Detecting method of ammonia (ammonium ion) and ammonia (ammonium ion) diagnosing/detecting 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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  • Detecting method of ammonia (ammonium ion) and ammonia (ammonium ion) diagnosing/detecting kit
  • Detecting method of ammonia (ammonium ion) and ammonia (ammonium ion) diagnosing/detecting kit
  • Detecting method of ammonia (ammonium ion) and ammonia (ammonium ion) diagnosing/detecting kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0132] Embodiment 1: Determination of ammonia (ammonia ion) content in plasma

[0133] 1) Preparation of standard samples

[0134] Dissolve a certain amount of ammonia salt in water or buffer, and then adjust the ammonia concentration to 100 μmol / L;

[0135] 2) Pretreatment of the sample to be tested

[0136] Plasma samples are used as samples to be tested without pretreatment;

[0137] 3) Blank sample

[0138] Described water or buffer is used as blank sample, and its ammonia concentration is 0 micromol / liter;

[0139] B. Preparation of reagent solution:

[0140] The ammonia diagnostic / assay reagent of this embodiment is a single-dose reagent, which contains:

[0141] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0142] Glycerol 1mol / L

[0143] NADPH 0.25mmol / L

[0144] Ammonia kinase 16000U / L

[0145] Carbamoyl phosphate synthase 18000U / L

[0146] Isocitrate dehydrogenase 12000U / L

[0147] Adenosine triphosphate 5mmol / L

[0148] Glutamate 5...

Embodiment 2

[0164] Example 2: Determination of ammonia (ammonia ion) content in plasma

[0165] 1) Preparation of standard samples

[0166] Dissolve a certain amount of ammonia salt in water, and then adjust the ammonia (ammonia ion) concentration to 100 μmol / L as a standard sample;

[0167] 2) Preparation of samples to be tested

[0168] Plasma samples are used as samples to be tested without pretreatment;

[0169] 3) Blank sample

[0170] Described water is used as blank sample, and its ammonia (ammonia ion) concentration is 0 micromol / liter;

[0171] B. Preparation of reagent solution:

[0172] Ammonia (ammonia ion) diagnostic / measurement reagent is a two-agent reagent, which contains:

[0173] Reagent 1 consisting of the following:

[0174] Phosphate buffer 100mmol / L

[0175] NADH 0.25mmol / L

[0176] Adenosine triphosphate 5mmol / L

[0177] Glutamate 5mmol / L

[0178] carbamoyl phosphate 5mmol / L

[0179] Monovalent phosphate 5mmol / L

[0180] Ketoglutaric acid 5mmol / L;

[01...

Embodiment 3

[0203] Example 3: Determination of ammonia (ammonia ion) content in plasma samples

[0204] The operation mode of this embodiment is the same as that of Embodiment 2, except that this embodiment:

[0205] B. Preparation of reagent solution:

[0206] Ammonia (ammonia ion) diagnostic / measurement reagent is a three-dose reagent, which contains:

[0207] Reagent 1 consisting of the following:

[0208] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0209] NADH 0.25mmol / L

[0210] Adenosine triphosphate 5mmol / L

[0211] Glutamate 5mmol / L

[0212] carbamoyl phosphate 5mmol / L

[0213] Monovalent phosphate 5mmol / L

[0214] Ketoglutaric acid 5mmol / L;

[0215] Reagent 2 consisting of the following:

[0216] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0217] Sodium chloride 2mol / L

[0218] Carbamoyl phosphate synthase 28000U / L

[0219] Isocitrate dehydrogenase 50000U / L;

[0220] Reagent 3 consisting of the following:

[0221] Tr...

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Abstract

The invention relates to a detecting method of ammonia (ammonium ion) content by utilizing an enzymatic colorimetry and an enzyme-linked method, and composition and ingredients of reagents. The technology principle of detection is that the detection is completed according to the serial catalytic reaction of ammonia kinase, carbamyl phosphate synthetase and isocitrate dehydrogenase. The invention also relates to an ammonia (ammonium ion) diagnosing/detecting kit. The detecting method has high sensitivity and small error, so that the detecting method and the kit can be widely applied to clinical medical/food inspection.

Description

technical field [0001] The present invention relates to the technical field of medical / food testing and determination, and more particularly, the present invention relates to an ammonia (ammonia ion) diagnosis / determination method and a kit thereof. Background technique [0002] The methods of ammonia determination include micro-diffusion method, ion exchange method, enzymatic method and ammonia electrode method. The most widely used methods are the enzymatic method and the ion-selective electrode-based blood ammonia analyzer analysis method. [0003] The diffusion method is that after the sample is alkalized, NH is released 3 , titrate the liberated ammonia with acid, or use the Nessler reaction to form a brown-yellow mercuric iodide for colorimetry. These methods require alkalization, which affects the accuracy and precision of endogenous ammonia formation, and is rarely used at present; the ion exchange method is more accurate than the diffusion method, with a CV of 8% ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/48C12Q1/25C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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