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Method for detecting 11 neurotransmitters in brain microdialysis fluid by ultra-high performance liquid chromatography-mass spectrometry

A neurotransmitter and microdialysate technology, applied in the field of analytical chemistry, can solve the problems of limited and single neurotransmitter types and numbers, and achieve the effects of reducing matrix effect, improving sensitivity and good peak shape

Active Publication Date: 2018-11-13
SHANGHAI UNIV OF T C M
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biological samples for the qualitative and quantitative detection of neurotransmitters by UPLC-MS / MS are all from tissue homogenate, and the types and numbers of neurotransmitters detected are also limited. For accurate quantification, amino acid and monoamine neurotransmitters still need Derivatized, and can only detect a single type of neurotransmitter
[0008] Methods for qualitative and quantitative detection of neurotransmitters using dialysate as a biological sample have also been reported, but they can only be used for the detection of a single type of neurotransmitter

Method used

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  • Method for detecting 11 neurotransmitters in brain microdialysis fluid by ultra-high performance liquid chromatography-mass spectrometry
  • Method for detecting 11 neurotransmitters in brain microdialysis fluid by ultra-high performance liquid chromatography-mass spectrometry
  • Method for detecting 11 neurotransmitters in brain microdialysis fluid by ultra-high performance liquid chromatography-mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065]The purpose of this example is to establish a detection method for the determination of 3 types of 11 neurotransmitters in brain dialysate based on liquid chromatography-mass spectrometry. 3 types of 11 neurotransmitters include: choline neurotransmitters include acetylcholine (Ach); amino acid neurotransmitters include glycine (Gly), glutamic acid (Glu), glutamine (Gln), γ-aminobutyric acid acid (γ-GABA), aspartic acid (Asp), and histamine (Hist); monoamine neurotransmitters include norepinephrine (NE), serotonin (5-HT), 5-oxindole Acetate (5-HIAA) and Dopamine (DA).

[0066] Preparation of standard solution: Precisely weigh 11 kinds of neurotransmitter reference substances, dissolve glutamic acid and aspartic acid in ultrapure water, and dissolve the remaining neurotransmitter reference substances in methanol, and prepare 10mM single-standard stock solutions respectively. Store at -20°C. Use acetonitrile-water (4:1, V / V) to prepare the total mixed standard solution (...

Embodiment 2

[0094] The purpose of this example is to use the established detection method to study the relationship between the in vitro probe recovery rate and perfusion velocity of 11 neurotransmitters.

[0095] Preparation of mixed standard solution: take the single standard stock solution of each neurotransmitter under the item "Preparation of Standard Solution" in Example 1, and use acetonitrile-water (4:1, V / V) to prepare each neurotransmitter with a concentration of 1.0 μM mixed standard solution.

[0096] Using the artificial cerebrospinal fluid prepared in Example 1 as the perfusion fluid, investigate the in vitro probes of the mixed standard solutions with the concentration of each neurotransmitter being 1.0 μM at the perfusion rates of 3.0 μL / min, 2.0 μL / min, and 1.0 μL / min respectively (Probe membrane length 1mm) Recovery rate (n=3). According to the formula R=C s / C m *100 calculates the probe recovery rate, and the results are shown in Table 5, C m Represents a mixed sta...

Embodiment 3

[0100] The purpose of this example is to use the established detection method to study the relationship between the in vitro probe recovery rate of 11 neurotransmitters and the concentration level of neurotransmitters.

[0101] Preparation of mixed standard solution: take the single standard stock solution of each neurotransmitter under the item "Preparation of Standard Solution" in Example 1, and prepare the concentration of each neurotransmitter with acetonitrile-water (4:1, V / V) to be 0.5 Mixed standard solution of μM, 1.0μM, 2.0μM.

[0102] The artificial cerebrospinal fluid prepared in Example 1 was used as the perfusate, and the investigation of each neurotransmitter concentration was respectively 0.5 μ M, 1.0 μ M, 2.0 μ M of the mixed standard solution at a perfusion speed of 1.0 μ L / min. Probe in vitro (probe membrane length 1mm ) recovery rate (n=3), a sample was collected every 30 min. According to the formula R=C s / C m *100 calculates the probe recovery rate, an...

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Abstract

The invention belongs to the field of analytical chemistry, and particularly relates to a method for detecting 11 neurotransmitters in brain microdialysis fluid by ultra-high performance liquid chromatography-mass spectrometry. The method comprises the following steps: preparing a brain microdialysis sample solution by diluting brain microdialysis fluid with an acetonitrile-water solution or an acetonitrile-formic acid solution, and performing qualitative and / or quantitative detection by using the ultra-high performance liquid chromatography-mass spectrometry / mass spectrometry. The method canrealize real-time qualitative and quantitative detection of 11 neurotransmitters by one time through an ultra-high performance liquid chromatography-mass spectrometry / mass spectrometry system only bysimply diluting the brain microdialysis fluid, the detection result is controllable, accurate and stable, and the method has good usability, and is short in analysis time. The method overcomes the defects of complicated processing, long time consuming, low accuracy and single data caused by various neurotransmitter types, low neurotransmitter content, large matrix effect and the like, and has a prospective clinical application prospect.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, and in particular relates to a method for detecting 11 kinds of neurotransmitters in brain microdialysate by ultra-high performance liquid mass spectrometry. Background technique [0002] Neurotransmitter refers to a class of chemical substances that are synthesized by presynaptic neurons and released at the end, can specifically act on receptors on postsynaptic neurons or effector cells, and can play a role in information transmission. Neurotransmitters mainly include amino acids such as γ-aminobutyric acid, glutamic acid, and glycine, monoamines such as dopamine, epinephrine, and norepinephrine, cholines such as acetylcholine and choline, and peptides. [0003] There are a large number of neurotransmitters in the central brain system, and changes in their content have an important impact on the behavior of organisms, and are closely related to many diseases, including depression, Alzheimer's...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/72G01N30/60
CPCG01N30/6039G01N30/72
Inventor 潘凌云贾益群王雨刘新华
Owner SHANGHAI UNIV OF T C M
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