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Ammonia (ammonia ion) content determination method and ammonia (ammonia ion) diagnosis/determination kit

A technology of ammonia ions and reagents, which is applied in the field of medical/food testing and determination, and can solve the problems of undiscovered blood ammonia determination methods, etc.

Inactive Publication Date: 2012-07-11
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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  • Ammonia (ammonia ion) content determination method and ammonia (ammonia ion) diagnosis/determination kit
  • Ammonia (ammonia ion) content determination method and ammonia (ammonia ion) diagnosis/determination kit
  • Ammonia (ammonia ion) content determination method and ammonia (ammonia ion) diagnosis/determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[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] Glycerol 1mol / L

[0142] NADPH0.25mmol / L

[0143] Ammonia kinase 16000U / L

[0144] Carbamyl phosphate synthase 18000U / L

[0145] Hydrogen cyanide synthase 12000U / L

[0146] Adenosine triphosphate 5mmol / L

[0147] Glutamic acid 5mmol / L

[0148] Carbamyl phosphate 5mmol / ...

Embodiment 2

[0164] 1) Preparation of standard samples

[0165] 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;

[0166] 2) Preparation of samples to be tested

[0167] Plasma samples are used as samples to be tested without pretreatment;

[0168] 3) Blank sample

[0169] Described water is used as blank sample, and its ammonia (ammonia ion) concentration is 0 micromole / liter;

[0170] B. Preparation of reagent solution:

[0171] Ammonia (ammonia ion) diagnostic / determination reagent is a two-dose reagent, which contains:

[0172] Reagent 1 with the following composition:

[0173] Phosphate buffer 100mmol / L

[0174] NADH0.25mmol / L

[0175] Adenosine triphosphate 5mmol / L

[0176] Glutamic acid 5mmol / L

[0177] Carbamyl phosphate 5mmol / L

[0178] Monovalent phosphate 5mmol / L

[0179] Hydrogen cyanide 5mmol / L;

[0180] Reagent 2 with the following composition:

[...

Embodiment 3

[0203] The mode of operation of this embodiment is the same as embodiment 2, but this embodiment:

[0204] B. Preparation of reagent solution:

[0205] Ammonia (ammonia ion) diagnostic / assay reagent is a three-dose reagent that contains:

[0206] Reagent 1 with the following composition:

[0207] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0208] NADH0.25mmol / L

[0209] Adenosine triphosphate 5mmol / L

[0210] Glutamic acid 5mmol / L

[0211] Carbamyl phosphate 5mmol / L

[0212] Monovalent phosphate 5mmol / L

[0213] Hydrogen cyanide 5mmol / L;

[0214] Reagent 2 with the following composition:

[0215] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0216] Sodium chloride 2mol / L

[0217] Carbamyl phosphate synthase 28000U / L

[0218] Hydrogen cyanide synthase 50000U / L;

[0219] Reagent 3 with the following composition:

[0220] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0221] Sodium chloride 2mol / L...

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PUM

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Abstract

The invention relates to an ammonia (ammonia ion) content determination method, and composition and ingredients of reagents adopted by the ammonia (ammonia ion) content determination method. The ammonia (ammonia ion) content determination method adopts an enzymatic colorimetry and an enzyme-linked immunosorbent assay technology. A technical principle of the ammonia (ammonia ion) content determination method comprises that determination is realized by a series of catalytic reactions of ammonia kinase, carbamyl phosphate synthetase and hydrogen cyanide synthetase. The invention also relates to an ammonia (ammonia ion) diagnosis / determination kit. The ammonia (ammonia ion) content determination method provided by the invention has high sensitivity and a small error. The ammonia (ammonia ion) content determination method and the ammonia (ammonia ion) diagnosis / determination kit can be widely utilized for clinical medicine and food examination.

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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IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/48C12Q1/25
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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