One-drop electrosynthesis of 3D Prussian blue analog@PEDOT on flexible porous graphene for portable NFC-enabled hydrogen peroxide determination in human semen.

Hydrogen peroxide (H2O2) is a relatively stable reactive oxygen species and an important biomarker of oxidative stress in human semen. Elevated H2O2 levels have been associated with impaired sperm function and male infertility. Therefore, the accurate determination of total H2O2 levels in semen is valuable for assessing semen oxidative status. In this work, a cost-effective and sensitive electrochemical sensing platform was developed for the determination of total H2O2 in human semen. A lab-made flexible porous graphene electrode was modified with a three-dimensional poly(3,4-ethylenedioxythiophene) film embedded with Prussian blue analog (3D PBA@PEDOT/F-PGE). The 3D PBA@PEDOT/F-PGE transducer was fabricated via a one-step electrodeposition process, providing high electrical conductivity, low charge-transfer resistance, excellent structural integrity, and stable electrocatalytic activity toward H2O2 reduction. The transducer exhibited a linear detection range from 0.50 μM to 6.0 mM, a sensitivity of 390 μA mM-1 cm-2, and a detection limit of 0.18 μM. The transducer was further integrated with a paper-based microfluidic device and a battery-free near-field communication (NFC) potentiostat to construct a portable electrochemical sensing platform capable of determining H2O2 over a concentration range of 1.0 μM to 4.0 mM with good precision and long-term stability. The smartphone-controlled sensing platform was successfully applied to the determination of total H2O2 levels in human semen samples, providing a convenient approach for assessing semen oxidative status. This platform shows considerable potential for point-of-care monitoring of oxidative stress associated with male reproductive health and for future applications in portable electrochemical sensing technologies.