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Reagent for HPLC detection of concentration of catecholamines in sample

A technology for detecting catecholamines and samples, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of deviation of test results and consumption of EDTA, and achieve the effects of cost reduction, accuracy improvement and detection accuracy

Active Publication Date: 2014-09-24
广州艾迪康医学检验所有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, blood and urine contain metal ions, which will combine with EDTA in the mobile phase and consume EDTA in the mobile phase, resulting in deviations in the detection results

Method used

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  • Reagent for HPLC detection of concentration of catecholamines in sample
  • Reagent for HPLC detection of concentration of catecholamines in sample
  • Reagent for HPLC detection of concentration of catecholamines in sample

Examples

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Embodiment 1

[0048] Example 1 Reagent for detecting the concentration of catecholamines in a sample by HPLC

[0049] The reagent that is used for HPLC detection catecholamine concentration in the sample, described reagent comprises mobile phase, is used for the activated alumina of sample pretreatment, 0.2M HCl solution and 0.01M EDTA-2Na solution:

[0050] In the mobile phase of every 1000ml, 4.1g of anhydrous sodium acetate, 0.05g of EDTA-2Na, 70~90ml of acetonitrile,

[0051] Sodium octane sulfonate 0.08~0.10g, glacial acetic acid 7.5ml, the rest is ddH 2 O, pH 4.0-4.3. Wherein the volume of acetonitrile is preferably 80ml, the amount of sodium octanesulfonate is preferably 0.08g, and the pH value of the mobile phase is preferably 4.0. When the sample is urine, the reagent also includes concentrated hydrochloric acid for sample pretreatment. The concentrated hydrochloric acid preferably has a mass fraction of 37.5% HCl solution.

[0052] In addition, the reagents may also include ca...

Embodiment 2

[0056] Embodiment 2 blood pretreatment method

[0057] (1) Add 20-25 mg of activated alumina (purchased from Sigma-Aldrich) into a test tube, and then add 400 μl of Tris-HCl buffer.

[0058] (2) Add 5 μl of 0.01M EDTA-2Na solution to 0.5ml of blood at a volume ratio of 100:1, centrifuge at 3500rpm for 10min, collect plasma, and store at 4°C until use.

[0059] (3) Add the plasma collected in (2) into the test tube in (1), shake it for 5 minutes, then centrifuge at 3500rpm for 10 minutes, and remove the supernatant;

[0060] (4) Add ddH to the test tube 2 O washing, then shake for 2 min, centrifuge at 3500 rpm for 3 min, remove the supernatant, repeat 4 times, then centrifuge at 3500 rpm for 5 min, and dry the water in the test tube with a cotton swab;

[0061] (5) Add 200 μl of 0.2M HCl to the test tube, shake for 5 minutes, centrifuge at 3500 rpm for 10 minutes, take 10 μl of the supernatant as the sample solution, and prepare for testing on the machine.

Embodiment 3

[0062] Example 3 Urine Pretreatment Method

[0063] (1) Add 20-25 mg of activated alumina (purchased from Sigma-Aldrich Company) into a test tube, and then add 400 μl of Tris-HCl buffer.

[0064] (2) Add 20ml of hydrochloric acid solution with a mass fraction of 37.5% to 0.2ml of urine at a volume ratio of 1:100, and at the same time add 2μl of 0.01M EDTA-2Na solution at a volume ratio of 100:1, centrifuge at 3500rpm for 10min, and collect supernatant.

[0065] (3) Take 200 μl of the supernatant collected in (2) and add it to the test tube in (1), shake it for 5 minutes, then centrifuge at 3500 rpm for 10 minutes, and remove the supernatant;

[0066] (4) Add ddH to the test tube 2 O washing, then shake for 2 min, centrifuge at 3500 rpm for 3 min, remove the supernatant, repeat 4 times, then centrifuge at 3500 rpm for 5 min, and dry the water in the test tube with a cotton swab;

[0067] (5) Add 200 μl of 0.2M HCl to the test tube, shake for 5 minutes, centrifuge at 3500 rpm f...

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Abstract

The invention discloses a reagent for the HPLC detection of concentration of catecholamines in a sample. The concentration of catecholamines in the sample can be rapidly, accurately and highly sensitively detected by optimizing a sample pretreatment method and regulating parameters such as a formula of a flowing phase, the flow velocity of the flowing phase, an elution process of the flowing phase, column temperature, reference voltage, working voltage and the voltage of a protection pool.

Description

technical field [0001] The invention relates to a reagent for detecting the concentration of catecholamines in a sample, in particular to a reagent for detecting the concentration of catecholamines in blood or urine by HPLC. Background technique [0002] Catecholamines (CAs) are a class of neurological substances containing catechol and amine groups, and also an important signal transduction substance in the human body, all of which are catalyzed by β-hydroxylase in the chromaffin cells of the adrenal medulla. Synthesized from phenylalanine and tyrosine. The most important catecholamines are dopamine (DOP), adrenaline (ADR) and norepinephrine (noradrenaline, NOR). Many psychostimulants are also analogs of catecholamines. [0003] 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 阳愿望霍玉美潘超单洪波
Owner 广州艾迪康医学检验所有限公司
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