LWT - Food Science and Technology 88 (2018) 152e158

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LWT - Food Science and Technology

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Development of silver/carbon screen-printed electrode for rapid

determination of vitamin C from fruit juices
Jaymin K. Jadav a, Valentina V. Umrania a, Khyati J. Rathod b, Baljibhai A. Golakiya b, *
a
Department of Microbiology, MVM Science & Home Science College, Saurashtra University, Rajkot, Gujarat, India
b
Department of Biotechnology, Junagadh Agricultural University, Junagadh, Gujarat, India

a r t i c l e i n f o a b s t r a c t

Article history: A silver/carbon screen-printed electrode (S/C-SPE) was fabricated using polymer-based conductive inks
Received 3 July 2017 blended with in-house synthesized nanoparticles and investigated for its application in electrochemical
Received in revised form analysis. Silver nanoparticles (~59 nm) and carbon nanoparticles (~76 nm) were synthesized having zeta
30 September 2017
potential (<±60 mV) and particle morphology were investigated for both the nanoparticles. These
Accepted 2 October 2017
Available online 5 October 2017
nanoparticles were blended with a thermoset epoxy resin to formulate different conductive inks viz.
silver (Ag) ink, carbon (C) ink and silver/silver chloride (Ag/AgCl) ink. The formulated inks were char-
acterized for their viscosity, which ranged from 1500 to 4500 cP making them ideal for screen printing.
Keywords:
Nanoparticles
Post printing and thermal curing, the sensing area of the S/C-SPE was characterized for its elemental
Conductive inks content by EDX, electrode thickness ranged from 0.3 to 0.8 mm and bulk resistance (r) (4.7
Screen-printed electrodes e13.4
104 U-cm). The level of vitamin C was determined in different fruit juices at the S/C-SPE using a
Cyclic-voltammetry developed voltammetric method and compared with that of standard biochemical method. The vitamin
Food analysis C content of different juices ranged from 5 to 60 mg/100 ml. The analytical characteristics of the S/C SPE
(linear range, reproducibility, stability, matrix effect, electrode saturation and response time) were
investigated. The results obtained by the two methods were in good agreement.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction and portability, sensitivity, selectivity, a wide linear range, minimal

In the last few years with the recent advancement in develop-
ment of printed electronics, there has been a growing interest in
replacing traditional silicon techniques with new and robust
methodologies to develop printed electronics as they offer low cost,
less tedious processing and better results. Development in the field
of analytical chemistry is aimed to meet the need of fast, easy and
on site analysis as well as improved analytical methods which offer
better result outputs with lesser environmental impact. A field
detection system for real time analysis is highly desirable to over-
come time consuming process such as sample collection, its
transfer to a laboratory as well as tedious conventional testing
methods. Such real time testing system put forward a rapid return
of the results with minimized errors and costs as compared to the
offsite laboratory-based analyses (Hayat & Marty, 2014; Wang,
2002). Current progress in analytical chemistry is to develop elec-
trochemical devices which include possibility of miniaturization
* Corresponding author.
E-mail address: bag@jau.in (B.A. Golakiya).
space and power requirement and cost effective instrumentation
(Honeychurch & Hart, 2003; Thiyagarajan, Chang, Senthilkumar, &
Zen, 2014). Such devices can be accomplished with printed elec-
tronics (PE) with various functional inks containing nanomaterials
such as metallic nanoparticles, organic electronics molecules/
polymers and carbon based nanomaterials, employed to bulk pro-
duction process for advanced electronic equipments (Lee, Jun, Kim,
& Joung, 2006). Use of silver nanoparticles based conductive inks,
has been found to be a very powerful tool for direct patterning of
desired schematic conductive tracks in electronic devices (Kim,
Jeong, Park, & Moon, 2006). When coupling such nanoparticles
with advanced polymer composites, printed schematics can be
annealed or cured at moderate temperatures to form conductive
films of low resistance with potential suitability for use in printed
electronics (Allen, Bayles, Gile, & Jesser, 1986; Buffat & Borel, 1976;
Jiang, Zhang, & Zhao, 2003).
Vitamin is a nutrient that does not provide energy to the body
but it is essential nutrients. It is necessary for the growth of the
body and also associated with the enzyme function in the body
(Okiei et al., 2009). Vitamin C minimum daily requirement for

