Ammonia (ammonia ion) measuring method and ammonia (ammonia ion) diagnosing/measuring 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: 2012-07-11
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 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) measuring method and ammonia (ammonia ion) diagnosing/measuring kit
  • Ammonia (ammonia ion) measuring method and ammonia (ammonia ion) diagnosing/measuring kit
  • Ammonia (ammonia ion) measuring method and ammonia (ammonia ion) diagnosing/measuring kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0129] Embodiment 1: the mensuration of ammonia (ammonia ion) content in blood plasma

[0130] 1) Preparation of standard samples

[0131] Dissolve a certain amount of ammonia salt in water or buffer solution, and then adjust the ammonia concentration to 100 micromole / liter;

[0132] 2) Pretreatment of samples to be tested

[0133] Plasma samples are used as samples to be tested without pretreatment;

[0134] 3) Blank sample

[0135] Described water or buffer solution are used as blank sample, and its ammonia concentration is 0 micromole / liter;

[0136] B. Preparation of reagent solution:

[0137] The ammonia diagnosis / measurement reagent of the present embodiment is a single-dose reagent, which contains:

[0138] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0139] Glycerin 1mol / L

[0140] NADPH0.25mmol / L

[0141] Ammonia kinase 16000U / L

[0142] Pyruvate carboxylase 18000U / L

[0143] Isocitrate dehydrogenase 12000U / L

[0144] Adenosine triph...

Embodiment 2

[0160] Embodiment 2: the mensuration of ammonia (ammonia ion) content in blood plasma

[0161] 1) Preparation of standard samples

[0162] 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;

[0163] 2) Preparation of samples to be tested

[0164] Plasma samples are used as samples to be tested without pretreatment;

[0165] 3) Blank sample

[0166] Described water is used as blank sample, and its ammonia (ammonia ion) concentration is 0 micromole / liter;

[0167] B. Preparation of reagent solution:

[0168] Ammonia (ammonia ion) diagnostic / determination reagent is a two-dose reagent, which contains:

[0169] Reagent 1 with the following composition:

[0170] Phosphate buffer 100mmol / L

[0171] NADH0.25mmol / L

[0172] Adenosine triphosphate 5mmol / L

[0173] Oxaloacetate 5mmol / L

[0174] Ketoglutarate 5mmol / L;

[0175] Reagent 2 with the following composit...

Embodiment 3

[0197] Embodiment 3: the mensuration of ammonia (ammonia ion) content in plasma sample

[0198] The mode of operation of this embodiment is the same as embodiment 2, but this embodiment:

[0199] B. Preparation of reagent solution:

[0200] Ammonia (ammonia ion) diagnostic / assay reagent is a three-dose reagent that contains:

[0201] Reagent 1 with the following composition:

[0202] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0203] NADH0.25mmol / L

[0204] Adenosine triphosphate 6mmol / L

[0205] Oxaloacetate 5mmol / L

[0206] Monovalent phosphate 5mmol / L

[0207] Ketoglutarate 4mmol / L;

[0208] Reagent 2 with the following composition:

[0209] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0210] Sodium chloride 2mol / L

[0211] Pyruvate carboxylase 28000U / L

[0212] Isocitrate dehydrogenase 50000U / L;

[0213] Reagent 3 with the following composition:

[0214]Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 10...

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Abstract

The invention relates to a method for measuring ammonia (ammonia ion) content through an enzymatic colorimetric method and an enzyme coupling reaction technique, and the composition and the constituents of a reagent. The measuring technical principle of the method is fulfilled through the serial catalytic reactions of ammonia kinase, pyruvate carboxylase, and isocitrate dehydrogenase. The invention further relates to an ammonia (ammonia ion) diagnosing / measuring kit. The method provided by the invention has the advantages of high sensitivity and small error, so that the method and the kit provided by the invention can be widely applied to clinical medicine / food inspections.

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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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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