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High performance liquid chromatography tandem mass spectrum detection method of free catecholamine and metabolites in human urine

A technology of high performance liquid chromatography and tandem mass spectrometry, which is used in measurement devices, material separation, instruments, etc., to simplify the operation process of pretreatment extraction and derivatization, reduce interference, and reduce interference and influence.

Pending Publication Date: 2019-05-03
杭州凯莱谱精准医疗检测技术有限公司
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Problems solved by technology

[0010] The purpose of the present invention is to solve some of the problems existing in the detection methods of catecholamines and their metabolites in human urine, and provide a high performance liquid chromatography tandem mass spectrometry detection method for free catecholamines and their metabolites in human urine. Strong specificity, high accuracy, high sensitivity, low detection cost, easy operation, high analysis efficiency and high throughput, etc.

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  • High performance liquid chromatography tandem mass spectrum detection method of free catecholamine and metabolites in human urine
  • High performance liquid chromatography tandem mass spectrum detection method of free catecholamine and metabolites in human urine
  • High performance liquid chromatography tandem mass spectrum detection method of free catecholamine and metabolites in human urine

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[0033] Example: High performance liquid chromatography tandem mass spectrometry detection method for free catecholamines and their metabolites in human urine

[0034] 1. Solution configuration:

[0035] Standard working solution configuration: Accurately weigh NE, E, DA, NMN and MN standard products, add 0.5% sodium metabisulfite solution to dissolve to obtain a primary stock solution with a concentration of 10 mg / mL, and then further dilute with 0.1% formic acid solution to obtain 1 mg / mL mL of secondary stock solution; put an appropriate amount of 0.1% formic acid solution into a 2mL centrifuge tube, then add appropriate amount of NE, E, DA, NMN and MN secondary stock solution to the centrifuge tube, vortex for 10s to mix uniformly, to obtain a mixed third-level stock solution with a concentration of NE of 160 μg / mL, a concentration of E of 8 μg / mL, a concentration of DA of 160 μg / mL, a concentration of NMN of 160 μg / mL, and a concentration of MN of 80 μg / mL. 0.1% formic ac...

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Abstract

The invention discloses a high performance liquid chromatography tandem mass spectrum detection method of free catecholamine and metabolites in human urine. The method comprises the following steps of(1) preprocessing a sample, successively adding urine samples in an internal standard working solution and a phosphoric acid solution, and carrying out vortex mixing; (2) carrying out weak cation exchange solid phase extraction 96-well plate (SPE96 well plate) activation; (3) carrying out solid phase extraction processing, after step (1) processing, loading the samples to the SPE 96-well plate after step (2) activation, carrying out formic acid methanol solution elution, and drying eluant with nitrogen; and (4) redissolving and using a high performance liquid chromatography tandem mass spectrometry to carry out analysis detection. The method has the advantages of high accuracy, high reproducibility, high sensitivity, high specificity, high efficiency, high flux, low cost, simple and rapidoperation and the like.

Description

technical field [0001] The invention relates to the technical field of analysis and detection, in particular to a high-accuracy, high-reproducibility, high-sensitivity, high-specificity, high-efficiency, high-throughput, low-cost, simple and quick operation of free catecholamines in human urine and its A method for the detection of metabolites by high-performance liquid chromatography tandem mass spectrometry. Background technique [0002] Catecholamines are catechol (catechol) amine compounds, including dopamine, norepinephrine, epinephrine and their derivatives. Norepinephrine and epinephrine are both hormones secreted by the adrenal medulla and neuromediators of noradrenergic fibers in the sympathetic and central nervous systems. Noradrenaline is widely distributed in the central nervous system, and its content is relatively large, while that of epinephrine is relatively small. Dopamine is mainly concentrated in the extrapyramidal system and is also a neurotransmitter. ...

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Application Information

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IPC IPC(8): G01N30/89
Inventor 马金飞冯加翼康洲阳刘华芬
Owner 杭州凯莱谱精准医疗检测技术有限公司
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