https://doi.org/10.1016/j.lwt.2017.10.005
0023-6438/© 2017 Elsevier Ltd. All rights reserved.
 J.K. Jadav et al. / LWT - Food Science and Technology 88 (2018) 152e158
adults is 60 mg. Vitamin C is potent anti-oxidant and free radical
scavenger, thus, helps in body tissue growth and repair (Cathcart,
1991). It is essential in synthesizing collagen, ligaments and blood
vessels, iron absorption, and immune response activation and is
involved in wound healing and osteogenesis. Furthermore, Vitamin
C has the properties of reducing agent and weak acid (Valente,
Sanches-Silva, Albuquerque, & Costa, 2014).
Electrochemical analysis such as voltammetry is an increasingly
popular methodology applied to the determination of vitamin C in
real samples (Wawrzyniak, Ryniecki, & Zembrzuski, 2005), because
it offers low detection limits with little or no sample preparation,
against more expensive techniques. Voltammetry offers an attrac-
tive alternative to the titrimetric or instrumental methods such as
spectroscopy and liquid chromatography, in particular in food
quality control due to its low cost as well as simplicity of the
employed procedures necessary to determine vitamin C. It is quick,
easy and does not require complicated, expensive equipment and
well-qualified personnel like the previously mentioned instru-
mental techniques.
Present work comprises synthesizing stable silver and carbon
nanoparticles and blending them to a thermoset clear epoxy resin
(Oldenzijl, Chen and Gunther 2015) to formulate different
conductive inks. The solid content of the said conductive inks was
maintained to make ink screen printable with viscosity in the range
of 1000e10,000 cP. A screen-printed three electrode electro-
chemical cell was manually screen-printed on to a polyester (PET)
sheet and was heat cured, which annealed nanoparticles with
epoxy polymer so as to get printed tracks. The S/C-SPE was char-
acterized for its electrochemical performance by cyclic voltamme-
try analysis of ferro-ferri couple at different scan rate followed by
the S/C-SPE was used to estimate ascorbic acid (vitamin C) from
different fruit juices.
2. Material and methods
Solid chemicals such as Silver Nitrate (AgNO3), Silver Chloride
(AgCl2), Tri-sodium citrate were purchased from Merck-KGaA
(Germany), the epoxy resin - Thermoset E-105 Epoxy resin and
Circalok 6055B Hardener were procured from LORD Corporation
(USA). Polymer dielectric (D2081009D6) was purchased from Gwent
Electronic Materials Ltd. (United Kingdom). Sodium dihydrogen
ortho-phosphate and ascorbic acid were procured from Merck
Chemical (Germany). 2,4-Dinitrophenylhydrazine (DNPH) dye pur-
chased from Sigma-Aldrich (Madrid, Spain). The deionized (DI)
water (resistivity ¼ 18.2 MU.cm, and conductivity ¼ 0.058 at 25  C)
for synthesis of nanoparticles was collected from Merck Millipore
water purification system (USA) installed in our laboratory.
2.1. Synthesis of nanoparticles for conductive inks formulation
2.1.1. Synthesis of silver nanoparticles by bottom-up technique
Silver nanoparticles (SNPs) were synthesized by bottom-up
techniques using chemical reduction of 100 ml 1 mM aqueous so-
lution of AgNO3 with quick addition of 10 ml 1.5% tri-sodium citrate
as reducing agent at 80e85  C under continuous stirring (Sodha,
Jadav, Gajera, & Rathod, 2015). The unfavorable agglomeration
was avoided with the AgNO3/PVP mass ratio of 1: 4 (Xiliang et al.,
2014). Following addition of reducing agent the temperature was
raised to boiling and maintained for 10 min. The reaction was given
a cold temperature shock by immediately placing it in ice bath.
Gryeish black powder of SNPs was obtained by high speed centri-
fugation at 12,000 rpm with twice intermediate washing with DI
water for 15 min and drying under vacuum at 70  C for 30 min.
These dried SNP powder was further used for formulation of
conductive inks.
153
2.1.2. Synthesis of carbon nanoparticles by top-down technique
Wooden charcoal pieces were burnt in fireplace for 3e4 h. The
small pieces of black mass so obtained with innate resistance be-
tween 50 and 80 U using two-probe multimeter were used for
synthesis of carbon nanoparticles (CNPs). These pieces were socked
in DI water overnight and blended using high speed homogenizer
as mentioned by (Yadav, Yadav, & Singh, 2012). The aqueous black
slurry so obtained was filtered under high vacuum through 0.2 mm
PTFE membrane filter. The filtrate containing the carbon nano-
particles was dried overnight at 60  C.
2.2. Formulation of conductive inks
2.2.1. Formulation of silver conductive ink
Silver conductive ink was prepared by dispersing SNP powder in
Thermoset E-105 clear epoxy resin at 10,000 rpm using high speed
homogenizer at 25  C in a temperature controlled water bath. The
percentage (%) silver content was kept as 90e94% w/v. Once the
nanoparticles and resin were homogeneously mixed, Circalok
6055B Hardener was added in 0.84:1 w/w ratio. The nanoparticles
dispersed epoxy resin and hardener was mixed at 25  C under high
vacuum to avoid formation of air bubbles. The final solid content in
the silver paste was 45e47% w/v (Jewell, Hamblyn, Claypole, &
Gethin, 2015).
2.2.2. Formulation of carbon conductive ink
Carbon conductive ink was prepared by uniformly dispersing
carbon nanoparticles in thermoset E-105 clear epoxy resin and
Circalok 6055B Hardener by process similar to silver ink. The final
solid content of the carbon ink was adjusted to 40e42% w/v
(Calixto et al., 2007).
2.2.3. Formulation of silver/silver chloride conductive ink
Silver/silver chloride (Ag/AgCl2) ink was prepared by initially
dissolving 80% w/v silver chloride ink E-105 epoxy resin followed
by dispersing SNPs by high speed homogenizer followed by addi-
tion of Circalok 6055B Hardener similar to that of silver ink
formulation. The final solid content was adjusted to 40e45% w/v
(Jewell et al., 2015).
2.3. Fabrication of screen-printed electrodes (SPEs)
Coral Draw X7 graphic suite was used to design the S/C-SPE. A
flat bed mono-filament polyester screen with average pore size of
45 mm was used to prepare separate screen for the four printing
steps as shown in Fig. 1(A). Manual screen printing process was
Fig. 1. A) Scheme for screen printing of silver/carbon screen-printed electrode (S/C-
SPE). (a) polyester substrate, (b) silver track, (c) carbon layer, (d) silver/silver chloride
track and (e) insulating layer; B) Screen printed silver/carbon screen-printed electrode
(S/C-SPE).
154 J.K. Jadav et al. / LWT - Food Science and Technology 88 (2018) 152e158
executed using polyurethane squeegee on to polyester film as
substrate (Numakura, 2008; Bhore, 2013; Van Osch, Perelaer, de
Laat, & Schubert, 2008). Firstly, silver conductive ink was used to
print the counter electrode (CE) and conducting track for working
electrode (WE). Followed by printing of carbon conductive ink at
the end of conducting track to form WE and lastly, Ag/AgCl2 was
used to print the reference electrode (RE) to complete the 3-
electrode system. The polyester film after each printing step was
heat cured at 150  C for 15 min in preheated box oven. Finally, a
transparent dielectric polymer (D2081009D6) was printed for
defining electrode sensing area as well as the connecter leads
forming protective layer over the electrode tracking during anal-
ysis. This layer was heat cured at 130  C for 10 min. Fig. 1 (B) shows
the final fabricated S/C-SPE which was further used for electro-
chemical analysis.
2.4. Characterization of nanoparticles, conductive inks and screen-
printed electrodes
The plasmon maximum of the silver nanoparticles was recor-
ded using UV-Visible spectrophotometer (Cary-60, Agilent Tech-
nologies, USA). The synthesized silver and carbon nanoparticles
were characterized for their particle size distribution along with
its colloidal stability i.e. zeta potential using Nanotrac wave
(Microtrac, USA). The morphology of the nanoparticles was
determined by scanning electron microscope (SEM) (EVO-18,
Zeiss, Germany). The viscosity of the conductive inks formulated
with thermoset resin was determined through Rheometer (DV-III
Ultra, Brookfiled, USA). The elemental spectrum of the conductive
inks was identified by energy dispersive X-ray spectroscopy
(EDAX, Amteck, USA). The heat cured screen-printed electrode was
characterized for its sheet and bulk resistivity using digital mul-
timeter with 0.1 U resolutions (DM-97, HTC Instruments, India).
The thickness of the each cured electrode was measured using
bright field research microscope (Axio Imager Z2, Zeiss, Germany).
The electrochemical performance of the screen-printed three
electrode electrochemical cell was evaluated from the cyclic vol-
tammetry analysis of potassium ferrocyanide using a potentiostat
(PGSTAT204, Autolab, Netharlands).
2.5. Application of fabricated S/C-SPE for vitamin C estimation from
fruit juices
A freshly prepared 10 mM aqueous solution of vitamin C was
