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Ultrasensitive detection of orthophosphate ions

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with reduced graphene oxide/ferritin field-effect

Cite this: DOI: 10.1039/c6en00661b
transistor sensors†
Shun Mao,ab Haihui Pu,a Jingbo Chang,*a Xiaoyu Sui,a Guihua Zhou,a Ren Ren,a
Yantao Chenac and Junhong Chen*a

Received 15th December 2016,
Accepted 8th February 2017
DOI: 10.1039/c6en00661b
rsc.li/es-nano
The phosphorus level is one of the major parameters in evaluating water eutrophication and there is an in-
creasing demand for accurate and real-time monitoring technology for phosphorus determination. As a
novel ion detection method, field-effect transistor (FET) sensors show a lot of potential in low-
concentration and real-time orthophosphate ion detection. Here we report a reduced graphene oxide
(rGO)-based FET platform for orthophosphate ion detection. The sensing platform was fabricated with rGO
nanosheets as the sensing channel and ferritin as the specific detection probe for orthophosphate ions.
The sensors hold great promise in real-time detection of orthophosphate ions with a lower limit of detec-
tion down to 26 nM (phosphorus level: 0.806 μg L−1), a response time on the order of seconds, and good
selectivity in the presence of other interfering anions. Compared with standard optical methods, this
electronic sensor could potentially be used for online and real-time monitoring of phosphorus in surface
water.

Environmental significance
The phosphorus level is one of the major parameters in evaluating water eutrophication. In order to provide field-deployable, real-time, inexpensive, and
environmentally- and energetically-sustainable sensors for phosphorus monitoring in surface water, accurate and accessible detection technologies are re-
quired. For phosphorus determination, the commonly used optical methods, e.g., stannous chloride and ascorbic acid methods, are inaccurate for concen-
trations lower than 0.01 mg L−1. In addition, these methods cannot be used for real-time monitoring. Therefore, there is a great demand for new sensor
technology for low-concentration and real-time monitoring of phosphorus in water. Field-effect transistor (FET) sensors have received significant attention
in sensing applications due to their high sensitivity, fast response, portability, and simple operation. In this study, we report a reduced graphene oxide/fer-
ritin FET platform for orthophosphate ion detection in water. The sensors achieved a lower limit of detection (LOD) of 26 nM (phosphorus concentration:
0.806 μg L−1), which is much lower than the traditional optical methods, and a good selectivity against other anions in water. The sensors have a response
time on the order of seconds, which enables their potential applications in real-time and online determination of phosphorus in surface water. We believe
that this study will promote the development of new sensor technology for nutrient monitoring in water and will benefit the water industry, agriculture,
and environmental regulators by providing real-time sensing capabilities for phosphorus monitoring to better evaluate and control the water quality.

Introduction harmful when they over fertilize aquatic plants and cause eu-
trophication (total phosphorus >0.01–0.02 mg L−1).1–3 Total
Phosphates enter waterways through human and animal
phosphorus in water is typically determined in two steps:
waste, phosphorus rich bedrock, laundry, cleaning, industrial
first, a digestion method to oxidize organic matter (algae,
effluents, and fertilizer runoff. These phosphates become
plant and animal tissue, waste solids, organophosphorus pes-
ticide or other organic matter) effectively to release phospho-
a
Department of Mechanical Engineering, University of Wisconsin-Milwaukee, 3200
rus as orthophosphate; second, orthophosphate determina-
N. Cramer Street, Milwaukee, WI 53211, USA. E-mail: jhchen@uwm.edu,
chang69@uwm.edu
tion. The vanadomolybdophosphoric acid method, stannous
b
State Key Laboratory of Pollution Control and Resource Reuse, College of chloride method, and ascorbic acid method are three com-
Environmental Science and Engineering, Tongji University, 1239 Siping Road, monly used methods for routine orthophosphate analysis,
Shanghai 200092, China which are based on optical signals using a color comparator
c
Tianjin Key Laboratory for Photoelectric and Devices, School of Materials Science
or field spectrophotometer/colorimeter.4 Although they work
and Engineering, Tianjin University of Technology, Tianjin 300384, China
† Electronic supplementary information (ESI) available: Sensing response of an-
accurately for phosphorous levels at 1 to 20 mg L−1 and 0.01
other sensor without a probe linker; control sensing test of the sensor without a to 6 mg L−1, respectively, they are inaccurate for concentra-
probe; pH impact study; sensor recycling study. See DOI: 10.1039/c6en00661b tions lower than 0.01 mg L−1.4 Other optical methods, e.g.,

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fluorescence and luminescence methods, were also reported
for the determination of orthophosphate.5–8 However, their
lower detection limits are still high and the setup is difficult
to adapt for online measurements because a lot of reagents
