Method for detecting blood ammonia content and blood ammonia diagnostic reagent kit

A diagnostic kit and blood ammonia technology are applied in the field of medical testing and determination, and can solve the problem of not finding a blood ammonia determination method and the like

Inactive Publication Date: 2006-03-22
王尔中
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Searching Chinese patents, only found that patent application No. 87105593.7 discloses a quick-frozen cup for blood ammonia measurement, but no ideal method for blood ammonia measurement was found

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Embodiment one (single agent)

[0087] The blood ammonia diagnostic kit of the present embodiment comprises:

[0088] Buffer 80mmol / l

[0089] Adenosine triphosphate 2mmol / l

[0090] Glutamic acid 6mmol / l

[0091] Pyruvate 5mmol / l

[0092] Phosphoenolpyruvate 2mmol / l

[0093] Reduced coenzyme 0.2mmol / l

[0094] Glutamine Synthetase 6000U / l

[0095] Pyruvate oxidase 6000U / l

[0096] Phosphoenolpyruvate Carboxylase 4000U / l

[0097] Malate dehydrogenase 8000U / l

[0098] Stabilizer 50% (total volume)

[0099] Set on the automatic biochemical analyzer: temperature 37°C, reaction time 10 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the measured blood ammonia sample to reagent is 1 / 25, and the reaction direction is negative reaction .

[0100] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0101] Ammonia + glutamic acid + adenosine triphosphate glutamine synthetase Glutami...

Embodiment 2

[0111] Embodiment two (two doses)

[0112] The blood ammonia diagnostic reagent of the present embodiment has:

[0113] Reagent I

[0114] Buffer 100mmol / l

[0115] Adenosine triphosphate 7mmol / l

[0116] Glutamic acid 8mmol / l

[0117] Pyruvate 10mmol / l

[0118] Phosphoenolpyruvate 5mmol / l

[0119] Reduced coenzyme 0.25mmol / l

[0120] Pyruvate oxidase 9000U / l

[0121] Phosphoenolpyruvate Carboxylase 7000U / l

[0122] Malate dehydrogenase 12000U / l

[0123] Stabilizer 50% (total volume)

[0124] Reagent II

[0125] Buffer 100mmol / l

[0126] Glutamine Synthetase 9000U / l

[0127] Stabilizer 50% (total volume)

[0128] When measuring blood ammonia content, the temperature is controlled at 30°C, the reaction time is 15 minutes, the main wavelength of the test is 340nm, the secondary wavelength of the test is above 405nm, the volume ratio of the blood ammonia sample to the reagent is 1 / 25, and the reaction direction is negative reaction.

[0129] The specific measurement s...

Embodiment 3

[0140] Embodiment three (three doses)

[0141] The blood ammonia diagnostic reagent of the present embodiment is three doses, has:

[0142] Reagent I

[0143] Buffer 120mmol / l

[0144] Adenosine triphosphate 12mmol / l

[0145] Glutamic acid 10mmol / l

[0146] Pyruvate 15mmol / l

[0147] Phosphoenolpyruvate 8mmol / l

[0148] Reduced coenzyme 0.3mmol / l

[0149] Stabilizer 20mmol / l

[0150] Reagent II

[0151] Buffer 120mmol / l

[0152] Pyruvate oxidase 12000U / l

[0153] Phosphoenolpyruvate Carboxylase 10000U / l

[0154] Malate dehydrogenase 16000U / l

[0155] Stabilizer 50% (total volume)

[0156] Reagent III

[0157] Buffer 120mmol / l

[0158] Glutamine Synthetase 12000U / l

[0159] Stabilizer 50% (total volume)

[0160] Set on the automatic biochemical analyzer: temperature 25°C, reaction time 20 minutes, test main wavelength 340nm, test sub-wavelength above 405nm, the volume ratio of the measured blood ammonia sample to reagent is 1 / 25, and the reaction direction is ne...

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Abstract

The present invention belongs to the field of medical detection technology. The reagent kit for blood ammonia diagnosis includes buffering solution, adenosine triphophate, glutamic acid, pyruvic acid, phosphoenolpyruvic acid, reduced coenzyme, glutamine synthetase, pyruvate oxidase, phosphoenolpyruvate carboxylase, malate dehydrogenase and stabilizer. Through mixing the sample and reagent in certain volume ratio to produce enzyme coupling reaction, and detecting in biochemical analyzer the main wavelength absorbency change speed, the blood ammonia content is measured. The present invention can obtain the measurement result in biochemical analyzer in high sensitivity, high precision and no contamination of various foreign and internal matters.

Description

technical field [0001] The invention relates to a method for measuring blood ammonia content, and at the same time, the invention also relates to a blood ammonia diagnosis kit, which belongs to the technical field of medical examination and measurement. Background technique [0002] Ammonia determination methods include micro-diffusion method, ion exchange method, enzymatic method and ammonia electrode method. The most widely used methods are enzymatic method and blood ammonia analyzer analysis method based on ion selective electrode. [0003] The diffusion method is that after the sample is alkalized, NH is released 3 , Titrate the released ammonia with acid, or use the Nessler reaction to form brown-yellow mercuric ammonium iodide for colorimetry. These methods require alkalization, which is affected by the formation of endogenous ammonia, which affects its accuracy and precision, and is rarely used at present; the ion exchange method is more accurate than the diffusion ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/31G01N21/33
Inventor 王尔中
Owner 王尔中
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