prepared and used in the experiment. For voltammetric experi-
ment, 20 ml (35 mg) of vitamin C in 100 mM phosphate buffer so-
lution (PBS) (pH 2.0 adjusted with orthophosphoric acid) was
pipetted onto the S/C-SPE and the potential was scanned from 0.0
to þ1.0 V with cyclic voltammetry (Wonsawat, 2014). pH is one of
the primary factors that would affects the stability of vitamin C in
fruit juices. Hence, the pH value of the matrix has an influence on
the stability of vitamin C. According to FAO/WHO Expert Consul-
tation on Human Vitamin and Mineral Requirements, Bangkok,
Thailand, 1998, the vitamin C will decay if the pH higher than 4
(Nagymate & Fodor, 2008). Vitamin C is unstable in neutral and
alkaline environments, therefore the higher the pH value and the
longer the exposure, the greater the loss of vitamin C. This is
because the higher the pH value, the faster the oxidation reaction of
vitamin C and causes the degradation of vitamin C. Then vitamin C,
increment volume of 20 ml upto final concentration of 350 mg; were
added to the electrode surface of the S/C-SPE and the potential was
scanned. Different commercial juice products orange, guava,
mango, lichi, sweet lime, mixed fruit, grape and apple were pur-
chased from the local super market of Junagadh, Gujarat-India. The
juice samples were clarified by centrifugation at 3200xg and 20  C.
After that the samples were diluted with PBS in 1:1 ratio and then
subjected for vitamin C determination by voltammetric analysis.
2.6. Statistical analysis
The results were analyzed using two-way analysis of variance
(ANOVA) with post-hoc Tukey's test (p < 0.05) (Tukey, 1949).
3. Results
3.1. UV-visible characterization
The nature of metal nanoparticles such as shape, size and level
of particle aggregation strongly affect the absorption maxima
featured due to the surface plasmon resonance. The spectral feature
of the silver nanoparticles relative to the particle size of 10e14 nm,
35e50 nm and 60e80 nm have plasmon maxima at 395e405 nm,
420 nm and 438 nm respectively (Link & El-Sayed, 2003; Solomon,
Bahadory, Jeyarajasingam, Rutkowsky, & Boritz, 2007) thus we can
state that SNPs synthesized by chemical reduction with tri-sodium
citrate gave plasmon maxima in visible range at 425 nm with
relative particle size in range of 50e60 nm.
3.2. Particle size distribution and colloidal stability
The particle size distribution of SNPs and CNPs was determined
by Dynamic Light Scattering (DLS) and colloidal stability by zeta
potential measurement. Fig. 2 is the computed histogram of particle
size distribution of SNPs and CNPs showed particle average diam-
eter about 59 ± 9 nm and 76 ± 10 nm respectively. The zeta po-
tential analysis showed that the surface potential of SNPs
was þ57 mV and were assumed to be moderately stable, while; the
surface potential of CNPs was 34 mV thus having good colloidal
stability (Zhang et al., 2008).
3.3. Scanning electron microscopy
Silver and carbon nanoparticle powder, were placed onto
separate aluminum stub (previously cleaned with 99% ethanol).
The SEM images were collected at an accelerating voltage of 20 kV
and at 15 kV for SNPs and CNPs respectively. The images depicted
the size of dried SNPs to be 33e55 nm while for dried CNPs it was
62e68 nm. Both SNPs and CNPs were spherical in shape and had
negligible aggregation.
3.4. Viscosity measurement of the conductive inks
The viscosity of the conductive inks was measured in centipoise
(cP) immediately after thorough homogenization of Circalok 6055B
Hardener with nanoparticle and thermoset epoxy resin E-105
mixture. T-Bar Helipath spindle “T-A” (range 156e187,460 cP)
equipped with LVDV-III Ultra Rheometer was used to measure the
viscosity (Willfahrt, 2014). 30 ml of each conductive ink maintained
at 25  C by temperature controlled water bath was subjected for
viscosity analysis. The rpm of the spindle for each ink was adjusted
to achieve Torque in above 60%. At 65 rpm, silver ink had viscosity
of 1540 cP, while carbon ink at 125 rpm, had viscosity of 2950 cP.
The silver/silver chloride ink had viscosity of 4480 cP at 195 rpm.
3.5. Energy dispersive X-ray spectroscopy (EDX) of screen-printed
electrode (SPEs)
The elemental analysis of heat cured conductive inks from the
sensing area of S/C-SPE was performed using EDX coupled with
high resolution SEM. Three random spots were selected from SEM
 J.K. Jadav et al. / LWT - Food Science and Technology 88 (2018) 152e158 155

