Accepted Manuscript

Title: Sensitive determination of caffeine by Copper Sulphide

nanoparticles Modified Carbon Paste Electrode

Author: Mallappa M.<ce:author id="aut0010"

biographyid="vt0010" orcid="0000-0002-7942-840X">
Shivaraj Y. Nagaraju Kottam Chiranjeevi Srinivasa Rao Vusa

PII: S0924-4247(16)30343-0
DOI: http://dx.doi.org/doi:10.1016/j.sna.2016.07.013
Reference: SNA 9757

To appear in: Sensors and Actuators A

Received date: 18-2-2016

Revised date: 7-7-2016
Accepted date: 14-7-2016

Please cite this article as: Mallappa M., Shivaraj Y., Nagaraju Kottam, Chiranjeevi
Srinivasa Rao Vusa, Sensitive determination of caffeine by Copper Sulphide
nanoparticles Modified Carbon Paste Electrode, Sensors and Actuators: A Physical
http://dx.doi.org/10.1016/j.sna.2016.07.013

This is a PDF file of an unedited manuscript that has been accepted for publication.
As a service to our customers we are providing this early version of the manuscript.
The manuscript will undergo copyediting, typesetting, and review of the resulting proof
before it is published in its final form. Please note that during the production process
errors may be discovered which could affect the content, and all legal disclaimers that
apply to the journal pertain.
Sensitive determination of caffeine by Copper Sulphide nanoparticles

Modified Carbon Paste Electrode

Mallappa M1, 2, 3, Shivaraj Y1*, Nagaraju Kottam2, Chiranjeevi Srinivasa Rao Vusa4
1Department of Chemistry, Government Science College, Bangalore-560 001
2Department of Chemistry, M.S. Ramaiah Institute of Technology, Bangalore - 560 054
3Visvesvarayya Technological University, Research Resource Centre, Belgaum-590 018
4Bio-sensors division, CSIR-Central Electrochemical Research Institute, Karaikudi-630003

*Correspondence author: shivaraj_y@rediffmail.com

Highlights
 Starch assisted synthesis of CuS NPs by simple co-precipitation method in aqueous
media.

 CuS NPs MCPE was used for caffeine sensor.

 CuS NPs MCPE exhibited better electrocatalytic activity towards the oxidation of
caffeine.

 The electrokinetic rate of CuS NPs MCPE towards caffeine oxidation was found to

be 2.174 × 103 s-1.

 The sensor showed lower detection limit, higher sensitivity and selectivity, wide

linear range, good reproducibility and stability.

ABSTRACT

Copper sulphide nanoparticles (CuS NPs) were synthesized by simple co-precipitation

method in aqueous media using starch as a biopolymer. The obtained product was

characterized by XRD, SEM, EDX, TEM, FTIR and UV-visible techniques. The particle size

was determined by TEM analysis and found to be in the range 20-50 nm. CuS NPs modified

1
carbon paste electrode (CuS NPs MCPE) was employed to determine caffeine (CAF) by

cyclic voltammetry (CV) and differential pulse voltammetry (DPV) methods. The CuS NPs

MCPE showed better electrocatalytic activity towards the oxidation of CAF, which was

confirmed by enhance in the oxidation peak current and decrease in oxidation peak potential.

Furthermore, in electrochemical impedance (EI) studies, the charge transfer resistance (RCT)

at CuS NPs MCPE (12.1 kΩ) decreases as compared to bare CPE (42.6 kΩ), which suggests

that CuS NPs improves the charge transfer capacity of CPE. Electron transfer kinetic

parameters, sensitivity and surface active area of the electrode were determined. Under

experimental conditions, the oxidation current of CAF was proportional to its concentration

from 2µM to 120µM. The detection limit was found to be 18 × 10-9 M. The developed

method was successfully applied to determine CAF content in food samples with appreciable

results.

Key words: CuS NPs, Co-precipitation method, CuS NPs MCPE, Electrocatalytic activity.

1. Introduction

In recent scenario, there is a growing interest in the development of the semiconductor

nanostructure materials owing to their distinctive physical and chemical properties. Among

the chalcogenides, copper sulphide (CuxS) has attracted extensive attention due to its unique

optical and electrical properties [1]. The attractiveness is also arises due to its low synthetic

cost, solution processing ability and the dependence of optoelectronic properties as a function

of size, shape, doping and surface chemistry. Copper sulphide exists in various stoichiometric

compositions, morphology and exhibits non-linear optical property [1-2]. In particular, the

band gap of CuxS can be varied in a wider range (1.2 to 2.5 eV) with different stoichiometric

compositions (x=1-2), making it a highly desirable material for solar cells, non-volatile

memory devices and gas sensing applications. Furthermore, CuxS is a potentially interesting

2
material for cold cathodes and lithium ion batteries [3-4]. Several methods have been

reported for the synthesis of CuS NPs such as sonochemical, sol-gel, hydrothermal method

etc [4-6].

Caffeine (CAF) is a natural alkaloid which belongs to a class of xanthenes derivatives

and found in various food and beverages such as coffee, tea, coca-cola, cocoa beans,

yerbamate, guarana berries and chocolates; and even in various drugs. CAF acts as a natural

pesticide in plants since it paralyses and kills certain insects which destroy the plants. CAF

has several physiological effects in humans such as stimulation of central nervous system,

gastric acid secretion, diuresis and cardiovascular systems [7-9]. Thus, the development of

sensitive, fast and cost effective method for the evolution and quantification of CAF is a

significant field of research.

