Anal. Chem.
2003, 75, 2075-2079
Carbon Nanotube/Teflon Composite
Electrochemical Sensors and Biosensors
Joseph Wang* and Mustafa Musameh
Department of Chemistry, New Mexico State University, Las Cruces, New Mexico 88003
The fabrication and attractive performance of carbon
nanotube (CNT)/Teflon composite electrodes, based on
the dispersion of CNT within a Teflon binder, are de-
scribed. The resulting CNT/Teflon material brings new
capabilities for electrochemical devices by combining the
advantages of CNT and “bulk” composite electrodes. The
electrocatalytic properties of CNT are not impaired by
their association with the Teflon binder. The marked
electrocatalytic activity toward hydrogen peroxide and
NADH permits effective low-potential amperometric bio-
sensing of glucose and ethanol, respectively, in connection
with the incorporation of glucose oxidase and alcohol
dehydrogenase/NAD+ within the three-dimensional CNT/
Teflon matrix. The accelerated electron transfer is coupled
with minimization of surface fouling and surface renew-
ability. These advantages of CNT-based composite devices
are illustrated from comparison to their graphite/Teflon
counterparts. The influence of the CNT loading upon the
amperometric and voltammetric data, as well as the
electrode resistance, is examined. SEM images offer
insights into the nature of the CNT/Teflon surface. The
preparation of CNT/Teflon composites overcomes a major
obstacle for creating CNT-based biosensing devices and
expands the scope of CNT-based electrochemical devices.
Carbon nanotubes (CNT) represent an important group of
nanomaterials with attractive electronic, chemical, and mechanical
properties.1,2 The unique properties of carbon nanotubes make
them extremely attractive for the task of chemical sensors, in
general, and electrochemical detection, in particular.3 Recent
studies demonstrated that CNT can impart strong electrocatalytic
activity and minimization of surface fouling onto electrochemical
devices. An improved electrochemical behavior of catecholamine
neurotransmitters,4 cytochrome c,5 ascorbic acid,6 NADH, 7 and
hydrazine compounds8 has thus been illustrated at carbon nano-
tube-modified electrodes. The ability of carbon nanotubes to
* Corresponding author. E-mail: joewang@NMSU.edu.
(1) Rao, C. N. Satishkumar, B. C.; Govindaraj, A.; Nath, M. ChemPhysChem.
2001, 2, 78.
(2) Baughman, R. H. Zakhidov, A.; de Heer, W. A. Science 2002, 297, 787.
(3) Zhao, Q.; Gan, Z.; Zhuang, Q. Electroanalysis 2002, 14, 1609.
(4) Wang, J.; Li, M.; Shi, Z.; Li, N.; Gu, Z. Electroanalysis 2002, 14, 225.
(5) Wang, J.; Li, M.; Shi, Z.; Li, N.; Gu, Z. Anal. Chem. 2002, 74, 1993.
(6) Wang, Z. H.; Liu, J.; Liang, Q. L.; Wang, Y. M.; Luo, G. Analyst 2002, 127,
653.
(7) Musameh, M.; Wang, J.; Merkoci A.; Lin, Y. Electrochem. Commun. 2002,
4, 743.
(8) Zhao, Y.; Zheng, W. D.; Chen, H.; Luo, Q. M. Talanta 2002, 58, 529.
10.1021/ac030007+ CCC: $25.00 © 2003 American Chemical Society
Published on Web 03/28/2003
promote the electron-transfer reactions of NADH and hydrogen
peroxide suggests great promise for dehydrogenase- and oxidase-
based amperometric biosensors. A major barrier for developing
such practical CNT-based biosensing devices is the insolubility
of CNT in most solvents. Previously reported CNT-modified
electrodes have thus commonly relied on casting a CNT/sulfuric
acid solution onto a glassy carbon surface,3 a procedure that is
not compatible with the immobilization of biocomponents. New
fabrication schemes are highly desired to broaden the application
of CNT-based electrochemical sensors.
