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 undiscovered blood ammonia determination method, high recovery rate, influence of accuracy and precision, etc.

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

[0078] Embodiment one (single agent)

[0079] The blood ammonia diagnostic kit of the present embodiment comprises:

[0080] Buffer 80mmol / l

[0081] 2-ketoglutarate 6mmol / l

[0082] Reduced coenzyme 0.2mmo / l

[0083] Phosphoenolpyruvate 2mmol / l

[0084] Glutamate dehydrogenase 6000U / l

[0085] Glutamate decarboxylase 6000U / l

[0086] Phosphoenolpyruvate Carboxylase 2000U / l

[0087] Malate dehydrogenase 6000U / l

[0088] Stabilizer 50% (total volume)

[0089] 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 .

[0090] After the sample and reagents are added and allowed to mix, the following reactions occur:

[0091] Ammonium ion + 2-ketoglutarate + reduced coenzyme glutamate dehydrogenase

[0092] Glutamic acid + coenzy...

Embodiment 2

[0100] Embodiment two (two doses)

[0101] The blood ammonia diagnostic reagent of the present embodiment has:

[0102] Reagent I

[0103] Buffer 100mmol / l

[0104] 2-ketoglutarate 18mmol / l

[0105] Reduced coenzyme 0.25mmo / l

[0106] Phosphoenolpyruvate 5mmol / l

[0107] Glutamic acid decarboxylase 8000U / l

[0108] Phosphoenolpyruvate Carboxylase 5000U / l

[0109] Malate dehydrogenase 8000U / l

[0110] Stabilizer 50% (total volume)

[0111] Reagent II

[0112] Buffer 100mmol / l

[0113] Glutamate dehydrogenase 8000U / l

[0114] Stabilizer 50% (total volume)

[0115] When measuring the 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.

[0116] The specific measurement steps are:

[0117] Ammonium ion + 2-ketoglutara...

Embodiment 3

[0126] Embodiment three (three doses)

[0127] The blood ammonia diagnostic reagent of the present embodiment is three doses, has:

[0128] Reagent I

[0129] Buffer 120mmol / l

[0130] 2-ketoglutarate 30mmol / l

[0131] Reduced coenzyme 0.3mmo / l

[0132] Phosphoenolpyruvate 8mmol / l

[0133] Stabilizer 20mmol / l

[0134] Reagent II

[0135] Buffer 120mmol / l

[0136] Glutamate decarboxylase 10000U / l

[0137] Phosphoenolpyruvate Carboxylase 8000U / l

[0138] Malate dehydrogenase 10000U / l

[0139] Stabilizer 50% (total volume)

[0140] Reagent III

[0141] Buffer 120mmol / l

[0142] Glutamate dehydrogenase 10000U / l

[0143] Stabilizer 50% (total volume)

[0144] 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 .

[0145] The specific measurement steps ...

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PUM

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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, 2ú¡ketoglutarate, reduced coenzyme, phosphoenolpyruvate, glutamate dehydrogenase, glutamate decarboxylase, phosphoenolpyruvate carboxylase, 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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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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