Molecular probes for Ascorbate detection

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

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

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