Microchimica Acta (2019) 186: 267

https://doi.org/10.1007/s00604-019-3364-2

ORIGINAL PAPER

A nanocomposite prepared from magnetite nanoparticles,

polyaniline and carboxy-modified graphene oxide for non-enzymatic
sensing of glucose
Razia Batool 1 & Muhammad Asim Akhtar 1 & Akhtar Hayat 1 & Dongxue Han 2,3 & Li Niu 2,3 & Muhammad Ashfaq Ahmad 4 &
Mian Hasnain Nawaz 1,2

Received: 28 February 2019 / Accepted: 12 March 2019 / Published online: 2 April 2019
# Springer-Verlag GmbH Austria, part of Springer Nature 2019

Abstract
The authors report on the synthesis of carboxy functionalized graphene oxide (fGO) decorated with magnetite (Fe3O4) nano-
particles. The resulting nanomaterial was used to prepare a composite with polyaniline (PANI) which was characterized by UV-
vis, Fourier transform-infrared and Raman spectroscopies. Its surface morphologies were characterized by atomic force and
scanning electron microscopies. A screen-printed carbon electrode was then modified with the nanocomposite to obtain an
enzyme-free glucose sensor. The large surface of fGO and Fe3O4 along with the enhanced charge transfer capability of PANI
warrant a pronounced electrochemical response (typically measured at 0.18 V versus Ag/AgCl) which is suppressed in the
presence of glucose. This reduction of current by glucose was used to design a sensitive method for quantification of glucose. The
response of the modified SPCE is linear in the 0.05 μM – 5 mM glucose concentration range, and the lower detection limit is
0.01 μM.

Keywords Screen printed carbon electrode . Functionalized graphene oxide . Iron oxide . Polyaniline . Electrochemical sensor .
Glucose detection . Serum sample . Enzyme free glucose sensor . Metal oxide decorated graphene

