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Ammonia (ammonia ion) determination method and ammonia (ammonia ion) diagnosis/determination reagent 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: 2012-07-04
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) determination method and ammonia (ammonia ion) diagnosis/determination reagent kit
  • Ammonia (ammonia ion) determination method and ammonia (ammonia ion) diagnosis/determination reagent kit
  • Ammonia (ammonia ion) determination method and ammonia (ammonia ion) diagnosis/determination reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] Embodiment 1: the mensuration of ammonia (ammonia ion) content in blood plasma

[0131] 1) Preparation of standard samples

[0132] Dissolve a certain amount of ammonia salt in water or buffer solution, and then adjust the ammonia concentration to 100 micromole / liter;

[0133] 2) Pretreatment of samples to be tested

[0134] Plasma samples are used as samples to be tested without pretreatment;

[0135] 3) Blank sample

[0136] Described water or buffer solution are used as blank sample, and its ammonia concentration is 0 micromole / liter;

[0137] B. Preparation of reagent solution:

[0138] The ammonia diagnosis / measurement reagent of the present embodiment is a single-dose reagent, which contains:

[0139] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0140] Glycerin 1mol / L

[0141] NADPH0.25mmol / L

[0142] Ammonia kinase 16000U / L

[0143] Carbamyl phosphate synthase 18000U / L

[0144] Malonate-semialdehyde dehydrogenase 12000U / L

[014...

Embodiment 2

[0161] Embodiment 2: the mensuration of ammonia (ammonia ion) content in blood plasma

[0162] 1) Preparation of standard samples

[0163] 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;

[0164] 2) Preparation of samples to be tested

[0165] Plasma samples are used as samples to be tested without pretreatment;

[0166] 3) Blank sample

[0167] Described water is used as blank sample, and its ammonia (ammonia ion) concentration is 0 micromole / liter;

[0168] B. Preparation of reagent solution:

[0169] Ammonia (ammonia ion) diagnostic / determination reagent is a two-dose reagent, which contains:

[0170] Reagent 1 with the following composition:

[0171] Phosphate buffer 100mmol / L

[0172] NADH0.25mmol / L

[0173] Adenosine triphosphate 5mmol / L

[0174] Carbamoyl phosphate 5mmol / L

[0175] Monovalent phosphate 5mmol / L

[0176] Acetyl-CoA 5mmol / L;

[...

Embodiment 3

[0199] Embodiment 3: the mensuration of ammonia (ammonia ion) content in plasma sample

[0200] The mode of operation of this embodiment is the same as embodiment 2, but this embodiment:

[0201] B. Preparation of reagent solution:

[0202] Ammonia (ammonia ion) diagnostic / assay reagent is a three-dose reagent that contains:

[0203] Reagent 1 with the following composition:

[0204] Tris(carboxymethyl)aminomethane-hydrochloric acid buffer 100mmol / L

[0205] NADH0.25mmol / L

[0206] Adenosine triphosphate 5mmol / L

[0207] Carbamoyl phosphate 5mmol / L

[0208] Monovalent phosphate 5mmol / L

[0209] Acetyl-CoA 5mmol / L;

[0210] Reagent 2 with the following composition:

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

[0212] Sodium chloride 2mol / L

[0213] Carbamyl phosphate synthase 28000U / L

[0214] Malonate-semialdehyde dehydrogenase 50000U / L;

[0215] Reagent 3 with the following composition:

[0216] Tris(carboxymethyl)aminomethane-hydr...

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Abstract

The invention relates to a method for determination of ammonia (ammonia ion) content by using an enzymatic colorimetric method and an enzyme couple reaction technique and composition and components of a reagent. A determination technology principle of the method lies in that the determination is completed according to series catalytic reactions of ammonia kinase, carbamyl phosphate synthetase and malonic acid-semialdehyde dehydrogenase. The invention further relates to an ammonia (an ammonia ion) diagnosis / determination reagent kit. The determination method is high in sensitivity and small in error. Therefore, the determination method and the reagent kit can be widely applied to 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% to 13...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N21/31G01N21/33C12Q1/48C12Q1/25C12Q1/32
Inventor 王尔中
Owner SUZHOU ANJ BIOTECHNOLOGY CO LTD
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