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366 ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28

New Electrochemical Sensors

The following are summaries of eleven of the papers presented at the Joint Analytical/
Faraday Symposium at the Annual Chemical Congress of the RSC held on April 8th-I2th,
1991, in Imperial College, London.

Solid-state Gas Sensors: Prospects for Selectivity

Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366

David E. Williams
Department of Chemistry, University College London, 20 Gordon Street, London WCIH OAJ
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Metal oxides, such as tin dioxide and zinc oxide, fabricated

either in the form of thin (=lo0 nm) films or as thicker porous Table 1 Range of materials investigated
bodies, show an electrical conductivity at temperatures of Perovskite-type Pyrochlore-type
about 300°C which is very sensitive to the presence of trace compounds compounds
amounts (ppm level) of reactive gases (hydrocarbons, hydro- of tin of tin Niobates
gen, carbon monoxide, methane, ammonia, oxides of sulphur Bronzes
and nitrogen, chlorine, hydrogen sulphide) in air. This Al - ,B,Snl - .CXO3- A2 - ,B,Sn2 - xCx07- Al - xBxNb206
phenomenon has been exploited for many years in warning A Ca, Sr, Ba Specific examples: Examples:
devices. Most commercial elements utilize porous, thick (==lo0 CaSnTi207 A, B-Pb, Ba, Sr
pm) films of tin dioxide. Typically, the layer might be deposited B Ca, Sr, Ba, CaCeSnzO7
onto the outside of a small alumina tube by successive dipping Pb Gd2Sn207 A6BxNblO- x o 3 0
into a tin salt solution and thermal decomposition. A heater is La, Y, Gd La2Sn207 Example:
y2sn207 Bi6Fe4Nb6030
threaded into the centre of the tube. These devices are simple,
robust and inexpensive. They are, however, if anything, too C Fe, Co Bi2Sn207
Ti, Zr, Ce
sensitive to too many things, although a certain degree of
selectivity can be obtained by control of the operating Transition metal
temperature. The question is, therefore, whether, by choice of Tantalates compounds
Specific examples: Fe, Co, Ni, Cu
oxide material or some other means, selectivity can be BaSn03 FeTa04 niobates
enhanced. 1,2 M ~ r r i s o n , in
~ discussing this question, has BaSn0.5Ti0.503 CoTa206
pointed out that although the gas sensing phenomenon appears BaSn0,9Zr0,103 NiTa206
to be intimately connected with the occurrence of a surface- BaSnO.8FeO.2 0 3 CUTa206
catalysed combustion, the notion of selectivity in a gas sensor is Ba0.9Gdo.1Sn03
different to that of the apparently related notion of selectivity
in a catalyst: for a catalyst, selectivity means a bias in favour of
a particular reaction product, whereas for a sensor it means a hydrogen s ~ l p h i d ebut
, ~ this was the exception rather than the
bias in favour of a particular reactant, often in a complex rule. A second conclusion was that there was a clear rz-typelp-
mixture of potential reactants. type classification of the oxides, even in the absence of a
As far as the second part of the question is concerned, it has response to varying oxygen partial pressure, and correspond-
been pointed out' that the surface-catalysed combustion causes ingly a classification of the gases into oxidizing and reducing
a gradient in the composition of reactant and product gases agents. Exceptions to this classification stood out clearly: Table
throughout a porous sensor structure, and that this can be used 2 shows a part of it, in which the response has been coded
to build in a degree of selectivity based on the differing simply in terms of its sign. The interpretation is that, in general,
reactivity of different gases. Hence, in a sufficiently thick the response of oxides to gases, being a change in the
structure, the outer layers of the device act as a kind of filter, concentration of the charge carriers giving rise to the conduc-
burning away the more reactive gases and leaving only the less tivity, is controlled by the surface concentration of a single,
reactive gases to affect the conductivity in the vicinity of reactive surface species, common to all oxides and presumed
electrodes buried within the structure. There seems to be (following studies on tin dioxide and zinc to be an
plenty of scope for using this kind of idea, the alteration of oxygen ion, 0 2 - 0-., Response mechanisms have been fully
relative response by alteration of the geometry of the sensor discussed elsewhere ,13235and, indeed, a model simply rational-
layer and of the measuring electrodes, to develop tailored izing the exceptions in the classification can be constructed.6
devices. The answer, therefore, to the first part of the question posed
In order to address the first part of the question, an above is that very strong selectivity seems rather rare, being
experimental programme was carried out2 to investigate the confined to a few materials and a few gases: if a generalization
response of several hundred different oxides to a range of is required, in the study these were those gases (NH3, H2S) that
gases. Table 1indicates the range of materials investigated; the could be considered to act as Lewis bases on materials in which
gases studied were: hydrogen, carbon monoxide, methane, it is presumed that oxygen surface species were unreactive at
propane, ethene, ammonia, hydrogen sulphide, nitrogen the measurement temperature. On the other hand, there was
dioxide, sulphur dioxide and chlorine, and changes in oxygen sufficient variation in the relative magnitude of the response of
partial pressure. The first conclusion of this work was strikingly different materials to different gases to allow ample scope for
simple: most oxides respond to most gases. A few materials the strategy of using arrays of differently responding sensors to
showed selectivity to some gases, most often ammonia and resolve the composition of mixtures.
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ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28 367

