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D-lactic acid diagnosis/determination reagent kit and method for determining D-lactic acid concentration

A kit, lactic acid technology, applied in the field of medical/food testing and determination, can solve problems such as microbial contamination

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

If D-lactic acid is present, microbial contamination may be the cause

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The D-lactic acid diagnosis / measurement reagent of this embodiment is a single reagent, including:

[0046] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0047] Stabilizer 500mmol / L

[0048] Reduced coenzyme 0.25mmol / L

[0049] 2 Ferricytochrome c 10mmol / L

[0050] Ammonia ion 20mmol / L

[0051] D-lactate dehydrogenase 10000U / L

[0052] Alanine dehydrogenase 10000U / L

[0053] Glycine oxidase 10000U / 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 secondary wavelength 405nm, the volume ratio of the measured D-lactic acid sample to the reagent is 1 / 25. The reaction direction is negative reaction (falling reaction), the delay time is about 1 minu...

Embodiment 2

[0058] The MMM diagnosis / measurement reagent of the present embodiment is double reagent, comprises:

[0059] Reagent 1

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

[0061]Stabilizer 50mmol / L

[0062] Reduced coenzyme 0.25mmol / L

[0063] 2 Ferricytochrome c 10mmol / L

[0064] Ammonia ion 20mmol / L

[0065] Reagent 2

[0066] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0067] Stabilizer 500mmol / L

[0068] D-lactate dehydrogenase 10000U / L

[0069] Alanine dehydrogenase 10000U / L

[0070] Glycine oxidase 10000U / L

[0071] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid double reagents, which can be used directly.

[0072] 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 of the measured D-lactic acid sample and reagent 1, reagent 2 Th...

Embodiment 3

[0075] The D-lactic acid diagnosis / measurement reagent of the present embodiment is three reagents, comprising:

[0076] Reagent 1

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

[0078] Stabilizer 50mmol / L

[0079] Reduced coenzyme 0.25mmol / L

[0080] 2 Ferricytochrome c 10mmol / L

[0081] Ammonia ion 20mmol / L

[0082] Reagent 2

[0083] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0084] Stabilizer 500mmol / L

[0085] Alanine dehydrogenase 10000U / L

[0086] Glycine oxidase 10000U / L

[0087] Reagent 3

[0088] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0089] Stabilizer 500mmol / L

[0090] D-lactate dehydrogenase 10000U / L

[0091] After all the reagents are dissolved and prepared, they are divided into bottles to make liquid three reagents, which can be used directly.

[0092] When measuring the concentration of D-lactic acid, set it on the automatic biochemical analyzer: temperature 37°C...

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Abstract

The invention relates to a kit for diagnosing / mensurating D-lactic acid by utilizing the technologies of the enzymatic cycling amplification method, the enzymic colorimetric method and the enzyme linked immunosorbent assay (ELISA) based on the specificity of the D-lactic acid. The invention further relates to a method, a principle and the composition and the components of a reagent for mensurating the concentration of the D-lactic acid, and belongs to the technology field of medical / food inspection and measurement. The main components of the kit include a buffer solution, reduced coenzyme, 2-ferricytochrome-c, ammonia ions, D-lactate dehydrogenase, alanine dehydrogenase, glycine oxidase 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 D-lactic acid.

Description

technical field [0001] The invention relates to a D-lactic acid diagnosis / measurement kit utilizing D-lactic acid specificity, and also relates to a method for measuring D-lactic acid concentration, which belongs to the technical field of medical / food inspection and determination. Background technique [0002] L-lactic acid is an organic acid that has attracted much attention and developed rapidly in recent years. At the same time, L-lactic acid has application prospects in many fields such as food, feed, medicine, plastics, feed, pesticide, daily chemical industry, papermaking and electronic industry. [0003] L-lactic acid is the end product of the metabolism of most organisms. Only a few microorganisms (such as Lactobacillus, Leuconostoc) can form D-lactic acid. Great care is taken in the production of yogurt to detect lactate to assess whether the microbes are producing L-lactic acid, D-lactic acid or both. If D-lactic acid is present, microbial contamination may be t...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N21/31C12Q1/26C12Q1/00
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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