are needed. Since phosphorus is one of the major reasons for
eutrophication, sensitive, online, and low-cost phosphorus
sensors are needed for monitoring the eutrophication pro-
cess, particularly well before the eutrophication incepts, i.e.,
a phosphorous level below 0.01 mg L−1.9
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In the past decade, there were many studies on new phos-
phate determination methods. For instance, the amperomet-
ric method, also known as the electrochemical sensor
method,10–14 utilizes the electrocatalytic activity of the work-
ing electrode in a redox system for orthophosphate determi-
nation. However, the lower detection limits of electro-
chemical sensors are still relatively high, e.g., 9.3 μg L−1 for
the Ni electrode,10 0.186 mg L−1 for the carbon black
electrode,13 and 0.31 mg L−1 for the Co electrode.14 In addi-
tion, the electrochemical method suffers from complicated
electrode fabrication, relatively low stability, and/or needs re-
action agents for the detection. The ion selective electrode
(ISE) method or the potentiometric sensor method is also
reported for orthophosphate determination and considerable
efforts have been directed to electrode membrane develop-
ment.15 For instance, a potentiometric sensor using a chiral
A2B2 macrocyclic compound as the ionophore could detect
10−6 to 10−2 M HPO42−;16 the one using an anthracene thio-
urea derivative as the ionophore achieved a 10−7–10−3 M
working range for HPO42−;17 and another one using tri-layer
maltose phosphorylase composite films worked for 1–20 μM
phosphate.18 Although ISE sensors show some potential in
orthophosphate detection, they have limitations such as rela-
tively high lower detection limit and short lifetime of the
membrane.
Recently, field-effect transistor (FET) sensors have
attracted great interest in the sensing community due to their
high sensitivity, fast response, portability, and simple opera-
tion. A typical FET sensor consists of source/drain electrodes
and the channel material, and works by monitoring the elec-
trical conductance change in the channel material upon the
adsorption of target analytes. Two-dimensional (2D) nano-
materials, due to their layer-dependent electronic properties
and high sensitivity to electronic perturbations from analyte
adsorption, have drawn significant attention in FET sensor
applications. For example, graphene (an atomic-thick carbon
layer)19 and reduced graphene oxide (rGO)-based nanostruc-
tures have been widely studied for gas sensors,20,21 biosen-
sors,22,23 and water sensors.24,25 Graphene FET sensors utilize
graphene nanosheets as the sensing channel. When the ana-
lyte attaches to the graphene surface, the charge transfer and
the adsorbed analyte-induced voltage (gating effect) will re-
sult in a conductivity change in graphene, which is measured
as the sensing signal. To make the sensor selective to the tar-
get analyte, usually a probe (chemical- or bio-material) is la-
beled on the surface of graphene to enable a high binding af-
finity to the specific target analyte.
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For example, graphene-based FET sensors have recently
been reported for heavy metal ion detection in water. In
2011, Chen et al. reported a label-free micropatterned rGO
film FET sensor for Cd2+ and Hg2+ ion detection.26 This sen-
sor can detect ions in a real-time fashion and the lower limit
of detection (LOD) is around 1 nM for both Cd2+ and Hg2+
ions with a good selectivity. Through surface modification of
graphene with Au nanoparticles (NPs) and a DNA probe, they
used a similar platform to detect Pb2+ ions, which achieved
an extremely low detection limit of 0.02 nM.27 Using
graphene-based FET sensors, many studies reported the de-
tection of heavy metal ions28–32 and bacteria33–35 in different
water systems and the sensors exhibited a lot of promise
compared with traditional methods.24
In this study, we report an rGO/ferritin FET platform for
the sensitive and selective detection of orthophosphate ions
(HPO42−) in water. To realize ultra-sensitivity and selectivity,
we use ferritin as the specific probe. On the one hand, ortho-
phosphate ions barely adsorb on rGO due to their closed
shell-induced chemical inertia, but they can uniquely react
with ferritin. On the other hand, ferritin is a relatively small
nanoparticle (12 nm), so that the gating effect from the
adsorbed orthophosphate ions can be preserved to a large ex-
tent without being screened by the ion solution, further lead-
ing to a high sensitivity. Owing to the designed rGO/ferritin
hybrid structure, the sensors achieved an LOD of 26 nM
(phosphorus concentration: 0.806 μg L−1), which is much
lower than those of the traditional optical methods (0.01 mg
L−1). The sensor selectivity was further confirmed by testing
the sensors against other common interfering anions (Cl−,
SO42− and CO32−) in water, and the sensors showed a much
higher sensitivity to HPO42− compared with other anions. In
addition to the good sensitivity and selectivity, the sensors
showed a response time on the order of seconds, which en-
ables their potential applications in real-time determination
of phosphorus for surface water quality control. We believe
that this study will open a new avenue for future develop-
ment of sensor technology for nutrient monitoring in water.