Table 2 Classification of materials and responses: the response is coded simply by the sign of the resistance change on exposure to the gas.
Exceptions to the general n-typelp-type classification of materials, showing opposite sign of response to oxidizing and reducing gases stand out
clearly. A plus sign indicates a resistance increase with respect to the value in air; a minus sign indicates a resistance decrease. In all atmospheres,
except for the first listed, the gases are introduced as mixtures (1'70 except where stated) in air
Low 0 2 C2H4 H2S
Material (100PPm) cH4 co H2 (0.7%) NH3 so2 (0.1%) Cl2 NO2
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
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2. Effectively p-type oxides-

Sr(SrNb)0 3 + + +
LiFeSn04 + +
Y2Ti20 + + + + + +
BaTi03 + + -
SrTiO3 + -
LaO.9CaO.lFe03 + - + -
La6W012 + +
NdzZ1-207 + + +
Y2BaZn05 + + +
References 3 Morrison, S. R., Sens. Actuators, 1987, 12, 425.
1 Williams, D. E., in Solid State Gas Sensors, eds. Moseley, P. T., 4 Moseley, P. T., and Williams, D. E., Sens. Actuators, 1990, B1,
and Tofield, B. C., Adam Hilger, Bristol and Philadelphia, 1987, 113.
pp. 71-123. 5 Heiland, G., and Kohl, D., in Chemical Sensor Technology,
2 Moseley, P. T., Stoneham, A. M., and Williams, D. E., Volume I , ed. Seiyama, T., Kodansha and Elsevier, Tokyo and
Techniques and Mechanisms in Gas Sensing, eds. Moseley, P. T., Amsterdam, 1988, pp. 15-38.
Williams, D. E., and Norris, J. 0.W., Adam Hilger, Bristol and 6 Williams, D. E., and Moseley, P. T., J. Muter. Chem., in the
Philadelphia, 1991, ch. 4. press.

Semiconductor Gas Sensors and Selectivity

Gary S. V. Coles
Department of Electrical and Electronic Engineering, University of Wales, Swansea SA2 8PP

The resistance of certain semiconductor materials has been

known to change in the presence of reducing gases since the
early 1950s. Gas sensors exploiting this observation, initially
based on zinc oxide but now usually based on the n-type 14
semiconductor tin dioxide, have been available commercially
for some two decade^.^.^ Early sensors received a great deal of
v)
criticism as they tended to be irreproducible and have poor m
selectivity, responding to a wide range of reducing gases. It $ 10
might also be true that industry and commerce expected too B
much of this type of sensor, despite the fact that they can
possess many of the characteristics desirable in a gas sensing
device, including high sensitivity, fast response speed, low 6
power consumption and cost. However, it is the problem of
selectivity that is addressed in this paper.
2
Selectivity in Tin Dioxide Gas Sensors 0 2 4 6
[Gas] (lo3 ppm)
Early workers in this area discovered that operating tin dioxide
sensors at different temperatures conferred a degree of
For example, operating a sensor at =300 "Cgave a
~electivity.~ Fig. 1 Variation of conductivity with gas concentration for a
device sensitive to the presence of carbon monoxide but hydrogen-selective sensor in hydrogen and carbon monoxide

