Liquid mass spectrometry method for detecting various amino acids in human blood plasma in underivatized mode

A technology for human body and amino acid detection, applied in the field of amino acid detection, can solve problems such as lack of time-consuming, complex operation of derivatization technology, inability to apply high-throughput analysis of clinical blood samples, etc., to reduce interference, short analysis time, meet high The effect of flux analysis

Inactive Publication Date: 2017-02-22
陈大为
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  • Claims
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Problems solved by technology

In order to solve this problem, some researchers use derivatization method to improve the ionization efficiency, so as to achieve the purpose of quantitative analysis. However,

Method used

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  • Liquid mass spectrometry method for detecting various amino acids in human blood plasma in underivatized mode
  • Liquid mass spectrometry method for detecting various amino acids in human blood plasma in underivatized mode
  • Liquid mass spectrometry method for detecting various amino acids in human blood plasma in underivatized mode

Examples

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Embodiment 1

[0057] Example 1 Liquid phase mass spectrometry method for non-derivatized detection of twenty amino acids in human plasma

[0058] Proceed as follows:

[0059] (1) Prepare the sample to be tested: Add an equal volume of 6% sulfosalicylic acid internal standard solution to the serum sample, vortex and shake at 13000rpm for 5 minutes, take 10ul of the supernatant and add 800ul of 2mM perfluoroheptane The internal standard solution of acid is uniformly mixed; the internal standard solution is mixed with 0.375uM glucosamine solution and 0.25uM S-2 aminoethylcysteine ​​solution, and the volume ratio of the two is 1:1. In this example, two samples of sample 1 and sample 2 were prepared.

[0060] (2) LC-MS / MS detection: The sample to be tested enters the chromatographic column through gradient elution mode, and the liquid chromatographic conditions are as follows:

[0061] Column: Pre-column: Thermo Fisher, 3um Hypercarb, 4.6mm*50mm, column temperature: 40℃; Analytical column: Avanced Mate...

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Abstract

The invention discloses a liquid mass spectrometry method for detecting various amino acids in human blood plasma in an underivatized mode. The method includes the following steps that firstly, the human blood plasma is obtained and subjected to deproteinization with sulfosalicylic acid, and supernate is obtained; secondly, the supernate is mixed with an internal standard solution containing a volatile ionized reagent, LC-MS/MS is used for analysis, wherein according to chromatographic conditions, 1 mM perfluoroheptanoic acid aqueous solution serves as a mobile phase A, a 1 mM perfluoroheptanoic acid acetonitrile solution serves as a mobile phase B, and gradient elution is conducted; according to mass spectrum conditions, multi-ion reaction monitoring of positive ion electrospray ionization is conducted. A volatile ionized reagent sample is added to a sample and mobile phase system, analysis time is short, analysis efficiency is improved, operation is easy, and accuracy of experiment results is improved. LC-MS/MS is used for detecting twenty amino acids in the human blood plasma, specificity, sensitivity and flux are high, operation is easy, sample processing is easy, detection time is short, and the method is particularly suitable for clinical application and popularization.

Description

Technical field [0001] The invention relates to a liquid phase mass spectrometry method for non-derivatized high-efficiency detection of multiple amino acids in human plasma, and belongs to the technical field of amino acid detection. Background technique [0002] Amines are the basis of human metabolism. They mainly represent amino acids and are the basic units of protein, which have important biochemical activities. Dipeptides are protein fragments or substances composed of amide bonds formed by the dehydration and condensation of two amino acids. They are closely related to nutrition, hormones, enzyme inhibition, immune regulation, antibacterial, antiviral, and antioxidant. In addition, studies have shown that amino acid methylation and acetylation products are closely related to human health and are an important feature of tumor occurrence and development. Therefore, the realization of metabolic profile analysis of amines has important research significance for studying the ...

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Application Information

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IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 陈大为
Owner 陈大为
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