Introduction variety of methods have been employed either to grow newer

Glucose sensors play a pivotal role in clinical detection, envi-
ronmental monitoring and in other fields. [1–3] In these lines,
* Akhtar Hayat
akhtarhayat@cuilahore.edu.pk
* Mian Hasnain Nawaz
mhnawaz@cuilahore.edu.pk
1
Interdisciplinary Research Centre in Biomedical Materials (IRCBM),
COMSATS University Islamabad Lahore Campus, Lahore 55150,
Pakistan
2
State Key Laboratory of Electroanalytical Chemistry, c/o
Engineering Laboratory for Modern Analytical Techniques,
Changchun Institute of Applied Chemistry, Chinese Academy of
Sciences, Changchun 130022, People’s Republic of China
3
Center for Advanced Analytical Science, c/o School of Chemistry
and Chemical Engineering, Guangzhou University,
Guangzhou 510006, People’s Republic of China
4
Department of Physics, COMSATS University Islamabad Lahore
Campus, Lahore 55150, Pakistan
protocols or to improve the analytical performances of
existing procedures. In past few decades, enzyme based elec-
trochemical glucose sensors are the primary choice for many
researchers due to their higher sensitivity and lower response
time. [4] However, enzymatic sensors possess limitations of
indirect transferring electrons, denaturing of enzymes and
high manufacturing cost. [5] To overcome these short com-
ings, enzyme free electrochemical sensors have been emerged
as promising replacement for glucose detection. In this direc-
tion, electro-catalysis of transition metal oxides proved to be
advantageous both in device performance and fabrication. [6,
7] On the other hands, discovery of reversible redox behavior
of Fe3O4 triggered its applications in mitigating the chemical
oxidation or reduction of different analyte molecules. [8]
These electroanalytical studies provide ample alternatives to
miniature device fabrication for enzyme-free glucose sensing.
Likewise, graphene oxide (GO) and its derivatives have been
under extensive research because of their diverse func-
tionalities and attractive physical and chemical proper-
ties. [9, 10] Many functionalized GO based metal oxide
nanomaterials, including cobalt oxide (CoO), manganese
267 Page 2 of 10
dioxide (MnO2), titanium dioxide (TiO2) and iron oxide
(Fe3O4) have been reported for the detection of enzyme
free glucose. [11] Fe3O4 nanoparticles offer better con-
ductivity by making composition with polymer as com-
pare to other metal oxides such as NiO, CoOOH, CuO,
MnO 2 . [12, 13] Similarly, graphene-based polymeric
composite materials exhibit ample thermal, electrical
and mechanical properties in comparison with pristine
polymer and their organic/inorganic counter parts. [14]
In these composite materials unlike conventional poly-
mers, electrically conducting polymers not only give di-
rect electric readout of analyte presence with high sensi-
tivity, but they also act as signal generating probe. This
feature makes conducting polymers excellent candidate
for integration in non-enzymatic sensors to generate an
electrochemical signal read out without any prerequisite
of additional electroactive species. [15] Among common-
ly used conducting polymers, polyaniline (PANI) pro-
duces great prospects with Fe3O4 anchored functional-
ized graphene oxide to enhance the electro active perfor-
mance. [16]
In this work, we describe a glucose sensor based on iron
oxide (Fe3O4) decorated functionalized graphene oxide (fGO)
and polyaniline (PANI) composite having tendency to form
complex while interacting with glucose. High density of
Fe3O4 was uniformly anchored on the basal plan of function-
alized graphene oxide. PANI was then introduced with fGO-
Fe3O4 to yield homogeneous fGO-Fe3O4-PANI nanocompos-
ite which functioned as electroactive as well as affinity based
material towards glucose detection. The nanocomposite was
characterized with UV-vis, FTIR, RAMAN, SEM and AFM.
The working interface of SPCE was modified with this com-
plex material resulting in increase of active surface area.
Moreover, remarkably convincing results were achieved in
case of real sample analysis. The present sensor system based
on catalytic potential of this nanocomposite material is a
promising candidate for the enzyme less glucose sensor.
Experimental
Chemicals and apparatus
Iron chloride hexahydrate (FeCl3.6H2O), iron chloride
