Determination method of ammonia (ammonia ion) and ammonia (ammonia ion) diagnosis / determination kit

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

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

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Problems solved by technology

[0004] Searching Chinese patents, only found 87105593.7 patent application which discloses a qui...

Method used

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  • Determination method of ammonia (ammonia ion) and ammonia (ammonia ion) diagnosis / determination kit
  • Determination method of ammonia (ammonia ion) and ammonia (ammonia ion) diagnosis / determination kit
  • Determination method of ammonia (ammonia ion) and ammonia (ammonia ion) diagnosis / determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0136] Embodiment 1: Determination of ammonia (ammonia ion) content in plasma

[0137] 1) Preparation of standard samples

[0138] Dissolve a certain amount of ammonia salt in water or buffer, and then adjust the ammonia concentration to 100 μmol / L;

[0139] 2) Pretreatment of the sample to be tested

[0140] Plasma samples are used as samples to be tested without pretreatment;

[0141] 3) Blank sample

[0142] Described water or buffer is used as blank sample, and its ammonia concentration is 0 micromol / liter;

[0143] B. Preparation of reagent solution:

[0144] The ammonia diagnostic / assay reagent of this embodiment is a single-dose reagent, which contains:

[0145] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0146] Glycerol 1mol / L

[0147] NADPH 0.25mmol / L

[0148] Glutamate oxidase 16000U / L

[0149] Carbamoyl phosphate synthase 18000U / L

[0150] Malonate-semialdehyde dehydrogenase 12000U / L

[0151] Hydrogen peroxide 10mmol / L

[0152] 2...

Embodiment 2

[0169] Example 2: Determination of ammonia (ammonia ion) content in plasma

[0170] 1) Preparation of standard samples

[0171] 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;

[0172] 2) Preparation of samples to be tested

[0173] Plasma samples are used as samples to be tested without pretreatment;

[0174] 3) Blank sample

[0175] Described water is used as blank sample, and its ammonia (ammonia ion) concentration is 0 micromol / liter;

[0176] B. Preparation of reagent solution:

[0177] Ammonia (ammonia ion) diagnostic / measurement reagent is a two-agent reagent, which contains:

[0178] Reagent 1 consisting of the following:

[0179] Phosphate buffer 100mmol / L

[0180] NADH 0.25mmol / L

[0181] Hydrogen peroxide 10mmol / L

[0182] 2-ketoglutaric acid 10mmol / L

[0183] Adenosine diphosphate 5mmol / L

[0184] Carbamoyl phosphate 10mmol / L

[0185] Acetyl-CoA 10mmol / L;

...

Embodiment 3

[0208] Example 3: Determination of ammonia (ammonia ion) content in plasma samples

[0209] The operation mode of this embodiment is the same as that of Embodiment 2, except that this embodiment:

[0210] B. Preparation of reagent solution:

[0211] Ammonia (ammonia ion) diagnostic / measurement reagent is a three-dose reagent, which contains:

[0212] Reagent 1 consisting of the following:

[0213] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0214] NADH 0.25mmol / L

[0215] Hydrogen peroxide 10mmol / L

[0216] 2-ketoglutaric acid 10mmol / L

[0217] Adenosine diphosphate 5mmol / L

[0218] Carbamoyl phosphate 10mmol / L

[0219] Acetyl-CoA 10mmol / L;

[0220] Reagent 2 consisting of the following:

[0221] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0222] Sodium chloride 2mol / L

[0223] Carbamoyl phosphate synthase 28000U / L

[0224] Malonate-semialdehyde dehydrogenase 50000U / L;

[0225] Reagent 3 consisting of the following...

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Abstract

The invention relates to a determination method of ammonia (ammonia ion) content utilizing enzymatic colorimetric method and couple reaction, and reagent composition and components. Technical principles of the determination method are in the light of a series of catalytic reactions of glutamate oxidase, carbamyl phosphate synthetase and malonate-semialdehyde dehydrogenase. The invention also relates to an ammonia (ammonia ion) diagnosis / determination kit. The determination method of the present invention has high sensitivity and little error, so the determination method and the kit of the invention can be widely applied to clinic medicine / food examination.

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...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/32C12Q1/26C12Q1/25
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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