This article reports on a new and simple avenue for preparing
effective CNT-based electrochemical sensors and biosensors using
CNT/Teflon composite materials. Carbon composites, based on
the dispersion of graphite powder within an insulator, have
received considerable attention.9,10 Such carbon composites offer
convenient bulk modification for the preparation of reagentless
and renewable biosensors. The use of Teflon as binder for graphite
particles has shown to be extremely useful for various electro-
chemical sensing applications.11,12 Unlike early CNT-modified
electrodes,3-8 the new composite devices rely on the use of CNT
as the sole conductive component rather than utilizing it as the
modifier in connection with another electrode surface. The bulk
of the resulting CNT/Teflon electrodes serves as a “reservoir” of
the enzyme, in a manner similar to their graphite-based counter-
parts. Such CNT/Teflon composites thus combine the major
advantages of CNT with those of bulk composite electrode and
open the door for wide-range sensing applications of CNT. For
example, the CNT/Teflon composites display a marked electro-
catalytic action of CNT toward hydrogen peroxide and NADH and,
hence, an effective biosensing of glucose and ethanol (in connec-
tion with the corresponding oxidase and dehydrogenase en-
zymes). These and other attractive features of CNT/Teflon
biocomposites, along with characterization of the new surfaces,
are illustrated in the following sections.
EXPERIMENTAL SECTION
Apparatus. Amperometric experiments were performed with
a Bioanalytical Systems (BAS) CV-27 voltammograph, in connec-
tion with a BAS X-Y recorder. The working electrode, the Ag/
AgCl reference electrode (model CHI111, CH Instruments, Austin,
(9) Kalcher, K.; Kauffmann, J. M.; Wang, J.; Svacara, I.; Vytras, K.; Neuhold,
C.; Yang, Z. Electroanalysis 1995, 7, 5.
(10) Tallman, D. E.; Peterson, S. Electroanalysis 1990, 2, 499.
(11) Wang, J.; Reviejo, A. J.; Angnes, L. Electroanalysis 1993, 5, 575.
(12) Del Cerro, M. A.; Cayuela, G.; Reviejo, A. J.; Pingarron, J. M.; Wang, J.
Electroanalysis 1997, 9, 1113.
Analytical Chemistry, Vol. 75, No. 9, May 1, 2003 2075
TX), and the platinum wire counter electrode were inserted into
the 20-mL cell (BAS, model VC-2) through holes in its Teflon
cover. A magnetic stirrer provided the convective transport during
the amperometric measurement. The flow injection system
consisted of a carrier solution reservoir, an injection valve with
250 µL of loop, interconnecting PTFE tubing, and a peristaltic
pump (FIAlab, Alitea US, Medina, WA). SEM images were
obtained using the Hitachi S-3200N unit.
Reagents. All solutions were prepared from double-distilled
water. NADH (β-nicotinamide adenine dinucleotide, reduced
form), NAD+ (β-nicotinamide adenine dinucleotide, sodium salt),
potassium dihydrogen phosphate, dipotassium hydrogen phos- Figure 1. Hydrodynamic voltammograms for 1 mM hydrogen
phate, glucose oxidase (GOx, EC 1.1.3.4, Type X-S from Aspergillus peroxide (A) and 1 mM NADH (B) using the graphite/Teflon (a) and
the MWCNT/Teflon (b) electrodes. Supporting electrolyte, phosphate
niger, 157 500 units/g of solid), alcohol dehydrogenease (ADH, buffer (0.05 M, pH 7.4); carbon/Teflon composition ratio, 60:40 wt
EC 1.1.1.1 360 000 units/g of solid from baker’s yeast), and β-D- %; stirring rate, ∼400 rpm.
(+)glucose were purchased from Sigma. Hydrogen peroxide (30
wt %) and potassium ferricyanide were purchased from Aldrich
(Milwaukee, WI) The Teflon granules (type 7A; average particle
size, 34 µm; density, 460 g L-1) were obtained from Dupont Inc.
(Wilmington, DE), while the graphite powder (grade 38) was
received from Fisher Scientific (Fair Lawn, NJ). Multiwall carbon
nanotubes (MWCNT), with ∼95% purity, were obtained from
NanoLab (Brighton, MA); further purification was accomplished
by stirring the CNT in concentrated nitric acid at 25 °C for 24 h.
Single-wall carbon nanaotubes (∼75-80% purity) were obtained
from Mer Inc. (Tuscon, AZ).
Electrode Preparation. CNT/Teflon composite electrodes
Figure 2. Current-time recordings obtained at the graphite/Teflon
were prepared in the dry state by hand-mixing (with a spatula)
(a) and MWCNT/Teflon (b) composite electrodes upon increasing the
the desired amounts of the carbon nanotubes with granular Teflon concentration of hydrogen peroxide (A) and NADH (B) in steps of 2
for 10 min (essential for complete coverage of the granules). The and 0.1 mM, respectively. Operating potential, +0.4 V; other condi-
composite biosensors were prepared by adding the desired tions, as in Figure 1.