tetrahydrate (FeCl2.4H2O), oxalic acid, ascorbic acid and
uric acid were purchased from Sigma Aldrich (https://
www.sigmaaldrich.com/technical-service-home/product-
catalog.html). Graphite powder, Ammonia, hydrobromic
acid (HBr), emeraldine form of polyaniline (PANI),
potassium ferrocyanide (K 4 Fe (CN) 6 ) and potassium
ferricyanide (K 3 Fe (CN) 6 ) were purchased from Alfa
Aesar (https://www.alfa.com/en/). All the solvents and
reagents were of analytical grade.
Microchim Acta (2019) 186: 267
UV-vis spectra were recorded on Perkin Elmer UV/Vis
spectrometer Lambda 35. FTIR spectroscopy was performed
on Thermo scientific Nicolet 6700 in ATR mode. RAMAN
spectroscopy was performed on InVia Raman Microscope
Renishaw UK at 514 nm wavelength. To observe the surface
morphology of modified electrode, Park Systems AFM XE7
in non-contact mode and TESCAN VEGA 3 were used for
AFM and SEM micrographs, respectively. For electrochemi-
cal studies, Palmsens Electrochemical sensor interface was
used. All the measurements were performed at room temper-
ature and screen printed carbon-based electrodes (SPCE) were
used as a three-electrode system, with Ag/AgCl as a reference
electrode and carbon-based counter and working interfaces.
Synthesis of carboxyl functionalized graphene oxide
(fGO)
Prior to functionalize graphene oxide (GO) with all-carboxylic
groups, [17] GO was synthesized via modified Hummers’
method from graphite flakes. [18] To get fGO, typically
150 mg of GO was dispersed into 60 mL distilled water under
ultra-sonication for 30 min at room temperature. To reduce
epoxides of basal plane, 10 mL of hydrobromic acid (HBr)
was then added and stirred vigorously for 12 h. To introduce
carboxylic functionality, 3 g of oxalic acid was added follow-
ed by continuous stirring for 4 h. The reaction mixture was
then filtrated, washed and dried at 50 °C in vacuum for 36 h.
In situ synthesis of fGO-Fe3O4 nanoparticles (NPs)
To decorate fGO with monodispersed Fe3O4 NPs, a synthetic
methodology [19] was used which resulted in growth of uni-
formly distributed fine nanoparticles onto the basal plane. To
synthesize fGO-Fe3O4 NPs, 48 mg of carboxylic functional-
ized graphene oxide was dissolved in 30 mL of distilled water
by temperature controlled ultra-sonication for about 30 min.
Afterward, 40 mg of FeCl3.6H2O was added and the mixture
was stirred for another 30 min under the nitrogen atmosphere.
Subsequently, 60 mg of FeCl2.4H2O was added under a nitro-
gen atmosphere with continuous stirring for 30 min followed
by dropwise addition of 4 mL of aqueous NH3 diluted with
20 mL of distilled water. The mixture was heated to 60 °C and
stirred for further two hours under nitrogen atmosphere.
Finally, the reaction mixture was centrifuged, washed with
distilled water and was dried to 50 °C for 36 h.
Synthesis of fGO-Fe3O4 NP-PANI nanocomposite
For the synthesis of nanocomposite via non-covalent interac-
tions, 70 mg of fGO-Fe3O4 NP were dissolved in 30 mL of
distilled water under stirring with subsequent addition of
140 mg of polyaniline. The reaction mixture was then stirred
for 30 min and cooled at 0–5 °C. Afterward, 0.5 g of
Microchim Acta (2019) 186: 267
FeCl3.6H2O dissolved into 7.5 mL of distilled water was
quickly added to the pre-cooled mixture and allowed to stir
for 15 h at 0–5 °C. It was added to get higher yield of nano-
composite. The final mixture was centrifuged, washed with
ethanol and distilled water and dried at 50 °C for 24 h.
Modification of electrodes with nanocomposite
and glucose detection
Prior to modification of screen-printed carbon electrodes
(SPCE) with nanocomposite, electrode surface was electro-
chemically cleaned to oxides the impurities by successive cy-
clic voltammetric scans with 0.5 M solution of sulfuric acid
(H2SO4) in the potential range of −1.5 – 1.0 V followed by
washing with distilled water. To deposit a thin layer of nano-
composite on working electrode surface, typically 5 μL of the
fGO-Fe3O4-PANI nanocomposite was drop cast on the work-
ing interface of SPCE followed by drying at room tempera-
ture. Three successive CV scans were run after casting 5 μL of
nanocomposite solution onto SPCE surface in potential range
of −0.7 – 0.9 V at scan rate of 50 mVs−1. Electrochemical
