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Kit for enzymatic determination of 1,5-anhydro-D-glucitol

A sorbitan and reagent technology, applied in the field of enzymatic determination 1, can solve the problems of troublesome operation, inability to store for a long time, difficult batch detection, etc., and achieve the effects of strong anti-interference ability, convenient use, and cost reduction

Active Publication Date: 2012-07-11
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are relatively accurate, but the operation is laborious and difficult to detect in batches, and these methods can only be prepared and used immediately, and cannot be stored for a long time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Take one liter of ultrapure water, heat the water to boiling before use, seal it for later use, add 60 mmoles of 3-hydroxy-methylaminomethane into the water, adjust the pH to 8.5, take 2.26 mmoles of oxidized coenzyme II (NADP+), protective agent Trehalose 1g, 2-methyl-4-isothiazol-3-one 0.1 active unit, Tween-80 0.1g, ADP-dependent hexokinase (ADP-HK) 1000U, AG-6-phosphate dehydrogenase 1000U and 0.1 mmol of adenosine diphosphate (ADP) were added to the reagent, and the resulting reagent solution was divided into 10 ml / bottles, and refrigerated for use at any time.

Embodiment 2

[0013] Take 1 liter of ultrapure water, heat the water to boiling before use, seal it for later use, adjust the pH to 9.5 with 140 mmol of 3-hydroxy-methylaminomethane, take 2.38 mmol of oxidized coenzyme II (NADP+), trehalose as a protective agent 5 grams, 2-methyl-4-isothiazol-3-one 0.6 activity units, Tween-80 0.6 grams, DP-dependent hexokinase (ADP-HK) 3000U, AG-6-phosphate dehydrogenase 80000U, Adenosine diphosphate (ADP) 0.3 mmol was added to the reagent, and the obtained reagent solution was divided into 5 ml / bottles, and kept refrigerated for future use.

Embodiment 3

[0015] Take one liter of ultrapure water, heat the water to boiling before use, and seal it for later use. Adjust the pH to 9.5 with 100 millimoles of 3-hydroxy-methylaminomethane, take 2.30 millimoles of oxidized coenzyme II (NADP+), 3 grams of protective agent trehalose, and 0.5 grams of 2-methyl-4-isothiazol-3-one Active unit, Tween-80 0.3 g, ADP-dependent hexokinase (ADP-HK) 2000 U, AG-6-phosphate dehydrogenase 50000 U, adenosine diphosphate (ADP) 0.2 mmol were added to the reagent, and the resulting reagent The solution was divided into 5ml / bottles, and kept refrigerated for use at any time.

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PUM

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Abstract

The invention discloses an reagent for enzymatic determination of 1,5-anhydro-D-glucitol. At present, determination operation has a lot of trouble and is difficult to be detected in bulk, and in the methods for the operation, the reagent only can be prepared when being used without long term preservation. The reagent of the invention comprises the following components with the contents in 1 literof water: 60-140 mmol of 3-hydroxyl methyl amino methane, 2.26-2.38 mmol of oxidized coenzyme II (NADP+), 1-5 grams of trehalose, 0.1-0.6 activity unit of 2-methyl-4-isothiazole-3-ketone, 0.1-0.6 gram of Twain, 1000-3000U of ADP-dependent hexokinase (ADP-HK), 1000-80000U of AG-6-phosphate dehydrogenase and 0.1-0.3 mmol of adenosine diphosphate (ADP). The invention is used for determining 1,5- anhydro-D-glucitol.

Description

Technical field: [0001] The invention relates to a reagent for the determination of 1,5-anhydrosorbitol by an enzymatic method. Background technique: [0002] Diabetes is a common and frequently-occurring disease in clinical practice. It is caused by insufficient insulin secretion. The level of AG in plasma is significantly negatively correlated with plasma glucose, HbAlc and fructosamine (FMN). According to data, there is no case of plasma glucose > 8.3mmol / L. Associated with high levels of plasma AG [12]. The plasma AG of diabetic patients is significantly lower than that of healthy people, especially in type 1 diabetic patients compared with type 2 diabetic patients. The plasma AG of new-onset and long-term patients with type 1 diabetes is generally below 1umol / L[13], and even reduced to undetectable out. Determination of 1,5-sorbitol has high performance liquid chromatography, micro-column chromatography, chemiluminescence and so on. These methods are relatively ac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/48C12Q1/32
Inventor 贺丽雅张颖于永光
Owner HARBIN MEDICAL UNIVERSITY