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Determination of blood ammonia content and blood ammonia diagnostic reagent kit

A diagnostic kit and blood ammonia technology, applied in the field of medical testing and determination, can solve the problems of accuracy and precision, no blood ammonia determination method, and high recovery rate.

Inactive Publication Date: 2006-05-31
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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 method, and the CV is 8%-13%; Selective electrode method is to use NH 3 Diffusion to the surface of the electrode, causing the pH of the electrode to change for measurement, the CV of this method is 3.5% - 4.8%, the recovery rate is high, but all need special instruments, not suitable for the specific reality of our country
[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

[0077] Embodiment one (single agent)

[0078] The blood ammonia diagnostic kit of the present embodiment comprises:

[0079] Buffer 80mmol / l

[0080] Glutamic acid 6mmol / l

[0081] Adenosine triphosphate 2mmol / l

[0082] Pyruvate 5mmol / l

[0083] Reduced coenzyme 0.2mmol / l

[0084] Glutamine Synthetase 6000U / l

[0085] Pyruvate oxidase 6000U / l

[0086] Malate dehydrogenase 8000U / l

[0087] Stabilizer 50% (total volume)

[0088] 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 .

[0089] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0090] Ammonia + glutamic acid + adenosine triphosphate glutamine synthetase Glutamine+

[0091] Adenosine diphosphate + phosphate

[0092] Phosp...

Embodiment 2

[0098] Embodiment two (two doses)

[0099] The blood ammonia diagnostic reagent of the present embodiment has:

[0100] Reagent I

[0101] Buffer 100mmol / l

[0102] Glutamic acid 8mmol / l

[0103] Adenosine triphosphate 7mmol / l

[0104] Pyruvate 10mmol / l

[0105] Reduced coenzyme 0.25mmol / l

[0106] Pyruvate oxidase 9000U / l

[0107] Malate dehydrogenase 12000U / l

[0108] Stabilizer 50% (total volume)

[0109] Reagent II

[0110] Buffer 100mmol / l

[0111] Glutamine Synthetase 9000U / l

[0112] Stabilizer 50% (total volume)

[0113] 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.

[0114] The specific measurement steps are:

[0115] Ammonia + glutamic acid + adenosine triphosphate glutamine synthe...

Embodiment 3

[0123] Embodiment three (three doses)

[0124] The blood ammonia diagnostic reagent of the present embodiment is three doses, has:

[0125] Reagent I

[0126] Buffer 120mmol / l

[0127] Glutamic acid 10mmol / l

[0128] Adenosine triphosphate 12mmol / l

[0129] Pyruvate 15mmol / l

[0130] Reduced coenzyme 0.3mmol / l

[0131] Stabilizer 20mmol / l

[0132] Reagent II

[0133] Buffer 120mmol / l

[0134] Pyruvate oxidase 12000U / l

[0135] Malate dehydrogenase 16000U / l

[0136] Stabilizer 50% (total volume)

[0137] Reagent III

[0138] Buffer 120mmol / l

[0139] Glutamine Synthetase 12000U / l

[0140] Stabilizer 50% (total volume)

[0141] 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 negative reaction .

[0142] The specific measurement steps are:

[0143] Ammonia + glut...

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Abstract

The invention is about a method of measuring the content of blood ammonia, and it also concerns the reagent box of blood ammonia diagnosis. This invention belongs to the field of medical testing and measuring technology. The reagent box is consisted of buffer solution, glutamate, adenosine triphosphate, acetonic acid, reduced coenzyme, glutamyl synzyme, pyruvate oxidase, malate dehydrogenase and stabilizer. Firstly, we cause an enzyme-coupled reaction through mixing the sample and the reagent according to a certain proportion of volume; secondly, put the final reactant under the biochemical analyzer and test the absorbance variational situation (speed) of dominant wavelength; then we can get the content ofblood ammonia. By using this invention, we can get the necessary measuring result with high sensitiveness and fine precision through biochemical analyzer, and the result would not be contaminated by material of internal and exogenous sources. Thus, this method can be conveniently promoted and applied.

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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IPC IPC(8): C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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