measurements were performed at various scan rates (25, 50,
100, 150 mVs−1), while an optimal output current was ob-
served at 50 mV/s. Hence, all the further experiments were
performed at scan rate of 50 mVs−1. The modified electrodes
were then washed with distilled water for further use.
Real sample analysis
To confirm the applicability of the fabricated sensor, human
blood serum sample studies were carried out. Human blood
serum of normal individual was collected from Services
Hospital, Lahore Pakistan. Freshly collected serum samples
were spiked with three different concentration of glucose
(0.78 μmol.L−1, 0.09 μmol.L−1). Cyclic voltammetry re-
sponse was measured at scan rate of 50 mV/s. Based on the
generated electrochemical signal in response to the spiked
concentrations, recovery percentages were calculated from
calibration curve for each concentration.
Results & discussions
Choice of materials
Graphene oxide (GO) and its derivatives have been under
extensive research due to their various functionalities and
smart physical and chemical properties. [9, 10] Various metal
oxides including Fe 3 O 4 , CoO, NiO, CoOOH, CuO,
MnO2,TiO2 and different sulfides (NiS; Ni3S2) in association
with functionalized GO have been used as transducing
nanomaterials for enzyme free detection of glucose. [11–13]
Higher conductivity and reversible redox behaviour of Fe3O4,
Page 3 of 10 267
trigger its performance in terms of electron transfer and
oxidation/reduction of different analyte molecules. [8]
Moreover, the current protocol of in situ synthesis of
Fe3O4 nanoparticles with uniform size distribution along
the basal plane of graphene provide more surface area and
enhanced catalytic performances. It also bears superior
performances in case of conductivity by making compos-
ites with conductive polymer. In this scenario the fGO-
Fe3O4-PANI nanocomposite may contribute in direct elec-
tric readout of analyte presence with high sensitivity and
may further perform as signal generating probe giving
better electro active performance in enzyme free detection
of glucose.
Surface characterizations
UV-vis absorption spectra of GO, fGO, fGO-Fe3O4NP and
fGO-Fe3O4NP-PANI nanocomposite are given in Fig. 1(a).
In curve a, the appearance of the absorption peak at 233 nm
shows the presence of GO which corresponds to C-C
stretching bonds as well as π → π* transitions. [20] The func-
tionalized graphene oxide is rich in carboxylic (-COOH)
groups on both basal planes and edges. The enhanced absorp-
tion peak at 236 nm evidenced the fGO synthesis which cor-
responds to π → π* transitions, curve b. [21] Furthermore, the
Fe3O4 anchoring on basal plane shifted and broadened the
absorption peak of GO which is related to the electronic inter-
action of metal nanoparticles with basal conjugation (c).
Finally, the formation of fGO-Fe3O4 NP-PANI was confirmed
from the peak appeared at 252 nm along with the appearance
of a new peak at 220 nm. The appearance of new absorbance
peak shows the presence of PANI which increased the con-
ductivity of nanocomposite as well as active surface area [22]
which were demonstrated by electrochemical characteriza-
tion. Similarly, FTIR was used to characterize the structural
changes at molecular level and the functionalization of GO
with carboxylic groups. The FTIR spectra of GO, fGO,
fGO-Fe3O4 NP and fGO-Fe3O4 NP-PANI nanocomposite is
shown in Fig. 1(b) confirming the successful functionalization
of graphene oxide, Fe3O4 NPs anchoring and their composite
formation with PANI. The absorption bands at 1562 cm−1
correspond to C=O stretching of COOH groups and the
C=C stretching of GO was found at 1704 cm−1 confirming
the functionalization of graphene oxide. [17] The anchoring of
Fe3O4 was confirmed by the stretching vibration of Fe-O,
recorded at 923 cm−1 (c). In case of fGO-Fe3O4 NP-PANI
nanocomposite (d), the peaks at 2355 cm − 1 and at
1562 cm−1 correspond to –OH stretching and C=O groups
and indicate that PANI protects the surface of functionalized
graphene oxide. Similarly, the appearance of C-O stretching at
1105 cm−1 and Fe-O-C at 783 cm−1 confirmed the presence of
iron oxide. Moreover, the C-O peak also indicated the proton-
ated conducting arrangements of PANI and the characteristic
267 Page 4 of 10 Microchim Acta (2019) 186: 267

0.45 0.9
(A) (B) (Fe-O-C)

Relative intensity (a.u)

(C-O)
Absorbance (C=O)
(-OH )
0.6
a d
0.30
(Fe-O)
b
0.3 c
c
d (C=C)

a
0.15 b
200 240 280 320 360 400 4000 3000 2000 1000
Wavelength (nm) Wave number (cm-1)
1200
(C) G- 1587
(N-H)

900 (C-H)
d
Intensity ( a.u)

D-1357
600

b
300 a

c
0

1200 1500 1800 2100 2400

Raman Shift (cm-1)
Fig. 1 UV-vis spectra (a), FTIR spectra (b) and Raman spectra (c) of (a) GO, (b) fGO, (c) fGO-Fe3O4 NP and (d) fGO-Fe3O4 NP-PANI nanocomposite.
For UV-vis and RAMAN spectra measurements aqueous suspensions of these materials were used

peak at 1288 cm−1 corresponded the benzenoid ring. [23] For
further insight into the structural changes of fGO-Fe3O4 NP-
PANI nanocomposite, the vibrational spectra of functionalized
graphene oxide and nanocomposite were recorded via
RAMAN spectroscopy, as shown in Fig. 1(c). The typical D
and G bands were observed in all samples. Due to the presence
of oxygen groups and SP3 bonded carbon atoms, D- band
offered to the disorder and defects. D-band recorded at
1357 cm−1 associated with the vibrations of dangling bonds
of carbon atoms and corresponds to k-point breathing mood.
[24] The G-band recorded at 1587 cm−1 corresponds to SP2
bonded carbon atoms in plane E2g stretching vibration mode
[25]. A change in intensity ratio of ID to IG corresponds to the
change in SP2 bonded carbon atoms which ultimately equates
the defects in GO basal plan. [26] In the given spectra of
functionalized GO, the C-H stretching was recorded at
1097 cm−1. [27] GO, fGO and fGO-Fe3O4NP show G and
D-band vibrations at the same point with same intensity, nano-
composite depicted both bands at a higher intensity. The ID/IG
intensity ratios 0.94, 0.82, 0.96, 0.8 were calculated from the
G-band and D-band peaks at different sites corresponding to
GO, fGO and fGO-Fe3O4NP and fGO-Fe3O4 NP-PANI
nanocomposite respectively. The change in intensity ratios
presented a decrease in the disorder and defects on the nano-
composite surfaces as well as change of average size of sp2
domains.
Figure 2 represents the scanning electron microscopic im-
ages of electrode surface at different stages of fabrication pro-
cess. The gradual changes in the surface morphology can be
envisioned in the corresponding micrographs. Disorderly dis-
tributed sheets of graphene oxide can be envisioned in the Fig.
2a. Nevertheless, in case of fGO well distributed and compact
architecture, nicely covering the electrode surface, was found
as shown in Fig. 2b.
Nanoparticles of Fe3O4 were augmented on graphene
sheets that are partially covered with nanoparticles. From
Fig. 2c it can be fancied that the nanoparticles are disorderly
distributed with lower homogeneity which was consequently
homogenised on addition of PANI, as shown in Fig. 3c.
Formation of nanocomposite increases the homogeneity and
capability of film formation resulting in the uniform distribu-
tion of fGO-Fe3O4 throughout the electrode surface which
ultimately enhances the electro catalytic properties of modi-
fied electrode.
Microchim Acta (2019) 186: 267 Page 5 of 10 267

Fig. 2 SEM micrographs of

electrodes modified with (a) GO,
(b) fGO, (c) fGO-Fe3O4 NP and
(d) fGO-Fe3O4 NP-PANI nano-
composite. The samples were
drop casted onto working inter-
face which was cut and pasted on
conducting adhesive tape prior to
microscopic analysis

Similar trends of surface morphology were observed in the process, as shown in Fig. 3a-d. The roughness of electrodes
topographic analysis of sequential sensor fabrication steps. modified with GO, fGO, fGO-Fe3O4 NP and fGO-Fe3O4 NP-
The topographic analysis revealed the decreasing roughness PANI were calculated to be 156 nm, 138 nm, 126 nm and
vogue during the successive steps of sensor fabrication 116 nm, respectively. This decrease in surface roughness

Fig. 3 Atomic force micrographs

of modified electrodes with (a)
GO, (b) fGO, (c) fGO-Fe3O4 NP
and (d) fGO-Fe3O4 NP-PANI
nanocomposite. The samples
were drop casted on working in-
terface of SPCE and dried at room
temperature
267 Page 6 of 10 Microchim Acta (2019) 186: 267

3
1.3
80 (B)
(A) b
b
c 2
40 c
0.0
Current (µA)

