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Molecular probes for Ascorbate detection

Molecular probes for Ascorbate detection
Field
Research Tools and Methods
Patent
IP00638

Technology Overview

 

  • Ascorbate or ascorbic acid, or Vitamin C, is extensively used as an additive in food products.  It is also an important cellular component that mediates many different biochemical/enzymatic reactions. Methods for detection and quantification of ascorbate in biological and food samples are therefore highly valuable bioanalytical tools.
  • The invention describes a method for designing and synthesizing various stimulus responsive materials that are activated by ascorbate.
  • It employs a biomimetic strategy using copper complexes to mimic the active site of some copper monooxygenases for selective ascorbate-triggered oxidative bond breaking reaction.
  • The fluorescence response is also highly selective to ascorbate over a wide range of potential interferences including other biological/inorganic/organic reducing agents, other vitamins, sugars, catecholamines, amino acids, organic acids, metal ions, reactive oxygen species and most importantly dehydroascorbate, the oxidized product of ascorbate.

 

Advantages

  • Direct detection of ascorbate/ascorbic acid.
  • The detection of ascorbate of the current probe (Cu(II) Compound #1) is specific and is not affected by reducing agents and radical/paramagnetic species in the sample.
  • Ascorbate induced fluorescence enhancement can be as high as 200-fold.
  • High selectivity of the trigger towards ascorbate over numerous potential interfering chemicals.
  • High modularity for developing different ascorbate-responsive materials.
  • Fast response time.

Existing devices or processes and their shortcomings

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Existing devices or processes and their shortcomings

HPLC (High performance liquid chromatography) method

  • Time consuming
  • Expensive maintenance for the instrument and skilled technician is required  

Fluorometric/colourmetric Assay : OPDA method 

 

  • Prone to interference from other proteins and serum components
  • Does not directly detect ascorbate Not applicable in live biological samples

Fluorometric/colourmetric Assay : FRASC method

 

  • An instable and expensive enzyme is used that needs fresh preparation.
  • The assay is not a direct detection of ascorbate but the total amount of antioxidants
  • The assay required 30 minutes for completion
  • Not applicable in live biological sample

Fluorometric/ colourmetric Assay : oxidation method

 

  • The ascorbate oxidase used is fairly instable and expensive which needs fresh preparation
  • thiol-containing compounds are known to interfere in this assay
  • The assay is not a direct detection of ascorbate but the H2O2 product from its reaction with ascorbate oxidase

 

   

Potential Product and Applications

  • Fluorescence sensor for detecting ascorbic acid in food, clinical, plant and pharmaceutical analysis.
  • Can be used on live biological samples.

Development and IP Status

  • US granted Patent – US 10018623

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