Determination method for ammonia (ammonia ions) and ammonia (ammonia ion) diagnostic/assay 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 for ammonia (ammonia ions) and ammonia (ammonia ion) diagnostic/assay kit
  • Determination method for ammonia (ammonia ions) and ammonia (ammonia ion) diagnostic/assay kit
  • Determination method for ammonia (ammonia ions) and ammonia (ammonia ion) diagnostic/assay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0126] Embodiment 1: the mensuration of ammonia (ammonia ion) content in blood plasma

[0127] 1) Preparation of standard samples

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

[0129] 2) Pretreatment of samples to be tested

[0130] Plasma samples are used as samples to be tested without pretreatment;

[0131] 3) Blank sample

[0132] Described water or buffer solution are used as blank sample, and its ammonia concentration is 0 micromole / liter;

[0133] B. Preparation of reagent solution:

[0134] The ammonia diagnosis / measurement reagent of the present embodiment is a single-dose reagent, which contains:

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

[0136] Glycerin 1mol / L

[0137] NADPH0.25mmol / L

[0138] Ammonia kinase 16000U / L

[0139] Pyruvate kinase 18000U / L

[0140] N-methylalanine dehydrogenase 12000U / L

[0141] Adenosine triph...

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] Enol pyruvate 5mmol / L

[0171] Methylamine 5mmol / L;

[0172] Reagent 2 with the following compositi...

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] Enol pyruvate 5mmol / L

[0203] Methylamine 4mmol / L;

[0204] Reagent 2 with the following composition:

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

[0206] Sodium chloride 2mol / L

[0207] Pyruvate kinase 28000U / L

[0208] N-methylalanine dehydrogenase 50000U / L;

[0209] Reagent 3 with the following composition:

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

[0211] Sodium chloride 2mol / L...

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Abstract

The invention relates to a determination method for the content of ammonia (ammonia ions) by using an enzymatic colorimetric technology and an enzymatic couple reaction and to composition and components of a reagent. A technical principle used in the method is that determination is realized through series reactions of ammonia kinase, pyruvate kinase and N-methylalanine dehydrogenase. The invention also relates to an ammonia (ammonia ion) diagnostic/assay kit. The determination method provided in the invention has high sensitivity and a small error; the determination method and the kit in 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 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% t...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/48C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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