Anal. Chem.
2005, 77, 5894-5901
Electrochemical Sensor for Organophosphate
Pesticides and Nerve Agents Using Zirconia
Nanoparticles as Selective Sorbents
Guodong Liu and Yuehe Lin*
Pacific Northwest National Laboratory, Richland, Washington 99352
An electrochemical sensor for detection of organophos-
phate (OP) pesticides and nerve agents using zirconia
(ZrO2) nanoparticles as selective sorbents is presented.
Zirconia nanoparticles were electrodynamically deposited
onto the polycrystalline gold electrode by cyclic voltam-
metry. Because of the strong affinity of zirconia for the
phosphoric group, nitroaromatic OPs strongly bind to the
ZrO2 nanoparticle surface. The electrochemical charac-
terization and anodic stripping voltammetric performance
of bound OPs were evaluated using cyclic voltammetric
and square-wave voltammetric (SWV) analysis. SWV was
used to monitor the amount of bound OPs and provide
simple, fast, and facile quantitative methods for nitroaro-
matic OP compounds. The sensor surface can be regener-
ated by successively running SWV scanning. Operational
parameters, including the amount of nanoparticles, ad-
sorption time, and pH of the reaction medium have been
optimized. The stripping voltammetric response is highly
linear over the 5-100 ng/mL (ppb) methyl parathion
range examined (2-min adsorption), with a detection limit
of 3 ng/mL and good precision (RSD ) 5.3%, n ) 10).
The detection limit was improved to 1 ng/mL by using
10-min adsorption time. The promising stripping vol-
tammetric performances open new opportunities for
fast, simple, and sensitive analysis of OPs in environ-
mental and biological samples. These findings can lead
to a widespread use of electrochemical sensors to detect
OP contaminates.
Organophosphates (OPs) are known to be highly neurotoxic;
they disrupt the cholinesterase enzyme that regulates acetyl-
choline,1-5 a neurotransmitter needed for proper nervous system
function. Because of their high neurotoxicity, the OPs are widely
used as pesticides and as nerve agents as part of chemical and
biological warfare agents. OP residuals in crop, livestock, and
* Corresponding author. Tel.: 01-509-376-0529. Fax: 01-509 376-5106.
E-mail: yuehe.lin@pnl.gov.
(1) Rosenberry, T. L. Advances in enzymology and related areas of molecular
biology; John Wiley & Sons: New York, 1975.
(2) Zhang, S.; Zhao, H.; John, R. Biosens. Bioelectron. 2001, 16, 1119-1126.
(3) Fennouh, S.; Casimiri, V.; Burstein, C. Biosens. Bioelectron. 1997, 12, 97-
104.
(4) Cremisini, C.; Disario, S.; Mela, J.; Pilloton, R.; Palleschi, G. Anal. Chim.
Acta 1995, 311, 273-280.
(5) Guerrieri, A.; Monaci, L.; Quinto, M.; Palmisano, F. Analyst 2002, 127, 5-7.
5894 Analytical Chemistry, Vol. 77, No. 18, September 15, 2005
poultry products are clearly dangerous to human health. The
related clinical signs include negative effects on the visual system,
sensory function, cognitive function, and nervous system. Specif-
ically, exposure to OPs has been shown to cause headache,
dizziness, profuse sweating, blurred vision, nausea, vomiting,
reduced heart beat, diarrhea, loss of coordination, slow and weak
breathing, fever, coma, and death.6 Infants and children may be
especially sensitive to health risks posed by pesticides: an
estimated 74 000 children were involved in common household
pesticide-related poisonings or exposures in the United States in
1994.7 Because of the high toxicity of OPs, the rapid detection of
these toxic agents in the environment, public places, or workplaces
and the monitoring of individual exposures to chemical warfare
agents have become increasingly important for homeland security
and health protection.8-11 Early detection of OPs may give an
indication of terrorist activity, allowing proper procedures to be
followed to mitigate dangers. It is still an extremely difficult
challenge to detect low concentrations of OPs accurately in
environmental samples. Soil and water samples are very likely to
contain OPs because of heavy urban and rural use of these
compounds. Military and terrorist activities may result in air,
water, and soil contamination with different chemical warfare
agents. Analysis of OPs in environmental and biological samples
is routinely carried out using analytical techniques, such as gas
or liquid chromatography and mass spectrometry.12 Such analysis
is generally performed at centralized laboratories, requiring
extensive labor and analytical resources, and often results in a
lengthy turnaround time. These analysis methods have a number
of disadvantages that limit their applications primarily to laboratory
settings and prohibit their use for rapid analyses under field
conditions. Biological methods, such as immunoassay, have also
been reported.13 Long analysis time and extensive sample handling
with multiple washing steps limit the applications. In recent years,
OP pesticide kits have become commercially available that offer
advantages, including portability, rapid turnaround time, and cost-
effectiveness.14 Drawbacks of these test kits include the compli-
(6) http://www.beyondpesticides.org/.