Several methods has reported for detection of CAF such as chromatographic and

spectroscopic techniques [10-11] but they have their own disadvantages such as low range of

detection and low selectivity, complexity in techniques, expensive instrumentation, requires

highly skilled technicians etc. Whereas electrochemical method is simple, shows high

sensitivity, good stability, less expensive etc [12] which makes it more promising technique

for the detection of CAF. The electrochemical detection of CAF at bare electrode is less

effective due to oxidation which occurs at high positive potential range and may overlap with

the electrochemical reactions, limiting potential window from the anodic side and often gives

low reproducible analysis [13]. Therefore, these limitations can be overcome by using

electrodes such as boron doped diamond electrode (BDDE), pencil graphite electrode (PGE)

and chemically modified electrodes [14-22].

In the present work, CuS NPs were synthesized by simple co-precipitation method at

low temperature. Electrochemical study of CAF at CuS NPs MCPE was investigated by

cyclic voltammetry (CV) and differential pulse voltammetry (DPV). CuS NPs MCPE shows

3
better electrocatalytic activity towards CAF oxidation. Electron transfer kinetics, sensitivity,

linear range, detection limit and interfacial properties of CuS NPs MCPE have been

discussed.

2. Experimental

2.1 Materials

CAF was purchased from Sigma-Aldrich. Cupric chloride, biopolymer starch and

other reagents were purchased from Merck and Sdfine chemicals. 0.1M acetate buffer

solution (ABS) was used as a supporting electrolyte throughout the experiment and different

pH was adjusted by adding acetic acid and sodium hydroxide in 0.1M sodium acetate. All the

reagents were prepared using doubly distilled water.

2.2. Synthesis of CuS nanomaterials

CuS NPs were prepared in solution phase by co-precipitation method. 0.1M solution

of cupric chloride was taken in a beaker, to which 3% starch solution (act as stabilizing as

well as capping agent, which controls the particle size) was added drop wise by constant

stirring. Then, 0.1M Na2S solution was added drop wise to the above mixture until a black

coloured precipitate of CuS was obtained. Stirring was continued for another 1 hour by

maintaining the temperature at 60 °C. The resulting precipitate was filtered, washed with

distilled water, 1:1 mixture of water-ethanol and dried at 170 °C for about 3 hours. The

obtained product was characterized by various techniques.

4
2.3 Reaction mechanism:

Starch is a natural biopolymer made up of two components amylose and amylopectin

which consists of α-glucose units in the carbon 1C4 conformation. The semicrystalline

structure of starch is lost when starch becomes soluble in water at 75 °C. The copper cations

are attracted to the O-H group of starch after the addition of starch to Cu (II) salt solution to

form Cu(OH)2. To this solution sodium sulphide is added to form a black precipitate of CuS.

On heating small amylose molecules forms a network that holds the water molecules and

increases the mixture viscosity. The obtained product forms covellite phase of CuS NPs [23].

The morphology and phase transformation in the present system could be described

according to XRD, SEM and other techniques.

CuCl2 Cu2+ + 2Cl-

Cu2+ + Starch +H2O [Cu (starch) 2 H2O] 2+

[Cu (starch) 2H2O] 2+ + S2- CuS (starch) + H2O

The overall chemical reactions involved can be described as given below

CuCl2 + Starch + Na2S CuS (starch) + 2NaCl

2.4 Characterization of copper sulphide nanoparticles

Powder XRD of CuS NPs were recorded on Philips X’pert PRO X-ray diffractometer

with graphite monochromatized CuKα radiation (k = 1.541 Å) in the range of 10°-80° with

2°/min scanning rate. The morphology of the CuS NPs were examined by JEOL-JSM–6490

LV scanning electron microscope and CM12 Philips transmission electron microscope

equipped with EDX (Kevex Sigma TM Quasar, USA). FT-IR spectra of pure starch and CuS

NPs in starch matrix were recorded using Agilent carry-630 FT-IR spectrophotometer

(Australia) over the range 400-4000 cm-1. In addition, the optical properties of CuS NPs were

analyzed by UV-1800 UV-Visible absorption spectrophotometer (Shimadzu, Japan).

5
 Cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical

impedance (EI) measurements were performed using a CH-Electrochemical analyzer (CHI

6039E, USA) connected with a three-electrode cell. A bare carbon paste electrode (BCPE)

and modified CPE were used as working electrode. A platinum wire was employed as

counter electrode while the reference electrode as saturated calomel electrode (SCE).

2.5. Fabrication of CuS nanoparticles modified carbon paste electrode (CuS NPs MCPE)

Bare carbon paste electrode (BCPE) was prepared according to the literature [24].

CuS NPs MCPE was prepared by mixing graphite powder, nujol oil and CuS NPs to form a

homogeneous paste. The resulting paste was then packed into the cavity of the Teflon tube

(internal diameter is 3.0 mm) and external electrical contact was made by copper wire. The

surface of the electrode was smoothened and then rinsed with distilled water thoroughly.

3. Results and Discussion

3.1. Crystal Structure and morphological analysis

3.1.1. Powder X-ray diffraction study

The phase purity and crystallinite of CuS NPs were confirmed by X-ray diffraction

studies. Fig. 1(a) shows the XRD pattern of starch assisted CuS NPs prepared in solution

phase and the diffraction patterns correspond to the hexagonal covellite phase of CuS NPs

(JCPDS no. 65-603; a = 3.8813 Å, c = 16.33307 Å) with a space group P63/mmc (No-194)

and cell parameters a = 3.7960 Å, c = 16.3820 Å. The broadening of the diffraction peaks and

high intensity indicate good crystalline nature of CuS NPs. The reflections correspond to

(100), (101), (102), (103), (006), (110), (108), (202) and (116) planes. The average crystallite

size calculated from the Debye-Scherer equation [25] was found to be 38 nm. The lattice and

6
the structural parameters of the nanocrystalline CuS NPs are summarized in Table 1 and

covellite hexagonal phase of CuS NPs as shown Fig.1 (b).