368
25
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
0 5 10 0 5
[Gas] ( l o 2 ppm)
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Fig. 2 Resistance versus contaminant gas concentration plots for a sensor fabricated from Sn02 pre-sintered at 1500 "C in air and maintained at
three different operating temperatures: ( a ) 400; ( b ) 280; and ( c ) 175"C
insensitive to the presence of methane. Producing a sensor with
the opposite selectivity required the addition of =l% platinum
to the oxide and operation at a higher temperature of -600 "C.
Initial work at Swansea produced two selective sensors.6 The
first, prepared from tin dioxide, aluminium silicate and
palladium chloride, gave a device sensitive to the presence of
the lower hydrocarbons but insensitive to the presence of
carbon monoxide. A second sensor, formed by the sintering of
tin dioxide and bismuth oxide, gave a sensor with the opposite
selectivity. The notable feature of these sensors is that the
selectivity is observed throughout the viable operating temper-
ature range (between =150 "C, below which resistances
become excessively high and surface reactions too slow, and
-700 "C, above which conductivity by thermal excitation
obscures the changes produced by the presence of a contami-
nant gas). A maximum sensitivity is observed at an operating
temperature of =250"C. In an attempt to understand the
properties exhibited by these sensors the carbon monoxide
selective device was studied in more detail.
Tin Dioxide-Bismuth Oxide System
Tin dioxide itself exhibits conductance changes in the presence
of many reducing gases. However, as bismuth oxide is added
the response to methane rapidly diminishes, falling to zero at
-15% m/m. The response to carbon monoxide remains
unaltered. Above a concentration of =17% m/m Biz03 the
response to CO also begins to fall and this reaches zero above
-23% m/m. The optimum composition of this sensor is
therefore 8345% m/m S n 0 2: 1517% m/m Bi203. Although
the term selective has been used, in reality these sensors, and
many other types of sensor, will also respond to certain other
gases, notably hydrogen. When the Bi203 content of these
devices is increased above that at which the carbon monoxide
response has fallen to zero, the response to hydrogen is
observed to persist undiminished until this also begins to fall to
zero above -25% m/m. It is therefore possible to produce a
third sensor which is sensitive to the presence of hydrogen but
shows no resistance changes in the presence of carbon
monoxide or the lower hydrocarbon^.^ A remarkable feature
of this sensor is its linear change in conductance with increasing
gas concentration even up to several thousand ppm, as shown
in Fig. 1. Sensors of this type usually tend towards saturation at
such high levels.
In the production of these sensors the oxide mixture is fired
at 800°C. It is known that bismuth oxide and tin dioxide
undergo a solid-state reaction above -600 "C producing