(7) http://www.epa.gov/pesticides/.
(8) Wang, J. Anal. Chim. Acta 2003, 507, 3-10.
(9) Sadik, O. A.; Land, W. H.; Wang, J. Electroanalysis 2003, 15, 1149-1159.
(10) Lin, Y.; Lu, F.; Wang, J. Electroanalysis 2004, 16, 145-149.
(11) Wang, J.; Pumera, M.; Collins, G.; Mulchandani, A.; Lin, Y.; Olsen, K. Anal.
Chem. 2002, 74, 1187-1191.
(12) Sherma, J. Anal. Chem. 1993, 65, 40R-54R.
(13) Miller J. K.; Lenz D. J. Appl. Toxicol. 2001, 21, S23-S26.
10.1021/ac050791t CCC: $30.25 © 2005 American Chemical Society
Published on Web 08/09/2005

Figure 1. Structure of nitroaromatic OP compounds. (A) Paraoxon;
(B) fenitrothion; (C) methyl parathion.
cated handling procedure and often a lack of sensitivity (ppm level)
and precision. Moreover, in most cases, these tests are qualitative
or semiquantitative and show false positive and negative results.
To meet the requirements of rapid warning and field deploy-
ment, more-compact low-cost instruments, coupled to smaller
sensing probes, are highly desirable for facilitating the task of
on-site monitoring of OP compounds. Various inhibition and
noninhibition biosensor systems, based on the immobilization of
acetylcholinesterase or OP hydrolase onto various electrochemical
or optical transducers, have been proposed for field screening of
OP neurotoxins.15-19 Specific antibodies against OP pesticides have
been recently developed for enzyme-linked immunoassay and
immunosensors.20,21 Although acetylcholinesterase is commercially
available, OP hydrolase and antibodies against OPs are still only
produced in laboratories, which limits wide applications of bio-
sensors. To avoid the use of enzymes and antibodies, molecular
imprint technologies with high selectivity toward specific OP
species have been developed and applied to the detection of
pesticides in environmental samples.22,23 Nitroaromatic OPs, such
as paraoxon, methyl parathion, and fenitrothion (Figure 1), exhibit
good redox activities at the electrode surface.11 Electrochemical
detection of nitroaromatic OPs showed great promise when it was
coupled with different separation technologies, such as high-
performance liquid chromatography24 or capillary electrophore-
sis.25 Surprisingly, little attention has been given to direct
electrochemical sensing of nitroaromatic OP compounds, despite
(14) The EnviroLogix Cholinesterase Screening Test (EP 014). EnviroLogix Inc.,
www.envirologix.com.
(15) La Rosa, C.; Pariente, F.; Hernandez, L.; Lorenzo, E. Anal. Chim. Acta 1994,
295, 273-282.
(16) Mulchandani, A.; Kaneva, I.; Chen, W. Anal. Chem. 1998, 70, 5042-5046.
(17) Wang, J.; Mulchandani, A.; Chen, L.; Mulchandani, P.; Chen, W. Anal. Chem.
1999, 71, 2246-2249.
(18) Mulchandani, A.; Mulchandani, P.; Kaneva, I.; Chen, W. Anal. Chem. 1998,
70, 4140-4145.
(19) Wang, J.; Mulchandani, A.; Chen, L.; Mulchandani, P.; Chen, W. Elec-
troanalysis 1999, 11, 866-869.
(20) Hu, S.; Xie, J.; Xu, Q.; Rong, K.; Shen, G.; Yu, R. Talanta 2003, 61, 769-
777.
(21) Marty, I.-L.; Leca, B.; Noguer, T. Analusis Mag. 1998, 26, M144-M149.
(22) Turiel, E.; Matin-Esteban, A.; Femandez, P.; Perez-Conde, C.; Camara, C.
Anal. Chem. 2001, 73, 5133-5141.
(23) Muldoon, M. T.; Stanker, L. H. Anal. Chem. 1997, 69, 803-808.
(24) Martinez, R. C.; Gonzalo, E. R.; Garcı́a, F. G.; Méndez, J. H. J. Chromatogr.
