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Quantitative analysis method of biogenic amine neurotransmitter

A neurotransmitter, quantitative analysis technology, applied in the field of detection, can solve the problems of high price of stable isotope labels, unfavorable promotion and application, no commercialization of metabolites, etc., to avoid matrix effects, low detection cost, and high sensitivity.

Active Publication Date: 2021-06-08
GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is expensive to equip each analyte with a corresponding stable isotope label, and most metabolites do not have commercial isotope labels, which is not conducive to the clinical application of the method

Method used

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  • Quantitative analysis method of biogenic amine neurotransmitter
  • Quantitative analysis method of biogenic amine neurotransmitter
  • Quantitative analysis method of biogenic amine neurotransmitter

Examples

Experimental program
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Effect test

preparation example Construction

[0110] In some preferred embodiments, the preparation method of the internal reference B includes the following steps:

[0111] To an aqueous solution containing homovanillic acid, 5-hydroxyindoleacetic acid, dopa, 3-methoxydopa, vanillylmandelic acid, dihydroxyphenylacetic acid, 5-hydroxytryptophan, and vanillyllactic acid, derivatization reagent C was sequentially added and derivatization reagent B, after mixing, extract with ethyl acetate, and the extract is the internal reference B;

[0112] The derivatization reagent C comprises D3-propanol and 3-picoline, and the volume ratio of D3-propanol and 3-picoline in the derivatization reagent C is (70-80):(20-30), preferably for 77:23.

[0113] Since each substance in the internal reference B is expensive, the present invention also provides a laboratory preparation method for the internal reference B. The preparation method is simple, low in cost, and the prepared isotopic internal reference is stable.

[0114] In some prefer...

Embodiment 1

[0129] A quantitative analysis method for biological amine neurotransmitters, comprising the following steps:

[0130] 1. Take 100 μL of cerebrospinal fluid sample, add 10 μL of stable isotope marker internal reference A solution, and 50 μL of saturated sodium tetraborate aqueous solution in sequence to obtain mixed solution 1.

[0131] Among them, the internal reference A is 0.5% hydrochloric acid aqueous solution, which contains the following stable isotopes of 250nmol / L: D4-dopamine, D4-3-methoxytyramine, D6-norepinephrine, D3-epinephrine, D3-metrenephrine , D3-norepinephrine.

[0132] 2. Add derivatization reagent A and derivatization reagent B to the mixed solution 1 in sequence and mix quickly to carry out the derivatization reaction to obtain the mixed solution 2;

[0133] Wherein, derivatization reagent A is the mixed solution of n-propanol and 3-picoline (volume ratio is 77:23), and derivatization reagent B is the mixed solution of dichloromethane, propyl chloroforma...

Embodiment 2

[0145] Example 2 Determination of the detection limit of 16 kinds of biological amine neurotransmitters

[0146] 1. Weigh a certain quality of 3-methoxy-4-hydroxy-phenylethylene glycol, 5-hydroxytryptamine, dopamine, 3-methoxytyramine, norepinephrine, epinephrine, metrepinephrine, normethyl Metanephrine, homovanillic acid, 5-hydroxyindoleacetic acid, dopa, 3-methoxydopa, vanillylmandelic acid, dihydroxyphenylacetic acid, 5-hydroxytryptophan and vanillyl lactic acid, each using 0.01mol / L The hydrochloric acid solution was dissolved and fixed to volume to obtain a sample solution with a concentration of 1mmol / L, and then each sample solution was diluted with 0.01mol / L hydrochloric acid solution to 0.1nmol / L, 0.2nmol / L, 0.5nmol / L, 1nmol / L, 2nmol / L, 5nmol / L to obtain sample solutions with different concentrations.

[0147] 2. Take 100 μL of the above dilutions of standard products with different concentrations, add 10 μL of 0.5% hydrochloric acid aqueous solution, and 50 μL of sa...

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Abstract

The invention provides a quantitative analysis method for biogenic amine neurotransmitters, and relates to the technical field of detection. According to the quantitative analysis method for the biogenic amine neurotransmitters, a sample is pretreated, and then the biogenic amine neurotransmitters in the sample are quantitatively analyzed by adopting a liquid chromatography-tandem mass spectrometry method. The pretreatment comprises derivatization and internal reference addition; the biogenic amine neurotransmitter is prepared from the following components: 3-methoxy-4-hydroxy-phenyl glycol, 5-hydroxytryptamine, dopamine, 3-methoxytyrosine, noradrenaline, epinephrine, methylephrine, noradrenaline, homovanillic acid, 5-hydroxyindoleacetic acid, dopa, 3-methoxydopa, vanillic mandelic acid, dihydroxyphenylacetic acid, 5-hydroxytryptophan and vanillic acid. The quantitative analysis method provided by the invention is small in required sample amount, simple, efficient, good in repeatability, low in detection cost and high in sensitivity.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a quantitative analysis method for biological amine neurotransmitters. Background technique [0002] Biogenic amine neurotransmitters and their metabolites play important roles in the central nervous system, including control of psychomotor functions, control of motor coordination, processing sensory input, maintaining emotional stability, regulating body temperature, and affecting memory, appetite, mood, and sleep. Similar to other neurotransmitters, biogenic amine neurotransmitters are released from presynaptic nerve endings in response to action potentials, bind to post-synaptic receptors and trigger biological responses (activation or inhibition of target cells). Neurotransmitters disappear in the synaptic cleft, may be hidden in vesicles and continue to function as neurotransmitters, or may be eliminated by metabolism. [0003] By detecting the concentration of biogenic a...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/02
CPCG01N30/06G01N30/02
Inventor 彭敏芝刘丽
Owner GUANGZHOU WOMEN AND CHILDRENS MEDICAL CENTER
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