Current (µA)
d 0.0 0.9
0 1
a d

-40 0
Blank

a
-80
-0.6 -0.3 0.0 0.3 0.6 0.9 -0.2 0.0 0.2 0.4 0.6 0.8
Potential (E/V) Potential (E/V)
Fig. 4 a Cyclic Voltammograms of each step modification of SPCE; (a) GO, (b) fGO, (c) fGO-Fe3O4 NP and (d) fGO-Fe3O4 NP- PANI nanocomposite
immobilized working electrode in 5 mM (K4Fe (CN)6) /(K3Fe(CN)6) and b PBS buffer with pH 7 at scan rate of 50 mV/s

comprehend the increased homogeneity of the fGO-Fe3O4 Glucose detection

NP-PANI nanocomposite which eventually increased the elec-
tron conductivity of the surface.
Electrochemical characterizations
Cyclic voltammetry (CV) has been widely used to probe the
electrolyte/electrode interfacial properties during fabrication
of various sensors. [28] The electrochemical output signal of
the fabricated electroactive transducer surface was dependent
on the following two phenomena; physical hindrance and
blocking of electroactive sites on the transducer surface in
the presence of glucose. All measurements were recorded
against 5 mM of ferri/ferro redox at scan rate of 50 mV/s as
shown in Fig. 4a. The change in redox current and peak to
peak separation in voltammograms relates electron transfer
rate constant for each fabrication step. These electrochemical
variations signify increase of electrons transfer between mod-
ified electrode surface and electrochemical probe. Moreover,
surface treatment by nanocomposite resulted in decrease of
electron transfer resistant that showing slightly increase in
peak shifting and redox peak potential. The redox current
was particularly greater for nanocomposite-modified elec-
trode as compared to the GO-modified electrode, escribing
Moreover, cyclic voltammograms of blank and modified
SPCE electrodes were recorded in the presence and absence
of glucose with concentration of 50 μM at scan rate of 50
mVs−1 is shown in Fig. 5. The CV measurement of blank
electrode shows no current peak. The CV response of the
SPCE modified with nanocomposite in the presence/absence
of glucose shows good conducting behaviour as compared to
bare electrode. Cyclic current response of modified SPCE was
decreased in the presence of glucose which shows that the
synthesized material was an electroactive material capable of
complexing with glucose. Because of glucose complex for-
mation with electroactive material of modified electrode the
electron transfer was blocked, and usable active surface area
was decreased. Consequently, the redox peak current was de-
creased, and the effect was proportional to the glucose con-
centration which was used as probe in quantitative detection
of glucose. Moreover, the peak to peak separation was also
decreased from 0.18 to 0.16 V.
1.0
Current ( µA)

the higher charge transfer of nanocomposite which influenced 0.5

Before Glucose
the redox performance.
After Glucose
In Fig. 4b, all measurements of modified electrodes were
recorded in phosphate buffer (PBS with pH 7) solution at scan 0.0 Blank
rate of 50 mVs−1. CV response of blank SPCE in the presence
of PBS buffer shows negligible current without any noticeable
-0.5
redox peak. Similarly, in the presence of (a-c) materials no -0.2 0.0 0.2 0.4 0.6 0.8
redox peak appeared. A prominent peak was recorded for Potential (E/V)
nanocomposite modified electrode at potential of 0.1 V attrib- Fig. 5 Cyclic Voltammetry response of modified SPCE in PBS pH 7
uted to the electroactive nature of nanocomposite as shown in before and after glucose detection, indicating the quenching of
Fig. 4b (d). electroactive material’s intrinsic peak at 0.1 V
Microchim Acta (2019) 186: 267 Page 7 of 10 267

0.9
(a) (b)
Current (µA) 0.0

0.8

-0.9
Increase Glucose
Concentration

-1.8

0.0

-2.7
0.0 0.8

-0.4 0.0 0.4 0.8

Potential (E/V)

Fig. 6 Cyclic Voltammetry (CV) (a) different concentrations of glucose on nanocomposite from up to down. b Calibration plot of different concentra-
tions of glucose corresponding to cyclic voltammetry at scan rate of 50 mV s−1. Error bar means ± standard deviation with N = 3

