Method of rapidly and quantitatively analyzing 48 amino acids

An amino acid, quantitative ion technology, applied in analytical materials, measuring devices, material separation, etc., can solve the problems of many test steps, high price, poor resistance to matrix interference, etc., to improve extraction efficiency, reduce analysis costs, and improve methods. The effect of sensitivity

Active Publication Date: 2019-07-16
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
View PDF11 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, each of these methods has some defects, which limit their popularization and application in clinical laboratories: the IEX method has a long data collection time (>120min) and cannot separate important biomarkers such as methionine and homocitrulline disadvantages; the OPA/FMOC-HPLC method can only analyze more than 20 common amino acids at a time, and the data acquisition time is faster than the IEX method, but it takes 30 minutes for a sample, and the ability to resist matrix interference and isomer separation is poor, resulting in The specificity of the method is not high; the iTraq-LC-MS/MS method has the advantages of rapidity and sensitivity, but the iTraq reagent used in the method is expensive, and the method is sensitive to sulfur-containing amino acids, such as methionine, cystine, The detection of cysteine, homocysteine, homocysteine, etc. has the disadvantages of poor repeatability and low recovery (<80%)
[0005] In the early 1990s, some scholars proposed to use chloroformyl ester derivatization-gas chromatography-mass spectrometry to analyze amino acids. This method significantly improved the sensitivity and detection throughput of the method, but this method could not detect amino acids containing guanidine groups, such

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method of rapidly and quantitatively analyzing 48 amino acids
  • Method of rapidly and quantitatively analyzing 48 amino acids
  • Method of rapidly and quantitatively analyzing 48 amino acids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0113] Example 1 A method for rapid quantitative analysis of 48 amino acids

[0114] 1. Reagents

[0115]Standards of pipemidic acid (purity≥97.0%) and α-aminobutyric acid (purity≥99.0%) were purchased from Anpu Company (Shanghai, China), and other standard amino acids, trihydroxypropylphosphine and ammonium formate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Chloroformyl propyl ester, 3-picoline, isooctane and n-propanol were purchased from Tokyo Chemical Industry Co. (Tokyo, Japan). 13 C 3 - Propanol, D 3 -Methionine and D 4 - Cystathionine was purchased from Cambridge Isotope Laboratories (Andover, MA, USA).

[0116] Solution preparation

[0117] Amino acid standard stock solution: Accurately weigh each amino acid standard, dissolve with ultrapure water or 1mol / L HCl solution to volume.

[0118] Standard 1: Accurately weigh the amino acid standard as shown in Table 2, add water or 1mL of 1mol / L hydrochloric acid aqueous solution to dissolve, and prepare a...

Embodiment 2

[0163] Example 2 A kit for detecting amino acid content by derivatization HPLC-MS / MS method

[0164] 1. Composition

[0165] Derivatization reagent kits, internal standard kits, and standards

[0166] Wherein, the combination of derivatization reagents is: derivatization reagent A and derivatization reagent B prepared in Example 1;

[0167] The combination of internal standard solution is: internal standard solution A and internal standard solution B prepared in Example 1;

[0168] The standard products are: standard product 1, standard product 2 and standard product 3.

[0169] 2. How to use

[0170] (1) Preparation of calibrator

[0171] According to the method of Example 1, calibrator 1 to calibrator 7 were prepared.

[0172] (2) Sample pretreatment

[0173] With embodiment 1.

[0174] (3) Sample data collection

[0175] With embodiment 1.

Embodiment 4

[0176] Embodiment 4 detects clinical sample

[0177] 1. Test samples

[0178] The plasma of volunteers without abnormal amino acid metabolism and patients with neonatal cholestasis syndrome was selected for detection.

[0179] 2. Detection method

[0180] As described in Example 1.

[0181] 3. Test results

[0182] Test results such as figure 2 shown. The content of methionine, citrulline and lysine in the plasma of patients with neonatal cholestasis syndrome was significantly increased, and the plasma amino acid profile of volunteers without abnormal amino acid metabolism was normal.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method of rapidly and quantitatively analyzing 48 amino acids. A derivatization reagent combination for quantitatively analyzing and detecting an amino acid content by a derivatization HPLC-MS/MS method is provided, and comprises a derivatization reagent A and a derivatization reagent B; the derivatization reagent A is a mixed solution of n-propanol and 3-methylpyridine;and the derivatization reagent B is a mixed solution of dichloromethane, chloroformyl propyl ester and isooctane. Furthermore, a stable isotope internal standard solution combination, a sample pretreatment method and a liquid chromatography-tandem mass spectrometry method based on the derivatization method are established. The derivatization reagent proposed by the invention has high derivatization efficiency and low toxicity; the method requires fewer samples, the processing is simple and quick, the repeatability is good, and the detection cost is low; absolute quantitative analysis on 48 amino acids is realized by only 22 minutes, the analysis time is greatly shortened, and the detection throughput is improved; and great promotion and application value are achieved.

Description

technical field [0001] The invention relates to the technical field of biological detection, more specifically, to a method for rapid quantitative analysis of 48 kinds of amino acids. Background technique [0002] Amino acids are a class of organic compounds containing amino and carboxyl functional groups. They are the basic constituent units and metabolites of proteins, and are an important class of biomarkers. The analysis of amino acid content in human body fluids (including blood, urine, cerebrospinal fluid, and interstitial fluid) can be used for screening and auxiliary diagnosis of amino acid metabolism disorders and evaluation and monitoring of human nutritional status. [0003] Amino acid metabolism disorders often lead to changes in the levels of multiple amino acids, which poses a challenge to traditional analysis methods. The ELISA method that analyzes one compound at a time is completely unsuitable for the analysis of such compounds. In addition, the presence of...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N30/89
CPCG01N30/89
Inventor 彭敏芝刘丽
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap