Bio-Electroanalysis Performance of Heme Redox-Center for π- π Interaction Bonding of a Methylene Blue-Graphene Modified Electrode

Porntip Khownarumit; Kanmanee Choosang; Rungtiva P Poo-Arporn; Yingyot Poo-Arporn; Narong Chanlek; Werasak Surareungchai

doi:10.3390/nano13040745

Bio-Electroanalysis Performance of Heme Redox-Center for π- π Interaction Bonding of a Methylene Blue-Graphene Modified Electrode

Nanomaterials (Basel). 2023 Feb 16;13(4):745. doi: 10.3390/nano13040745.

Authors

Porntip Khownarumit¹, Kanmanee Choosang¹, Rungtiva P Poo-Arporn², Yingyot Poo-Arporn³, Narong Chanlek³, Werasak Surareungchai^{4

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Affiliations

¹ Sensor Technology Laboratory, Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi, Bangkok 10150, Thailand.
² Biological Engineering Program, Faculty of Engineering, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand.
³ Synchrotron Research and Applications Division, Synchrotron Light Research Institute, 111 University Avenue, Nakhon Ratchasima 30000, Thailand.
⁴ Faculty of Science and Nanoscience & Nanotechnology Graduate Program, King Mongkut's University of Technology Thonburi, Bangkok 10140, Thailand.
⁵ Bangkhuntein Campus, School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok 10150, Thailand.
⁶ Analytical Sciences and National Doping Test Institute, Mahidol University, Bangkok 10400, Thailand.

Abstract

Hemeprotein detection has motivated extensive research on the direct reaction of a heme molecule and a redox dye. The present study used methylene blue as both donor and acceptor for a redox reaction. First, the solid phases of methylene blue (MB) and graphene (GP) formed a π-π interaction bond at the aromatic rings. The conductivity of GP was better than that of carbon in a carbon electrode (CE). Then, the working CE was modified using strong adsorption of MB/GP on the electrode surface. The surface of the electrode was investigated using a modified and an unmodified electrode. The electrode's properties were studied using voltammograms of redox couple K₃[Fe(CN)₆]^3-/4-. Its reaction was used to find the active area of the modified electrode, which was 1.76 times bigger than that of the unmodified electrode. The surface coverage values of the modified and unmodified electrodes were 8.17 × 10^-6 and 1.53 × 10^-5 mol/cm², respectively. This research also studied the application of hemeprotein detection. Hemoglobin (Hb), myoglobin (Mb), and cytochrome c (Cyt. C) were studied by the reaction of Fe (III/II) at the heme-redox center. The electrocatalytic reaction between MB/GP and hemeproteins produced an anodic peak at 0.35 V for Hb, Mb, and Cyt. C. This nanohybrid film enhanced electron transfer between protein molecules and the modified carbon electrode. The amperometric measurements show that the limit of detection was 0.2 µM, 0.3 µM, and 0.1 µM for Hb, Mb, and Cyt. C, respectively. The measurement spanned a linear range of 0.2 µM to 5 µM, 0.3 µM to 5 µM, and 0.1 µM to 0.7 µM for Hb, Mb, and Cyt. C, respectively. Hb showed the lowest sensitivity compared with Mb and Cyt. C due to the role of steric hindrance in the hemeprotein specificity structure. This study offers a simple and efficient fabrication platform for electrochemical sensors for hemeproteins. When compared to other complex immobilization processes, the fabrication method for this sensor has many benefits, including no need for special chemicals and easy preparation and electrode modification-both of which are crucial for the development of electrochemical sensing devices.

Keywords: electrochemical sensor; graphene; heme; methylene blue; modified electrode.

Abstract

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