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Method for identifying amino acid

An amino acid, the first technology, applied in the field of identifying amino acids, can solve the problems of inability to distinguish and identify natural amino acids, increase costs, etc., and achieve the effect of simple detection method, low cost and mild reaction conditions

Active Publication Date: 2021-07-20
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has certain deficiencies: first, this method needs to introduce additional fluorine-containing reactants, namely 3-(trifluoromethyl)thiophenol, which increases the cost; second, this method only identifies 15 natural Amino acids, the four natural amino acids glutamine, asparagine, tryptophan, and cysteine ​​cannot be identified; finally, this method cannot distinguish phenylalanine Phe from serine Ser under optimal conditions. 19 F-NMR signal peaks completely overlap

Method used

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  • Method for identifying amino acid
  • Method for identifying amino acid
  • Method for identifying amino acid

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0044] Step 1): Select 19 test tubes, respectively mix target amino acids (glycine Gly, alanine Ala, valine Val, leucine Leu, isoleucine Ile, methionine Met, phenylalanine Phe, Tryptophan Trp, serine Ser, threonine Thr, cysteine ​​Cys, tyrosine Tyr, asparagine Asn, glutamine Gln, aspartic acid Asp, glutamic acid Glu, lysine Lys, essence Amino acid Arg, histidine His) and o-trifluoromethylbenzaldehyde (a) were mixed in 500 μL of 20 mM sodium carbonate-sodium bicarbonate buffer at pH 10, and then 50 μL of D 2 O, wherein the solution concentration of each amino acid is 2.5 mM, and the concentration of o-trifluoromethylbenzaldehyde (a) is 5 mM, and each mixed solution is transferred to an NMR tube for 19 F NMR signal acquisition, respectively, to obtain 19 corresponding o-trifluoromethylbenzaldehyde-labeled target amino acids 19 F-NMR spectrum.

[0045] Step 2): Select a test tube, mix 19 kinds of amino acid mixtures (including: glycine Gly, alanine Ala, valine Val, leucine Leu,...

Embodiment 2

[0052] All the other are identical with embodiment one, and difference is:

Embodiment 1

[0053] In step 2) of Example 1, the pH of the mixed solution after the reaction was adjusted to 12.0 with NaOH solution, and then nuclear magnetic resonance was carried out 19 F-NMR signal collection, obtain the mixed solution under the condition of pH=12.0 19 F-NMR spectrum, as attached image 3 Shown, the chemical shift table is shown in Table 2.

[0054] Table 2 The concentration of amino acids labeled with o-trifluoromethylbenzaldehyde at pH=12.0 19 Chemical shift table of F-NMR spectral signals

[0055] o-Trifluoromethylbenzaldehyde labeled amino acid 19 F-NMR chemical shift (ppm)

[0056] Experimental results: From attached image 3 As can be seen from Table 2, under the condition that the pH is 12.0, the chemical shift of asparagine Asn and glutamine Gln can be clearly distinguished, indicating that the present invention can be mixed solution after adding o-trifluoromethylbenzaldehyde through simple adjustment The pH value is used to achieve the purpos...

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Abstract

The invention discloses a method for identifying amino acids, which comprises the following steps: 1) preparing a target amino acid standard solution, respectively mixing the target amino acid standard solution with o-trifluoromethyl benzaldehyde, adjusting the pH value of the mixed solution to 10-12, adding a deuterated reagent, and carrying out nuclear magnetic resonance <19>F-NMR signal acquisition to obtain a first <19>F-NMR spectrum; 2) preparing a sample solution containing amino acid to be detected, mixing the sample solution with o-trifluoromethylbenzaldehyde, adjusting the pH value of the mixed solution to 10-12, adding a deuterated reagent, and performing nuclear magnetic resonance second <19>F-NMR signal acquisition to obtain a second <19>F-NMR spectrum; 3) performing superposition comparative analysis on the first <19>F-NMR spectrum and the second <19>F-NMR spectrum, and identifying each amino acid to be detected in the sample solution in the step 2) through each <19>F-NMR chemical shift signal corresponding to the first <19>F-NMR spectrum and the second <19>F-NMR spectrum. The detection method has the advantages that the detection method is low in cost, simple and easy to implement, samples can be prepared and detected immediately, and qualitative and quantitative detection of various free amino acids in the samples is achieved.

Description

technical field [0001] The invention belongs to the field of amino acid detection, in particular to a method for identifying amino acids. Background technique [0002] Free amino acids in biological samples are the most important analytical indicators for diagnosing various diseases, especially amino acid index cancer screening, which uses the concentration of amino acids in plasma as a sign of cancer detection, and is a tool for screening early cancers and reducing life-threatening A promising technique, therefore, the identification of amino acids is essential for basic diagnosis. [0003] At present, there are various methods for identifying amino acids, including: high performance liquid chromatography-mass spectrometry (HPLC-MS), nuclear magnetic proton spectroscopy, based on 13 C isotopically labeled 1 H- 13 C HSQC spectrum, 19 F-NMR spectrum, etc. The HPLC-MS method can complete the amino acid analysis of samples in less than 10 minutes. Although the method based...

Claims

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Application Information

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IPC IPC(8): G01N24/08
CPCG01N24/08
Inventor 黄碧玲黄少华赵玉芬
Owner NINGBO UNIV