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Method for rapidly detecting activity of catechol-oxygen-methyltransferase (COMT) in biological sample by capillary electrophoresis

A technology of catecholoxymethyl and capillary electrophoresis, which is applied in the direction of measuring devices, material analysis through electromagnetic means, instruments, etc., to achieve the effects of good separation, high analysis speed and fast analysis speed

Inactive Publication Date: 2015-05-27
JIANGSU INST OF NUCLEAR MEDICINE
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  • Abstract
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  • Application Information

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Problems solved by technology

[0006] The present invention uses the established capillary electrophoresis method to study the change of PQQ on the COMT activity in the brain of depression model mice caused by chronic stress-induced oxidative damage, and there is no literature report

Method used

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  • Method for rapidly detecting activity of catechol-oxygen-methyltransferase (COMT) in biological sample by capillary electrophoresis
  • Method for rapidly detecting activity of catechol-oxygen-methyltransferase (COMT) in biological sample by capillary electrophoresis
  • Method for rapidly detecting activity of catechol-oxygen-methyltransferase (COMT) in biological sample by capillary electrophoresis

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Experimental program
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Embodiment 1

[0040] 1. Sample preparation:

[0041] Brain Tissue Processing:

[0042] Accurately weigh the brain tissue, add lysate (brain tissue weight: tissue lysate = 1g:7.5mL), homogenate, centrifuge at 14000g at 4°C for 15min, take supernatant A and centrifuge at 14000g for 15min, take the supernatant after the second centrifugation Liquid B-80 ℃ refrigerator reserved.

[0043] Before the test, thaw the supernatant B at low temperature, and then add an equal volume of PCA precipitant (that is, containing 1.2M K 2 HPO 4and 2mM EDTA disodium solution), placed in an ice bath for 10min, centrifuged at 14000g at 4°C for 15min, and the supernatant C was taken. Afterwards, the enzyme reaction system was incubated, 2mM magnesium chloride 100μL + 0.2mM SAM 100μL + 0.2mM 3,4-dihydroxybenzoic acid 150μL + the final centrifuged supernatant C 150μL, the total volume of the reaction system was 500μL, vortexed at 37 Incubate at ℃ for 60min. Afterwards, 80 μL of ice-cold 2M PCA was added to term...

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Abstract

The invention relates to a method for rapidly detecting the activity of catechol-oxygen-methyltransferase (COMT) in a biological sample by capillary electrophoresis, belonging to the technical fields of capillary electrophoresis detection and COMT detection. The method is used for analyzing the change of the activity of the COMT in cerebral tissues of a mouse which has chronic stress-induced oxidative damage depression and is treated by intraperitoneal injection with folic acid and different doses of pyrroloquinoline quinone (PQQ), is not reported in literature, and provides a new basis for the pathogenesis and drug action target point of the depression. The capillary electrophoresis adopted by the method is higher in analysis speed and better in separation degree, and is capable of rapidly detecting the active concentration of the COMT in the cerebral tissues; compared with the existing method for detecting the COMT, the method has the obvious advantage of not needing derivatization; compared with the methods in the literature, the method is better in reliability and selectivity and has the characteristics of high sensitivity, high analysis speed and the like.

Description

technical field [0001] The invention discloses a method for quickly detecting the activity of catecholoxymethyltransferase COMT in biological samples by capillary electrophoresis, which belongs to the technical fields of capillary electrophoresis detection and catecholoxymethyltransferase detection. Background technique [0002] Catechol Oxymethyltransferase (COMT) widely exists in the nervous system and body tissues, and is one of the main metabolic enzymes of catecholamine neurotransmitters. COMT is the main regulator of dopamine levels in the prefrontal cortex of the brain. It has been reported that depression is related to the dysfunction of dopamine in the brain. One of the methods to evaluate the function of dopamine in the brain is to measure the active concentration of COMT. [0003] With the rapid development of society and the increase of work, life and mental pressure, the incidence of depression is increasing year by year. According to the statistics of the Wor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
Inventor 周杏琴毛师师钦晓峰
Owner JIANGSU INST OF NUCLEAR MEDICINE
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