amount of the enzyme (GOx or ADH) and of the NAD+ cofactor,
to the 30/70 wt % CNT/Teflon composite. A portion of the
resulting composite was packed firmly into the electrode cavity potentials lower than 0.6 (A) and 0.5 V (B). A small gradual
(2 mm diameter, 2 mm deep) of a glass sleeve. This was increase of the response is observed at higher potentials. In
accomplished by multiple dipping of the glass sleeve onto the contrast, the CNT/Teflon electrode responds favorably to both
composite material (placed on a weigh paper). The electrical analytes over the entire (0.0-1.0 V) potential range. Significant
contact was established via a copper wire. The composite surface oxidation and reduction currents, starting around +0.20 V, are
was smoothed on a weighing paper and rinsed carefully with thus observed for hydrogen peroxide. The anodic signal of NADH
double-distilled water prior to each measurement. increases rapidly between 0.0 and 0.6 V and levels off at higher
Procedure. Measurements were carried out in a phosphate potentials. The substantial lowering of the detection potential
buffer (0.05 M, pH 7.4) supporting electrolyte medium. Ampero- observed for both analytes at the CNT-based composite is coupled
metric detection proceeded under forced-convection batch and to significantly larger current signals. Overall, the data of Figure
flow conditions. The desired working potential was applied, and 1 indicate that the association of CNT with the Teflon binder does
transient currents were allowed to decay to a steady-state value. not impair their strong electrocatalytic properties.
All measurements were performed at room temperature. Such electrocatalytic action facilitates low-potential ampero-
metric measurements of hydrogen peroxide and NADH. Figure
RESULTS AND DISCUSSION 2 compares the amperometric response (at +0.40 V) of the
The attractive behavior of the new CNT/Teflon composite graphite/Teflon (a) and CNT/Teflon (b) electrodes to successive
electrodes has been illustrated in connection with the detection additions of 2 mM hydrogen peroxide (A) and 0.1 mM NADH
of hydrogen peroxide and NADH owing to the involvement of (B). As expected from the voltammetric data, the graphite/Teflon
these compounds in a wide range of biosensing applications. electrode is not responsive to these concentration changes using
Control experiments, using graphite-based Teflon composites, this low-detection potential. The CNT/Teflon electrode, in con-
were used to demonstrate the various advantages of the new trast, responds very rapidly to the changes in the level of hydrogen
CNT/Teflon sensors. Figure 1 compares hydrodynamic voltam- peroxide and NADH, producing steady-state signals within 8-10
mograms (HDV) for 1 mM hydrogen peroxide (A) and 1 mM s. The favorable signals are accompanied by a low noise level.
NADH (B) at the graphite/Teflon (a) and CNT/Teflon (b) The hydrogen peroxide response was not affected by polishing
composite electrodes. No redox activity is observed for either (regenerating) the surface; such renewability was indicated from
analyte at the conventional graphite/Teflon electrode using a series of six successive measurements, each recorded on a
2076 Analytical Chemistry, Vol. 75, No. 9, May 1, 2003

Figure 3. Calibration plots for potassium ferricyanide using CNT/
Teflon electrodes of different compositions. MWCNT/Teflon loading
ratios, 10:90 (a), 30:70 (b), 40:60 (c), 60:40 (d), and 80:20 (e).
Operating potential, +0.1 V; other conditions, as in Figure 1.
Figure 4. Influence of the CNT loading upon the electrode
resistance (a) and amperometric response to 1mM ferricyanide (b).
Operating potential, +0.1 V; other conditions, as in Figure 1.
freshly polished surface, that yielded highly reproducible results
(RSD ) 4%; not shown). Similar reproducibility was observed upon
packing different surfaces from the same batch or for different
batches with the same composition.
The CNT content of the new composites has a profound effect
upon their electrochemical behavior. Figure 3 compares calibra-
tion plots for potassium ferricyanide obtained at electrodes
containing different MWCNT loadings. All electrode compositions
yield highly linear calibration plots over the entire concentration
range. The sensitivity increases with the CNT loading between
10 and 60 wt % and decreases thereafter. The influence of the
CNT content was also examined using cyclic voltammetry (CV)
experiments. Such experiments led to a well-defined peak-shaped
Figure 5. SEM images of the surfaces of the graphite/Teflon (A) and MWCNT/Teflon (B) electrodes. Accelerating voltage, 14 kV; carbon/
Teflon composition ratio, 80:20 wt %.
response for composites containing higher than 20 wt % CNT (scan
rate, 10 mV s-1; 100 mM ferricyanide; not shown). The peak-
shaped CV response indicates overlap of the diffusion layers of
adjacent CNT sites that leads to a linear diffusional flux. The CV
ferricyanide cathodic and anodic peaks increased linearly with the
CNT content between 30 and 70 wt %. Nearly identical peak
separations (∆Ep) of ∼0.15 V were observed upon raising the CNT
content between 30 and 70 wt %; a larger peak separation of 0.31
V was observed in connection with a CNT content of 80 wt %.