The analyte concentration has better influence in device effect was proportional to the glucose concentration which
performance particularly in selectivity and sensitivity. was used as a probe in quantitative detection of glucose. The
During the fabrication of enzyme-less sensor cyclic voltamm- detection limit of 0.01 μM was calculated for a signal-to-noise
etry with different concentration of glucose was performed at ratio of these data. Similarly, limit of quantification was found
50 mV s−1 is shown in Fig. 6a. The oxidation anodic peak to be 0.03 μM. These results suggest that synthesized
current decreased significantly upon increasing the glucose electroactive material are very much useable for the enzyme-
concentration (0.05 μM-5 mM). The oxidation peak of glu- less sensor.
cose positioned at 0.18 V as shown in Fig. 6a which is linearly To assess the repeatability, reproducibility, and stability of
associated with the glucose concentration. [29] Analytical re- the prepared fGO–Fe3O4–PANI nanocomposite based elec-
sponse of designed glucose sensor was confirmed by making trodes, several electrodes were primed and tested under the
linear curve between oxidation current and glucose concentra- similar environmental conditions. For repeatability, a single
tion as shown in Fig. 6b. To establish the calibration plot as a modified electrode was used to sense glucose for ten times,
line graph the glucose concentrations in the range of followed by thoroughly washing with deionized water after
(0.05 μM-5 mM) were incubated on modified electrodes in- each test. The standard deviation of the electrochemical cur-
dependently. A linear curve was showing a gradual decrease rent response was found to be just 0.3% which displays the
in Δratio with increasing the glucose concentration in the linear excellent repeatability, as shown in Fig. 7a.
range of 0.05 μM to 5 mM. Electrochemical response of the The sensor showed to be reproducible, as 6 electrodes were
electroactive transducer surface was decreased in the presence primed individually and used for glucose detection, as shown
of glucose, demonstrating the complex formation between in Fig. 7b. A satisfactory reproducibility in the electrochemi-
transducer surface and glucose. This complex formation not cal response of the sensor was attained with a standard devi-
only hindered the electron transfer flow but also covered the ation of 4%. Likewise, the long-term stability of the sensor
available electroacitve sites on the transducer surface. was estimated by determining the current response of three
Consequently, the redox peak current was decreased, and the electrodes against glucose for 16 days. The modified

(a) (b) (c)

1.2 1.2 1.2
Current (uA)
Current (uA)

Current (uA)

0.8 0.8 0.8

0.4 0.4 0.4

0 0 0
1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 0 4 8 12 16
Repeats Sensors Days

Fig. 7 a Repeatability of fGO–Fe3O4–PANI nanocomposite modified environment for their response towards glucose. c Stability of fGO–
electrodes for their response towards glucose for ten times. b Fe3O4–PANI nanocomposite modified electrodes towards glucose
Reproducibility assessment of 6 electrodes primed under similar tested after every 4 days for 16 days. (N = 3)
267 Page 8 of 10 Microchim Acta (2019) 186: 267

1.5 Table 2 Recovery percentages attained with the designed

electrochemical sensor for glucose monitoring in human blood serum
sample
1
Glucose added Glucose found R.S.D. % R.E % R%
(μmol/L) (μmol/L)
0.5
0.09 0.091 4.3 1 101
0 0.78 0.74 4.7 5.2 94.8
50 48 4.5 4 96

R.S.D. % = relative standard deviation percentage; R.E % = relative error

Fig. 8 Interference studies of different analytes indicating the selectivity
of fabricated enzyme less sensor towards glucose detection. Error bars
indicating the ± SD based on three independent measurements (N = 3)
electrodes were stored underneath ambient condition and were
electrochemically tested after every 4 days. The current re-
sponse was slightly different, and 93% current was engaged
with a standard deviation of 2.3%, as shown in Fig. 7c, which
determined the long-term stability and practice of the designed
sensor.
percentage; R% = recovery percentage
was negligible as compared to glucose even for much higher
concentrations. These experiments show the absence of any
cross reactions during glucose detection with much superior
selectivity. Ultimately, to certify the usage of our glucose sen-
sor in real sample analysis the protocol was demonstrated with
human blood serum sample. Table 1 indicates the comparision
of the fabricated sensor with the already reported nanocom-
posites for glucose detection (Table 1).

