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Method for detecting catecholamines in blood plasma by using liquid chromatography-tandem mass spectrometry

A technology of tandem mass spectrometry and liquid chromatography, which is applied in the field of blood detection and analysis, can solve problems such as low accuracy, poor anti-interference ability, and easy to be affected by external unfavorable factors, so as to reduce polarity, eliminate potential negative ions, and improve ionization efficiency. The effect of chemical efficiency

Pending Publication Date: 2020-09-29
浙江湖州三体生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has poor anti-interference ability and low accuracy, and the measurement results are easily affected by external adverse factors.

Method used

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  • Method for detecting catecholamines in blood plasma by using liquid chromatography-tandem mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Preparation of test blood samples: collect the whole blood to be tested and healthy whole blood, centrifuge with a low-temperature centrifuge, and store the separated plasma in the dark at -80°C to obtain the plasma to be tested, healthy plasma, and low-temperature centrifuge The centrifugal force is 1000g, the temperature during centrifugation is 4°C, and the centrifugation time is 10min;

[0020] (2) Prepare internal standard working solution: dilute the mixed solution of DHBA (dihydroxybenzoic acid), MN-D3, VMA-D3 mother solution with acetonitrile, vortex and mix for 30s, the concentration of internal standard working solution is 35.51nmol / L, after dilution Then put it into 4°C to obtain the internal standard working solution, and keep it in the refrigerator for later use;

[0021] (3) Take the standard mother solution of DA (which can be purchased on the market), dilute the standard mother solution with standard diluent, and dilute it into standard working solut...

Embodiment 2

[0029] (1) Preparation of test blood samples: collect the whole blood to be tested and healthy whole blood, centrifuge with a low-temperature centrifuge, and store the separated plasma in the dark at -80°C to obtain the plasma to be tested, healthy plasma, and low-temperature centrifuge The centrifugal force is 1200g, the temperature during centrifugation is 4°C, and the centrifugation time is 8min;

[0030] (2) Prepare internal standard working solution: dilute the mixed solution of DHBA (dihydroxybenzoic acid), MN-D3, VMA-D3 mother solution with acetonitrile, vortex and mix for 30s, the concentration of internal standard working solution is 42.84nmol / L, after dilution Then put it into 4°C to obtain the internal standard working solution, and keep it in the refrigerator for later use;

[0031] (3) Take the standard mother solution of epinephrine (which can be purchased in the market), dilute the standard mother solution with standard diluent, and dilute it into standard worki...

Embodiment 3

[0038] (1) Preparation of test blood samples: collect the whole blood to be tested and healthy whole blood, centrifuge with a low-temperature centrifuge, and store the separated plasma in the dark at -80°C to obtain the plasma to be tested, healthy plasma, and low-temperature centrifuge The centrifugal force is 1100g, the temperature during centrifugation is 4°C, and the centrifugation time is 9min;

[0039] (2) Prepare internal standard working solution: dilute the mixed solution of DHBA (dihydroxybenzoic acid), MN-D3, VMA-D3 mother solution with acetonitrile, vortex and mix for 30s, the concentration of internal standard working solution is 40nmol / L, after dilution Place at 4°C to obtain the internal standard working solution, and store in the refrigerator for later use;

[0040] (3) Take the standard mother solution of norepinephrine (which can be purchased on the market), dilute the standard mother solution with standard diluent, and dilute it into standard working solutio...

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Abstract

The invention provides a method for detecting catecholamines in blood plasma by using liquid chromatography-tandem mass spectrometry, which belongs to the technical field of blood detection and analysis. The method comprises the steps of (1) preparing a test blood sample; (2) preparing an internal standard working solution; (3) taking the standard mother liquor of the to-be-detected substance, anddiluting the standard mother liquor with a standard diluent; (4) taking out the internal standard working solution, and preparing the following solutions: a standard curve sample, a substrate samplesolution, a test sample and a quality control sample; (5) transferring the supernatant of the standard curve sample, the substrate sample solution, the test sample and the quality control sample intoan EP tube by using a pipette, and carrying out freeze drying; (6) adding a NaOH solution and a dansyl chloride acetonitrile solution, and carrying out a derivative reaction; and (7) adding a reactionterminator, and carrying out liquid chromatography-mass spectrometry in a pore plate. According to the invention, potential anions are eliminated, the ionization efficiency in a positive ion mode isimproved, potential solvents or other interfering substances can be reduced, meanwhile, the polarity of an analyte is reduced, and the detection sensitivity is improved.

Description

technical field [0001] The invention belongs to the technical field of blood detection and analysis, and in particular relates to a method for detecting catecholamines in plasma by using liquid chromatography tandem mass spectrometry. Background technique [0002] Mass spectrometry uses electric and magnetic fields to move moving ions (charged atoms, molecules or molecular fragments, including molecular ions, isotopic ions, fragment ions, rearranged ions, multiply charged ions, metastable ions, negative ions and ion-molecular The ions produced by the interaction) are separated according to their mass-to-charge ratio and then detected. The exact mass of the ion is determined to determine the compound composition of the ion. This is due to the fact that the exact mass of a nuclide is one decimal place, there will never be two nuclides with the same mass, and there will never be a nuclide whose mass is exactly an integer multiple of the mass of another nuclide. Analysis of th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/7233G01N30/34G01N2030/042G01N2030/067
Inventor 薛雪陈虎诚陈效张桉瑜郑国艳
Owner 浙江湖州三体生物科技有限公司
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