1993, 644, 49-58.
(25) Wang, J.; Chatrathi, M.; Mulchandani, A.; Chen, W. Anal. Chem. 2001,
73, 1804-1808.
their inherent redox activity and the compact nature of electro-
chemical instruments.
Zirconia (ZrO2) is an inorganic oxide with thermal stability,
chemical inertness, and lack of toxicity.26-29 Researchers have
demonstrated that zirconia has a strong affinity for the phosphoric
group. This has been used to prepare multiple films by self-
assembly,27-29 or a DNA probe30,31 was attached with the phosphate
group at the 5′ end to develop a DNA biosensor. Zirconia films
or microcrystals were prepared by electrodeposition of ZrOCl2 at
bare or functionalized gold surfaces.32-34 In this paper, we describe
the electrochemical sensing nitroaromatic OPs based on a gold
electrode modified with zirconia nanoparticles (Figure 2). The new
ZrO2 nanoparticle-based electrochemical sensing protocol involves
electrodynamically depositing ZrO2 nanoparticles onto a gold
electrode surface (A), followed by OP adsorption (B), and
electrochemical stripping detection of adsorbed electroactive OPs
(C). The electrochemical characterization and anodic stripping
voltammetric performance of bound nitroaromatic OP compounds
were evaluated using cyclic voltammetric and square-wave vol-
tammetric (SWV) analysis. The promising stripping voltammetric
performances open new opportunities for fast, simple, and sensi-
tive analysis of OPs. A disposable screen-printed gold electrode
and portable electrochemical instrument would benefit the field
monitoring of OPs.
EXPERIMENTAL SECTION
Reagents. Paraoxon, methyl parathion, and fenitrothion were
purchased from Sigma-Aldrich, and their 10 000 mg/L stock
solutions were prepared in acetonitrile. Stock solutions of 5 mg/L
trinitrotoluene (TNT) were prepared from a 1000 mg/L standard
solution of TNT in acetonitrile (Cerilliant, Austin, TX) in 0.1 M
potassium chloride, which was used as the supporting electrolyte
and also served as the adsorption medium during the adsorption
experiments. Zirconium oxychloride (ZrOCl2), nitrobenzene, and
p-nitrophenol were obtained from Sigma-Aldrich (St. Louis, MO)
and used without further purification. Other reagents were
commercially available and were of analytical reagent grade.
Solutions were prepared with ultrapure water from a Millipore
Milli-Q water purification system (Billerica, MA).
Instruments. Cyclic voltammetric and SWV measurements
were performed using an electrochemical analyzer CHI 660 (CH
Instruments, Austin, TX) connected to a personal computer. A
three-electrode configuration was employed, consisting of a
zirconia nanoparticle-modified gold electrode (3-mm diameter)
serving as a working electrode, while Ag/AgCl/3 M KCl and
platinum wire served as the reference and counter electrodes,
respectively. Electrochemical experiments were carried out in a
(26) Thomas Buscher, C.; McBranch, D.; Li, D. J. Am. Chem. Soc. 1996, 118,
2950-2953.
(27) Fang, M.; Kaschak, D. M.; Sutorik, A. C.; Mallouk T. E. J. Am. Chem. Soc.
1997, 119, 12184-12191.
(28) Lee, H.; Kepley, L. J.; Hong, H.; Mallouk T. E. J. Am. Chem. Soc. 1988,
110, 618-620.
(29) Hong, H.; Sackett, D. D.; Mallouk, T. E. Chem. Mater. 1991, 3, 521-527.
(30) Zhu, N.; Zhang A.; Wang, Q.; He, P.; Fang, Y. Anal. Chim. Acta 2004, 510,
163-168.
(31) Liu, S.; Xu, J.; Chen, H. Bioelectrochemistry 2002, 57, 149-154.
(32) Yu, H.; Rowe, A.; Waugh, D. M. Anal. Chem. 2002, 74, 5742-5747.
(33) Aslam, M.; Pethkar, S.; Bandyopadhyay, K.; Mulla, I. S.; Sainkar, S. R.;
Mandale, A. B.; Vijayamohanan, K., J. Mater. Chem. 2000, 10, 1737-1743.
(34) Bandyopadhyay, K.; Vijayamohanan, K. Langmuir 1998, 14, 6924-6929.