Experimental
Sensor device fabrication
Monolayer GO solution (10 mg mL−1) dispersed in deionized
water (DI water) was ordered from ACS Materials (Single
Layer Graphene Oxide Dispersion, SKU# GNO1W001). In the
FET sensor, gold interdigitated electrodes with both finger-
width and inter-finger spacing of about 2 μm and a thickness
of 50 nm were fabricated using an optical lithography process
on a silicon wafer with a top layer of thermally-formed SiO2
(thickness of 200 nm). To deposit rGO sheets between gold
electrodes, a self-assembly method was used, in which a di-
luted GO suspension was drop-cast on the electrode and
annealed in an argon flow for 1 h at 400 °C to reduce GO to
rGO. After that, ferritin (Sigma-Aldrich, ferritin from equine
spleen, #F4503) was labeled on the rGO surface by incubating
the sensor device in the ferritin solution (10 mg mL−1) for 2
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hours. To enhance the stability of ferritin on the rGO surface,
some of the sensors were treated with a linker
(1-pyrenebutyric acid N-hydroxysuccinimide ester/C24H19NO4)
first before the incubation in the ferritin solution. After the
ferritin incubation, the sensors were washed with DI water
several times and used for the ion sensing test.
Sensor structure characterization and sensing test
A Hitachi S-4800 scanning electron microscope (SEM) was
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used for SEM characterization at an acceleration voltage of 10
kV. Electrical measurements on the rGO sensor were
performed using a Keithley 4200 semiconductor analyzer at
room temperature. In the I–V measurement, the drain-source
current (Ids) was recorded when the drain-source voltage (Vds)
was swept from −2.0 to +2.0 V. In the transistor measure-
ment, a fixed Vds (0.05–0.5 V) was applied to the electrode,
and a back gate potential (Vg) was swept from −40.0 to +40.0
V. In the ion sensing experiment, the dynamic responses of
the sensors were recorded by monitoring the Ids in the sen-
sors under a given Vds when the sensors were exposed to dif-
ferent ions with various concentrations. In our experiment,
Na2HPO4 was used as the source of orthophosphate ions and
dissolved in DI water. The Na2HPO4 solution was in a weakly
basic condition with a pH range from 7.5 (5 nM Na2HPO4) to
8.9 (100 μM Na2HPO4). For the recycling test, the sensor de-
vice was immersed in a 2% NaOH and 2% NaCl solution for
30 minutes and then washed with DI water several times.
Results and discussion
Fig. 1 left panel shows a schematic of the rGO/ferritin
sensing platform for orthophosphate ion detection. As we
discussed, for graphene-based FET sensors, a probe is needed
for the sensitive and selective detection of target ions. Be-
sides, better sensor performance requires that the probe be
firmly attached to the surface of rGO nanosheets without
degrading rGO's electronic properties. In this study, ferritin
is selected as the specific probe for orthophosphate ions
since it has been proved to have a high binding affinity/fast
reaction rate to orthophosphate ions in phosphate ion re-
moval applications.36,37 For example, a previous study
reported that ferritin was sensitive and could trace low levels of
orthophosphates with an effective phosphate removal capacity
Fig. 1 Schematic of the rGO/ferritin sensing platform for HPO42− detection. In the sensing process, HPO42− binds with the ferritin core (ironIJIII)
oxide-hydroxide groups), leading to a change in the electrical conductivity of the rGO sheet.
