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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

[0127] Embodiment 1: the mensuration of ammonia (ammonia ion) content in blood plasma

[0128] 1) Preparation of standard samples

[0129] Dissolve a certain amount of ammonia salt in water or buffer solution, and then adjust the ammonia concentration to 100 micromole / liter;

[0130] 2) Pretreatment of samples to be tested

[0131] Plasma samples are used as samples to be tested without pretreatment;

[0132] 3) Blank sample

[0133] Described water or buffer solution are used as blank sample, and its ammonia concentration is 0 micromole / liter;

[0134] B. Preparation of reagent solution:

[0135] The ammonia diagnosis / measurement reagent of the present embodiment is a single-dose reagent, which contains:

[0136] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0137] Glycerin 1mol / L

[0138] NADPH0.25mmol / L

[0139] Ammonia kinase 16000U / L

[0140] Formate-dihydrofolate ligase 18000U / L

[0141] Dihydrofolate reductase 12000U / L

[0142] Adenosin...

Embodiment 2

[0157] Embodiment 2: the mensuration of ammonia (ammonia ion) content in blood plasma

[0158] 1) Preparation of standard samples

[0159] 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;

[0160] 2) Preparation of samples to be tested

[0161] Plasma samples are used as samples to be tested without pretreatment;

[0162] 3) Blank sample

[0163] Described water is used as blank sample, and its ammonia (ammonia ion) concentration is 0 micromole / liter;

[0164] B. Preparation of reagent solution:

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

[0166] Reagent 1 with the following composition:

[0167] Phosphate buffer 100mmol / L

[0168] NADH0.25mmol / L

[0169] Adenosine triphosphate 5mmol / L

[0170] 10-Formyldihydrofolate 5mmol / L

[0171] Monovalent phosphate 5mmol / L;

[0172] Reagent 2 with the f...

Embodiment 3

[0194] Embodiment 3: the mensuration of ammonia (ammonia ion) content in plasma sample

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

[0196] B. Preparation of reagent solution:

[0197] Ammonia (ammonia ion) diagnostic / assay reagent is a three-dose reagent that contains:

[0198] Reagent 1 with the following composition:

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

[0200]NADH0.25mmol / L

[0201] Adenosine triphosphate 6mmol / L

[0202] 10-Formyldihydrofolate 5mmol / L

[0203] Monovalent phosphate 4mmol / L;

[0204] Reagent 2 with the following composition:

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

[0206] Sodium chloride 2mol / L

[0207] Formate-dihydrofolate ligase 28000U / L

[0208] Dihydrofolate reductase 50000U / L;

[0209] Reagent 3 with the following composition:

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

[021...

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Abstract

The invention relates to an ammonia (ammonia ion) content determination method adopting an enzymatic colorimetric method-couple reaction united technology, and composition and ingredients of reagents adopted by the ammonia (ammonia ion) content determination method. 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, formate-dihydrofolate ligase and dihydrofolate reductase. 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): G01N33/52G01N21/31G01N21/33C12Q1/48C12Q1/25C12Q1/26
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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