Analytical Chemistry, Vol. 77, No. 18, September 15, 2005 5895
Figure 2. Scheme of electrochemical sensing nitroaromatic OP compounds. (A) Electrodeposition ZrO2 nanoparticle to gold electrode surface;
(B) nitroaromatic OP compounds adsorb to ZrO2 nanoparticle surface; (C) electrochemical stripping detection of nitroaromatic OP compounds;
X ) O or S and R ) nitroaromatic OP group.
2-mL voltammetric cell at room temperature (25 °C). All potentials
are referred to the Ag/AgCl reference electrode (CH Instru-
ments). Scanning electron microscopy (SEM) was carried out
using a JEOL JSM-5900 LV machine. All samples were imaged
under vacuum conditions using secondary electron imaging. The
typical accelerating voltage of the electron beam used was 10 kV.
All samples were grounded with a piece of copper tape to curtail
specimen charging.
Preparation of Gold Electrode Modified with Zirconia
Nanoparticles. A gold electrode (3-mm diameter) from CH
instruments was polished carefully to a mirrorlike surface with
0.3- and 0.05-µm alumina slurry and sequentially sonicated for 2
min in 6 M nitric acid, acetone, and water. Before the experiment,
the bare gold electrode was cyclic-potential scanned within the
potential range 0.5-1.5 V in freshly prepared 0.2 M H2SO4 until
a voltammogram characteristic of the clean polycrystalline gold
was established. Then it was washed with distilled water and dried
by nitrogen. Zirconia nanoparticles were deposited onto bare gold
electrodes in an aqueous electrolyte of 5.0 mM ZrOCl2 and 0.1 M
KCl by cycling the potential between -1.1 and + 0.7 V (versus
Ag/AgCl) at a scan rate of 20 mV/s for 10 consecutive scans.32
The gold electrodes modified with zirconia nanoparticles (ZrO2/
Au) were rinsed with water and dried with N2 for further
experiments.
Electrochemical Stripping Detection. A ZrO2/Au electrode
was dipped into a stirring 0.1 M KCl solution containing the
desired concentration of OP pesticides for 2 min, washed with
distilled water carefully, and transferred to a 2-mL electrochemical
cell containing 0.1 M KCl solution. Before electrochemical
measurements, the electrolyte solution was purged with nitrogen
for 5 min. SWV measurements were performed from -0.4 to +0.3
V with a step potential of 4 mV, an amplitude of 20 mV, and a
frequency of 25 Hz (unless otherwise stated). Baseline correction
of the resulting voltammogram was performed using the “linear
baseline correction” mode of the CHI 660 (CH Instruments,
Austin, TX) software. Cyclic voltammetric measurements were
performed under batch conditions. The cyclic voltammogram was
recorded between -0.8 and +0.5 V at a scan rate of 100 mV/s.
All measurements were performed at room temperature.
5896 Analytical Chemistry, Vol. 77, No. 18, September 15, 2005
Regeneration of Electrode Surface. After the electrochemi-
cal stripping measurement, multiple successive SWV scanning was
used to remove the bound OPs until the anodic stripping peak
disappeared. The electrode was washed with distilled water for
the next measurement.
Safety Considerations. OP pesticides are highly toxic and
should be handled in a fumehood. Skin and eye contact and
accidental inhalation or ingestion should be avoided.
RESULTS AND DISCUSSION
In the current study, the ZrO2 nanoparticles were electrody-
namically deposited onto a cleaned gold electrode surface in an
aqueous electrolyte of 5.0 mM ZrOCl2 and 0.5 M KCl by cycling
the potential scanning between -1.1 and +0.7 V (versus Ag/AgCl)
for 10 consecutive scans at a scan rate of 20 mV/s (unless
otherwise stated). Figure 3A shows a representative cyclic
voltammogram of the formation processes of ZrO2 nanoparticles
on the cleaned gold electrode surface (curve a, red line). A normal
electropolymerization growth, with increasing current upon repeti-
tive scanning, is observed during the electrodeposition processes.