Composites containing more than 75 wt % CNT were too dry and
porous and had poor mechanical stability. A poorly defined and
negligible CV response was observed on the other extreme using
a 10 wt % CNT content.
The above trend in the sensitivity can be attributed, in part, to
the effect of the CNT content on the electrode resistance. Figure
4 shows the sensitivity-loading (b) and resistance-loading (a)
profiles. As expected, “Teflon-rich” composites (containing less
than 30 wt % CNT) lead to high resistance (i.e., a nearly insulating
matrix) and low sensitivity. Increasing the CNT content beyond
30 wt % results in a low resistance (5-10 Ω) and higher sensitivity
(except of a lower sensitivity at 80 wt % CNT). The latter reflects
an operation below the mass-limiting plateau associated with the
shift of the voltammetric signal (in accordance with the CV data).
SEM microscopy was employed to gain insights into the nature
of the new carbon/Teflon composites. Figure 5 compares the SEM
images for graphite/Teflon (A) and MWCNT/Teflon (B) electrode
surfaces. It can be seen that the two electrodes have distinctly
different morphologies. The CNT/Teflon surface is characterized
with bundles of MWCNT covering the Teflon granules. The
carbon nanotube fibers have a diameter ranging from 30 to 50
nm. In contrast, distinct Teflon “mountains”, without such fibrous
bundles, are observed at the graphite/Teflon electrode surface.
Different morphologies are expected for different composite
compositions.
Similar to common carbon composite electrodes, the CNT/
Teflon composite can be used as effective detector for flowing
streams. The attractive performance of such a detector was
demonstrated for flow injection measurements of NADH. For
example, Figure 6A displays the flow injection response for NADH
solutions of increasing concentrations (0.2-1.0 mM, b-f). Well-
Analytical Chemistry, Vol. 75, No. 9, May 1, 2003 2077
Figure 6. (A) Flow injection amperometric response of the CNT/
Teflon detector to NADH solutions of increasing concentrations (0.2-
1.0 mM, b-f), along with the response to the blank solution (a) and
the resulting calibration plot (inset). (B, C) Flow injection response to
14 successive injections of a 1.0 mM NADH solution at the MWCNT/
Teflon and bare glassy-carbon electrodes, respectively. Operating
potential, +0.4 V; flow rate, 1.5 mL min-1; carbon/Teflon composition
ratio (A, B), 50:50 wt %. Other conditions, as in Figure 1.
defined peaks, proportional to the NADH concentration, are
observed along with a low noise level. The resulting calibration
plot (shown in the inset) is highly linear (slope, 4.375 µA/mM;
correlation coefficient, 0.999). The CNT/Teflon composite detector
greatly minimizes surface passivation effects (common to am-
perometric detection of NADH). Figure 6 compares the stability
of the response for repetitive flow injection measurements of 1.0
mM NADH at the CNT/Teflon (B) and conventional glassy-carbon
(C) detectors. The glassy-carbon detector displays a gradual
decrease of the response (with a 70% decrease and a RSD of 39%;
n ) 14). In contrast, a stable signal is observed over the entire
operation using the CNT-coated electrode (RSD ) 1.2%). Notice
also the significantly higher sensitivity of the CNT-based detector
(from the 10-fold different current scales), although this comes
with the cost of a slower response time (peak width of 70 vs 45 s
for the GC detector).
The attractive low-potential detection of hydrogen peroxide and
NADH, along with the minimal surface fouling, makes the CNT
extremely attractive for amperometric biosensing in connection
with oxidase or dehydrogenase enzymes. The new CNT-based
composites permit convenient and controlled incorporation of the
desired enzyme (and its cofactor or mediator), in a manner
analogous to other bulk-modified bioelectrodes.11-13 The bulk of
these biosensors thus serves as a source for the biocatalytic
activity. The favorable behavior of the resulting CNT/Teflon-based
2078 Analytical Chemistry, Vol. 75, No. 9, May 1, 2003
Figure 7. Current-time recordings for successive 2 mM additions
of glucose at the graphite/Teflon/GOx (a) and the MWCNT/Teflon/
GOx (b) electrodes measured at +0.6 (A) and +0.1 V (B). Electrode
composition, 30:69:1 wt % carbon/Teflon/GOx. Other conditions, as
in Figure 1.
composites (in comparison to graphite/Teflon composites) was
illustrated in connection with amperometric biosensing of glucose
and ethanol through the incorporation of GOx or ADH/NAD+
within the three-dimensional electrode matrix.
