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Detection method of ammonia (ammonia ion) and diagnosis/detection kit of ammonia (ammonia ion)

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: 2012-05-23
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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  • Detection method of ammonia (ammonia ion) and diagnosis/detection kit of ammonia (ammonia ion)
  • Detection method of ammonia (ammonia ion) and diagnosis/detection kit of ammonia (ammonia ion)
  • Detection method of ammonia (ammonia ion) and diagnosis/detection kit of ammonia (ammonia ion)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0128] Example 1: Determination of ammonia (ammonia ion) content in plasma

[0129] 1) Preparation of standard samples

[0130] Dissolve a certain amount of ammonia salt in water or buffer, and then adjust the ammonia concentration to 100 micromol / l;

[0131] 2) Pretreatment of the sample to be tested

[0132] The plasma sample is used as the sample to be tested, without pretreatment;

[0133] 3) Blank sample

[0134] The water or buffer is used as a blank sample, and its ammonia concentration is 0 micromol / L;

[0135] B. Preparation of reagent solution:

[0136] The ammonia diagnostic / measurement reagent in this embodiment is a single-dose reagent, which contains:

[0137] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0138] Glycerin 1mol / L

[0139] NADP + 0.25mmol / L

[0140] Ferredoxin nitrite reductase 16000U / L

[0141] Nitrite reductase 18000U / L

[0142] Nitric oxide synthase 12000U / L

[0143] Oxidized ferredoxin 5mmol / L

[0144] Ferrous c...

Embodiment 2

[0158] Example 2: Determination of the content of ammonia (ammonia ion) in plasma

[0159] 1) Preparation of standard samples

[0160] 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;

[0161] 2) Preparation of samples to be tested

[0162] The plasma sample is used as the sample to be tested, without pretreatment;

[0163] 3) Blank sample

[0164] The water is used as a blank sample, and its ammonia (ammonia ion) concentration is 0 micromol / L;

[0165] B. Preparation of reagent solution:

[0166] Ammonia (ammonia ion) diagnostic / determination reagent is a two-dose reagent, which contains:

[0167] Reagent 1 consisting of:

[0168] Phosphate buffer 100mmol / L

[0169] NAD + 0.25mmol / L

[0170] Oxidized ferredoxin 5mmol / L

[0171] Ferrous cytochrome C 5mmol / L

[0172] Citrulline 5mmol / L;

[0173] Reagent 2 consisting of:

[0174] Phosphate buffer 100mmol / L

[0175] Et...

Embodiment 3

[0195] Example 3: Determination of ammonia (ammonia ion) content in plasma samples

[0196] The operation mode of this embodiment is the same as that of embodiment 2, except that this embodiment:

[0197] B. Preparation of reagent solution:

[0198] Ammonia (ammonia ion) diagnostic / determination reagent is a three-agent reagent, which contains:

[0199] Reagent 1 consisting of:

[0200] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0201] NAD + 0.25mmol / L

[0202] Oxidized ferredoxin 6mmol / L

[0203] Ferrous cytochrome C 5mmol / L

[0204] Citrulline 4mmol / L;

[0205] Reagent 2 consisting of:

[0206] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0207] Sodium chloride 2mol / L

[0208] Nitrite reductase 28000U / L

[0209] Nitric oxide synthase 50000U / L;

[0210] Reagent 3 consisting of:

[0211] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0212] Sodium chloride 2mol / L

[0213] Ferredoxin nitrite reduc...

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Abstract

The invention relates to a detection method of the ammonia (ammonia ion) content by enzymatic colorimetry and enzyme coupled reaction technology and composition and components of a reagent. The technology principle of the detection method is based on a series of catalytic reactions of ferredoxin-nitrite reductase, nitrite reductase and nitric oxide synthetase. The invention also relates to an ammonia (ammonia ion) diagnosis / detection kit. The detection method provided by the invention has advantages of high sensitivity and small error. Therefore, the detection method and the kit provided by the invention can be widely applied in clinical medicine / 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%; i...

Claims

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Application Information

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