3.1.2. Scanning electron microscope and Energy-dispersive spectroscopy

The surface morphology of CuS NPs were investigated by SEM, as shown in Fig.2

(a). The difference in particle size can be easily observed from the morphology. The

diameters of the particles grow larger with uneven size distribution and irregular shape. The

elemental composition and purity of CuS NPs were investigated by EDX analysis.

3.1.3. Transmission electron microscope

Fig. 2(b) shows the typical TEM images of CuS NPs. TEM images has revealed the

formation of quasi-spherical nanoparticles with non-uniform particle size distribution which

can be attributed to the agglomeration of nanoparticles. The particles size of CuS NPs were

found to be in the range of 20-50 nm.

3.1.4. FT-IR spectra of CuS nanoparticles

The structure and chemical composition of pure starch and CuS NPs in starch matrix

were examined in the 400-4000 cm−1 region using FT–IR spectroscopy as shown in

Fig. 3 (a). A relatively broad and intense band is observed at 3,314 cm-1, indicating the

presence of free hydroxyl group of starch, whereas the absorption band at 3,452 cm-1

represents the interaction of CuS NPs with hydroxyl group of starch due to well incorporation

of nanoparticles into the starch matrix. The absorption bands occurring at 2,901 cm-1 and

2,917 cm-1 are attributed to anti-symmetric CH2 stretching and C-H stretching of CH2 groups,

respectively. The new absorption band observed at 2358 cm-1 corresponds to -S-H interaction

which may be due to the interaction of CuS NPs with the hydroxyl group of starch whereas

no such peak was observed in case of pure starch [26]. The absorption bands at 1050 cm-1 and

7
1150 cm-1 corresponds to C-O bond stretching of C-O-C and C-O-H groups in starch,

respectively. The absorption band at 470 cm−1 is due to the vibration of Cu-S bond. The

above results confirm the presence of starch which acts as capping agent and also increases

the stability of the nanoparticles.

3.1.5. Optical properties of copper sulphide nanoparticles

The optical absorption spectra of the CuS NPs which was dispersed in 1:1

water/ethanol was recorded at room temperature in the range of 200 to 800 nm as shown in

Fig. 3 (b). It is well known that smaller particles have a larger surface to volume ratio, that

results in the distribution of more surface defects. Thus, the lower the particle size, the

nanomaterials exhibit strong and broad absorption bands [27]. The characteristic absorption

maxima of CuS NPs observed at 378 nm may be assigned to the band excitonic absorption.

The absorption of CuS NPs shift towards lower wavelength (blue shift) as compared with that

of bulk covellite phase (625 nm) [28] of copper sulphide. The optical band gap energy (Eg) of

CuS NPs estimated by using the relation, Eg = 1240/ λ [29] was found to be 3.28 eV. This

evidently shows that the blue shift and corresponding increase in band gap may be attributed

to the size effect of the synthesized CuS NPs.

3.2. Electrochemical characterization of the CuS NPs modified CPE

Electrochemical impedance studies (EIS) is a useful technique to investigate the

interfacial properties of surface modified electrodes. Fig. 4 shows the Nyquist plots observed

for BCPE and CuS NPs MCPE in presence of 5 mM [Fe(CN)6]3−/4− containing KCl in the

frequency range from 10−1 to 104 Hz and equivalent circuit (inset of Fig. 4) was chosen to fit

the obtained impedance data. There is a considerable decrease in the diameter of the

semicircle for CuS NPs MCPE due to the high interfacial electron transfer process and low

8
RCT. The smaller semicircle indicates that, the RCT is decreased for CuS NPs MCPE (12.1

kΩ) as compared with the BCPE (42.6 kΩ). Hence, the enhanced charge transfer capacity is

due to the large surface area, good conductivity of CuS NPs which accelerates the electron

transfer rate [30, 31]. The above results evidences that the presence of mediator (CuS NPs)

on CPE plays an important role in increasing the charge transfer capacity of the CPE.

3.3. Electrocatalytic oxidation of caffeine at CuS NPs modified CPE

Fig. 5 (a) & (b) shows the CV’s and DPV’s for 0.12 mM CAF at BCPE and CuS NPs

MCPE in 0.1 M ABS pH 7.0 with scan rate 0.05 Vs-1. At BCPE (dashed line), the oxidation

peak of CAF was observed at 1.457 V (CV) and 1.363 V (DPV) with miniature current

response. On the other hand, under identical conditions CuS NPs MCPE (solid line) showed

significant enhancement in the current signal and oxidation peak was observed at 1.38 V

(CV) and 1.332 V (DPV). It is noticed that the minimization of over potential and

enhancement of peak current indicates the electrocatalytic activity of CuS NPs MCPE

towards the oxidation of CAF. The enhancement in the peak current of CAF at CuS NPs

MCPE may be attributed to several possible reasons; (i) electrostatic attractions between CAF

and CuS NPs, (ii) high surface area and (iii) high conductivity of CuS NPs [32]. The above

results suggest that the irreversibility of CAF is significantly improved by using CuS NPs

MCPE.

