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All-isotope internal standard mass spectrometry quantitative method for neurotransmitter metabolite

A neurotransmitter and isotope labeling technology, applied in the field of neurotransmitter metabolite mass spectrometry, can solve the problems of low accuracy and low sensitivity, achieve high specificity, mild reaction conditions, and improve the effect of detection and quantification

Inactive Publication Date: 2017-07-18
上海谱领生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for quantifying neurotransmitter metabolites with full isotope internal standard mass spectrometry to solve the existing multiple problems related to the metabolism of neurotransmitters such as tryptophan pathway, γ-aminobutyric acid pathway, and catecholamine pathway. There are problems of low sensitivity and low accuracy in compound detection and quantification

Method used

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  • All-isotope internal standard mass spectrometry quantitative method for neurotransmitter metabolite
  • All-isotope internal standard mass spectrometry quantitative method for neurotransmitter metabolite
  • All-isotope internal standard mass spectrometry quantitative method for neurotransmitter metabolite

Examples

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Effect test

Embodiment 1

[0031] The purpose of this embodiment is to establish a UPLC-MS / MS quantitative method for isotope reagent derivatization of neurotransmitter compounds, such as Figure 1-3 shown.

[0032] Prepare 50% acetonitrile solution (mixed standard mother solution) of compound standard substances of tryptophan pathway, γ-aminobutyric acid pathway and catecholamine pathway, dilute, take two parts of 100 microliters of dilution, blow dry with nitrogen, add 30 microliters respectively (The range is 20-50 microliters, the value of the following range is expressed by this method) The concentration is 100mM (50-200mM) sodium tetraborate buffer solution, and then add 30 microliters (20-50 microliters) of 5% benzyl Stable isotope-labeled derivatization reagents and non-labeled derivatization reagents of the acid chloride solution (volume ratio), reacted at 30°C for 25 minutes, and obtained isotope-labeled derivatization products and non-labeled derivatization products of tryptophan pathway, γ-a...

Embodiment 2

[0036] The purpose of this example is to use the UPLC- / MS / MS quantitative method of the established tryptophan pathway, γ-aminobutyric acid pathway, and catecholamine pathway compounds to conduct semi-quantitative analysis of mouse brain tissue, providing important information for scientific research. research data.

[0037] Take neurotransmitter metabolites mixed standard mother liquor acetonitrile solution, dilute, take 50 microliters, blow dry with nitrogen, first add 100 microliters (50-200 microliters) of sodium tetraborate buffer solution (pre-cooled), then add 100 microliters Microliter (50-200 microliter) stable isotope-labeled derivatization reagent D 5 -Benzoyl chloride, conduct derivatization reaction, heat treatment at 50°C for 30 seconds to obtain isotope-labeled derivatives of neurotransmitter metabolites, dilute the isotope-labeled derivatives of neurotransmitter metabolites by 15 times to obtain stable isotopes Labeled internal standard stock solution.

[003...

Embodiment 3

[0041] The purpose of this example is to quantitatively analyze human plasma samples using the established UPLC- / MS / MS quantitative method for compounds in the tryptophan pathway, γ-aminobutyric acid pathway, and catecholamine pathway.

[0042] Take 60 microliters of the acetonitrile solution of the mother solution of the mixed standard of neurotransmitter metabolites, first add 100 microliters (50-200 microliters) of sodium tetraborate buffer solution, and then add the stable isotope-labeled derivatization reagent 13 C 6 - Benzoyl chloride, carry out acylation derivatization reaction, heat treatment at 10°C for 30 minutes, and obtain neurotransmitter metabolites 13 C 6 - Benzoyl chloride acylation derivative product, the isotope-labeled derivative product of neurotransmitter metabolites is diluted 15 times to obtain a stable isotope-labeled internal standard mother solution.

[0043] Take cryopreserved human plasma samples, thaw, shake and mix for a short time, take 30 micr...

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Abstract

The invention discloses an all-isotope internal standard mass spectrometry quantitative method for a neurotransmitter metabolite. The method is an all-isotope derivative internal standard quantitative method for the neurotransmitter metabolite by adopting a neurotransmitter metabolite standard substance and a derivative of a stable isotope labeling reagent as quantitative internal standard references and a liquid chromatography-tandem quadrupole mass spectrometry as detection means. Qualitative and quantitative analysis can be carried out on a plurality of neurotransmitter metabolites at the same time, so that the all-isotope internal standard mass spectrometry quantitative method has the advantages of being simple in sample preparation, accurate in qualitative and quantitative analysis, high in throughput and low in cost, and has an expectable clinical application prospect. According to the method disclosed by the invention, the defects that a current neurotransmitter metabolite test is low in qualitative and quantitative accuracy and high in cost and only a few of substances can be analyzed at the same time are compensated.

Description

technical field [0001] The invention relates to a method for quantifying neurotransmitter metabolite mass spectrum, and furthermore, to a method for quantifying neurotransmitter metabolite all-isotope internal standard mass spectrum. Background technique [0002] Neurotransmitters are an important class of biologically active molecules that maintain various physiological functions in the brain. Changes in their content are closely related to many neurological disorders such as depression, Parkinson's disease, schizophrenia and other diseases. The metabolism of such substances mainly involves tryptophan pathway, γ-aminobutyric acid pathway and catecholamine pathway. Quantitative research on the composition and content changes of such substances is of great significance for neuroscience research and clinical diagnosis. At present, the high performance liquid chromatography-electrochemical detection method is mainly used to detect neurotransmitter substances. This method can ...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N27/62
CPCG01N27/62G01N30/02G01N30/06G01N30/72
Inventor 占同文
Owner 上海谱领生物科技有限公司
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