The steep rise in the cathodic and anodic current at the potential
range of -0.6 to -1.1 V corresponds to the complex redox
behavior of ZrOCl2 on gold.32 Such redox behavior was not
observed in the absence of ZrOCl2 (curve b, blue line). Note that
the increasing cathodic and anodic current is different from the
results reported by Yu et al. The cathodic current decreased with
a thiol self-assembled monolayer modified gold electrode.32 The
observed difference may come from electrode material or a
different electrodeposition mechanism of zirconia. Different cy-
cling potential ranges (between -1.1 V to varying high potential
from 0.7 to 1.2 V) were used to prepare ZrO2 nanoparticles on
the gold electrode surface. Experimental results showed there
was no significant difference observed including the shape, density
of formed ZrO2 nanoparticle, and stripping voltammetric charac-
teristics of bound OPs. A cycling potential range between -1.1 V
and +0.7 V was used to prepare the ZrO2/Au electrode. A SEM
image (Figure 3B) confirms the distinct ZrO2 nanoparticle forma-
tion on the gold electrode surface. The ZrO2 nanoparticles formed
by 10 consecutive potential cycling possess an average size of 50-

Figure 3. (A) Cyclic voltammograms of gold electrode (curve a,
red line) during electrodeposition process in 5.0 mM ZrOCl2 and 0.1
M KCl aqueous solution at a scan rate of 20 mV/s. Potential cycles,
10; curve b (blue line) is the cyclic voltammograms of gold electrode
in 0.1 M KCl aqueous solution in the absence of ZrOCl2 under the
same conditions. (B) Typical SEM image of zirconia nanoparticles
formed with 10 consecutive potential cycling on a gold electrode.
150 nm with a small interparticle distance that is much smaller
than the ZrO2 microcrystalline (7-15 µm), which has a 17-µm
interparticle distance, formed on the dithiol functionalized gold
surface or bare vacuum-deposited 2000-Å gold surface.34 The
formed nanosize ZrO2 particles in our experiments may benefit
from the polycrystalline gold seeds that were formed during the
electrochemical cleaning step in the 0.2 M H2SO4 solution. The
amount of zirconia nanoparticle on the gold electrode surface
increases with the increase in the number of potential cycles. An
average of 300 zirconia nanoparticles/µm2 was observed on the
prepared ZrO2/Au electrode surface (based on counting at six
different locations) after 10 potential cycles.
Zirconia has a strong affinity to the phosphoric group and
provides a facile method to attach OPs to an electrode surface.
Nanosize ZrO2 particles offer a large electrode surface area and
increase the interacting opportunities of OPs. Figure 4 shows the
cyclic voltammograms of methyl parathion/ZrO2/Au electrode (a)
and ZrO2/Au (b) in a 0.1 M potassium chloride solution. A pair
of rather well-defined redox peaks (Epa, 0.093 V and Epc, 0.037)
and an irreversible reduction peak (Epc, -0.61 V) were observed
with a methyl parathion/ZrO2/Au electrode in the potential range
of -0.8 to +0.5 V (Figure 4 a). The irreversible reduction peak
corresponds to the reduction of the nitro group to the hydroxyl-
amine group (reaction 1), and the reversible redox peaks are
attributed to a two-electron-transfer process (reactions 2 and 3),
as shown below:
These profiles are consistent with those described elsewhere
for nitroaromatic OP pesticides and nitrophenyl derivates.25,35-37
A control experiment (Figure 4b) was performed under the same
conditions with the ZrO2/Au electrode in the absence of methyl
parathion; no redox peak appeared at the selected potential range
(the standard reduction potential of ZrO2 is -1.544 V). SWV
analysis has a higher sensitivity than other electrochemical
technologies, such as cyclic voltammetry and differential pulse
voltammetry. The inset of Figure 4 shows corresponding SWV
voltammograms of methyl parathion /ZrO2/Au electrode (a) and
ZrO2/Au electrode (b) in 0.1 M KCl. There is no anodic stripping
peak observed at the ZrO2/Au electrode (inset, curve b). The
methyl parathion/ZrO2/Au electrode exhibits a very sharp and
well-defined stripping peak at the potential range from -0.8 V to
+0.4 V (inset, curve a). The peak potential of the oxidation peak
(0.06 V) shifts 20 mV to a negative potential direction compared
with that in the cyclic voltammogram.
The attracting voltammetric characteristics of adsorbed methyl
parathion on the ZrO2/Au show that ZrO2 nanoparticles have a
strong affinity to the OP compound, which possesses a phosphate
group. To confirm that the affinity occurred between ZrO2 and
methyl parathion instead of the nonspecific adsorption between
the exposed gold surface and methyl parathion, Figure 5 shows
a comparison of the SWV signals of a cleaned bare gold electrode
(a) and a ZrO2/Au electrode (b) after incubating 2 min in 0.1 M
KCl containing 200 ng/mL methyl parathion. A substantially
smaller signal (35 times less compared to the ZrO2/Au electrode)
is observed for a bare electrode. Such a big difference in SWV
signals is attributed to the specific adsorbing between ZrO2
(35) Lin, Y.; Zhang, R. Electroanalysis 1994, 6, 1126-1131.
