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Monoamine oxidase diagnostic kit and method for measuring monoamine oxidase concentration

A technology of monoamine oxidase and pyruvate oxidase, which is applied in the field of medical inspection and measurement, and can solve the problems of inability to apply automatic biochemical instruments, measurement difficulties, and limited practicability

Inactive Publication Date: 2009-12-23
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, benzylamine and p-benzylamine-β-azonaphthol are mostly used as substrates for the determination of monoamine oxidase, but the p-benzylamine-β-azonaphthol benzylamine method needs to be extracted with cycloethane, which limits the practicality of the method The principle of the benzylamine method is that benzylamine generates benzaldehyde under the action of monoamine oxidase, and then reacts with dinitrophenylhydrazine under the condition of strong alkaline sodium hydroxide to generate aldehyde benzene Hydrazone is also difficult to measure on a biochemical analyzer

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The monoamine oxidase diagnostic reagent of this embodiment is a single reagent, comprising:

[0043] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0044] Stabilizer 500mmol / L

[0045] Reduced coenzyme 0.25mmol / L

[0046] Pyruvate oxidase 10000U / L

[0047] Alanine dehydrogenase 10000U / L

[0048] Amine compounds 10mmol / L

[0049] Sodium bicarbonate (carbon dioxide) 15mmol / L

[0050] Acetyl-CoA 8mmol / L

[0051] After the reagents are all dissolved and prepared, they are divided into bottles and freeze-dried to make dry powder reagents; before use, add purified water and reconstitute for use.

[0052] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test subwavelength 405nm, volume ratio of tested monoamine oxidase sample to reagent is 1 / 25, The reaction direction is negative reaction (decreasing reaction), the delay time is about 1 minute, the detect...

Embodiment 2

[0055] The monoamine oxidase diagnostic reagent of this embodiment is a double reagent, comprising:

[0056] Reagent 1

[0057]Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0058] Stabilizer 50mmol / L

[0059] Reduced coenzyme 0.25mmol / L

[0060] Amine compounds 10mmol / L

[0061] Sodium bicarbonate (carbon dioxide) 15mmol / L

[0062] Acetyl-CoA 8mmol / L

[0063] Reagent 2

[0064] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0065] Stabilizer 500mmol / L

[0066] Pyruvate oxidase 10000U / L

[0067] Alanine dehydrogenase 10000U / L

[0068] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0069] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test secondary wavelength 405nm, the volume ratio of the tested monoamine oxidase sample to reagent 1 a...

Embodiment 3

[0072] The monoamine oxidase diagnostic reagent of this embodiment is three reagents, comprising:

[0073] Reagent 1

[0074] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0075] Stabilizer 50mmol / L

[0076] Reduced coenzyme 0.25mmol / L

[0077] Amine compounds 10mmol / L

[0078] Sodium bicarbonate (carbon dioxide) 15mmol / L

[0079] Acetyl-CoA 8mmol / L

[0080] Reagent 2

[0081] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0082] Stabilizer 500mmol / L

[0083] Alanine dehydrogenase 10000U / L

[0084] Reagent 3

[0085] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0086] Stabilizer 500mmol / L

[0087] Pyruvate oxidase 10000U / L

[0088] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0089] When measuring the activity concentration of monoamine oxidase, set on the automatic biochemical analyzer: temperature 3...

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Abstract

The invention relates to a monoamine oxidase diagnostic kit utilizing technologies of an enzymatic-colorimetric method and an enzyme-link method, also relates to a method and a principle of measuring monoamine oxidase and compositions and components of reagents, and belongs to the technical field of testing and measuring of medical science. The kit mainly comprises the following compositions: buffer solution, reduced coenzyme, amine compounds, sodium bicarbonate (carbon dioxide), acetylcoenzyme A, pyruvate oxidase, alanine dehydrogenase and stabilizer; samples are mixed with the reagents in certain volume ratio to perform a series of enzymatic reactions; then reactants are placed under a UV / visible analyzer; and the descending speed of absorbance is tested at the position where dominant wave length is 340nm so as to measure and calculate the inorganic phosphorus (phosphate radical) concentration.

Description

technical field [0001] The invention relates to a diagnostic kit for monoamine oxidase, and at the same time, the invention also relates to a method for measuring the activity concentration of monoamine oxidase, which belongs to the technical field of medical examination and measurement. Background technique [0002] The determination of monoamine oxidase (MAO) can be divided into chemical spectrophotometry, fluorescence method, immunosuppressive method and enzymatic method which is now applying for a patent. Generally, chemical spectrophotometry is used. The substrates of monoamine oxidase are benzylamine and p-benzylamine-β-azonaphthol. Monoamine oxidase can also be used to catalyze the release of H from amines. 2 o 2 Oxidative chromogens such as 10-N-methylcarbamoyl-3,7-dimethylamino-10-hydro-phenothiazine (MCDP). In addition, the reaction substrates of monoamine oxidase include butylamine, amylamine, b-phenylethylamine, tyramine (3-p-hydroxyphenylethylamine 3 -parahdr...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/25C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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