bismuth stannate, Bi2Sn207, which has a pyrochlore struc-
ture.' In our sensors all of the bismuth oxide reacts and the
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ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
10 0 5 10
final sensor element is composed of bismuth stannate and tin
dioxide only. There are several other stannates of general
formula M2Sn207 which also possess the pyrochlore structure .9
These were also studied in the hope that they would yield new
selective sensors and further elucidate the properties exhibited
by these devices. An immediate difference is the high
temperatures required to produce these other stannates , being
typically in the region of 1500 "C. Sensors produced from these
materials in an analogous way to the tin dioxide-bismuth oxide
sensors, but fired at higher temperatures, showed various
degrees of selectivity and sensitivity but none compared
favourably with the original devices. As a control, sensors were
also produced from pure tin dioxide fired at 1500°C. These
devices showed remarkable characteristics as shown in Fig. 2.
At high operating temperatures ( ~ 4 0 "C) 0 the sensor shows
conventional decreases in resistance in the presence of all three
gases as would be expected for this n-type material. At low
operating temperatures (-175 "C) the device shows no re-
sponse to carbon monoxide or methane but exhibits an increase
in resistance in the presence of hydrogen as would be expected
in a p-type material. At intermediate temperatures (-280 "C)
no response is observed for methane; a conventional response
is observed in the presence of hydrogen but a resistance
increase is observed in the presence of carbon monoxide.
Conclusions
It is possible to produce tin dioxide based sensors with marked
selectivity towards the common toxic and/or flammable gases
methane, carbon monoxide and hydrogen. These sensors are
selective throughout the viable operating temperature range. It
is also possible to produce a single sensor in which the selectivity
and mode of operation can be switched and tuned by modulation
of the operating temperature. The exact reasons for these
observations are as yet not fully understood but are currently
being studied and will form the basis of a subsequent paper.
References
1 Brattain, W. H., and Bardeen, J., Bell Syst. Tech. J . , 1953,32,1.
2 Heiland, G., 2. Phys., 1954, 138, 459.
3 Seiyama, T., Kato, A., Fukiishi, K., and Nagatini, M . , Anal.
Chem., 1962, 34, 1502.
4 Taguchi, N., Br. Pat., 1280809, 1288009, 1282993, 1970.
5 Firth, J. G., Jones, A . , and Jones, T. A., Environmental Sensors
and Applications, IERE Conference Proceedings, 1974, p. 57.
6 Coles, G. S. V., Watson, J., and Gallagher, K. J., Sens.
Actuators, 1985, 7 , 89.
7 Coles, G. S. V., Williams, G . , and Smith, B., Sens. Actuators,
1991, B3, 7.
8 Roth, J., J. Res. Natl. Bur. Stand., 1956, 56, 17.
9 Brisse, F., and Knop, O., Can. J. Chem., 1968, 46, 859.

ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
Novel Solid-state Potentiometric Sensors
Michel Kleitz
Laboratoire d’lonique et d’Electrochimie des Solides de Grenoble, LIESG/ENSEEG, Domaine Universitaire,
BP 75, 38402 Saint Martin-d’Heres, France
Solid-state potentiometric sensors can be described as being
composed of sensitive surfaces and an electrolytic reference
transmission line.’ These functions can be separated to a large
extent which has not yet been fully explored.
A good transmission line is composed of elements through
which the electrolytic reference is constant and possibly
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
interphases where the electrolytic reference steps are well
defined by local chemical buffers. The differential zirconia
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sensor,2 the zirconia-tip sensor3 and the sulphate-zirconia
contact4 illustrate the different behaviour of such interphases.
Some of these interphases, such as the sulphate-zirconia
interface can be used as sensing elements. This is also true of
the conventional ion-sensitive electrodes and of the Nasicon
membrane5 investigated for Na+ sensing.
In order to be of interest, a sensitive surface must be ionically
anchored to the electrolytic reference and must exhibit a redox
reaction which equilibrates rapidly with the species to be
analysed.6 Generally, this latter condition implies that the
electron conduction associated with the redox system is not
very high. Therefore, an additional potential-collecting com-
pound must be suitably incorporated. Electrode reactions, in
the conventional electrochemical sense, are the natural choice.
As in aqueous electrolytes, an efficient ionic anchoring does
not require that the counter ion of the species to be analysed be
the predominant ionic ~ a r r i e rCatalysts
.~ known to be active
for the species to be analysed can also be good candidates for
the redox function. It has been demonstrated with 02*- and
Design and Synthesis of Receptors for Sensor Use
Joyce C. Lockhart
Chemistry Department, The University of Newcastle upon Tyne, Newcastle upon Tyne NEI 7RU
The availability of crystal structure information on biological
receptor sites’ for metal ions has provided a blueprint for the
design of synthetic receptor molecules, causing the deliberate
synthesis of thousands of new molecules, many of which have
provided analytical reagents for sensors. There is much less
information available on the receptor sites for anions. Natural
strategies for anions, as at present understood, involve two
main types, one using sites of opposite charge (provided
principally by the proximity of protonated Arg and Lys
residues) with hydrogen bonding as a directional force, and
hydrophobic interactions, the other apparently constructed
without the obvious Coulombic interactions. In each instance
the receptor site is a close fit to the contours of the anionic
substrate, whether achieved by perfect size match or by
induced fit. The first type is found in proteins which bind to
DNA,’ and the second in the sulphate receptor protein2 of
Salmonella typhimurium, for example.
The synthetic approach to coordinating anions essentially
turns the inorganic chemist’s ideas of coordination inside out.
A straight comparison of the coordination of 18-crown-6 to
potassium ions with the coordination of hexaprotonated l8-aza
crown-6 to chloride ions indicates that the ion-dipole interac-
tion of the crown molecule has no direct analogue in the second
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369
02-that a catalysed partial redox reaction can give better
sensor performance than a conventional complete reaction. At
low temperatures, the transition to the peroxide reduction can
result in shorter response times.8
References
Kleitz, M., Siebert, E., Fabry, P., and Fouletier, J., in Chemical
and Biochemical Sensors, eds. Gopel, W., Hesse, J., and Zemel,
J. N., VCH Weinheim, 1991, in the press.
Fouletier, J., Seinezia, H., and Kleitz, M., J . Appl. Electro-
chem., 1975, 5 , 177.
Fouletier, J . , Fabry, P., and Kleitz, M., J . Electrochem. SOC..
1976, 123, 204.
Kleitz, M., Pelloux, A . , and Gauthier, M.. in Fast Ion Transport
in Solids, eds. Vashishta, et al., Elsevier, Amsterdam, 1979, pp.
69-73.
Fabry, P., Gros, J. P., Million-Brodaz, J. F., and Kleitz, M.,
Sens. Actuators, 1988, 15, 33.
Kleitz, M., and Siebert, E., in Chemical Sensor Technology, ed.
Seiyama, T., Kodansha Elsevier, Tokyo and Amsterdam, 1989,
V O ~ .2, pp. 151-171.
Pelloux, A., Quessada, J. P., Fouletier, J . , Fabry, P.. and Kleitz,
M., Solid State lonics, 1980, 1, 343.
Siebert, E., Fouletier, J., and Kleitz, M., 1. Electrochem. Soc.,
1987, 134, 1573.
situation, as the directional force, in that instance hydrogen
bonding, is unlikely to involve as many as six interactions to
chloride simultaneously; if one N-H on each nitrogen points
into the cavity of the ring, the other must point outwards. In
fact the chloride binds exo3 to the ring, while the potassium
goes into the ether cavity. Macrocycles which are oligomers of
ethyleneimine also tend to have the nitrogens anti on each
ethane segment and to have a large open cavity. It is also
noteworthy that polyamines in which the linkers have three or
more methylene links between nitrogens (such as the biological
amines spermidine and putrescine) are much more basic than
the ethylenediamine oligomers ubiquitous in inorganic
chemistry, and thence more suitable for coordination to
anions. In some crystal structures, anions are found inside the
cavity of polyaza macro cycle^.^
Basic Design
We used another approach with the intention of building a
receptor site which would have an induced fit to anions of
choice. Molecules containing two crown ether moieties,
separated by a linking group, will sandwich potassium cations,