3.4. Influence of scan rate

The effect of scan rates for 0.12 mM CAF at CuS NPs MCPE in 0.1 M ABS pH 7.0

was studied by CV technique as shown in Fig. 6(a). According to Randles-Sevcik equation,

the experimental results obtained at CuS NPs MCPE showed increase in the oxidation peak

current with increase in scan rate. As the scan rate increased from 0.05 to 0.4 Vs−1, the peak

9
potential shifted to more positive and peak current increased linearly as represented by the

graph of peak current (Ipa) vs. scan rate (ν) (Ipa (µA) = -1.43 + 6.04ν (Vs-1); r2= 0.9836). In

order to confirm the electrode reaction process, a graph of peak current (Ipa) vs. square root of

scan rate (ν½) was plotted which (inset of Fig. 6 (a)) yielded a straight line; linear regression

equation, Ipa (µA) = -3.42621 + (-1.69336) ν1/2 (Vs−1)1/2; r2 = 0.9979. Further, the graph of

log Ipa vs. log ν, yielded a straight line (log Ipa (µA) = -2.13 + 0.56 log ν (Vs-1): (r2 = 0.9930))

as shown in Fig. 6 (b) with a slope value of 0.56. This slope value is very close to the

theoretical value of 0.5, which is expected for an ideal reaction condition for diffusion

controlled electrode process [33]. Hence, the above results suggest that the electrochemical

oxidation of CAF at CuS NPs MCPE is a diffusion-controlled process.

The surface area of the electrode was obtained by CV method using 0.12 mM CAF at

different scan rates. The surface area available for reaction species in the solution can be

estimated by the Randles-Sevcik equation [24]. This equation relates the peak current for an

electron-transfer controlled process to the square root of the scan rate:

Ip = 2.99 ×105 n (αn) 1/2 A C D1/2 ν1/2

Where Ip is the peak current, A is electroactive area (cm2), C is concentration of

electroactive species (mol cm-3), n is number of electrons transferred, α is the electrons

transferred co-efficient, D is diffusion coefficient (cm2 s-1) and υ is scan rate (Vs-1). The

value of diffusion co-efficient for CAF is obtained by plotting Ipa vs. υ1/2. As compared to

BCPE (0.02082 cm2), CuS NPs MCPE provides more electroactive surface area (0.04099

cm2).

Irreversible oxidation process of CAF at CuS NPs MCPE and BCPE in ABS pH-7.0

involves four protons and four electron oxidation [34] as shown in Scheme 1.

The electron transfer coefficient ‘α’ is calculated from the difference between peak

potential (Ep) and half wave potential (Ep/2).

10
 ΔEp = E-Ep/2 = (47.7/αn) mV (for irreversible reaction at 298 K)

The value of α was calculated from the above equation [35] and it was found to be

0.539.

For an irreversible oxidation reaction, the following equation is used to calculate the

standard rate constant (k0) [36, 37]

Ep = E0 + (RT/αn) [ln (RTk0/αnF)-lnν]

Where, E0 is formal potential, R, T, α and F are universal gas constant, absolute

temperature, electron transfer co-efficient and Faradays constant, respectively. The value of

E0 was obtained from the intercept of Ep vs. ln ν plot by the extrapolation to the vertical axis

at ν = 0 and the linear regression equation, Ep (V) = 1.46198 + 0.08398 log ν (Vs-1); r2=

0.9580. The value of k0 was determined by plotting Ep vs. ln ν and it was found to be 2.174 ×

103 s-1.

3.5. Effect of Modifier

Fig. 7 shows the CVs of 0.12 mM CAF at CuS NPs MCPE with different amount of

CuS NPs with a scan rate 0.05 Vs-1. The amount of CuS NPs mixed in the carbon paste would

influence the performance of the modified electrode. According to the literature, the electron

transfer behaviour of the CAF oxidation depends on the electrode surface and microstructure

as well as the density of electronic states of electrode materials near the Fermi level [38].

Modified CPEs showed improved electrochemical response towards CAF oxidation with the

increasing amount of CuS NPs (2 to 10 mg), indicating the fast electron transfer kinetics. The

tremendous conducting property of CuS NPs contributed to the enhanced electrochemical

response of modified CPEs towards CAF oxidation. As the amount of CuS NPs was

increased to 12 mg, the peak current minimised due to the decrease in conductivity with

11
decrease in surface area of the electrode. Therefore, based on the maximum current response,

10 mg of CuS NPs were used for the subsequent electrochemical study.

3.6 Influence of supporting electrolyte and pH

The choice of supporting electrolytes contributes a significant role in the

electrochemical oxidation of CAF at CuS NPs MCPE. The current response of 0.12 mM CAF

was examined in a series of supporting electrolytes such as Bitton Robinson, acetate and

phosphate buffer as shown in supplementary Fig. S1. It was found that peak with high

sensitivity and better current response was observed in acetate buffer as compared to other

electrolytes.

The pH of the acetate buffer has a significant contribution on the electrocatalytic

oxidation of CAF at CuS NPs MCPE by influencing both on peak current and peak potential.

The effect of pH of ABS on the determination of CAF at CuS NPs MCPE was examined over

the pH range of 3.0-8.0, shown in Fig. 8. A significant current response was observed at pH

7.0 with lower peak potential. Hence, ABS of pH 7.0 was chosen for the electrochemical

study of CAF.

3.7. Optimization of the determination conditions

In order to obtain more sensitive peak current, DPASV was employed for the

determination of CAF. The optimum instrumental parameters such as accumulation potential,

accumulation time, scan increment and pulse height were studied for the determination of

CAF at CuS NPs MCPE was investigated [39, 40]. Fig. 9 (a) represents the effect of

oxidation peak current with respect to accumulation potential. The oxidation peak current of

CAF gradually increased and peak potential remain almost constant when the accumulation

potential was increased from -0.1 to -0.4 V. Further, increase in accumulation potential

resulted the rapid increase in peak current, whereas the peak potential shifted towards higher

12
potential. This deviation was observed because of the complete reduction of CAF as

accumulation potential became more negative [11]. The maximum oxidation peak current

was observed at -0.4 V, hence this accumulation potential was selected for further studies.

