New approach to biosensing of co-enzyme nicotinamide adenine dinucleotide (NADH) by incorporation of neutral red in aluminum doped nanostructured ZnO thin films

Fidal V T; Chandra T S

doi:10.1016/j.bbagen.2017.01.001

New approach to biosensing of co-enzyme nicotinamide adenine dinucleotide (NADH) by incorporation of neutral red in aluminum doped nanostructured ZnO thin films

Biochim Biophys Acta Gen Subj. 2017 Jun;1861(6):1559-1565. doi: 10.1016/j.bbagen.2017.01.001. Epub 2017 Jan 4.

Authors

Fidal V T¹, Chandra T S²

Affiliations

¹ Bhupat and Jyoti Mehta School of Biosciences, Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, India.
² Bhupat and Jyoti Mehta School of Biosciences, Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, India. Electronic address: chandrasainathan@gmail.com.

PMID: 28062235
DOI: 10.1016/j.bbagen.2017.01.001

Abstract

Biosensing of NADH on bare electrodes has drawbacks such as high over-potential and poisoning during the oxidation reaction. To overcome this challenge a different approach has been undertaken by incorporating neutral red (NR) in Al doped ZnO (AZO) thin films using one-pot chemical bath deposition (CBD). The surface morphology of the films was hexagonal nanorods along the c-axis, perpendicular to the substrate. The thickness of the thin films were ranging from 400 to 3000nm varying dependent on time of deposition (30 to 150min). The average diameter of the nanorods was larger in the presence of neutral red (NR-AZO) with ~300nm in contrast to its absence (AZO) with ~200nm. The density of the packing of nanorods was dependent on the citrate concentration used during deposition. Control over the dopant concentration in the films was achieved by varying the area of Al foil used in the deposition solution. The selected area diffraction (SAED) and X-ray diffraction (XRD) indicated 002 plane of orientation in the nanorods. FTIR and FT-Raman analysis revealed conserved structure of NR and AZO. Chronoamperometric (CA) analysis showed a sensitivity of 0.45μAcm^-2mM^-1 and LoD of 22μM within the range 0.075-4mM of NADH. The biological sensing of NADH was validated by physical adsorption of NAD⁺ dependent-lactate dehydrogenase (LDH) on NR-AZO. CA showed sensitivity of 0.56μAcm^-2mM^-1 and LoD for lactate was 27μM in the range of 0.1-1mM of lactate. Further validation with real-time serum sample shows that LDH/NR-AZO correlates with the clinical values. The distinction in this study is that the organic mediator like neutral red has been incorporated into the grain structure of the ZnO thin film whereas other study with the mediators have only attempted surface functionalization. This article is part of a Special Issue entitled "Recent Advances in Bionanomaterials" Guest Editor: Dr. Marie-Louise Saboungi and Dr. Samuel D. Bader.

Keywords: Al and Ga doped ZnO films; Lactate dehydrogenase; NADH; Neutral red.

MeSH terms

Adsorption
Aluminum Compounds / chemistry*
Biosensing Techniques / instrumentation*
Citric Acid / chemistry
Electric Conductivity
Electrodes*
Equipment Design
Gallium / chemistry*
Humans
L-Lactate Dehydrogenase / chemistry
L-Lactate Dehydrogenase / metabolism
Lactic Acid / chemistry
Lactic Acid / metabolism
Microscopy, Electron, Scanning
Microscopy, Electron, Transmission
NAD / blood
NAD / chemistry*
Nanotechnology / instrumentation*
Nanotechnology / methods
Nanotubes / chemistry*
Neutral Red / chemistry*
Particle Size
Spectroscopy, Fourier Transform Infrared
Spectrum Analysis, Raman
Surface Properties
Time Factors
X-Ray Diffraction
Zinc Oxide / chemistry*

Substances

Aluminum Compounds
NAD
Neutral Red
Citric Acid
Lactic Acid
Gallium
L-Lactate Dehydrogenase
Zinc Oxide