(36) Roston, D. A.; Kissinger, P. T. Anal. Chem. 1982, 54, 429-434.
(37) Kastening, B., Zuman, P., Meites, L., Kolthoff, I. M, Eds. Progress in
Polarography; Wiley-Interscience: New York, 1972; Vol. 3, p 259.
Analytical Chemistry, Vol. 77, No. 18, September 15, 2005 5897
Figure 4. Cyclic voltammograms of methyl parathion/ZrO2/Au (a) and ZrO2/Au electrode (b) in 0.1 M KCl solution (pH 7.0). Potential scanning
rate, 100 mV/s. Methyl parathion/ZrO2/Au electrode was prepared by dipping the ZrO2/Au electrode in stirring 0.1 M of KCl solution containing
800 ng/mL methyl parathion for 2 min and carefully washing with distilled water before electrochemical measurement. Inset is corresponding
stripping voltammograms. SWV conditions: scanning potential range, -0.8 to +0.4 V; frequency, 25 Hz; increasing potential, 4 mV.

(Figure 1), exhibit similar voltammetric characteristics after they

Figure 5. Stripping voltammograms of bare gold electrode (a) and
ZrO2/Au electrode (b) after 2-min adsorption in stirring 0.1 M of KCl
solution containing 200 ng/mL methyl parathion. Potential scanning
potential. -0.4 to +0.3 V; other conditions, same as Figure 4.
nanoparticles and methyl parathion. Also to be noted is that the
peak potentials for the stripping voltammograms shift to positive
potential direction with the increase of concentration of methyl
parathion on the ZrO2/Au electrode surface. The peak potential
for the stripping voltammogram (Figure 5, 200 ng/mL methyl
parathion) is shifted around 100 mV from that observed in Figure
4 (800 ng/mL methyl parathion). Similar behavior was observed
in the stripping voltammograms of different concentrations of
methyl parathion (see Figure 9).
Another two electroactive OPs, paraoxon and fenitrothion,
which possess a structure similar to that of methyl parathion
5898 Analytical Chemistry, Vol. 77, No. 18, September 15, 2005
are adsorbed on the ZrO2/Au electrode surface. The mixture of
three identical concentrations of OP compounds shows a big
stripping response, which almost equals the sum of the individual
OPs. To confirm that the affinity occurred between ZrO2 and the
phosphate group instead of the nitro group, trinitrotoluene was
used to perform the comparison experiment. A negligible signal
was obtained even though the concentration of TNT (1 µg/mL)
is 5 times more than methyl parathion (not shown).
One of the most important issues in the development of a
chemical biosensor is the regeneration of the sensor surface.
Electrochemical stripping analysis includes built-in preconcentra-
tion and stripping steps. The target analyte is normally ac-
cumulated on the working electrode by applying a constant
potential followed with a stripping step, which can also be regarded
as a cleaning step to remove the target from the electrode surface.
In our experiments, the adsorption process of methyl parathion
corresponds to the stripping step to obtain the electrochemical
stripping signal of the analyte, which automatically removes the
adsorbed OPs. Figure 6 presents a typical successive SWV
voltammogram of a methyl parathion/ ZrO2/Au electrode. It was
found that the stripping peak currents decreased rapidly with the
increase of scanning times, and the anodic stripping peak
disappeared completely after multiple scanning, indicating that
the methyl parathion-ZrO2 complex is dissociated. The electrode
was washed carefully with distilled water and measured again in
fresh 0.1 M KCl solution; no stripping peak was obtained. Also to
be noted is that the SWV scanning times depended on the
concentration of adsorbed OPs. More scanning times are neces-
sary for a higher amount of bound OPs.
Figure 6. Stripping voltammograms (without baseline correction) of the regeneration process of methyl parathion/ZrO2/Au electrode; other
conditions, same as Figure 5.

The attracting stripping voltammetric characteristics of methyl

parathion on the ZrO2/Au provide a facile electrochemical
quantitative method for analyzing OPs. Parameters of the assay
procedure would affect the stripping response of OPs. The amount
of ZrO2 nanoparticles influences the amount of OPs bound to the
surface of the ZrO2/Au electrode. The cycles of cyclic-potential
scanning were used to control the amount of ZrO2 nanoparticle
on the gold electrode surface. Figure 7A shows the effect of the
cycles of cyclic-potential scanning on the adsorption of methyl
parathion. The stripping current of methyl parathion rises with
the cycles at first up to 10 cycles and then decreases. The increase
of the stripping current indicates that the amount of adsorbed
methyl parathion is increasing with the increase of the amount of
zirconia nanoparticle on the electrode surface. The decrease of
the stripping current can be understood by considering the
continual buildup of zirconia nanoparticles, which consequently
cause aggregation of zirconia nanoparticles and generate higher
resistance for the electrochemical stripping processes, leading to
a change of sensing characteristics of the electrode. The change
of electrochemical sensing characteristics of ZrO2/Au was inves-
tigated by cyclic voltammetry of a 5 mM Fe(CN)63-/0.1 M KCl.