The effect of accumulation time on the oxidation peak current and peak potential is

shown in Fig. 9 (b). As the accumulation time increased from 0 to 180s, the oxidation peak

current of CAF increased gradually upto 120s, above which a steady current was observed

which could be due to the saturation of CAF on the surface of CuS NPs MCPE. Experimental

DPASV parameters were optimized to study pulse height (10-70 mV) and scan increment (2-

14 mV). The more pronounced peaks with better shape was obtained by maintaining the

following parameters, 8 mV and 60 mV for scan increment and pulse height, respectively as

shown in supplementary Fig. S2.

3.8. Calibration curve and analytical parameters

Differential pulse voltammetry (DPV) was chosen as a more sensitive voltammetric

technique in comparison with cyclic voltammetry to investigate the dependence between

peak current and CAF concentration. To construct the calibration curve using the BCPE and

CuS NPs MCPE, different aliquots of the standard solution of CAF were added to the acetate

buffer (pH 7.0) solution (Fig. 10). The analytical parameter obtained from the calibration

curve is shown in Table 2.

An average of nine consecutive measurements was used for calibration curve

construction. The dependence of peak current on CAF concentration shows a good linearity

in the concentration range from 2 × 10-6 to 1.2 ×10-4 M solutions for CuS NPs MCPE and

8 × 10-6 to 1.2×10-4 M solutions for BCPE as depicted in the inset of Fig. 10. The linear

regression equation is given by, Ip = 7.39614 µA + 22.3447 (μA mol-1) C; r2 = 0.99424 for

CuS NPs MCPE and Ip = 4.8657µA + 7.750 (μA mol-1) C; r2 = 0.9292 for BCPE. The

13
following equations were used to determine the limit of detection (LOD) and the limit of

quantification (LOQ): LOD = 3σ/m and LOQ = 10σ/m, where σ is the standard deviation of

the y-coordinate from the line of best fit (linear coefficient) and ‘m’ is the slope (angular

coefficient). The detection limit and quantification were determined and results are shown in

Table 2. A comparison of results obtained in the present work with previous reported

literature is mentioned in Table 3.

3.9. Reproducibility, stability and interference study

The reproducibility of CuS NPs MCPE was studied using CV and DPV techniques.

The repeatability of the modified electrode was estimated by using 0.12 mM CAF for nine

consecutive measurements which resulted in no significant change in peak currents. The RSD

for nine successive determinations of 0.12 mM CAF on CuS NPs MCPE was 2.3% which

indicated that the electrode showed good reproducibility.

The stability of the CuS NPs MCPE was also studied for a period of 90 days. The

analytical performance of CAF biosensor showed no significant change in the current

response, indicating that the electrode was stable (RSD ≤5 %) and also had acceptable

storage stability.

Interference study of CAF sensor with different species was investigated by DPV in

0.1 M ABS pH 7.0 by the addition of Na+, K+, Ba2+, Ca2+, Cl-, NO3- ions and sugars like

glucose, fructose, sucrose which are usually found in beverages and drugs together with CAF.

There was no significant change in the oxidation peak of CAF which implies that CuS NPs

MCPE exhibited better electrocatalytic activity even in the presence of interfering species.

3.10. Real sample analysis

In order to examine the practical application of the proposed method, CuS NPs MCPE

was used to determine the CAF in some commercial tea and coffee samples. The samples for

14
the study were used according to the literature [22]. 1 mL sample solution was injected into

9 mL of 0.1M ABS and DPASVs were recorded under the optimal experimental conditions.

Meanwhile, in order to determine the percentage of recovery, standard solution of CAF was

added. The results are provided in Table 4. The recovery was in the range of 94-106 %,

indicating that the sensor can be successfully employed for the detection of CAF in real

samples.

4. Conclusion:

CuS NPs were synthesized by a simple co-precipitation method in aqueous media

using biopolymer starch. The particle size was determined from TEM analysis and was found

to be in the range 20-50 nm. The CuS NPs were used to modify the surface of CPE for

electrochemical detection of CAF. CuS NPs MCPE exhibited better electrocatalytic activity,

sensitivity and selectivity towards the oxidation of CAF as compared with BCPE. The limit

of detection was found to be 18×10-9 M concentration of CAF. According to EI studies, the

charge transfer resistance is decreased for CuS NPs MCPE (12.1 kΩ) as compared with

BCPE (42.6 kΩ), suggesting that the presence of mediator (CuS NPs) on CPE plays an

important role in increasing the charge transfer capacity of CPE. CuS NPs MCPE can be used

as a good electrochemical sensor for various electroactive species in the field of

electroanalysis.

5. Acknowledgement

The authors are grateful to SERB [No. SB/FT/CS-009/2012], DST New Delhi, India

for the financial assistance and also thankful to Prof. S. Sampath, Department of Inorganic &

15
Physical Chemistry, Indian Institute of Science, Bangalore for valuable suggestions and

discussion.

References

[1] M. Nafees, S. Ali, S. Idrees, K. Rashid, M. A. Shafique, A simple microwave assists

aqueous route to synthesis CuS nanoparticles and further aggregation to spherical

shape, App. Nano. Sci., 3 (2013)119-124.

[2] N. Khaorapapong , A. Ontam, M. Ogawa, Very slow formation of copper sulfide and

cobalt sulfide nanoparticles in montmorillonite, App. Clay Sci., 51 (2011) 182–186.

[3] H. Lee, S. W. Yoon, E. J. Kim, J. Park, In-situ growth of copper sulfide nanocrystals

on multiwalled carbon nanotubes and their application as novel solar cell and

amperometric glucose sensor materials, Nano. Lett., 7 (2007) 778-784.

[4] J. Kundu, D. Pradhan, Influence of precursor concentration, surfactant and

temperature on the hydrothermal synthesis of CuS: structural, thermal and optical

properties, New. J. Chem., 37 (2013) 1470-1478.

[5] R.S. Christy, J. T. T. Kumaran, Phase transition in CuS nanoparticles, J. Non-Oxide

Glass., 6 (2014)13-22.

[6] S. Radhakrishnan, H.Y. Kim, B. S. Kim, A novel CuS microflower superstructure

based sensitive and selective nonenzymatic glucose detection, Sens. Actu. B: Chem.,

233 (2016) 93–99.

[7] M. Aklilu, M. Tessema, M. R. Abshiro, Indirect voltammetric determination of

caffeine content in coffee using 1,4-benzoquinone modified carbon paste electrode,

Talanta., 76 (2008) 742-746.

16
[8] A. Carolina Torres, M. M. Barsan, C. M.A. Brett, Simple electrochemical sensor for

caffeine based on carbon and Nafion-modified carbon electrodes, Food Chem.,149

(2014) 215–220.

[9] L. Svorc, P. Tomc ik, J. Svitkova, M. Rievaj, D. Bustin, Voltammetric determination

of caffeine in beverage samples on bare boron-doped diamond electrode, Food Chem.,

135 (2012) 1198-1204.

[10] Y. Tadesse, A. Tadese, R. C. Saini, R. Pal, Cyclic voltammetric investigation of

caffeine at anthraquinone modified carbon paste electrode, Int. J. Electrochem., 2013

(2013)1-7.

[11] J. Zhang, L. P Wang, W. Guo, X. D. Peng, M. Li, Z. B. Yuan, Sensitive differential

pulse stripping voltammetry of caffeine in medicines and cola using a sensor based on

multi-walled carbon nanotubes and Nafion, Int. J. Electrochem. Sci., 6 (2011) 997-

1006.

[12] S. Yang, R. Yang, G. Li, L. Qu, J. Li, L. Yu, Nafion/multi-wall carbon nanotubes

composite film coated glassy carbon electrode for sensitive determination of caffeine,

J. Electroanal. Chem., 639 (2010) 77-82.

[13] F. Zhao, F. Wang, W. Zhao, J. Zhou, Y. Liu, L. Zou, B. Ye, Voltammetric sensor

for caffeine based on a glassy carbon electrode modified with Nafion and graphene

oxide, Microchim. Acta, 174 (2011) 383-390.

[14] M. Amare, S. Admassie, Polymer modified glassy carbon electrode for the

electrochemical determination of caffeine in coffee, Talanta., 93 (2012) 122–128.

[15] B. Brunetti, E. Desimoni, P. Casati, Determination of caffeine at a nafion covered

glassy carbon electrode, Electroanal., 19 (2007) 385-388.

17
[16] G. A. M. Mersal, Experimental and computational studies on the electrochemical

oxidation of caffeine at pseudo carbon paste electrode and its voltammetric

determination in different real samples. Food Anal. Meth., 5 (2012) 520-529.

[17] J. I. Gowda, S. T. Nandibewoor, Simultaneous electrochemical determination of 4-

aminophenazone and caffeine at electrochemically pre-treated graphite pencil

electrode, Anal. Meth., 6 (2014) 5147-5154.

[18] C. A. Martinez-Huitle, N. S. Fernandes, S. Ferro, A. de Battisti, M. A. Quiroz,

Fabrication and application of Nafion-modified boron-doped diamond electrode as

sensor for detecting caffeine, Diam. Rel. Mater., 19 (2010) 1188-1193.

[19] J. Y Sun, K. J. Huang, S. Y. Wei, Z. W. Wu, F. P. Ren, A graphene-based

electrochemical sensor for sensitive determination of caffeine, Coll. Sur. B, 84(2011)

421-426.

[20] T. Alizadeh, M. R Ganjali, M. Zare, P. Norouzi, Development of a voltammetric

sensor based on a molecularly imprinted polymer (MIP) for caffeine measurement,

Electrochim. Acta, 55 (2010) 1568-1574.

[21] L. Svorc, P.Tomcik, J. Svitkova, M. Rievaj, D. Bustin, Voltammetric determination

of caffeine in beverage samples on bare boron-doped diamond electrode, Food Chem.,

135 (2012) 1198-1204.

[22] S. Guo, Q. Zhu, B. Yang, J. Wang, B. Ye, Determination of caffeine content in tea

based on poly (safranine T) electroactive film modified electrode, Food Chem., 129

(2011) 1311–1314.

[23] A. K. Zak, W.H. A. Majid, M.R. Mahmoudian, M. Darroudi, R. Yousefi, Starch-

stabilized synthesis of ZnO nanopowders at low temperature and optical properties

study, Adv. Powd. Techn., 24 (2012) 1-7.

18
[24] K. R. Mahanthesha, B.E. K. Swamy, Pretreated/Carbon paste electrode based

voltammetric sensors for the detection of dopamine in presence of ascorbic acid and

uric acid, J. Electroanal. Chem., 703 (2013) 1-8.

[25] V. D. Mote, Y. Purushotham, B. N. Dole, Williamson-Hall analysis in estimation of

lattice strain in nanometer-sized ZnO particles, J. Theoret. Appl. Phys., 6(2012) 1-8.