370
X = polyaza
Fig. 1 Structural formula of bis(crowns)
and it has been postulated that the linker between the two
. ~occurred to us that if the
crown rings will form a p ~ c k e t It
linker was formed from a polyamino residue, as for example
formula (I) (Fig. l),then in hydroxylic solvents at neutral pH,
it should be capable of complexing the counter ion of its
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
potassium ‘filling’.
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Discussion
Adenosine triphosphate (ATP) is a favourite anion for
complexation studies. Suitable receptors for the phosphate
residues have been detected previously by examining the time-
averaged 31P NMR signals for the a,P,y phosphorus atoms,
which are shifted relative to those of the free ATP in solutions
containing receptor in a suitable mole ratio to the ATP.6,7
Molecular graphics studies indicated that diethylenetriamine
in the extended, protonated conformation was an excellent
sideways fit to the phosphates of a triphosphate chain, while a
slightly longer chain, tetraethylenepentamine, likewise
extended and protonated, could wrap simultaneously round
the y and residues; small cyclic polyamines (four nitrogens)
were more likely to interact with the terminal y phosphate
only. The 31P shifts observed for 1 : 1 ratios of polyamine to
ATP were shifted for these situations in accordance with this
prediction. The size of cavity required to complex various
anions from chloride to ATP was also estimated by using
molecular graphics for the bis(crown) designs.
The bis(crowns) (I) (Fig. 1) were shown by ‘H NMR to
sandwich potassium ions.5 The Schori-Grodzinski NMR
method8 of determining rates of exchange of quadrupolar
nuclei between solvated and complexed environments was
applied with limited success, to show that the coreceptor is
clearly binding potassium ions plus counter ions
simultaneously.
Control of Ion Transport Through Lipid Membranes
Ulrich J. Krull, Dimitrios P. Nikolelis, John D. Brennan, R. Stephen Brown, Michael Thompson,
Vida Ghaemmaghami and Krishna M . Kallury
Chemical Sensors Group, Department of Chemistry, Erindale Campus, University of Toronto, 3359
Mississauga Road North, Mississauga, Ontario L5L IC6, Canada
The perturbation of the structure of artificial lipid membranes
can be monitored by electrochemical methods, and offers
opportunities for development of chemically selective biosen-
sors. An important advantage of such an electrochemical
sensing system is the increased sensitivity which is derived from
an intrinsic amplification process. A single selective binding
event between a receptor and a target molecule can result in an
increase of the transmembrane conduction that involves
thousands of ions.’
A large number of biochemical reactions based on enzyme-
substrate , antibody-antigen, lectin-saccharide , hormone-
receptor and avidin-biotin interactions have been monitored
by observation of the transmembrane ion current. A further
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ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
A synthetic route, and the separation and characterization of
the bis(crowns) (I) , the first multi-receptors to exhibit multiple
binding to cation and counter ion simultaneously, will be
described elsewhere. Further evaluation of the modified
potential of the assembly for recognition and binding of anionic
substrates in the presence of potassium ions is proceeding,
prior to development of the new materials as anion sensors.
Thanks are due to the SERC for financial support, and to Dr.
N. P. Tompkinson, Dr. K. I. Kinnear, Dr. E. Arafa and D. P.
Mousley for their help in this project.
References
Brookhaven Protein Data Bank, Bernstein, F. C., Koetzle,
T. F., Williams, G. J. B., Meyer, E. F., Jr., Brice, M. D.,
Rodgers, J. R., Kennard, O., Shimanouchi, T., and Tasumi, M.,
J. MoI. Biol., 1977, 112, 535.
Pflugrath, J. W., and Quiocho, F. A., Nature (London), 1985,
314, 257.
Cullinane, J., Gelb, R. I., Margulis, T. N., and Zompa, L. Z.,
J. Am. Chem. SOC., 1982, 104, 3048.
Lehn, J.-M., Meric, R., Vigneron, J.-P., Bkouche-Waksman, I.,
and Pascard, C., J. Chem. SOC., Chem. Commun., 1991, 62.
Handyside, T. M., Lockhart, J. C., McDonnell, M. B., and
Subba Rao, P. V., J. Chem. SOC.,Dalton Trans., 1982, 2331.
Hosseini, M. W., Lehn, J.-M., and Mertes, M. P., Helv. Chim.
Acta, 1983, 66,2454.
Hosseini, M. W, Lehn, J.-M., and Mertes, M. P., Helv. Chim.
Acta, 1985, 68, 818.
Shchori, E., Jagur-Grodzinski, J., Luz, Z., and Shporer, M.,
J. Am. Chem. SOC., 1971, 93, 7133.
extension of artificial lipid membranes for electrochemical
sensing was the construction of ‘biomimetic ion-channel
sensors’.* These devices were based on Langmuir-Blodgett
deposition of multilayers of acidic lipid membranes on glassy
carbon electrodes, More than three layers of lipid were
necessary to block the permeation of Fe(CN)64-, which was
used as a marker ion. In the presence of the stimulant Ca2+, a
change in the alignment of the lipid occurred on the glassy
carbon surface owing to electrostatic complexation of calcium
ions with acidic phosphate head groups of the lipids, resulting
in an increase of the marker ion penetration to the electrode.
When ethylenediaminetetraacetic acid was added to the
solution in slight excess of that required to complex the calcium

ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
ions, the electrode signal for Ca2+ was eliminated. This
indicates that a conductive zone can be opened reversibly by
interaction of an analyte at a membrane in the absence of ion-
channel proteins; however, the nature of such a conductive
zone is poorly understood.
Determination of the mechanism of ion permeation and
establishment of control of the energetics associated with
conductive pathways would permit optimization of the struc-
ture of lipid membranes for biosensor development. Further
development must include mechanical stabilization of a lipid
membrane onto a solid electrode , where covalent linkages
between the support and the lipidreceptor molecules are
desirable. In order to avoid the effects caused by accumulation
of ions on one side of such a stabilized membrane, the
measurements must be made by driving the system with an a.c.
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
voltage and by monitoring admittance with a phase-sensitive
amplifier. Progress towards a surface-stabilized a.c. admittance
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modulation lipid membrane-based biosensor that operates on
the basis of control of ion permeation by artificial ion channels
is described.
Control of Ion Permeability Through Planar Bilayer
Lipid Membranes
Ion conductivities through solvent-free planar bilayer lipid
membranes (BLMs) formed from mixtures of egg phosphatidyl
choline and dipalmitoylphosphatidic acid were evaluated to
determine the effect of surface charge and phase domain
formation on the process of ion translocation. Ion conductivity
in a small d.c. voltage field was controlled by the surface
distribution of potassium ions (0.1 mol dmP3 KC1 electrolyte)
at the membrane-solution interface as predicted from electri-
cal double layer theory. It was found that the conductivity of
the membranes could be approximated as a linear function of
the per cent. mass composition of the charged lipid. The
conductivity was observed to alter drastically at a lipid
composition containing a minimum of 25% phosphatidic acid
as this component within the membrane was increased. This
was attributed to the presence of a phase transition induced by
the phosphatidic acid. At compositions of acid less than 25%,
ion conduction occurred through zones that were enriched in
the charged lipid. At concentrations of the acid above 25%, the
average surface charge was the dominant factor which deter-
mined the magnitude of ion conductivity. The adjustment of
pH to control the degree of ionization of the phosphatidic acid
had a similar effect to the variation of the amount of the acidic
phospholipid within the membrane for experiments carried out
at fixed pH. The ion conductivity could be used to measure
accurately the pK, value associated with the acidic lipid within
a planar BLM, and suggests a general method for evaluation of
pK, for charged species in lipid membranes.
Based on results of studies of cystic fibrosis which implicated
hydroxystearic acid (HSA) as a contributing factor in altered
biomembrane function, solvent-free planar BLMs and mono-
layer films were prepared from a lipid mixture (by mass)
containing 34% phosphatidylcholine, 19% dipalmitoylphos-
phatidylserine , 47% cholesterol and variable amounts of 10-
and 12-hydroxystearic acid (0-50%). The structures of mono-
layer films at the air-water interface of a Langmuir-Blodgett
trough were studied by pressure-area correlations and by
further correlations with microscopic phase separation as
revealed by fluorescence microscopy. In order to elucidate the
role of the hydroxyl moieties in ion permeability the transmem-
brane ion current was corrected for the effect of the negative
surface charge of the carboxylic acid, and determined by
replacement of the HSA component with stearic acid. The ion
currents were found to increase approximately exponentially
with the mole ratio of HSA. Two models of ion conduction
through BLMs were considered: ‘hopping’ via hydrophilic sites
within the hydrophobic zone of BLMs introduced by the
hydroxyl moiety of 10- or 12-HSA; and transport through
interfacial regions between phase domains which represent
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371
areas of low steric density and structural order within
monolayers. While the two mechanisms are not distinct, the
ion permeability and monolayer compression results did
indicate that HSA had a large effect at low concentrations. The
