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Synthesis of p-nitrobenzene-beta-D- glucoside and use thereof

A technology of p-nitrobenzene and p-nitrophenol, which is applied in the direction of material analysis, sugar derivatives, and microbial determination/inspection through observation of the impact on chemical indicators, can solve problems such as synthesis that has not been reported, and achieve The effect of simple method

Inactive Publication Date: 2009-01-07
天津市医药科学研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the synthesis of β-D-glucosidase substrate p-nitrophenyl-β-D-glucoside has not been reported

Method used

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  • Synthesis of p-nitrobenzene-beta-D- glucoside and use thereof
  • Synthesis of p-nitrobenzene-beta-D- glucoside and use thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0028] Substrate: Synthesis of p-Nitrophenyl-β-D Glucoside

[0029] ——Preparation of β-pentaacetylglucose: Take 200g of dried D-glucose, 40g of anhydrous sodium acetate, 1200g of acetic anhydride, warm to dissolve, stir and reflux on a water bath for 2 hours, pour into 6000ml of ice water while stirring, and stir 1 After hours, a large amount of white crystals precipitated, filtered, washed with water, and dried to obtain crude β-pentaacetylglucose, which was recrystallized with 7 times the amount of 95% ethanol to obtain 220 g of β-pentaacetylglucose crystals with a melting point of mp130-132°C;

[0030] ——Preparation of Bromine Reagent: Add 3g of red phosphorus to 30ml of glacial acetic acid, gradually add 18g of bromine dropwise under ice bath, let it stand for half an hour after completion, filter to obtain Bromine Reagent, seal and store in the refrigerator for later use.

[0031] ——Preparation of bromo-α-D-tetraacetylglucose: 50g of β-pentaacetylglucose, 75ml of bromine ...

Embodiment 2

[0048] For the rapid identification of Candida albicans and Candida dublinis, and identification of enterococci.

[0049] The steps are as follows:

[0050] --Material:

[0051] 1. Production of substrate paper: 6mm circular filter paper, then put the filter paper in a glass dish and sterilize at high temperature. Accurately weigh 4mg of p-nitrophenyl-β-D-glucoside and dissolve it in 1ml of normal saline, stir continuously to evenly distribute the substrate on the filter paper, and dry it at 35°C for later use.

[0052] 2. Buffer: Phosphate buffer with different pH (pH5.0~pH7.0).

[0053] 3. Stop solution: 0.1mol / L NaOH solution.

[0054] 4. Strains: 82 clinically isolated Candida albicans, Streptococcus, Enterococcus and Enterobacteriaceae. Standard strains for quality control: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Pseudomonas aeruginosa ATCC27853.

[0055] --operate:

[0056] (1) Add pH 5.5 phosphate buffer solution dropwise on the filter paper ...

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Abstract

The invention discloses the synthesis and the application of p-nitrophenyl beta-D-glucuronide. The synthesis method comprises the following steps: selecting D-glucose as the raw material, acetylating with acetic anhydride, brominating glycosyl group, condensating with p-nitrophenol, hydrolyzing in sodium methoxide solution to obtain p-nitrophenyl beta-D-glucuronide, which is the substrate of beta-D-glucosidase. The invention successfully synthesizes p-nitrophenyl beta-D-glucuronide, which is the substrate of beta-D-glucosidase, and provides a method for determining beta-D-glucosidase by using the substrate to conveniently and rapidly differentiate Candida albicans and Candida dubliniensis and rapidly identify enterococcus. The substrate is made to paper sheet, and the optimal substrate concentration is 4 mg / mL to achieve remarkable color change of the substrate. The optimal pH value of a phosphate buffer solution is 5.5, a 0.1 mol / L NaOH solution is selected as a stop solution to differentiate Candida albicans and Candida dubliniensis in 20 min and to identify enterococcus in 20 min. The invention provides a novel method for differentiating Candida albicans and Candida dubliniensis for clinical laboratory and for rapidly detecting enterococcus at a short time.

Description

Technical field: [0001] The present invention relates to the synthetic method of the substrate p-nitrophenyl-β-D-glucoside of β-D-glucosidase, and use this substrate to measure bacterial β-D-glucosidase to distinguish Candida albicans and Dublin Candida, and a rapid method for the identification of enterococci. Background technique: [0002] Both Candida albicans and Candida dublini are highly pathogenic. Candida dublinis is a new pathogenic candida named in 1995. In recent years, Candida dublini has been isolated from HIV-infected patients, and its virulence is stronger than that of Candida albicans. The hymenospore capture test was all positive, and the two bacteria could not be separated, which would affect the pertinence of later-stage medication. Furthermore, Enterococcus is an important pathogen of nosocomial infection, which can not only cause urinary tract infection, skin and soft tissue infection, but also life-threatening abdominal infection, sepsis, pericarditi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H15/203G01N21/78C12Q1/04
Inventor 孙家莉王金良陈丽宇
Owner 天津市医药科学研究所