[26] A. K. Thottoli, A. K. Achuthanunni, Effect of polyvinyl alcohol concentration on the

ZnS nanoparticles and wet chemical synthesis of wurtzite ZnS nanoparticles, J.

Nanostru. Chem., 3 (2013)1-9.

[27] R. H. Krishna, B. M. Nagabhushana, H. Nagabhushana, N. S. Murthy, S. C. Sharma,

C. Shivakumara, R. P. S. Chakradhar, Effect of calcination temperature on structural,

photoluminescence and thermoluminescence properties of Y2O3:Eu3+ nanophosphor,

J. Phys. Chem. C., 117 (2013) 1915-1924.

[28] E. J. Silvester, F. Grieser, B. A. Sexton, T.W. Healy, Spectroscopic studies on

copper sulfide sols, Longmuir, 7 (1991) 2917-2922.

[29] U. Sirimahachai, S. Phongpaichit, S. Wongnawa, Evaluation of bactericidal activity

of TiO2 photocatalysts: a comparative study of laboratory-made and commercial TiO2

samples, Songklanakarin, J. Sci. Technol., 31 (2009) 517-525.

[30] T. P. See, A. Pandikumar, H. N. Ming, L. H. Ngee, Y. Sulaiman, Simultaneous

electrochemical detection of dopamine and ascorbic acid using an iron oxide/reduced

graphene oxide modified glassy carbon electrode, Sensor., 14 (2014) 15227-15243.

[31] Y. Yu, W. Su, M. Yuan, Y. Fu, J. Hu, Electrocatalytic oxidation of formaldehyde on

nickel ion implanted-modified indium tin oxide electrode, J. Pow. Sour., 286 (2015)

130-135.

19
[32] M. Roushania, M. Shamsipur, H. R. Rajabi, Highly selective detection of dopamine

in the presence of ascorbic acid and uric acid using thioglycolic acid capped CdTe

quantum dots modified electrode, J. Electroanal. Chem.,712 (2014) 19-24.

[33] S. Pruneanu, F. Pogacean, A. R. Biris, S. Ardelean, V. Canpean, G. Blanita, E.

Dervishi, A. S. Biris, Novel graphene-gold nanoparticle modified electrodes for the

high sensitivity electrochemical spectroscopy detection and analysis of

carbamazepine., J. Phys. Chem. C., 115 (2011) 23387-23394.

[34] R. S. Nunes, E. T. G. Cavalheiro, Caffeine determination at a carbon fiber

ultramicroelectrodes by fast-scan cyclic voltammetry, J. Braz. Chem. Soc., 23 (2012)

670-677.

[35] M. L. Yola, N. Ozaltın, Electrochemical studies on the interaction of an antibacterial

drug nitrofurantoin with DNA, J. Electroanal. Chem., 653 (2011) 56-60.

[36] J. I. Gowda, S. T. Nandibewoor, Electrochemical behavior of paclitaxel and its

determination at glassy carbon electrode, Asi. J. Pharma. Sci., 9 (2013) 8.

[37] E. Laviron. General expression of the linear potential sweep voltammogram in the

case of diffusion less electrochemical systems, J. Electroanal. Chem., 101 (1979) 19-

28.

[38] F. Li, J. Li, Y. Feng, L. Yang, Z. F. Du, Electrochemical behavior of graphene

doped carbon paste electrode and its application for sensitive determination of

ascorbic acid, Sens. Actu. B., 157 (2011) 110-114.

[39] H. Ibrahim, Y. Temerk, Novel sensor for sensitive electrochemical determination of

luteolin based on In2O3 nanoparticles modified glassy carbon paste electrode, Sens.

Actu. B: Chem., 206 (2015) 744–752.

20
[40] M. Ibrahim, Y. Temerk, H. Ibrahim, M. Kotb, Indium oxide nanoparticles modified

carbon paste electrode for sensitive voltammetric determination of aromatase inhibitor

formestane, Sens. Actu. B: Chem., 209 (2015) 630–638.

Shivaraj Yellappa received his Master’s degree in Industrial

Chemistry from Kuvempu University, Karnataka, India in 2002.
Ph.D. degree in Physical Chemistry from Kuvempu University,
Karnataka, India in 2007. He is currently working as Assistant
Professor in Chemistry at Government Science College, Bengaluru,
India. His research interests include design and development of smart
materials based resonant sensors, modified electrodes.

Mallappa M received his Master’s degree in Chemistry from

Davanagere University, Karnataka, India in 2012. He is currently
working as Project Assistant in Government Science College,
Bengaluru, India. His research interests include design and
development of smart materials based resonant sensors, modified
electrodes.

Nagaraju Kottam received his Master’s degree in Chemistry from

Bangalore University, Karnataka, India in 1998. Ph.D. degree in
Inorganic Chemistry from Bangalore University, Karnataka, India in
2005. He is currently working as Associate Professor in Chemistry at
M.S. Ramaiah Institute Of Technology, Bengaluru, India. His
research interests include Material Science-Nano materials Carbon
dots, Photocatalysis, Waste Water Treatment.

21
Figures caption:

Fig.1 (a) Powder XRD pattern and (b) Crystal structure (covellite hexagonal) of CuS NPs.

Fig.2 (a) SEM and (b) TEM images of CuS Nanoparticles

Fig.3 (a) FT-IR Spectrum of pure starch (dashed line) and CuS NPs-Starch (solid line) and

(b) UV-Visible Spectrum of CuS NPs.

Fig.4 Electrochemical impedance curves of (a) BCPE and (b) CuS NPs MCPE in 0.1 M KCl

containing 5 mM [Fe(CN)6] 3-/4- (Amplitude: 5 mV).

