Blood ammonia reagent kit for enzyme detection

A kit and blood ammonia technology, applied in the field of kits, can solve the problems of poor stability, cumbersome measurement, difficult long-term use in large quantities in laboratories, etc.

Inactive Publication Date: 2005-05-18
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] There are diffusion method, ion exchange method, ion selective electrode method and enzymatic method for the determination of blood ammonia. The first three methods are difficult to carry out on a large scale in clinical practice due to the cumbersome determination, while the enzymatic method has the characteristics of high specificity, trace amount, rapidity and accuracy. However, the enzymatic method for measuring blood ammonia introduced in the current literature is only suitable for a small amount of scientific research, because the enzymes and coenzymes used in this method are only suitable for short-term use and have poor stability, making it difficult to use them in large quantities in various hospital laboratories for a long time

Method used

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specific Embodiment approach 1

[0004] Specific embodiment one: the kit of this embodiment is composed of trishydroxymethylaminomethane, Tween-20, sodium azide, lactose, glutamate dehydrogenase, α-ketoglutarate, sodium pyruvate, reduced Nicotinamide adenine dinucleotide phosphate (NADPH) components, the content of each component is: Tris-hydroxymethylaminomethane 50-150mmol, Tween-200.1-2ml, sodium azide 0.5-2g, lactose 5 ~50g, glutamate dehydrogenase 500~1500u (u is the international unit of enzyme activity), α-ketoglutarate 2~20mmol, sodium pyruvate 1~10mmol, reduced nicotinamide adenine dinucleotide phosphate 0.1 ~ 1mmol, take distilled water, heat the distilled water to boiling before use, seal it for later use; dissolve Tris, Tween-20, sodium azide, lactose, α-ketoglutarate and sodium pyruvate in the above Adjust the pH to 8.0 in distilled water, then add glutamic acid dehydrogenase and NADPH to it, constant volume, sub-package, and then freeze-dry to obtain the finished product, with a validity period ...

specific Embodiment approach 2

[0005] Specific embodiment 2: The difference between this embodiment and specific embodiment 1 is that the content of each component is: trishydroxymethylaminomethane 55~90mmol, Tween-200.2~0.8ml, sodium azide 0.6~1.2g, Lactose 6-25g, glutamate dehydrogenase 600-900u, α-ketoglutarate 5-10mmol, sodium pyruvate 2-5mmol, reduced nicotinamide adenine dinucleotide phosphate 0.2-0.5mmol.

specific Embodiment approach 3

[0006] Specific embodiment three: the difference between this embodiment and specific embodiment one is that the content of each component is: trishydroxymethylaminomethane 100~140mmol, Tween-201.0~1.8ml, sodium azide 1.3~1.9g, Lactose 30-45g, glutamate dehydrogenase 1000-1400u, α-ketoglutarate 12-18mmol, sodium pyruvate 6-9mmol, reduced nicotinamide adenine dinucleotide phosphate 0.6-0.9mmol.

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Abstract

The reagent kit for enzyme detection of blood ammonia consists of tromethamine in 50-150 mmol, Tween-20 in 0.1-2 ml, sodium azide om 0.5-2 g, lactose in 5-50 g, glutamate dehydrogenase in 500-1500 u, alpha-ketoglutaric acid in 2-20 mmol, sodium acetonate in 1-10 mmol, and reducing nicotin-amide-adenine dinuclectide phosphate in 0.1-0.5 mmol. After regulating pH to optimal value of 8.0, the components are dissolved, packed and freeze dried and may be cold stored for two years. After being re-dissolved, the components are stabilized at room temperature for 1 week and cold stored to stabilize for 1 month. The reagent kit has powerful interference resistance, medium hemolysis and chylemia, blood sugar below 20 mmol / L, bilirubin below 0.1 mmol / L, enzyme activity loss less than 5 %, MADPH loss less than 10 %, linearity reaching 0.2 mmol / L, CV inside batch of 3.8 % and CV between batches of 5.1 %, and recovering rate of 96.1 %.

Description

Technical field: [0001] The invention relates to a kit, in particular to a kit for measuring blood ammonia by enzymatic method. Background technique: [0002] There are diffusion method, ion exchange method, ion selective electrode method and enzymatic method for the determination of blood ammonia. The first three methods are difficult to carry out on a large scale in clinical practice due to the cumbersome determination, while the enzymatic method has the characteristics of high specificity, trace amount, rapidity and accuracy. However, the enzymatic method for measuring blood ammonia introduced in the current literature is only suitable for a small amount of scientific research, because the enzymes and coenzymes used in this method are only suitable for short-term use and have poor stability, making it difficult to use them in large quantities in various hospital laboratories for a long time. Invention content: [0003] The purpose of the present invention is to provide ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/32
Inventor 李晓光于永光
Owner HARBIN MEDICAL UNIVERSITY
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