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of approximately 11 mg PO43− g−1 ferritin.37 Ferritin is an iron
storage protein, which has a roughly spherical structure with
an outer diameter of approximately 12 nm and an inner cav-
ity diameter of 8 nm. In its nanocage, ferritin can form a hy-
drated iron oxide mineral core, which acts as a sorbent for
orthophosphates, forming an iron-oxyhydroxide-phosphate
nanoparticle (as shown in Fig. 1, right panel).
Fig. 2a and b show the SEM images of the rGO nanosheets
and the hybrid rGO and ferritin structure on the sensor
electrode, respectively. The image shows that rGO sheets
bridge the sensor electrode, and ferritin nanoparticles are
uniformly decorated on the surface of the rGO sheets. The
size of the ferritin nanoparticle is around 10 nm estimated
from the magnified image shown in Fig. 2b inset, which is
close to its theoretical value (12 nm). Before the ion sensing
test, the electrical properties of the rGO device were tested by
I–V and transistor measurements, and the results are shown
in Fig. 2c and d. As shown in Fig. 2c, the I–V curve of the
rGO device is nearly linear, indicating a small Schottky bar-
rier between the rGO nanosheets and electrodes. In addition
to the I–V measurement, transistor measurements on the
rGO FET device are also needed to understand its transistor
characteristics, e.g., the type of FET and the on–off current ra-
tio. Based on the results shown in Fig. 2d, the rGO shows a
p-type semiconducting characteristic and its on–off current
ratio is around 1.64. The pristine graphene is a semimetal
(e.g., zero bandgap) and typically shows a bipolar characteris-
tic in the FET curve with a low on–off current ratio. However,
because of the structural defects, residual oxygen groups in
the carbon lattice, and the surface adsorbed oxygen mole-
cules from air, the rGO shows a p-type semiconducting char-
acteristic when the FET test is carried out in an ambient envi-
ronment. After the ferritin labeling (without linker), very
small changes are found in the FET curve, which indicates
that the ferritin probes have a negligible impact on the elec-
trical properties of rGO.
Generally, a linker is required to firmly link the sensing
probe onto the rGO surface. However, ferritin nanoparticles
are relatively small and could be firmly attached onto the rGO
surface due to the van der Waals force. As a result, a linker
might not be necessary. To test the hypothesis, we compared
the performance of the sensor with and without a linker
(1-pyrenebutyric acid N-hydroxysuccinimide ester). The linker
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Fig. 2 SEM images of the (a) rGO and (b) rGO/ferritin hybrid structure on the sensor electrode. The dotted line shows the edges of rGO
nanosheets in the electrode gap. The magnified inset in (b) shows the ferritin particles. (c) I–V measurement results of the rGO/ferritin sensor. The
sensor shows a nearly linear I–V curve. (d) Transistor measurement results of the rGO/ferritin sensor. The rGO sensor shows p-type semiconduct-
ing characteristic and the ferritin labelling brings very small changes in the FET curve.

has a pyrene group that can bond with rGO through π–π inter-
action and an N-hydroxysuccinimide (NHS) group that links
with ferritin. The orthophosphate ion tests were carried out
by a drop casting method. Before adding the orthophosphate
ion solution, DI water was added to the sensor surface to cre-
ate a baseline. Fig. 3a and b show the dynamic responses of
the rGO/ferritin sensors to HPO42− ions of different concentra-
tions with and without the probe linker, respectively. After in-
jection of HPO42− ions, the current increased abruptly then de-
creased slowly. The abrupt current increase is due to the
adsorption of HPO42− ions onto ferritin, while the slow de-
crease arises from the adsorption competition induced detach-
ment of HPO42− ions until the adsorption equilibrium is
reached. And this is more pronounced at high concentrations
since more HPO42− ions are adsorbed instantly upon injection
to ferritin. Surprisingly, with the linker, the sensor showed very
small responses to HPO42− ions with concentrations of 5, 33
and 250 nM; and when the concentration increased to 2 μM,
the sensor started to show a sensing response. In contrast,
without the probe linker, the sensor started to show a signifi-
cant response at a much lower concentration level; e.g., the sen-
sor can respond to HPO42− ions with a concentration of 50 nM.
