Synthesis of robust electrochemical substrate and fabrication of immobilization free biosensors for rapid sensing of salicylate and β-hydroxybutyrate in whole blood

Chen TY 1 , Mani V 1 , Huang ST 2 , Chang PC 3 , Huang CH 3 , Huang NM 4

Affiliations 

  • 1 Graduate Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, Taipei, Taiwan, ROC
  • 2 Graduate Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, Taipei, Taiwan, ROC; Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, Taiwan, ROC. Electronic address: ws75624@ntut.edu.tw
  • 3 Graduate Institute of Biochemical and Biomedical Engineering, National Taipei University of Technology, Taipei, Taiwan, ROC; Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, Taiwan, ROC
  • 4 Low Dimensional Materials Research Centre, Department of Physics, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia; Center for Printable Electronics, University of Malaya, 50603 Kuala Lumpur, Malaysia
Anal Chim Acta, 2017 Oct 16;990:78-83.
PMID: 29029745 DOI: 10.1016/j.aca.2017.08.051

Abstract

An electrochemical latent redox probe, SAF 5 was designed, synthesized and characterized. A rapid and sensitive solution-based assay was demonstrated for salicylate hydroxylase (SHL). In presence of NADH at aerobic conditions, SHL catalyzed the decarboxylative hydroxylation of SAF and released a redox reporter amino ferrocene (AF 6). The release of AF 6 was monitored at interference free potential region (-50 mV vs. Ag|AgCl) using differential pulse voltammetry as signal read-out. The current signal generated by this process is highly specific, and insensitive to other biological interfering compounds. Next, the SAF incorporated SHL assay was extended to fabricate immobilization-free biosensors for rapid sensing of salicylic acid (SA) and β-hydroxybutyrate (β-HB) in whole blood. The described method rapidly detects SA in a linear range of 35-560 μM with detection limit of 5.0 μM. For β-HB determination, the linear range was 10-600 μM and detection limit was 2.0 μM. Besides, the assay protocols are simple, fast, reliable, selective, sensitive and advantageous over existing methods. The whole blood assay did not required cumbersome steps such as, enzyme immobilization, pre-treatments and holds great practical potential in clinical diagnosis.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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