AN IMMUNOSENSOR UTILIZING REDUCED GRAPHENE OXIDE AS A POTENTIAL SENSING MATERIAL FOR CORTISOL DETECTION: OPTIMIZED SCREEN-PRINTED GOLD ELECTRODE DESIGN FOR STRESS HORMONE MONITORING

Authors

  • Ahmad Farid Mohd Azmi Department of Chemistry & Biology, Centre for Defence Foundation Studies, National Defence University of Malaysia, Sungai Besi Camp, 57000 Kuala Lumpur, Malaysia
  • Vayithiswary Kannan Centre for Tropicalization, Defence Research Institute, National Defence University of Malaysia, Sungai Besi Camp, 57000 Kuala Lumpur, Malaysia
  • Muhammad Hafiz Ahmad Centre for Tropicalization, Defence Research Institute, National Defence University of Malaysia, Sungai Besi Camp, 57000 Kuala Lumpur, Malaysia
  • Jahwarhar Izuan Rashid Department of Chemistry & Biology, Centre for Defence Foundation Studies, National Defence University of Malaysia, Sungai Besi Camp, 57000 Kuala Lumpur, Malaysia & Centre for Tropicalization, Defence Research Institute, National Defence University of Malaysia, Sungai Besi Camp, 57000 Kuala Lumpur, Malaysia

Keywords:

cortisol; immunonsensor; graphene oxide; electrochemical; screen printed gold electrode

Abstract

Monitoring cortisol levels enables us to assess the physiological stress on our body and mind. An electrochemical immunosensor was fabricated by modifying a screen-printed gold electrode (SPGE) with reduced graphene oxide (rGO) and immobilizing cortisol monoclonal antibodies via EDC/NHS coupling chemistry. The surface morphology and electrochemical behavior of the fabricated rGO-modified SPGE were characterized using Field Emission Scanning Electron Microscopy (FESEM) and cyclic voltammetry (CV), respectively. The utilization of rGO as a sensing material significantly improved the electrochemical current signal, both in the presence and absence of cortisol. We optimized four key parameters to enhance the sensor's performance: rGO concentration, EDC: NHS solution ratio, C-Mab concentration, and cortisol incubation time. Through optimization, we reduced the rGO concentration from 2 mg/mL to 0.5 mg/mL, adjusted the EDC:NHS ratio from 0.6:1 to 0.2:1, decreased C-Mab concentration from 0.7 mg/mL to 0.5 mg/mL, and shortened the cortisol incubation time from 15 minutes to 3 minutes. These improvements led to a significant enhancement in sensor performance, with the reduction of peak current increasing from 39.03% to 66%, representing a 69.1% overall performance improvement. Under these optimized conditions, the developed sensor exhibited good selectivity among various hormones and demonstrated reasonable within 0.001–10 μg/mL, encompassing typical human levels (0.1–0.3 μg/mL under normal conditions and up to 1–2 μg/mL under stress with a detection limit of 1.9677 μg/mL. The sensor shows promise as a potential platform for future stress-related cortisol monitoring in various body fluids.

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Published

23-05-2026

How to Cite

Ahmad Farid Mohd Azmi, Vayithiswary Kannan, Muhammad Hafiz Ahmad, & Rashid, J. I. (2026). AN IMMUNOSENSOR UTILIZING REDUCED GRAPHENE OXIDE AS A POTENTIAL SENSING MATERIAL FOR CORTISOL DETECTION: OPTIMIZED SCREEN-PRINTED GOLD ELECTRODE DESIGN FOR STRESS HORMONE MONITORING. Zulfaqar Journal of Defence Science, Engineering & Technology, 9(1). Retrieved from https://zulfaqarjdset.upnm.edu.my/index.php/zjdset/article/view/190

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Vol. 9 No. 1 (2026)

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