Fig. 2. Particle size distribution of (a) silver nanoparticle; 5e142 nm and (b) carbon nanoparticle; 8e199 nm.

image of each electrode to determine the percent elemental con- two probe digital multimeter. Sheet resistance of the S/C-SPE, which
tent. The EDX spectrum of CE (Fig. 3a) i.e. silver ink showed a strong is measure of thin film; here layer of conductive ink of nominal
signal at 2.9 keV (99.97 ± 0.04%) which corresponds to binding thickness was using the below equation (Jewell et al., 2015).
energy of silver (AgL) as mentioned by Echegoyen and Nerín (2013)
reported that SNPs usually exhibit typical optical absorption signals L
Rs ¼ R (1)
at ~3 keV. Similarly, for RE (Fig. 3b) i.e. silver/silver chloride ink two W
signals one at 2.9 KeV (86.67 ± 0.94%) and other at 2.7 keV
where: Rs is sheet resistivity (U/sq), R is resistance (U), L is length
(13.40 ± 0.88%) were observed, which corresponds to the presence
(mm), W is width (mm).
of elemental silver (AgL) and chlorine (ClK) in the ink respectively.
Bulk resistance is the ohmic resistance of the conductive ma-
The EDX spectrum WE (Fig. 3c) showed intense signal at 0.27 keV
terial and was calculated using below equation (Jewell et al., 2015);
(99.45 ± 0.49%) which referred to Carbon (CK) atoms. The annota-
tion of L and K refers to the transition of electrons from L-shell and P ¼ Rs
t (2)
K-shell of atom due to incident electrons during electron micro-
scopy analysis (Magudapatty, Gangopaghyayrans, Panigrahi, Nair, & where: Р is bulk resistivity (U ecm* mil), Rs is sheet resistivity (U/
Dhara, 2011; Kanmani & Lim, 2013). sq), t is thickness (cm).
The thickness of each electrode determined by microscopy was
3.6. Thickness of heat cured screen-printed electrodes (SPEs) used to calculate bulk resistance. The results of resistance, sheet
resistivity and bulk resistivity are given in Table 1 makes the
A longitudinal cross section of the heat cured S/C-SPE was fabricated S/C-SPEs to be of good conductivity and ideal for
taken from the sensing area for each electrode to measure its detection of electroactive species or analyte.
thickness. The section was placed on a glass slide and observed
under bright field microscope at 45X, equipped with AxioVision 3.8. Electrochemical performance of SPE by cyclic voltammetry
SE64 software. The thickness of the CE, WE and RE as measured
were 3.3 ± 0.06
105 cm, 3.5 ± 0.02
105 cm and Cyclic voltammetry (CV) is a versatile electroanalytical
7.5 ± 0.26
105 cm. The dielectric polymer coated as insulating
layer had a thickness of 3.8 ± 0.02
105 cm. The polyester film
used as SPE support had the thickness of 49.2 ± 0.01
105 cm. Table 1
The results obtained were further used to determine bulk re- The resistance, sheet resistivity and bulk resistivity of sensing area of S/C-SPEs.
sistivity (Numakura, 2009). S/C-SPEs Resistance (U) Sheet resistivity Rs (U/sq) Bulk resistivity r (U-cm)
Mean ± SD
3.7. Sheet and bulk resistivity of heat cured screen-printed CE 1.3 ± 0.1 20.45 6.7
104
electrodes WE 38.4 ± 2.6 38.4 13.4
104
RE 1.7 ± 0.1 6.21 4.7
104

