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Application of phthalic acid in amino acid analysis

A phthalic acid and amino acid technology, applied in the field of amino acid analysis, can solve problems such as affecting detection sensitivity, and achieve the effects of improved analysis effect, easy quantitative analysis, and improved detection sensitivity

Active Publication Date: 2014-12-17
国科新研国际技术转移有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since amino acids are amphoteric compounds, in order to facilitate their separation and improve peak shape, it is usually necessary to add a buffer solution formed by a higher concentration of formic acid and ammonium formate or acetic acid and ammonium acetate in the mobile phase, and a higher concentration of buffer solution in the mobile phase Salt will inhibit the response of the analyte and affect the detection sensitivity

Method used

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  • Application of phthalic acid in amino acid analysis
  • Application of phthalic acid in amino acid analysis
  • Application of phthalic acid in amino acid analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034]The purpose of this example is to investigate the relationship between the presence or absence of phthalic acid in the mobile phase and the signal response of amino acids, so as to verify the feasibility of using phthalic acid as a mobile phase additive in amino acid analysis.

[0035] 1. Accurately weigh phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, proline, glutamine, cysteine, alanine, glycine , threonine, serine, arginine, glutamic acid, asparagine, histidine, γ-aminobutyric acid, lysine, aspartic acid and citrulline, hydroxyproline, 2-amino Butyric acid, β-alanine 10mg and tyrosine 5mg were placed in a 100mL volumetric flask, acetonitrile / water (v / v, 1:1) was added for ultrasonic dissolution, and then mixed solvent was added to dilute to the mark to prepare Standard mixture to be tested;

[0036] 2, use mobile phase of the present invention: A phase (being aqueous phase) is formed by 0.08mmol / L phthalic acid, 10mmol / L ammonium formate, 0.2V% f...

Embodiment 2

[0039] The purpose of this example is to use the mobile phase added with phthalic acid to establish a standard curve for the determination of amino acids by the internal standard method, and to investigate the quantitative feasibility of this method.

[0040] a) Accurately weigh phenylalanine, leucine, isoleucine, tryptophan, methionine, valine, proline, glutamine, cysteine, alanine, glycine , threonine, serine, arginine, glutamic acid, asparagine, histidine, γ-aminobutyric acid, lysine, aspartic acid and citrulline, hydroxyproline, 2-amino 10 mg each of butyric acid, β-alanine, and 5 mg tyrosine were placed in a 100 mL volumetric flask, and were ultrasonically dissolved with 100 mL of acetonitrile and water in a volume ratio of 1:1, and then diluted to scale, and then diluted into standard solutions of different concentrations as required;

[0041] b) Accurately prepare an internal standard stock solution containing four isotopic amino acid internal standards, wherein: the c...

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Abstract

The invention discloses application of phthalic acid in amino acid analysis. The application comprises adding the phthalic acid in a mobile phase for performing amino acid liquid chromatography-mass spectrum combined analysis, wherein concentration of the added phthalic acid is 0.005-0.1mmol / L. According to the application of phthalic acid in amino acid analysis, detection sensitivity of the amino acid can be significantly improved, and meanwhile chromatography separating effect can be improved; the application is convenient that only adding proper amount of the phthalic acid into the mobile phase is needed, and thereby analysis result of the amino acid can be significantly improved without changing other analysis conditions; in addition, experiments show that a chromatographic column and a mass spectrum ion source are prevented from being seriously damaged by using the mobile phase added with the phthalic acid for a long time, reproducibility is good, and quantitative analysis is easy to implement, and good application and promotion values are provided.

Description

technical field [0001] The invention relates to the application of phthalic acid in amino acid analysis, in particular to the application of phthalic acid as a mobile phase additive in amino acid liquid chromatography-mass spectrometry analysis, and belongs to the technical field of amino acid analysis. Background technique [0002] Amino acids are not only the basic constituent units of proteins in living organisms, but also important precursors for the synthesis of nitrogen-containing compounds such as neurotransmitters, porphyrins, and polyamines in vivo. Recent studies have shown that amino acids also play an important regulatory role in the body, such as gene expression, protein phosphorylation, and hormone synthesis and secretion. There are two main forms of amino acids in the body, one exists in physiological fluids in a free state, such as plasma, urine, etc., and the other exists in polypeptides and proteins in a bound state. Amino acids are an important part of th...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 郭寅龙祁婉舒张立张菁吴梦茜范若静
Owner 国科新研国际技术转移有限公司
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