This comparison obviously shows that the probe linker is in-
deed not necessary for the ferritin probe. Moreover, the linker
could even decrease the sensor sensitivity, probably due to the
degradation of carrier mobility resulting from the π–π interac-
tion between the aromatic rings in the linker and the rGO
nanosheets. Based on the dynamic test results, it is also found
that the sensor response (∼seconds) is much faster than that
of the conventional optical method, which takes minutes or
even longer to respond.4
Fig. 3c summarizes the sensor sensitivities to HPO42− at dif-
ferent concentrations. The sensitivity error bars were obtained
by testing 5–8 sensors for each concentration (Fig. S1† shows
the sensing response of another typical sensor without a probe
linker, ESI†). The sensor sensitivity is defined as the ratio of
the current change after ion addition (ΔI = I − I0) over the ini-
tial current (I0). The sensors without probe linker have much
higher sensitivities compared to those with probe linker. We
could see that the sensor sensitivity is directly proportional to
the concentration of HPO42−, which indicates that the sensor
response is indeed related to the binding of HPO42− to the sen-
sor sensing channel. The noise level of the sensor is deter-
mined to be 0.05–0.07%, and a reliable sensor signal should be
larger than three times the noise level, which is 0.21%. By ana-
lyzing the sensor sensitivity vs. concentration, the LOD of the
sensor is determined to be 0.026 μM by using the fitting equa-
tion of the fitted line (y = 3.1213 logIJx) + 5.1347). To further un-
derstand the sensor responses to the HPO42− ions, we have
tested control sensors without the ferritin probe. As shown in
Fig. S2 (ESI†), without the ferritin probe, the sensor current
only shows very small fluctuations to HPO42− with no steady in-
crease, due to the weak nonspecific binding of HPO42− ions to
rGO. The control experiment thus further confirms that the la-
beling of the ferritin probe on the rGO surface is critical to the
specific binding and the stable current response in the sensor;
and the increase in the sensor current is directly related to the
binding of HPO42− ions with the ferritin probe.

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Fig. 3 (a) and (b) Dynamic responses of the rGO/ferritin sensors to HPO42− of different concentrations with and without the probe linker. (c)
Sensitivity (%) plots of the sensors to HPO42− of different concentrations with and without the probe linker. The error bars were obtained by
testing 5–8 sensors for each concentration. (d) Transistor characteristics of an rGO/ferritin sensor exposed to DI water (black), 33 nM HPO42− (red),
and 250 nM HPO42− (blue) solutions. The dash lines indicate the linear response region. The intercepts with the gate potential are the threshold
voltages for the sensor exposed to different concentrations of HPO42−.

An FET sensor works by monitoring the analyte-induced
conductance change. To study the sensing mechanism, we
measured the transistor characteristics (Fig. 3d) of the rGO/
ferritin sensor exposed to DI water (black), 33 nM HPO42−
(red), and 250 nM HPO42− (blue) solutions. It is found that
the threshold voltage of rGO increases upon HPO42− ion ad-
sorption, implying that the p-type rGO is hole doped, which
can be due to either direct hole charge transfer from HPO42−
ions into rGO or the negative gating effect induced increase
of hole concentration in rGO. For our rGO/ferritin sensor, or-
thophosphate ions can be detected as they migrate into the
nanocage of ferritin NPs and attach on the surface of the iron
mineral core. Chemically, a reaction takes place between the
hydroxyl groups in both the orthophosphate ion and the iron
mineral core, giving rise to the production of water molecules
(cf. reaction (1)):
FeO–OH + HPO42− → FeO–O–PO32− + H2O.
The iron mineral core (i.e., ferritin) is then negatively
charged due to the adsorbed orthophosphate ions. On the
one hand, electron transfers from ferritin into rGO, leading to
the conductance decrease due to the decrease of hole concen-
tration in rGO. However, in our case, the charge transfer effect
is negligible. This is possibly because the iron mineral core of
ferritin rather than the surface protein is charged and the size
of ferritin is relatively large for charge transfer (∼10 nm in di-
ameter as analyzed above). Thus, the electron transfer from
the iron mineral core into rGO is greatly mitigated/blocked
by the long transfer path induced by the surface proteins of
ferritin itself. On the other hand, negatively charged ferritin
NPs collectively exert an effective negative gate voltage to the
p-type rGO, and thus the hole concentration is increased by
this voltage and the conductance in rGO correspondingly in-
creases. Therefore, the threshold voltage increase arises from
the gating effect of the adsorbed HPO42− ions.
