Method for determining amino acid concentration and amino acid diagnosis/determination reagent kit

An amino acid and reagent technology, applied in the field of amino acid concentration determination and amino acid diagnosis/determination kits, can solve the problems of poor specificity, complicated operation methods, and high cost of instruments and equipment

Inactive Publication Date: 2008-12-17
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), spectropho...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The amino acid diagnosis / measurement reagent of this embodiment is a single reagent, including:

[0041] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0042] Stabilizer 500mmol / L

[0043] Reduced coenzyme 0.25mmol / L

[0044] Pyruvate 30mmol / L

[0045] Amino acid oxidase 10000U / L

[0046] Alanine dehydrogenase 12000U / L

[0047] 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.

[0048] 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 about 0 minutes, and the detection time is about 5 minutes.

[0049] After adding samples and reagents, let them m...

Embodiment 2

[0051] The amino acid diagnosis / measurement reagent of this embodiment is a double reagent, including:

[0052] Reagent 1

[0053] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0054] Stabilizer 50mmol / L

[0055] Reduced coenzyme 0.25mmol / L

[0056] Pyruvate 30mmol / L

[0057] Reagent 2

[0058] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0059] Stabilizer 500mmol / L

[0060] Amino acid oxidase 10000U / L

[0061] Alanine dehydrogenase 12000U / L

[0062] After the reagents are all dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0063] 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 be tested and reagent 1 and reagent 2 is 2 / 20 / 5, the reaction direction is negative reaction (downward reacti...

Embodiment 3

[0066] The amino acid diagnosis / measurement reagents of this embodiment are three reagents, including:

[0067] Reagent 1

[0068] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0069] Stabilizer 50mmol / L

[0070] Stabilizer 500mmol / L

[0071] Pyruvate 30mmol / L

[0072] Reagent 2

[0073] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0074] Stabilizer 500mmol / L

[0075] Alanine dehydrogenase 12000U / L

[0076] Reagent 3

[0077] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0078] Stabilizer 500mmol / L

[0079] Amino acid oxidase 10000U / L

[0080] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0081] When measuring amino acid concentration, 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 405...

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PUM

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Abstract

The invention relates to a kit for diagnosing/mensurating amino acid by utilizing the technologies of the enzymic colorimetric method and the enzyme linked immunosorbent assay (ELISA). The invention further relates to a method, a principle and the composition and the components of a reagent for mensurating the concentration of the amino acid, and belongs to the technology field of medical/food/industrial/agricultural/environmental inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, pyruvic acid, amino acid oxidase, alanine dehydrogenase and a stabilizer. Through mixing a sample and the reagent by a certain volume ratio, a series of enzymatic reactions occur, then the reactant is placed under an ultraviolet/visible light analyzer, and the degree/velocity of the decrease in absorbance at 340nm of the dominant wavelength is detected, thereby mensurating the concentration of the amino acid.

Description

technical field [0001] The invention relates to an amino acid diagnosis / measurement kit, and at the same time, the invention also relates to a method for determining amino acid concentration, which belongs to the technical field of medicine / food / industry / agriculture / 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 / environm...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/31C12Q1/26C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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