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Purification method of electron mediator azure c

A technology of electronic medium and purification method, which is applied in the field of purification of electronic medium, can solve the problems of low sample loading, low HPLC purification efficiency, and high cost, and achieve the effects of improving production efficiency, large output, and large processing capacity

Active Publication Date: 2022-06-24
GENCLONN BIOTECH HANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the solubility of azure C and the low amount of sample prepared by HPLC, generally about 5 mg of a single sample is loaded, and 1-2 mg of azure C can be obtained in a single purification, resulting in a very low purification efficiency of HPLC. High cost

Method used

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  • Purification method of electron mediator azure c
  • Purification method of electron mediator azure c
  • Purification method of electron mediator azure c

Examples

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Embodiment 1

[0026] Example 1 Purification of Azure C (Azure C) with Fmoc-Cl as an amino protecting reagent

[0027] Fluorene methoxycarbonyl chloride (Fmoc-Cl) as an amino protecting agent to purify azurine C.

[0028]

[0029] Weigh 15g (141.5mmol) of sodium carbonate, add 150ml of water and stir to dissolve, then add 150ml of dioxane, 10g (36mmol) of commercially available Azure C with a content of 40%, and cool to 10°C in an ice-water bath. 23.28 g (90 mmol) of fluorene methoxycarbonyl chloride was added in batches, reacted in an ice-water bath for 1 hour, and then transferred to room temperature to react for 2 hours, resulting in a viscous product. The supernatant was poured off, the viscous product was dissolved in dichloromethane, insoluble impurities were filtered off, and the filtrate was washed with water; dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 3.4 g of intermediate (Fmoc) 2 -Azure C. (Fmoc) 2 -Azure C is amino-protected azure C.

[0030]...

Embodiment 2

[0031] Example 2 Commercially available Azure C was directly prepared and purified by HPLC (comparative example)

[0032] As a comparative example of the present invention, the method for directly purifying commercially available Azure C by HPLC is as follows.

[0033] Instrument: Waters Prep150 preparative liquid chromatography, chromatographic column: SunFire® Prep C18 OBD™ 5μm 19×150mm Column.

[0034] Mobile phase: acetonitrile / water (0.05% trifluoroacetic acid), acetonitrile %=15~55% (10min).

[0035] Flow rate: 20ml / min.

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Abstract

The invention provides a method for purifying an electron mediator azure C, comprising the following steps: taking low-purity azure C, using fluorenylmethoxycarbonyl chloride as an amino-protecting reagent to react with azure C to obtain amino-protected electrons After deprotection of the medium, high-purity azure C can be obtained. The purification method of the present invention has a large amount of single treatment, and the output of the obtained high-purity product is also large, which can effectively meet the preparation requirements of biosensors for blood sugar detection and the like, and improve production efficiency.

Description

technical field [0001] The invention relates to a purification method of an electronic mediator, in particular to a purification method of the electronic mediator azure C used for the preparation of biosensors. Background technique [0002] Diabetes is a disease caused by defective insulin secretion or reduced biological effects. At present, there is no medical method that can completely cure diabetes. Actively passing early screening and corresponding preventive treatment is an effective means to control blood sugar elevation. The blood glucose meter using the enzyme electrode method for daily monitoring of blood glucose levels is a commonly used method at present. [0003] In the blood glucose biosensor (also known as blood glucose test strip) of the enzyme electrode method, the enzyme used is a macromolecule, and its redox active center is coated in the enzyme protein molecule, and the direct connection between it and the electrode surface is Electron transfer is diffic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D279/18G01N27/327
CPCC07D279/18G01N27/3275
Inventor 陆晓峰徐俊杰
Owner GENCLONN BIOTECH HANGZHOU
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