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Method for determining ammonia (ammonium ion) and ammonia (ammonium ion) diagnosis/determination kit

An ammonia ion and reagent technology, which is applied in the field of medical/food testing and determination, and can solve the problem of not finding blood ammonia determination methods, etc.

Inactive Publication Date: 2012-05-23
SUZHOU ANJ BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Searching Chinese patents, only found 87105593.7 patent application which discloses a quick-frozen cup for measuring blood ammonia, but did not find an ideal method for measuring blood ammonia

Method used

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  • Method for determining ammonia (ammonium ion) and ammonia (ammonium ion) diagnosis/determination kit
  • Method for determining ammonia (ammonium ion) and ammonia (ammonium ion) diagnosis/determination kit
  • Method for determining ammonia (ammonium ion) and ammonia (ammonium ion) diagnosis/determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Embodiment 1: the mensuration of ammonia (ammonia ion) content in blood plasma

[0132] 1) Preparation of standard samples

[0133] Dissolve a certain amount of ammonia salt in water or buffer solution, and then adjust the ammonia concentration to 100 micromole / liter;

[0134] 2) Pretreatment of samples to be tested

[0135] Plasma samples are used as samples to be tested without pretreatment;

[0136] 3) Blank sample

[0137] Described water or buffer solution are used as blank sample, and its ammonia concentration is 0 micromole / liter;

[0138] B. Preparation of reagent solution:

[0139] The ammonia diagnosis / measurement reagent of the present embodiment is a single-dose reagent, which contains:

[0140] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0141] Glycerin 1mol / L

[0142] NADP + 0.25mmol / L

[0143] Ammonia kinase 16000U / L

[0144] Citric acid coenzyme A ligase 18000U / L

[0145] Coenzyme A-glutathi...

Embodiment 2

[0162] Embodiment 2: the mensuration of ammonia (ammonia ion) content in blood plasma

[0163] 1) Preparation of standard samples

[0164] A certain amount of ammonia salt is dissolved in water, and then the concentration of ammonia (ammonia ion) is adjusted to 100 micromole / liter, as a standard sample;

[0165] 2) Preparation of samples to be tested

[0166] Plasma samples are used as samples to be tested without pretreatment;

[0167] 3) Blank sample

[0168] Described water is used as blank sample, and its ammonia (ammonia ion) concentration is 0 micromole / liter;

[0169] B. Preparation of reagent solution:

[0170] Ammonia (ammonia ion) diagnostic / determination reagent is a two-dose reagent, which contains:

[0171] Reagent 1 with the following composition:

[0172] Phosphate buffer 100mmol / L

[0173] NAD + 0.25mmol / L

[0174] Adenosine triphosphate 5mmol / L

[0175] Monovalent phosphate 5mmol / L

[0176] Citroyl-CoA 5mmol / L

[0177] G...

Embodiment 3

[0200] Embodiment 3: the mensuration of ammonia (ammonia ion) content in plasma sample

[0201] The mode of operation of this embodiment is the same as embodiment 2, but this embodiment:

[0202] B. Preparation of reagent solution:

[0203] Ammonia (ammonia ion) diagnostic / assay reagent is a three-dose reagent that contains:

[0204] Reagent 1 with the following composition:

[0205] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0206] NAD + 0.25mmol / L

[0207] Adenosine triphosphate 6mmol / L

[0208] Monovalent phosphate 5mmol / L

[0209] Citroyl-CoA 5mmol / L

[0210] Glutathione 4mmol / L;

[0211] Reagent 2 with the following composition:

[0212] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0213] Sodium chloride 2mol / L

[0214] Citric acid coenzyme A ligase 28000U / L

[0215] Coenzyme A-glutathione reductase 50000U / L;

[0216] Reagent 3 with the following composition:

[0217] Tris(carboxy...

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Abstract

The invention relates to a method for determining the content of ammonia (ammonium ion) by technologies of an enzyme colorimetric method and an enzyme linking method, and composition and components of a reagent. The invention adopts technical principle that the determination is completed according to a series of catalytic reactions of ammonia kinase, citric acid coenzyme A ligase, and coenzyme A-glutathione reductase. The invention also relates to an ammonia (ammonium ion) diagnosis / determination kit. The determination method has high sensitivity and small errors, so that the determination method and the kit can be widely applied to clinical medicine / food inspection.

Description

technical field [0001] The present invention relates to the technical field of medical / food inspection and determination, more specifically, the present invention relates to ammonia (ammonia ion) diagnosis / measurement method and kit thereof. 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 need to be alkalized, and the endogenous ammonia formation will affect the accuracy and precision, so they are rarely used at present; the ion exchange method is more accurate than the diffusion method, and the CV is 8% to 13...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/48C12Q1/26C12Q1/25
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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