We can also see from Fig. 3d that the current in both hole
and electron channels increases at the same gate voltage
upon HPO42− ion adsorption with increased slope at higher
concentration. It had been reported that such an increase
could result from the carrier mobility enhancement and
Schottky barrier reduction.38 For our sensor structure, ferritin
is negatively charged upon exposure to HPO42− ions and it acts
as a scattering center of the charge carriers in rGO, indicating
that the mobility should be reduced and thus cannot give rise
(1) to the current increase. However, the local work function of
the Au electrode (∼5.4 eV)39 can decrease because of the ad-
sorption of HPO42− ions.40 This means that the work function
difference between Au and rGO (∼4.7 eV)41,42 decreases. As a
result, the Schottky barrier as shown in Fig. 2c between the
Au and rGO contact should decrease, and thus lead to the cur-
rent increase in both the hole and electron transport regimes.
Consequently, the sensing mechanism of our sensor is domi-
nated by both the gating effect and Schottky barrier reduction.
Note that for the gating effect, screening from positive
ions in water may also influence the sensor response;

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therefore, we further evaluate the stray positive ions (e.g., Na+
from the source of phosphate solution) in screening the gat-
ing effect. We could quantify this screening effect in terms of
the Debye length λD of the ion solution,43,44
(2)
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where λB (=0.7 nm at 300 K) is the Bjerrum length in water,
and CiIJZi) is the concentration (valence charge) of ion species
i, respectively. Using eqn (2), the Debye length λD is estimated
to be 5.6 μm, 0.18 μm, and 18 nm corresponding to 1 nM, 1
μM, and 0.1 mM Na2HPO4 solution, respectively. In our sensor
test, the orthophosphate ion concentration is in the range of
nM–μM, indicating that the Debye length λD is much larger
than the radius r of the ferritin NPs (∼5 nm), namely, lim r/λD
= 0. Therefore, the gating effect from the ion solution is quan-
titatively screened in the magnitude of [1 − expIJ−r/λD)], reveal-
ing that the screening can be reasonably ignored. For example,
in 1 μM Na2HPO4 solution, the gating effect from the adsorbed
orthophosphate ions on ferritin is degraded by only 2.7% com-
pared with the case without the ionic screening effect from the
orthophosphate solution considered. Consequently, the accu-
mulated orthophosphate ions on the surface of the iron min-
eral core inside the ferritin nanocage work dominantly as a
negative gating potential and increases the hole concentration
in rGO, thereby leading to a current increase.
Since the sensor signal depends on the ionic strength of
the sample, the sensor response could be affected by the pH
of the sample. To experimentally study how pH influences
the sensor sensitivity, control tests were carried out by testing
the sensor exposed to DI water prepared with a pH from 7 to
9 (pH of HPO42− solution is 7.5 to 8.9, depending on the con-
centration). As shown in Fig. S3 (ESI†), the sensor has no re-
sponses in this pH range, which indicates that the sensor is
stable under weakly basic conditions for HPO42− sensing. To
demonstrate the selectivity of the rGO/ferritin sensor, we
studied its sensing behavior when it was exposed to solutions
containing other common interfering anions such as Cl−,
Fig. 4 (a) Dynamic responses of the rGO/ferritin sensors to HPO42−, Cl−, SO42− and CO32− ions. (b) Sensitivity summary of the sensors to different
anions in water. The error bars were obtained from three sensors. The sensor shows a much higher response to HPO42− even with a lower
concentration (2 μM) compared with those of Cl− (5 μM), SO42− (3.3 μM) and CO32− ions (2.5 μM).
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SO42− and CO32− ions. The experiment was carried out by
adding the interfering ions one by one and finally adding the
HPO42− ions. As shown in Fig. 4a, the sensor shows very weak
responses to Cl− (5 μM, current increase), SO42− (3.3 μM, cur-
rent increase) and CO32− ions (2.5 μM, current decrease).
Compared with Cl−, SO42− and CO32−, which only have coulom-
bic interaction, HPO42− has both coulombic interaction and
Lewis acid–base interaction with the hydrated iron oxide of
ferritin, which leads to highly specific binding affinity. Be-
cause of this, the sensor shows a much larger response to
HPO42− even with a lower concentration (2 μM) than the inter-
fering ions. Fig. 4b summarizes the sensitivities of HPO42−
and interfering ions. The sensitivity of HPO42− (1.90%) is sev-
eral times higher than those of Cl− (0.10%), SO42− (0.30%)
and CO32− (0.35%), which indicates that the sensor could
differentiate HPO42− from other interfering anions in water.