The resistance of each electrode of S/C-SPE was measured using Mean of n ¼ 4electrodes.

Fig. 3. EDX spectrum of conductive inks formulated from silver and carbon nanoparticles (a) silver ink, (b) silver/silver chloride ink and (c) carbon ink. (Inset) SEM image of heat
cured conductive inks for EDX spectroscopy.
156 J.K. Jadav et al. / LWT - Food Science and Technology 88 (2018) 152e158

technique for the study of electroactive species. In CV a triangular
voltage is imposed on the electrochemical cell and the current
through the cell is analyzed. The current versus voltage curve of an
electro-active chemical species that undergoes red-ox behavior
displays two prominent peaks, one in each direction of the sweep
voltage. To access the electrochemical performance of S/C-SPE, CV
of ferrocyanide/ferricyanide couple (Joshi, 2011) was carried out by
connecting the S/C-SPE to a potentiostat (PGSTAT204, Autolab). The
effect of varying the scan rate at 5, 10, 40, 60, 80, 100, 120, 150, and
200 mV/s was studied at unmodified SPEs in 1.0 M KCl solution
containing 10 mM potassium ferricyanide [K3Fe(CN)6] (Frag, Ali,
Mohamed, & Awad, 2012) (Table 2). For each scan rate, the ferro-
cyanide/ferricyanide couple showed that the reaction was quasi-
reversible at the SPEs which can be observed in Fig. 4 (A). With
an increase in scan rate, the peak current separation increased and
also the peak potential shifted slightly with the anodic peak to
positive and the cathodic peak to negative potential directions.
Similar, ferro-ferri reaction was carried out on unmodified glassy
carbon electrode DS-110 from DropSens Inc. which also showed
quasi-reversible behavior (Fig. 4(B)) .
The mean values of the anodic and cathodic peak currents for
the two electrodes obtained at variable scan rates were compared
by two-way ANOVA. The p-values obtained for anodic and cathodic
peak current were 1.758
105 and 2.152
105. The results were
found to be significant for p < 0.05 which showed that the peak
currents produced were in association with the scan rates
employed for CV analysis on S/C SPE as well as DS-110.
3.9. Determination vitamin C from different fruit juices
Cyclic voltammetry of the fruit juice sample was conducted as
per the procedure mentioned by Wonsawat (2014) to estimate the
vitamin C content. After the parameters were optimized for vitamin
C determination using cyclic voltammetry, a calibration plot of
anodic peak current as a function of vitamin C concentration
ranging from 35 to 280 mg was prepared (Fig. 5). The oxidation peak
potential was observed to shift positively from 0.24 V to 0.42 V with
the increase in concentration of vitamin C because oxidation of
vitamin C at working electrode is generally believed to be totally
irreversible (Zhao et al., 2006). Based on such peak shifting, the
anodic current corresponding to each concentration was calculated
using automated peak search tool of Nova 1.11 software of poten-
tiostat instrument used for the analysis. The calibration curve was
obtained in the linearity corresponded with the coefficient of
determination (R2) of 0.9983. The limit of detection was 35 mg or
0.2 mM (S/N ¼ 3). The developed S/C-SPE had a wide linearity range
for detection of vitamin C from juice samples. Vitamin C from
different fruit juices was determined from the anodic peak current
generated by CV analysis and also through spectrophotometric
analysis wherein proportionate coupling of the 2,4-dinitrophenyl
hydrazine (DNPH) dye with vitamin C in the juice samples. The
results for vitamin C determination through both the analytical

