Tanin Tangkuaram. Development of electrochemical sensors for analysis of medical and environmental analytes. Doctoral Degree(Analytical Chemistry). Mahidol University. : Mahidol University, 2007.
Development of electrochemical sensors for analysis of medical and environmental analytes
Abstract:
Four different electrochemical sensors were developed for analysis of some selected
analytes, namely: (i) insulin, (ii) glycosidases, (iii) hydrogen peroxide (H2O2), and (iv)
phosphate (PO4
3-). These are very important in health care and environmental monitoring. The
improvement of sensitivity, selectivity, detection limit and stability of these sensors were
investigated.
(i) Insulin is a pancreatic endocrine hormone for controlling body cells adsorbing and
using glucose. The insulin sensor was developed by using a bilayer surface coating of
ruthenium oxide (RuOx) onto a carbon nanotube (CNT) layer. RuOx/CNT offers dramatic
improvement in the stability and sensitivity of voltammetric and amperometric measurements
of insulin. A wide linear dynamic range (10 - 800 nM) was achieved with a detection limit of
1 nM insulin.
(ii) Glycosidases are index enzymes for cancer screening. The mechanism of the
glycosidases sensor involved the enzymes converting a probe substrate, 4-nitrophenol-β-Dgalactopyranoside,
to an electrochemical active form as p-nitrophenol. The detection signal of
p-nitrophenol oxidation is a proportion of the glycosidases content. The electrochemical assay
for glycosidases enzymes is better than the conventional spectrophotometric techniques.
(iii) Hydrogen peroxide is an intermediate species in many enzymatic reactions. The
H2O2 biosensor was designed on a screen printed carbon electrode (SPCE). A bare SPCE was
modified by horseradish peroxidase (HRP)/gold nanoparticles (AuNP) in the matrix of
chitosan (HRP/AuNP/CHIT). The HRP/AuNP incorporated with CHIT yielded the higher
performance. It retained amperometric response over 95% of the initial current of the first day
up to 30 days of storage at 4 °C. The biosensor showed a linear range of 0.01 – 11.3 mM
H2O2, with a detection limit of 0.65 μM H2O2.
(iv) Phosphate is an important nutrient form of phosphorus related to the algae bloom
phenomenon. A phosphate biosensor was designed by immobilizing pyruvate oxidase on
H2O2 biosensor. Pyruvate associated phosphate via a phosphorylation reaction to generate
H2O2 is an important step of phosphate biosensor. The detectable H2O2 is proportional to the
phosphate content in the samples. The practical use of this biosensor for phosphate
determination in real samples was demonstrated and validated with UV-Vis
spectrophotometry.