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Determination method of ammonia (ammonia ions) and diagnosis/determination kit for ammonia (ammonia ions)

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Example 1: Determination of ammonia (ammonia ion) content in plasma

[0127] 1) Preparation of standard samples

[0128] Dissolve a certain amount of ammonia salt in water or buffer, and then adjust the ammonia concentration to 100 micromol / l;

[0129] 2) Pretreatment of the sample to be tested

[0130] The plasma sample is used as the sample to be tested, without pretreatment;

[0131] 3) Blank sample

[0132] The water or buffer is used as a blank sample, and its ammonia concentration is 0 micromol / L;

[0133] B. Preparation of reagent solution:

[0134] The ammonia diagnostic / measurement reagent in this embodiment is a single-dose reagent, which contains:

[0135] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0136] Glycerin 1mol / L

[0137] NADPH 0.25mmol / L

[0138] Aminokinase 16000U / L

[0139] Biotin carboxylase 18000U / L

[0140] Formate dehydrogenase 12000U / L

[0141] Adenosine triphosphate 5mmol / L

[0142] Carboxybiotin-carboxy-carrying protein 3mmol / L

[0143] ...

Embodiment 2

[0157] Example 2: Determination of the content of ammonia (ammonia ion) in plasma

[0158] 1) Preparation of standard samples

[0159] Dissolve a certain amount of ammonia salt in water, then adjust the concentration of ammonia (ammonia ion) to 100 micromol / l, as a standard sample;

[0160] 2) Preparation of samples to be tested

[0161] The plasma sample is used as the sample to be tested, without pretreatment;

[0162] 3) Blank sample

[0163] The water is used as a blank sample, and its ammonia (ammonia ion) concentration is 0 micromol / L;

[0164] B. Preparation of reagent solution:

[0165] Ammonia (ammonia ion) diagnostic / determination reagent is a two-dose reagent, which contains:

[0166] Reagent 1 consisting of:

[0167] Phosphate buffer 100mmol / L

[0168] NADH 0.25mmol / L

[0169] Adenosine triphosphate 5mmol / L

[0170] Carboxybiotin-carboxy-carrying protein 3mmol / L

[0171] Monovalent phosphate 3mmol / L;

[0172] Reagent 2 consisting of:

[0173] Phosphate buffer 100mmol / L

[0174] Ethylene...

Embodiment 3

[0194] Example 3: Determination of ammonia (ammonia ion) content in plasma samples

[0195] The operation mode of this embodiment is the same as that of embodiment 2, except that this embodiment:

[0196] B. Preparation of reagent solution:

[0197] Ammonia (ammonia ion) diagnostic / determination reagent is a three-agent reagent, which contains:

[0198] Reagent 1 consisting of:

[0199] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0200] NADH 0.25mmol / L

[0201] Adenosine triphosphate 5mmol / L

[0202] Carboxybiotin-carboxy-carrying protein 3mmol / L

[0203] Monovalent phosphate 3mmol / L;

[0204] Reagent 2 consisting of:

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

[0206] Sodium chloride 2mol / L

[0207] Biotin carboxylase 28000U / L

[0208] Formic acid dehydrogenase 50000U / L;

[0209] Reagent 3 consisting of:

[0210] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0211] Sodium chloride 2mol / L

[0212] Ammonia kinase 26000U / ...

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Abstract

The invention relates to a determination method of ammonia (ammonia ions) content by means of technologies of enzymatic colorimetry and enzyme-linked assay, and composition and components of a reagent. The technical principle of determination is based on a series of catalytic reactions of ammonia kinase, biotin carboxylase and formate dehydrogenase. The invention further relates to a diagnosis / determination kit for ammonia (ammonia ions). The determination method provided by the invention has high sensitivity and small error. Therefore, the determination method and the kit provided by the invention can be widely applied to clinical medical / food inspection.

Description

Technical field [0001] The present invention relates to the technical field of medical / food inspection and measurement. More specifically, the present invention relates to a diagnosis / measurement method of ammonia (ammonia ion) and a kit thereof. Background technique [0002] Ammonia determination methods include trace diffusion method, ion exchange method, enzymatic method and ammonia electrode method. At present, the most widely used methods are enzymatic method and blood ammonia analyzer analysis method based on ion selective electrode. [0003] The diffusion method is to release NH after alkalization of the specimen 3 , Use acid to titrate the released ammonia, or use Nessler reaction to form brown-yellow amalgam iodide for color comparison. These methods require alkalization, and the formation of endogenous ammonia affects their accuracy and precision. They are rarely used at present; ion exchange methods are more accurate than diffusion methods, with a CV of 8% to 13%; ion s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/48C12Q1/527C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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