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Method for accurately measuring content of primary amine group

A technology for accurate determination of primary amine groups, applied in analytical materials, instruments, etc., can solve the problems of not meeting the reaction time of macromolecules and the inability to collect gas by gas chromatography, avoiding incomplete gas washing, accurate temperature control accuracy, and improving The effect of testing precision

Inactive Publication Date: 2014-01-15
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The gas chromatography detection method utilizes the Van Slyke method and detects the N2 content by gas chromatography. However, the gas chromatography cannot collect gas and requires the reaction to be completed within 5-10 minutes. Although this method can accurately measure the amino content in small molecules, it does not meet the requirements. The need for long reaction times for macromolecules

Method used

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  • Method for accurately measuring content of primary amine group
  • Method for accurately measuring content of primary amine group
  • Method for accurately measuring content of primary amine group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The method of the present invention measures the content of primary amino groups in the gelatin molecule and compares the result with the traditional Van Slyke device.

[0032] The specific operation is as follows: Prepare the primary amino quantitative instrument, apply vacuum silicone grease to each cock, and seal each interface. Among them, the drain port (26) of the hemispherical gas washing bottle and the drain port (17) of the reactor are clamped with water-stop clips, and Prevent liquid leakage.

[0033] First, slowly pour the washing liquid from the buffer bottle (23), fill the hemispherical washing bottle (20), and make the washing liquid reach 1 / 3 of the volume of the buffer bottle (23). Adjust the inclined hole three-way cock (29) to be filled with lotion to the inclined hole three-way cock (29), and turn the inclined hole three-way cock (29) so that the inclined hole three-way cock (29) does not communicate with all parties. Inject deionized water from the ...

Embodiment 2

[0039] The method of the present invention measures the primary amino group content result in gelatin molecule and contrasts with the determination result of TNBS colorimetric method, setting measurement temperature is 45 ℃, measurement time is 40 minutes, other is the same as embodiment 1.

[0040] The results of primary amino group content in gelatin molecules determined by the primary amino group quantifier Figure 4 . At the same time, use the TNBS colorimetric method to measure the results for comparison, see Figure 4 .

[0041] Determination, the data stability of the primary amino group quantifier for gelatin primary amino content determination is good, and the relative error is 0.5%. However, the data stability of gelatin primary amino content determined by TNBS colorimetry is poor, and the relative error reaches 7%.

[0042] The results can be used to show that under the conditions of determined reaction time and temperature, the accuracy of the data measured by t...

Embodiment 3

[0044] Test the influence of temperature and time on the determination result of primary amino group content in amino acid in amino acid.

[0045] Since there are many kinds of amino acids that make up gelatin, we selected representative alanine, glycine, glutamic acid, arginine, and lysine for determination. Before the determination, the above amino acids were purified according to their differences in properties. For alanine, glycine, lysine, and arginine, the differences in the solubility of amino acids in different solvents were mainly used, and they were precipitated several times with absolute ethanol, and then vacuum-dried for later use. As for glutamic acid, the difference in solubility in water of different pH is used to carry out multiple recrystallizations by adjusting the pH of the solution, and vacuum-dry it for future use.

[0046] The purified amino acids were formulated into 0.1 mol / L solutions, and then 2 ml were taken and measured with a primary amino quanti...

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Abstract

The invention provides a method for accurately measuring content of primary amine group on the basis of a Van method and particularly relates to a method for accurately measuring the content of the primary amine group in molecules of macromolecular biological compounds such as gelatin, proteins and the like. The method comprises the following steps: a primary amine group quantometer is prepared; the air in a system is emptied; testing conditions of concentration of nitrous acid, a ratio of glacial acetic acid to nitrous acid, concentration of a solution to be measured, a testing temperature, testing time and the like are fixed; the problems of great limitation to the environmental conditions and long macromolecular reaction balancing time of measurement on the content of the primary amine group in biomacromolecules are solved; the method has a calibration error of 0.5 percent on amino acid and has a measurement deviation of 1 percent on the gelatin.

Description

technical field [0001] The invention relates to a method for accurately measuring the content of primary amino groups, in particular to a method for accurately measuring the content of primary amino groups in biomacromolecules such as gelatin and protein. Background technique [0002] Formaldehyde titration, ninhydrin colorimetric method, TNBS colorimetric method, Van Slyke method and gas chromatography are commonly used methods for determining the content of primary amino groups. Among them, the formaldehyde titration method calculates the content of primary amino groups by titrating the H+ dissociated after the combination of formaldehyde and free primary amino groups. The color method uses the alkylation reaction between trinitrobenzenesulfonic acid and the primary amino group to determine the content of the primary amino group. However, the hydrogen on the α-carbon of the carbonyl group and the active methylene group can also participate in the alkylation reaction. The s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N7/18
Inventor 李天铎许静姜青伟唐小龙
Owner QILU UNIV OF TECH
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