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Reagent (kit) for diagnosing/determining amino acid and method for determining concentration of amino acid

A technology of amino acids and reagents, which is applied in the field of determination of amino acid concentration and amino acid diagnosis/determination reagents, which can solve the problems of complicated operation methods, high cost of instruments and equipment, and poor specificity

Inactive Publication Date: 2010-06-23
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing determination methods include methods such as chromatography, electrochemical method, instrumental analysis method (amino acid analyzer), spectrophotometer, etc., and these methods have disadvantages such as comparatively complicated operation methods, poor specificity, or high cost of instruments and equipment

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The amino acid diagnosis / measurement reagent of this embodiment is a single reagent, including:

[0045] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0046] Stabilizer 500mmol / L

[0047] Reduced coenzyme 0.25mmol / L

[0048] Amino acid oxidase 6000U / L

[0049] Pyruvate oxidase 8000U / L

[0050] N-methylalanine dehydrogenase 10000U / L

[0051] Sodium bicarbonate 6mmol / L

[0052] Acetyl-CoA 9mmol / L

[0053] Methylamine 15mmol / L

[0054] 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.

[0055] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, initial absorbance 1.8±0.7, test main wavelength 340nm, test sub-wavelength 405nm, the volume ratio of the amino acid sample to be tested is 1 / 25, The reaction direction is negative reaction (decreasing reaction), the delay time is abou...

Embodiment 2

[0058] The amino acid diagnosis / measurement reagent of this embodiment is a double reagent, including:

[0059] Reagent 1

[0060] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0061] Stabilizer 50mmol / L

[0062] Reduced coenzyme 0.25mmol / L

[0063] Sodium bicarbonate 6mmol / L

[0064] Acetyl-CoA 9mmol / L

[0065] Methylamine 15mmol / L

[0066] Reagent 2

[0067] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0068] Stabilizer 500mmol / L

[0069] Amino acid oxidase 6000U / L

[0070] Pyruvate oxidase 8000U / L

[0071] N-methylalanine dehydrogenase 10000U / L

[0072] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0073] 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 amino acid sample to b...

Embodiment 3

[0076] The amino acid diagnosis / measurement reagents of this embodiment are three reagents, including:

[0077] Reagent 1

[0078] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0079] Stabilizer 50mmol / L

[0080] Reduced coenzyme 0.25mmol / L

[0081] Sodium bicarbonate 6mmol / L

[0082] Acetyl-CoA 9mmol / L

[0083] Methylamine 15mmol / L

[0084] Reagent 2

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

[0086] Stabilizer 500mmol / L

[0087] Pyruvate oxidase 8000U / L

[0088] N-methylalanine dehydrogenase 10000U / L

[0089] Reagent 3

[0090] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0091] Stabilizer 500mmol / L

[0092] Amino acid oxidase 6000U / L

[0093] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0094] When measuring amino acid concentration, set on the automatic biochemical analyzer: temperatu...

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Abstract

The invention relates to a reagent (kit) for diagnosing / determining amino acid by using an enzyme colorimetric method and an enzyme link method, and also discloses a method for determining the concentration of the amino acid, a composition and components of the reagent, belonging to the technical field of medicine / food / environmental test determination. The reagent (kit) comprises the main components: a buffer solution, a reduced coenzyme, sodium bicarbonate (carbon dioxide), an acetoacetyl coenzyme A, an amino acid oxidase, a pyruvic oxidase, an N-methyl alanine dehydrogenase and a stabilizing agent. A sample and the reagent are mixed according to a certain volume ratio to generate a series of enzymatic reactions, and reactants are placed under an ultraviolet / visible light analyzer for detecting the degree of absorbance decrease at the dominant wavelength of 340 nm, thereby measuring and calculating the concentration of the amino acid.

Description

technical field [0001] The invention relates to an amino acid diagnosis / determination reagent (box), and simultaneously the invention also relates to a method for determining amino acid concentration, which belongs to the technical field of medicine / food / environmental inspection and determination. Background technique [0002] The determination of amino acid content is an important determination item in medicine / food / industry / agriculture / environment. Existing determination methods include chromatography, electrochemical method, instrumental analysis method (amino acid analyzer), spectrophotometer and other methods. These methods have disadvantages such as relatively complicated operation methods, poor specificity, or high equipment costs. [0003] Amino acid is not a simple substance, 17 kinds of amino acids can be directly measured with an amino acid analyzer (the instrument is expensive and cannot be widely used). For medicine / food / industry / agriculture / environment, in most...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/17G01N33/48C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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