier-Edwards ThermaFix Process AMethod for Extracting Calcium Binding Sites from Pericardial Tissue), it was mentioned that aldehyde groups were oxidized to carboxyl groups in a mildly acidic solution, and the content of aldehyde groups was indirectly reflected by measuring the content of carboxyl groups , since biological tissue itself contains a certain amount of carboxyl groups, this method cannot truly reflect the content of aldehyde groups
In 2016, Edwards Lifesciences published an article (The association of bound aldehyde content with bioprosthetic tissue calcification) to provide a new method for the determination of aldehyde groups in biological tissues: first react with cystamine dihydrochloride and aldehyde groups under certain conditions to form the first intermediate, then the first intermediate reacts with 5,5-dithiobis(2-nitrobenzoic acid) to form a second intermediate, and finally tris(2-carboxyethyl)phosphine is added to decompose the second intermediate to release The substance to be tested is 5-thio-2-nitrobenzoic acid. This method has many reaction steps, so the detection accuracy is limited.

Method used

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  • Method for detecting content of aldehyde groups contained in biological tissue
  • Method for detecting content of aldehyde groups contained in biological tissue
  • Method for detecting content of aldehyde groups contained in biological tissue

Examples

Experimental program
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Embodiment 1

[0045] (1) Establish a standard curve

[0046] A 0.1 mmol / L DNPH solution was precisely prepared, wherein the pH of the DNPH solution was adjusted to 6.0 with acetic acid.

[0047] Select respectively 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, 7ml, 8ml, 9ml, 10ml of DNPH solutions with a concentration of 0.1mmol / L, dilute to 10ml with purified water, and use a UV spectrophotometer to measure its concentration at 218nm. Absorbance value, with the concentration of DNPH solution as the abscissa and the absorbance value as the ordinate, draw a standard curve. The obtained standard curve is as figure 1 As shown, the regression equation of the curve is: y=10.248x+0.0273, R2=0.9991, wherein x represents the concentration of DNPH solution in mmol / L, and y represents the absorbance value.

[0048] (2) Detection of aldehyde content contained in biological tissue

[0049] Get the DNPH solution of 5ml 0.1mmol / L and use the ultraviolet spectrophotometer to test its absorbance value A1 at 218nm place...

Embodiment 2

[0054] (1) Establish a standard curve

[0055] Precisely prepare 0.3mmol / L DNPH solution, wherein phosphoric acid is used to adjust the pH of DNPH solution to 2.0.

[0056] Select respectively 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, 7ml, 8ml, 9ml, 10ml of DNPH solutions with a concentration of 0.1mmol / L, dilute to 10ml with purified water, and use an ultraviolet spectrophotometer to measure its concentration at 264nm. Absorbance value, with the concentration of DNPH solution as the abscissa and the absorbance value as the ordinate, draw a standard curve. The obtained standard curve is as figure 1 As shown, the regression equation of the curve is: y=10.293x+0.0227, R 2 =0.9998, wherein x represents the concentration of DNPH solution in mmol / L, and y represents the absorbance value.

[0057] (2) Detection of aldehyde content contained in biological tissue

[0058] Get 10ml of 0.3mmol / L DNPH solution and dilute to 20ml, as DNPH reaction solution, use ultraviolet spectrophotometer to te...

Embodiment 3

[0065] (1) Establish a standard curve

[0066] A 1.0 mmol / L DNPH solution was precisely prepared, and the pH of the DNPH solution was adjusted to 4.0 with perchloric acid.

[0067] Select respectively 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, 7ml, 8ml, 9ml, 10ml of DNPH solution with a concentration of 1.0mmol / L, dilute to 10ml with purified water, and measure its concentration at 323nm using a UV spectrophotometer. Absorbance value, with the concentration of DNPH solution as the abscissa and the absorbance value as the ordinate, draw a standard curve. The obtained standard curve is as figure 1 As shown, the regression equation of the curve is: y=10.329x+0.034, R2=0.9998, wherein x represents the concentration of DNPH solution in mmol / L, and y represents the absorbance value.

[0068] (2) Detection of aldehyde content contained in biological tissue

[0069] Take 1ml of 1.0mmol / L DNPH solution and dilute it to 10ml, and use an ultraviolet spectrophotometer to test its absorbance value A...

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Abstract

The invention provides a method for detecting the content of aldehyde groups contained in a biological tissue, and the method comprises the following steps: preparing a plurality of 2, 4-dinitrophenylhydrazine standard solutions with different concentration gradients, measuring the absorbance values of the plurality of standard solutions at a specific wavelength, and drawing a standard curve of which the concentrations and the absorbance values are in a functional relationship; preparing a 2, 4-dinitrophenylhydrazine reaction solution, and determining the absorbance value A1 of the reaction solution at the specific wavelength; reacting the 2, 4-dinitrophenylhydrazine reaction solution with the biological tissue, and measuring the absorbance value A2 of the solution at the specific wavelength after the reaction; and calculating the content of the aldehyde groups contained in the biological tissue based on the A1, the A2 and the standard curve. According to the detection method, the biological tissue is soaked in the 2, 4-dinitrophenylhydrazine solution, the change of the ultraviolet absorbance value of the 2, 4-dinitrophenylhydrazine before and after the reaction is determined, andthe aldehyde group content of the biological tissue is indirectly determined, so that the detection method has the advantages of simplicity in operation, rapidness in detection, high detection accuracy and the like.

Description

technical field [0001] The invention belongs to the detection field, and in particular, the invention relates to a method for detecting the content of aldehyde groups contained in biological tissues. Background technique [0002] Artificial heart valves are divided into mechanical valves and biological valves. Biological valves are mainly composed of alloy brackets and leaflets. The leaflets carry the opening and closing function of the artificial heart valve and realize the one-way movement of blood flow. The valve leaflets are usually made of bovine pericardium, pig pericardium and other biological tissues. At present, one of the main reasons for limiting the durability of biological valves is the calcification of the valve leaflets. After calcification, the opening and closing function of the valve leaflets will be weakened or even lost. Biological tissues are mainly composed of collagen fibers, elastic fibers, and connective tissue, which contain functional groups such a...

Claims

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Application Information

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IPC IPC(8): G01N21/33
CPCG01N21/33
Inventor 尚大鹏于吉温贤涛
Owner SHANGHAI NEWMED MEDICAL CO LTD
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