Figure 7 compares the amperometric response to successive
additions of 2 mM glucose at the graphite/Teflon/GOx (a) and
the MWCNT/Teflon/GOx (b) electrodes using operating poten-
tials of +0.6 (A) and +0.1 (B) V, along with the resulting
calibration plots (insets). Both electrodes respond to the glucose
additions at +0.6 V. Yet, the CNT-based bioelectrode offers
substantially larger signals reflecting the electrocatalytic activity
of CNT (A, a vs b). Such electrocatalytic action is even more
pronounced from comparison of the response at +0.1 V, where
the conventional graphite/Teflon biosensor is not responding (B,
a). Such low-potential operation of the CNT-based biosensor
results in a highly linear response (over the entire 2-20 mM
range) and a slower response time (∼1 min vs 25 s at +0.6 V).
As expected from the HDV (of Figure 1A), higher sensitivity is
observed at +0.6 V (note the different scales), along with currents
in the opposite directions. The +0.6-V operation is coupled to good
linearity up to 12 mM, with a slight curvature at higher levels. An
analogous glucose biocomposite based on single-wall rather than
multiwall CNT resulted in a more sensitive and slower response
(not shown). The low-potential detection leads also to high
selectivity, i.e., effective discrimination against coexisting elec-
(13) Pena, N.; Ruiz, G.; Reviejo, A. J.; Pingarron, J. M. Anal. Chem. 2001, 73,
1190.

Figure 8. Current-time recordings for successive 1 mM additions
of ethanol at the graphite/Teflon/ADH/NAD+ (a) and the MWCNT/
Teflon/ADH/NAD+ (b). Operating potential, +0.2 V; electrode com-
position, 28.5:65:1.5:5 wt % carbon/Teflon/ADH/NAD+. Other condi-
tions, as in Figure 1.
troactive species. Despite the absence of external (permselective)
coating, the glucose response at +0.1 V was not affected by the
addition of the common acetaminophen and uric acid interferences
(at 0.2 mM; not shown). A similar addition of ascorbic acid
resulted in a large interference, reflecting the accelerated oxidation
of this compound at the CNT surface.6 Reproducible activity was
observed after two weeks of dry storage at 4 °C (not shown),
which is in good agreement with the high stability of enzymes in
Teflon-based carbon composites.12
A sensitive low-potential detection is observed in Figure 8 for
the biosensing of ethanol in connection with the coimmobilization
of ADH and NAD+ within the CNT/Teflon matrix. The resulting
reagentless biocomposite responds favorably to the 1 mM ethanol
additions using a low detection potential of +0.20 V (b). The
response is relatively fast (∼60 s for steady state) and nonlinear.
No response is observed in analogous measurements at the
graphite-based biocomposite (a). The greatly enhanced biosensing
of ethanol at the CNT-based device reflects the accelerated
oxidation of NADH, in accordance with the HDV data of Figure
1B. Such low-potential detection of ethanol commonly requires a
redox mediator (to shuttle the electrons from the NADH product
to the surface).
CONCLUSIONS
The experiments described above indicate that CNT can be
used to prepare attractive composite electrodes for amperometric
biosensing. The preparation of such CNT/Teflon composites
overcomes a major obstacle for creating CNT-based biosensing
devices. The resulting composite CNT/Teflon material brings new
capabilities for electrochemical devices by combining the advan-
tages of CNT and bulk composite electrodes. The electrocatalytic
properties of CNT toward hydrogen peroxide and NADH are not
impaired by their association with the Teflon binder. Various
chemical and biological moieties could be readily incorporated
into the bulk of the CNT/Teflon composite. While most of our
data have been obtained with multiwall CNT, our preliminary
results indicate similar improvements using single-wall CNT. By
providing a useful avenue for preparing renewable CNT-based
biosensors and CNT-modified electrodes, such fabrication scheme
expands the scope of CNT-based electrochemical devices and
holds great promise for routine biosensing applications.
ACKNOWLEDGMENT
This work was supported by grants from the U.S. EPA (Grant
RD830900) National Science Foundation (Grant CHE 0209707).
M.M. acknowledges a fellowship from the Islamic Development
Bank (IDB) (Jeddah, SA).
Received for review January 2, 2003. Accepted February
20, 2003.
AC030007+
Analytical Chemistry, Vol. 75, No. 9, May 1, 2003 2079