Figure 7B presents cyclic voltammograms of a bare Au electrode,
and different amounts of ZrO2 nanoparticles modified the Au
electrodes. We can see the redox peak currents of Fe(CN)63-
decrease with the increase of potential cycle times (from top to
bottom, 0, 2, 4, 6, 8, 10, 15, and 20 cycles). Although more potential
cycles increase the amount of ZrO2 nanoparticles on the electrode
surface, aggregations of nanoparticles increase the electron- Figure 7. (A) Effect of the amount of zirconia nanoparticle on methyl
transfer distance, which leads to the decrease of redox peak parathion adsorption. ZrO2/Au electrodes were prepared by different
current of Fe (CN)63- and decreases the sensitivity of the potential scanning cycles (2, 4, 6, 8, 10, 15, 20). The adsorption
experiments were performed for 2 min in 0.1 M KCl solution containing
electrode. So 10 potential cycles were used to prepare the ZrO2
200 ng/mL methyl parathion. Stripping detection conditions, same as
nanoparticle modified gold electrode. Figure 5. (B) Cyclic voltammograms of corresponding electrodes in
The effect of adsorption time on the stripping peak current 5 mM Fe(CN)63-/0.1 M KCl solution (from top to bottom, 0, 2, 4, 6, 8,
was investigated (Figure 8A). The peak currents increase rapidly 10, 15, 20 cycles), potential scanning rate. 100 mV/s.

Analytical Chemistry, Vol. 77, No. 18, September 15, 2005 5899

Figure 8. Effect of adsorption time (A) and the pH of adsorption
medium (B) on the adsorption of methyl parathion. The concentration
of methyl parathion in adsorption medium was 200 ng/mL. The
adsorption experiments of (A) were performed in a pH 7.0 of 0.1 M
KCl. Electrochemical stripping detection conditions, same as
Figure 5.
with the accumulation time at first and then more slowly from 2
min. The resulting current versus time plot displays a curvature
consistent with adsorption processes. No such surface adsorption
is indicated in analogous measurements at the cleaned bare gold
Figure 9. Stripping voltammograms of increasing methyl parathion concentration, from bottom to top, 5, 10, 20, 40, 60, 80, 100, and 200
ng/mL, respectively. The inset shows the calibration curve. Electrochemical stripping detection conditions, same as Figure 5.
5900 Analytical Chemistry, Vol. 77, No. 18, September 15, 2005
electrode surface (not shown). We also observed the adsorption
of methyl parathion under constant potential conditions (not
shown); there is no significant increase of the stripping peak
current. Two minutes of adsorption time under open-circuit
conditions was thus employed.
An additional parameter that affected the adsorption of methyl
parathion was the pH of the adsorption medium. The pH of the
adsorption solution (0.1 M KCl) was adjusted with 1.0 M NaOH
or HCl solution and varied from 3.0 to 9.0. Figure 8B presents
the pH effect of the adsorption solution on the adsorption of
methyl parathion. We can see that the stripping signal increases
with an increase of pH up to 7.0, and then it decreases at higher
pH. It indicates that ZrO2 has the maximum adsorption to methyl
parathion in a neutral environment. The loss of signal at acidic or
basic environment may be attributed to the effect of H+ or OH-
on adsorption. The mechanism of the pH effect is under investiga-
tion in our laboratory. So a pH 7.0 of 0.1 M KCl solution was used
as the adsorption medium in most experiments.
Analytical Performance. Figure 9 displays the SWV response
of the ZrO2/Au electrode incubated in increasing concentrations
of methyl parathion solution under optimum experimental condi-
tions. Well-defined peaks, proportional to the concentration of the
corresponding methyl parathion, were observed. A linear relation-
ship between the stripping current and the methyl parathion
concentration was obtained covering the concentration range from
5 to 100 ng/mL, the linear regression equation being I (nA) )
1.0696C + 5.4453, with a correlation coefficient of 0.9939. A wide
linear range will be realized by increasing the amount of zirconia
nanoparticle on the gold electrode surface. A detection limit of 3
ng/mL (based on signal-to-noise ratio equal to 3) was obtained
under the optimum experimental conditions. The detection limit
was improved significantly by increasing the accumulation time.
