Abstract:
In this work, we aim to develop a simple, and rapid, nonenzymatic and
homogeneous biosensor for sensitive and rapid quantification of hydrogen peroxide.
Methods: In this technique, hemoglobin was used as a bioreceptor, where heme groups act as
electroactive centers to catalyze hydrogen peroxide reduction. The chemiluminescence reagent; luminol
is also a peroxidase substrate and can be oxidized by hemoglobin thus generating a CL signal. The
working principle was based on the competition between hydrogen peroxide and luminol towards
hemoglobin. A 96-well microplate was used to perform this assay. For thatat, 25 µL of Hb solution (3
µg mL−1) was added to each well, followed by 25 µL of different concentrations of H2O2. After
incubation, 25 µL of luminol solution was also added. Then, the chemiluminescence intensity was
measured at an emission wavelength of 425 nm.
Results and discussion: The detection principle is mainly based on the chemiluminescence
signal diminution in the presence of H2O2. Because this latter will react with Hb that is essential for
luminol reaction, thus leading to a lower chemiluminescence signal. AlsoUnder optimized conditions,
the chemiluminescent signal decreased with increasing hydrogen peroxide concentrations within the
linear range of 0.5 to 12 mM, with a correlation coefficient R2 of 0.99762. The limit of detection was
calculated to be as low as 0.308 mM. The selectivity of the biosensor was successfully demonstrated
against different interferents.
Conclusion: In this work, an homogenous chemiluminescent assay was developed using Hb as
a bioreceptor. The developed strategy provides a one step, simple, and low-cost bioanalytical method
which can be applied for the monitoring of hydrogen peroxide