Fig.5 (a) Cyclic and (b) differential pulse voltammograms of 0.12 mM of CAF on BCPE

(dashed lines) and CuS NPs MCPE (Solid line) in 0.1M ABS pH-7.0 at scan rate 0.05

Vs-1.

Fig.6 (a) CVs of 0.12 mM CAF in 0.1M ABS (pH 7.0) at CuS NPs MCPE at different scan

rates (0.050 to 0. 4 Vs-1), (inset of the plot of variation of peak current (Ipa) vs. υ1/2) and

(b) The plot of log Ipa vs. logυ.

Fig.7 CVs of 0.12 mM CAF in 0.1M ABS (pH 7.0) recorded at different amount CuS NPs [

a) 2, b) 4, c) 6, d) 8and f) 12 mg (solid line) and e) 10 mg (dashed line)] on the anodic

peak current and inset plots of the peak current vs. different values of CuS NPs).

Fig.8 DPVs of 0.12 mM CAF at CuS NPs MCPE in different values of pH (from a to e: 3.0,

4.0, 5.0, 6.0, 7.0 and 8.0) (Inset plots of the peak current of CAF vs. different values of

pH.

Fig. 9 (a) Effect of accumulation potential and (b) Effect of accumulation time on the

oxidation peak current of CuS NPs MCPE.

Fig.10 DPVs of CuS NPs MCPE in 0.1M ABS pH 7.0 for various concentration of CAF

(0.002 to 0.12 mM) at optimized parameters (inset the calibration plots peak current

(Ipa) vs. concentration of CAF for BCPE and CuS NPs MCPE).

Scheme 1. Possible mechanism for CAF oxidation.

22
Figures

Fig. 1 (a)

Fig. 1(b)

23
Fig. 2(a)

Fig. 2(b)

24
Fig.3 (a)

Fig.3 (b)

25
Fig. 4

26
Fig.5 (a)

Fig. 5 (b)

27
Fig. 6 (a)

Fig.6 (b)

28
Fig. 7

Fig. 8

29
Fig. 9 (a)

Fig. 9 (b)

30
 Fig.10

O O

H3C N H3C N
N N
-
+ H2O O + 2H+ + 2e
O N N O N N
H

CH3 CH3

-H+
-2e-

O O
OH
H3C H3C N
N
N 2H2O N
O + H+ O

N O N N
O N H
OH
CH3 CH3

Scheme 1

31
Table 1

Crystal parameters of the nanocrystalline CuS

Atoms Oxidation Wyckoff X/a Y/b Z/c Occupancy

state notation

Cu1 2 2d 1/3 2/3 3/4 -6m2

Cu2 2 4f 1/3 2/3 0.1072 3m

S1 -2 2c 1/3 2/3 1/4 -6m2

S2 -2 4e 0 0 0.0637 3m.

Crystal system: covellite hexagonal; space group: P63/mmc (No. 194) Lattice parameters: a = b =
3.7960 Å, c=16.3820 Å α = β = 90.0, γ = 120.0, Z = 6.

Table 2

Analytical data obtained for CAF using bare CPE and CuS NPs MCPE

Electrode
Parameter
Bare CPE CuS NPS MCPE

Peak potential (V) 1.363 1.31

Linear range (μmol L-1) 8 - 120 2 - 120

Correlation coefficient (r) 0.9292 0.99424

Slope (μA L mol-1) 7.750 22.3447

Standard deviation of slope (μA L mol-1) 0.591 1.2415

Intercept (μA) 4.8657 7.394

Standard deviation of intercept (μA) 0.4039 0.139

Detection limit (μmol L-1) 0.1563 0.0186

Quantification limit (μmol L-1) 0.5212 0.0622

32
Table 3
Comparison of analytical parameters of several modified electrodes for caffeine determination with caffeine sensors in the literature.
Linear range
Solution Sensitivity/ Peak Potential LOD/
Electrode Type upper limit/ References
pH µAcm-2mM-1 (Ep in V) µM
mM
BDD1 0.5 0.025 - 1.55 0.15 7
AQMCPE2 0.56 0.8 - 1.527 0.143 10
BQMCPE 6.0 8.0 28.8 1.45 5.1 7
Nafion/MWCNT 2.0 0.4 125.2 1.33 0.23 11
MWCNT-Nafion/ GCE 4.1 2.4 491.1 1.34 0.51 12
GO-Nafion/GCE3 ~2.0 0.08 232.7 1.45 0.20 13
Poly(AHNSA) /GCE4 5.0 0.04 638.4 1.34 0.14 14
Nafion/ GCE 1.0 0.011 - 1.45 0.79 15
CPE5 2.7 1.0 255.8 1.5 0.35 16
PPGE6 3.0 11 - ~1.5 0.0098 17
Nafion-GCE 7.0 7.0 169.7 1.32 0.04 8
CuS NPs MCPE 7.0 0.12 545.12 1.31 0.018 This work
BDD(Boron doped diamond electrode)1, AQMCPE(Anthroquinone modified carbon paste electrode) 2, BQMCPE (1, 4-benzoquinone modified carbon paste electrode) 3
GO(Graphene oxide)3. AHNSA (4-Amino-3-hydroxynaphthalene sulfonic acid) 4, CPE (Carbon paste electrode)5, PPGE(Pre treated pencil graphite electrode)6

33
Table 4

Determination results of caffeine in some tea and coffee samples

Samples Detection results Standard added Total found Recovery

(mg) (mg) (%)
(mg/g)

Green tea 19.86 20.00 40.23 101.6

Black tea 32.69 20.00 54.94 106.8

Bru coffee 41.71 20.00 58.81 94.65

Nescafe 32.58 20.00 51.54 96.80

34