Table 2
Comparison of ferro-ferri scan rate analysis of S/C-SPE and DS-110 electrode.a

Scan rate S/C SPE DS-110

(mV/s) A1 C1
Anodic peak current Cathodic peak current Anodic peak currentA2 Cathodic peak currentC2
(mA) (mA) (mA) (mA)

5 79.36 ± 0.44 84.27 ± 1.28 97.15 ± 0.96 102.37 ± 0.98

10 103.42 ± 0.63 98.17 ± 0.88 120.34 ± 0.97 118.57 ± 0.99
40 142.24 ± 0.35 143.37 ± 1.27 173.82 ± 0.94 171.99 ± 0.94
60 164.37 ± 0.37 164.03 ± 0.26 197.60 ± 0.67 197.53 ± 0.69
80 194.27 ± 0.14 198.58 ± 0.35 221.87 ± 0.87 227.18 ± 0.92
100 214.36 ± 0.42 217.57 ± 0.29 252.14 ± 0.51 249.13 ± 0.90
125 239.04 ± 0.70 237.18 ± 0.23 274.16 ± 0.46 281.34 ± 0.92
150 269.22 ± 0.39 274.18 ± 0.26 312.67 ± 0.42 319.24 ± 0.99
200 324.64 ± 0.46 331.54 ± 0.98 374.15 ± 0.63 382.14 ± 0.97
Linearity Equation 1.2151x þ 89.039 1.246x - 88.415 1.3708x þ 108.36 1.4103x - 107.85
Coefficient of Determination (R2) 0.9913 0.9941 0.9902 0.9944
a
Data expressed as the mean ± SD, n ¼ 3. Values for A1 and A2 as well as C1 and C2 were compared and are significantly different (p < 0.05).

Fig. 4. Quassi-reversible duck curves of S/C-SPE (A) and DS-110 (B) of 10 mM potassium ferricyanide in 1.0 M KCl solution at different scan rates 5, 10, 40, 60, 80, 100, 125, 150 and
200 mV/s.
 J.K. Jadav et al. / LWT - Food Science and Technology 88 (2018) 152e158 157

Fig. 5. Cyclic voltammograms of vitamin-C (35 mge280 mg) in 100 mM PBS (pH 2.0) on S/C-SPE. (Inset) calibration curve of the current responses against vitamin-C concentration
ranging from 35 mg to 280 mg in PBS on S/C-SPE.

methods were comparatively similar, with highest content of
vitamin C in sweet lime, followed by lichi. The least amount of
vitamin C was estimated in apple juice. The vitamin C content of all
the fruit juice samples as determined by both the analytical
methods is displayed in Table 3, where the mean values of vitamin
C estimated by CV and DNPH were compared and p-value obtained
for CV analysis and biochemical method was 0.586 and thus results
obtained are statistically not significant at p < 0.05.
3.10. Reproducibility, stability, matrix effect (interference study),
electrode saturation and response time of S/C SPE for detection of
vitamin C
The reproducibility of S/C SPE was characterized by performing
cyclic voltammetry (CV) of vitamin C at varying concentration from
35 to 280 mg in PBS. The cyclic voltammogram data obtained from
triplicate analysis of vitamin C standard showed an average of
relative standard deviation (RSD) value of 0.14% proving an excel-
lent reproducibility of the developed S/C SPE for vitamin C deter-
mination. The stability of the fabricated S/C SPE was investigated by
running 20 cycles of CV at 100 mV/s scan rate in 100 mg of vitamin C
in 100 mM phosphate buffer solution. A well defined qussi-
reversible redox peaks was observed for each cycle. A RSD value
of 0.31% showed that the electrode was stable enough for the
electrochemical estimation of vitamin C.
From observations derived by the previous studies conducted by
Sun, Lian, Liang, & Liu, 2004 and Pisoschi, Danet, & Kalinowski,
2008 for cyclic voltammetric estimation of vitamin C it can be
concluded that compounds like citric acid, tartaric acid, phenylal-
anine, glutamic acid, aminoacetic acid and glucose are commonly
found in fruit juices which did not interfere in determination when
the working electrode is carbon based. As in the present study, the
fabricated S/C SPE comprises carbon working electrode there was
no need to check the effect of interference species on vitamin C
determination. Upon continuous determination of vitamin C from
any of the same juice samples the S/C SPE showed saturation
leading to reduced anodic peak current. This saturation of the
electrode was nullified by intermittent washing of the electrode
surface with DI water. The anodic current of ascorbic acid was
measured as function of scan rate of 0.1 V/s. The current intensity
peak of ascorbic acid is obtained at a potential range of 0.25e0.35 V,
thus the response corresponding to a time of <4 s was taken into
account.
4. Conclusions
Nanoparticles and related materials when applied to fabrication
and development of electrochemical devices have brought down
tedious and cumbersome analytical and electrochemical analysis.
The results of the cyclic voltammetric analysis when compared