Real sample analysis

Effect of interfering analytes on detection of glucose
Moreover, for sensors the analyte specificity is an important
point for device fabrication. In order to validate the specificity
of as fabricated glucose sensor many interacting molecules
including uric acid (UA), dopamine (DA), cholesterol, fruc-
tose, maltose, saccharose, carrageenan and ascorbic acid (AA)
were studies using the same protocol as in case of glucose. As
can be seen from the Fig. 8 the response for all these analytes
Similarly, to validate the selectivity and sensitivity of the fab-
ricated glucose sensor for real life applications the detection of
known concentrations of glucose in human serum samples
were also carried out. Three different concentrations of glu-
cose (0.09 μmol.L−1, 0.78 μmol.L−1 and 50 μmol.L−1) were
spiked with human blood serum at room temperature. The
satisfied recoveries of 96% - 101% with negligible deviations
shows acceptable accuracy of the protocol. The recoveries of

Table 1 Comparison of
electroanalytical response of Sr# Materials Used Methodologies LOD Linear Range Ref
different nanocomposite materials (μM) (mM)
towards glucose detection
Au/LDH-CNTs-G Co-precipitation 1.00 0.01 to 6.1 [1]
ZnO NRs Hydrothermal method 0.40 8.45 [7]
Fe2O3 nanowire arrays Solution phase – 0.015–8 [8]
PtFe/C GCE Co-reduction 3.00 0.01 to 18.9 [12]
PtxFe alloy NP Chemical 3.00 0.01 to 18.9 [13]
AuNPs/GC Seed Mediated growth 05.0 0.1 to 25 [29]
Fe3O4 nanorod array (NRA) In situ annealing 0.10 0.765 to 3.7 [30]
GCE/PANI/GNPs Physical mixing 100 0.3 to 10 [31]
CuFe2O4/PPy Chemical oxidative 0.10 0.02 to 5.6 [32]
polymerization
CNFs-Ni(OH)2/GCE hydrothermal route 0.76 0.001 to 1.2 [33]
Fe3O4-nanoparticles/Ag/GCE Reduction method 1.20 Up to 3.5 [34]
Pt/PANI/Au porous electrodes electrodeposition – 0.009 to 5 [35]
Cobalt oxide NPs/r-GOPE Chemical reduction 1.40 0.04 to 4 [36]
NiNP/MWCNTs/copper Precipitation 0.70 0.002 to 10 [37]
fGO-Fe3O4 NP In situ anchoring of NPs 0.01 0.00005 to 0.050 This Work
Microchim Acta (2019) 186: 267
the method are excellent. This shows that the sensor can be
applied to glucose detection (Table 2).
Conclusions
A new non-enzymatic glucose based on fGO-Fe3O4 NP-PANI
nanocomposite has been synthesized and characterized under
different physiological conditions. The enhanced electro ac-
tivity, surface area and conductivity of the fGO-Fe3O4 NP-
PANI nanocomposite consequently enabled the fabricated
sensor to monitor the glucose at lower potential with outstand-
ing sensitivity and selectivity. A linear relationship between
electron transfer resistance and different analytes concentra-
tion was found. The selectivity of the fabricated glucose sen-
sor was further explained with various interfering analytes and
was validated with human blood serum samples. The present
sensor system based on superior performances is a promising
candidate for the enzyme less glucose sensor. The future work
may focus to improve the immobilization efficiency of fGO-
Fe3O4 NP-PANI on the transducer surface. It is highly desir-
able to obtain uniform and homogenous layer of composite
material on the transducer surface. The reproducibility and
repeatability of the method can be further improved with ex-
ploration of novel immobilization methodologies.
Acknowledgments M. H. N thanks the Higher Education Commission
of Pakistan for financial assistance under start-up research grant and na-
tional research program for universities (21-329/SRGP/R&D/HEC/2014
and 20-4993/R&D/HEC/14/614). R. B acknowledges the HEC supported
studentship under NRPU.
Compliance with ethical standards The author(s) declare that
they have no competing interests.
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