In addition to the selectivity, the multiple-time deploy-
ment or reuse of the sensor is worthy of special attention for
its practical applications. The sensor will lose its sensitivity
after one test (especially testing with a high-concentration
sample) due to the occupation of the binding sites on the
probe. A previous study showed that a solution containing
2% NaOH and 2% NaCl was effective to desorb phosphate
from the exhausted hydrated iron oxide after the phosphate
removal.45 By adopting this idea, we studied the possibility of
recovering the mineral core of ferritin by treating the sensor
device in an NaOH and NaCl solution after each sensing test.
Fig. S4 (ESI†) shows the recycling results of the sensor for
five-time use in HPO42− sensing. The sensor showed strong
responses to 250 nM and 2.5 μM HPO42− in the first test.
With the recovery treatment, although the sensitivity of the
sensor still decreased in the repeating tests, the sensor
showed good responses to 2.5 μM HPO42− and recovered 71%
and 57% sensitivity in the 4th and 5th tests, respectively. It is
found that the recovery treatment is more efficient for high
concentrations and the sensor sensitivities were much higher
in the 4th and 5th tests compared with the 2nd and 3rd tests.
We believe that this is due to the recovery recipe and a longer
treatment time may be needed for the efficient desorption of
orthophosphate ions from the ferritin core. Further
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refinement of the recovery recipe is needed to improve the re-
covery efficiency.
Conclusions 5 Q.-L. Li, N. Bao and S.-N. Ding, Electroanalysis, 2014, 26,
In summary, we have demonstrated an rGO/ferritin FET sen- 6 J. Liu, K. Wu, X. Li, Y. Han and M. Xia, RSC Adv., 2013, 3,
sor for ultrasensitive detection of orthophosphate ions
(HPO42−) with an LOD of 26 nM (corresponding to a phospho- 7 G.-L. Wang, H.-J. Jiao, X.-Y. Zhu, Y.-M. Dong and Z.-J. Li,
rus concentration of 0.806 μg L−1), which is one order of mag-
Published on 13 February 2017. Downloaded by Freie Universitaet Berlin on 21/02/2017 13:06:42.
nitude lower than the US EPA recommended total phospho- 8 A. Wild, A. Winter, M. D. Hager and U. S. Schubert, Analyst,
rus level (25 μg L−1) for water reservoirs. The performance of
the rGO/ferritin sensor is very encouraging for low- 9 U. S. EPA, Ecoregional Criteria Documents, http://www2.epa.
concentration and real-time monitoring of orthophosphate
ions and could be further enhanced by tuning the sensor 10
structures. The reliability of the sensor is critical to its real
application. To improve the sensor reliability, the device fab- 11
rication should be precisely controlled to minimize the
device-to-device variations; e.g., the rGO sheet density on the 12
electrode and probe density on the rGO sheet should be con-
trolled. To this direction, using thickness and size controlled 13
rGO films is one choice. Second, a calibration method should
be developed to precisely correlate the sensor sensitivity with 14
the HPO42− concentration. In the calibration, the sensor elec-
trical characteristics, e.g., initial resistance and transistor on– 15
off current ratio, should be taken into consideration. For this
type of sensor, clinical diagnostics is another field where low- 16
concentration phosphate measurement is in demand. For ex-
ample, hyperparathyroidism, vitamin D deficiency, and 17
Fanconi syndrome can be diagnosed based on specific phos-
phate concentrations in body fluids. The sensor platform 18
reported here opens up a new avenue for designing
nanomaterial-based FET sensors by combining semiconduct-
ing nanomaterials with ion adsorption agents and offers a 19
new sensing modality for low-concentration monitoring of
nutrients in water. 20
Acknowledgements 21
This work was financially supported by the U.S. National Sci-
ence Foundation through the NSF Industry/University Cooper-
ative Research Center on Water Equipment & Policy led by 22
the University of Wisconsin-Milwaukee and Marquette Univer-
sity (IIP-1540032) and a GOALI grant (CBET-1606057). The au- 23
thors thank Dr. H. A. Owen for technical support with SEM
analyses. The SEM imaging was conducted at the Electron 24
Microscope Laboratory of UWM.
25
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