A detection limit of 1 ng/mL was estimated on the basis of a
s/n ) 3 characteristic of the 3 ng/mL data points in connection
with a 600-s incubating time. The detection limit obtained is

Figure 10. Electrochemical stripping signals of methyl parathion/
ZrO2/Au electrodes. Adsorption experiments were performed with pH
7 0.1 M KCl containing 100 ng/mL methyl parathion in the absence
and presence of 100 ng/mL p-nitrophenol, 100 ng/mL nitrobenzene,
100 ng/mL TNT, 0.1 M PO43-, 0.1 M SO42-, and 0.1 M NO3-,
respectively. Electrochemical stripping detection conditions, same as
Figure 5.
comparable with that reported so far with an enzyme-based
biosensor.18 A series of 10 repetitive measurements of a solution
containing 20 ng/mL yielded reproducible peak currents with
relative standard deviations of 5.3.
Interferences arising from the other electroactive nitrophenyl
derivatives and oxygen-containing inorganic ions (PO43-, SO42-,
NO3-) that are expected to coexist in solution were used to
evaluate the selectivity of the ZrO2/Au electrode to nitroaromatic
OPs. Separate adsorbing experiments were performed with 100
ng/mL methyl parathion in 0.1 M KCl solution in the absence
and presence of 100 ng/mL of p-nitrophenol, 100 ng/mL nitroben-
zene, 100 ng/mL TNT, 0.1 M of PO43-, 0.1 M of SO42-, and 0.1 M
of NO3-. Figure 10 shows the electrochemical stripping signals
of methyl parathion at different experimental conditions. One can
see that electroactive nitrophenyl derivatives and oxygen-contain-
ing inorganic ions do not interfere with the adsorption of methyl
parathion, and the stripping peak current varies slightly. Also note
that it was reported in the literature that zirconia has a good affinity
to PO43-,33, 34 but in this case, it does not interfere with the
adsorption of methyl parathion. The reason may be that the
adsorption capability of methyl parathion to zirconia is much
stronger than PO43-. Further experiments are being conducted
in our laboratory. The stripping peak potential of OPs is ∼0 mV,
which also avoids the interferences from other phenol compounds
and electroactive species, whose oxidation potentials are more
than 0.3 V. An electrochemical stripping analysis used in conjunc-
tion with a zirconia nanoparticle modified gold electrode thus
holds great promise for direct analysis of relevant water samples
without any prior separation or pretreatment.
CONCLUSION
We have demonstrated a sensitive electrochemical sensing
protocol for nitroaromatic OPs based on the use of zirconia
nanoparticles as selective sorbents. The strong affinity of zirconia
nanoparticles for the phosphoric group and the promising SWV
characteristics of nitroaromatic OPs provide a facile quantitative
method for a group of electroactive OPs. Other electroactive
nitrophenyl derivatives do not interfere with the adsorption of OPs.
An anodic stripping analysis with a very low stripping peak
potential avoids the interferences from other electroactive species.
The results obtained from this work imply that the combination
of a disposable screen-printed gold electrode with a portable
electrochemical instrument would benefit the field monitoring of
OPs. Current methods are limited to the detection of a group of
nitroaromatic OPs. Nonelectroactive OPs can be monitored by
combining zirconia nanoparticles (selective absorbents) with
enzyme or metal nanoparticle-labeled antibodies against OPs
(recognition elements) and electrochemically measuring the
enzymatic product or dissolved metal ions.
The proposed electrochemical sensing technology is thus
expected to open new opportunities for detecting OP pesticides
and nerve agents in the environment, public places, or workplaces
and for monitoring the exposures of individuals to chemical
warfare agents.
ACKNOWLEDGMENT
The work is supported by a laboratory-directed research and
development program at Pacific Northwest National Laboratory
(PNNL). The research described in this paper was performed at
the Environmental Molecular Sciences Laboratory, a national
scientific user facility sponsored by the U.S. Department of
Energy’s (DOE’s) Office of Biological and Environmental Research
and located at PNNL. PNNL is operated by Battelle for DOE under
Contract DE-AC05-76RL01830.
Received for review May 6, 2005. Accepted July 12, 2005.
AC050791T
Analytical Chemistry, Vol. 77, No. 18, September 15, 2005 5901