Development of Cu-Laser Induced Graphene Nanocomposite-based

Spectroelectrochemical Sensor for Detection of Vanillin in Food Products

Vanillin (3-methoxy-4-hydroxybenzaldehyde), extracted from vanilla seeds, is a fragrant

substance. It is used as a flavor enhancer in soft drinks, cosmetics, medicines, and foods. It
has antioxidant properties that reduce the risk of heart attacks, oxidation of low-density
lipoproteins. However, the regular intake of vanillin containing food can establish a
detrimental pattern, as the fascinating aroma of vanillin may trigger a persistent appetite. The
WHO and Food and Agriculture Organization set a 7 mg per 100 g limit for vanillin in food
products for adults, while it is strictly prohibited in infant foods. Therefore, it is important to
quantify vanillin in food items for food safety.
In the last few decades simple and efficient electrochemical sensors have been
emerged for efficient sensing of vanillin and other biomolecules. Ruzgas et al. (1995) used
horseradish peroxidase-modified graphite electrode to convert phenolic compound in vanillin
to phenoxy radical. The current produced during electrochemical reduction of phenoxy
radical is proportional to the concentration of vanillin [1]. Subsequently, metal nanoparticles
with larger specific surface area and electrocatalytic activity were used to improve the
stability and sensitivity of the electrode material. The research group of Zheng et al. (2010)
immobilized Au-Ag bimetallic nanoparticles on glassy carbon electrode. The modification
enhanced the peak oxidation current five times in comparison to bare glass carbon electrode.
The sensor showed linear response over 0.2 - 50 μM [2]. Later on, Li et al. (2015) used the
composite of graphene oxide and Ag-Pd bimetallic nanoparticles. The electrode showed
relatively wide range of detection (0.02 - 45 μM) [3]. Recently, Chen et al. (2023)
synthesized chitosan based molecularly imprinted gel and combined it with CuS-modified
multiwalled carbon nanotubes [4]. The functionalized materials synergistically improved the
selectivity and sensitivity of the sensor. It is challenging to avoid interference from the
readily oxidizable electroactive molecules which are already present in real samples. The
UV-Vis spectroelectrochemical (SEC) approach can effectively handle the problem of
interference.
Material synthesis
A phenol-formaldehyde copolymeric film dispersed with Cu nanoparticles (Cu-P) is prepared
using suspension polymerization [5]. The laser ablation of Cu-P is performed using a fiber
laser with the optimized working parameters: 25% power (30 W), 5% speed, 50% frequency,
and 1200 dots per inch. Post-laser ablation, the polymeric surface of the Cu-P film is
transformed to graphene, termed as Cu-LIG. The electrocatalyst powder are prepared by
scrapping the Cu-LIG layer from the film. A homogeneous ink of Cu-LIG is prepared by
mixing 10 mg of the Cu-LIG powder and 2 mg of polyvinylidene fluoride in 10 mL of
isopropyl alcohol, followed by the sonication of the mixture for 10 min. An ITO electrode
(15 mm × 8 mm) is properly cleaned by sequentially sonicating the electrode in acetone,
water, and IPA, each for 10 min. The Cu-LIG ink is then spray coated over the indium tin
oxide (ITO) electrode.
Materials characterization
The morphology of the synthesized materials is determined by scanning electron microscopy
(SEM). Fig. 1a shows the SEM image of Cu-LIG film. The groove like structure aligned in
the direction of laser ablation is observed. Fig. 1b shows the SEM image of Cu-LIG powder.
The highly porous structures of powder indicate the large specific area which is desirable
property for an electrocatalyst. Fig. 1c and 1d show the EDX spectra of Cu-P film and Cu-
LIG powder, respectively. After laser ablation the content of C and Cu is increased from 75.9
to 83.4% and 2.3 to 10.2%, respectively. X-ray diffraction patterns of the synthesized
materials are used to determine the phase composition and crystal structure (Fig. 1e). The
broad peak in the spectra of Cu-P at 2θ value of 19.8° shows the presence amorphous
polymeric material. The presence of sharp peak at 2θ value of 26.3° in the spectra of Cu-LIG
attributes to the (0 0 2) plane of graphene. The peaks at 2θ values of 43.7, 50.6, 74.4, and
90.1° show the presence of (1 1 1), (2 0 0), (2 2 0) and (3 1 1) crystal planes of Cu
nanoparticles, respectively. Fig. 1f shows the Raman spectroscopy analysis of Cu-LIG. The
peaks corresponding to 1345, 1590, and 2680 cm-1 wavenumber attributed to the presence of
D, G and 2D bands, respectively in Cu-LIG. The calculated I D/IG ratio is 0.864, implying
higher amount of sp2 hybridized planar carbon or graphene. The 2D band shows the presence
of multilayered graphene sheets.
Fig. 1. SEM images of Cu-LIG (a) film (b) powder, EDX spectra of (c) Cu-P film, (d) Cu-
LIG powder, (e) XRD spectra of Cu-P and Cu-LIG, and (f) Raman spectra of Cu-LIG.
Spectroelectrochemical workstation
Fig.2(a-c) shows the digital photograph of spectroelectrochemical workstation. The
electrochemical potentiostat and UV-Vis spectrophotometer are connected in parallel to the
quartz cuvette. The working (Cu-LIG/ITO), reference (Ag/AgCl), counter electrodes are
placed in the quartz cuvette in which electrochemical reaction take place. Optical fibers from
the opposite sides of the cuvette are attached to the light source and detector. The UV-Vis
spectra are recorded at different applied potentials during the cyclic voltammetry. The light
source as well as detector are attached to the Potentiostat. The input signal is transmitted by
potentiostat to light source and the output signals are received at the detector.
Spectroelectrochemical analysis of vanillin
The spectroelectrochemical behaviour of vanillin over ITO and Cu-LIG/ITO is determined
from cyclic voltammetry (CV) analysis of vanillin solution. Fig. 4d and 4e show the CV and
UV-Vis spectra of vanillin, respectively. Vanillin solution is prepared in the acetate buffer.
No peaks are observed in the ITO electrode with no modification. The oxidation peaks at
0.33 and 0.72 V are observed in the forward scan while the peak at 0.6 V is observed in the
backward scan. In UV-Vis spectra the peak in the spectra would start appearing when the
applied potential exceeds 0.7 V. The peak intensity increases with increase in the applied
potential.
 Fig. 2. Digital photographs of the spectroelectrochemical workstation (a) potentiostat, (b)
spectroelectrochemical three-electrode assembly and (c) UV-Vis light source and detector,
(d) Cyclic voltammetry profile of vanillin over ITO and Cu-LIG electrode, and (e) UV-Vis
spectra of vanillin in acetate buffer at different applied potentials.

Future Work
The future work will include TEM and XPS analysis of the synthesized material.
Electrochemical characterization (electrochemical impedance spectroscopy, electrochemical
surface area, etc). Optimizing the parameters of spectroelectrochemical reaction study of
vanillin. Interference and selectivity study of vanillin over Cu-LIG electrode. Determination
of vanillin in the real food samples.
References

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