Table 3
Vitamin C determined using cyclic-voltammetry and biochemical method from commercial fruit juice samples.*

Juice samples Cyclic voltammetry analysisa Biochemical method (DNPH coupling)b

Vitamin C (mg/100 ml) Vitamin C (mg/100 ml)

Orange 45.58 ± 0.27 48.34 ± 1.20

Guava 26.83 ± 0.22 24.13 ± 0.16
Mango 7.65 ± 0.10 6.67 ± 0.29
Lichi 50.21 ± 0.37 54.58 ± 0.47
Sweet Lime 59.28 ± 1.29 58.87 ± 0.30
Mixed fruit 40.26 ± 2.13 42.64 ± 0.74
Grape 29.04 ± 0.57 27.18 ± 0.67
Apple 5.51 ± 0.43 5.97 ± 0.63

*Data expressed as the mean ± SD, n ¼ 3. Values of column-a and column-b are not significantly different (p > 0.05).
158 J.K. Jadav et al. / LWT - Food Science and Technology 88 (2018) 152e158
with standard biochemical analysis of vitamin C showed signifi-
cantly similar results. The voltammetric response of ferro-ferri
couple on our S/C-SPE and commercially available Dropsens DS-
110 electrode were compared and it showed that fabricated SPE
almost similar electrochemical signals to DS-110.
Modifications of the SPE such as, electrochemical immuno-
sensors utilizing SPEs are developed for applications in various
fields including medical, environmental and food (Vig, Mun ~ oz-
Berbel, Radoi, Cortina-Puig, & Marty, 2009). Another attractive
feature of SPEs is that they can be surface-modified in the same way
as conventional electrodes, thus enabling increased sensitivity
when using superior electro-catalytic properties of nanoparticles.
More, they can be used with redox mediators to enhance the
catalysis of targets that may otherwise not be easily analyzed due to
their poor redox activity (Lien, Takamura, Tamiya, & Vestergaard,
2015; Khan, Gorski, Garcia. 2011; Prieto-Simon, Macanas, Munoz,
& Fabregas, 2007). The fabrication process mentioned in this paper
would help many researchers to develop smaller, portable, in-filed
analytical tools and sensor devices for food industry, medical fields,
chemical sciences and many more.
Acknowledgment
We acknowledge Dr. B. A. Golakiya, Professor & Head, Depart-
ment of Biotechnology, Junagadh Agricultural University (JAU),
Junagadh, Gujarat-India; for allowing the access to the laboratory
and instrumentation facilities of Food Testing Laboratory (FTL), a
unit of Department of Biotechnology, JAU, Junagadh. We also
acknowledge the support provided by team of GFQA Cell, FTL; (Dr.
H. P. Gajera, Mr. Kishan H. Sodha, Mr. Samir A. Anuj and Ms. Radhika
P. Gondaliya) for nanoparticle synthesis and characterization and
the team of the Microbial Cell, FTL; (Mr. M. V. Parakhiya and Mr.
Rohit L. Bhalara) for microscopy, SEM and EDAX facilities.
Appendix A. Supplementary data
Supplementary data related to this article can be found at
https://doi.org/10.1016/j.lwt.2017.10.005.
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