results suggested that the physical location of ion permeation
through BLMs containing low concentrations of HSA was
predominantly at interfacial zones at the edges between
domains, and that artificial ‘ion channels’ had been generated
by virtue of the edge activity of HSA.
Electrochemistry of Surface-immobilizedMembranes
A portable admittance modulation measurement device was
constructed, and was designed to measure both the in-phase
and out-of-phase signal components for determination of
‘effective’ ion current and membrane capacitance. The sensiti-
vity and detection limit for the a.c. system were tested by
studying the interaction of the ionophore valinomycin with
planar BLMs.’ The electrochemical effects were observed as
conductance changes of the membrane, with the limit of
detection for valinomycin being at the 1 nmol dm-3 concen-
tration level.
Metal electrodes were used as supports for covalent attach-
ment of amphiphiles. Platinum electrodes with a surface area
of 1 cm2 were oxidized to provide a high density of surface
hydroxyl sites.3 Immobilization of amphiphiles was carried out
by reaction of the hydroxyl sites with ~ i l a n eGold
. ~ electrodes
were prepared by vacuum deposition of a 250 nm layer of the
metal onto a 30 nm layer of chromium that covered borosilicate
glass slides. Surface attachment of amphiphiles to this metal
was achieved by sulphur-gold interaction^.^
A wide variety of amphiphiles differing in hydrocarbon chain
length, number of chains (one or two), chain polarity, head
group charge and head group size ( e . g . , acidic phosphate,
carboxylic acid and ester, phosphatidylcholine) were attached
to both the platinum and gold surfaces. Two-step attachment
procedures, as exemplified by the initial deposition of amino-
propyltriethoxysilane (APTES) onto platinum followed by
linkage of a 10-carbon phosphatidylcholine through an amide
bond to the amino group of APTES, often provided greater
surface coverage than if an equivalent 16-carbon phosphatidyl-
choline had been deposited directly by the reaction of hydroxyl
moieties with silane at the metal surface. Surface coverage for
most experiments was in the range 40-85% as determined by
X-ray photoelectron spectroscopy, and was dependent on the
deposition procedure, the chain length and the number of
chains. The electrochemical results indicated that the best
blockage of ion conductivity occurred when the amphiphiles
contained long hydrocarbon chains. Species such as trichloro-
octadecylsilane and octadecylthiol provided the best blockage,
reducing the in-phase signal component by 95% for both types
of metal electrode. Analogues of natural lipids such as
dimyristoylphosphatidylcholine reduced the in-phase compo-
nent by values of about 50%. Subsequent incubation of lipid-
coated electrodes in solutions containing membrane-soluble
species such as cholesterol greatly reduced the magnitude of
the in-phase component of the signal to values approaching
those found for BLMs. These latter electrodes were tested for
response to the presence of valinomycin, and achieved
detection limits in the range 10-100 nmol dm-3.
A further series of analytical experiments were carried out to
develop electrodes that were sensitive to pH variations to
extend the concept of control of phase domain structure to
immobilized membranes. Linear 10-carbon silane and thiol
carboxylic acids were immobilized onto platinum and gold
surfaces, and both systems showed good sensitivity to pH (e.g. ,
a pH change from 7 to 8 caused a conductivity change of 25% ).
Modification of this system to evaluate a biosensing strategy
was then carried out. Enzymically active urease has been
adsorbed onto these acidic surfaces, and has also been
covalently immobilized through the acidic functional group.

372
The intent was to use the urease-urea reaction to generate a
local transient pH gradient which would perturb the membrane
structure. Tentative electrochemical results, and definitive
spectroscopic results, indicate that this has been accomplished
and that sub-micromolar detection levels can be achieved for
the substrate with good reversibility and longevity.
We are grateful to the Canadian Defense Research Establish-
ment-Suffield and the Natural Sciences and Engineering
Research Council of Canada for financial support of this work.
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
Gas Sensors Using the Work Function of Organic Semiconductors
Downloaded by University of Missouri-Columbia on 21 September 2010
Jiii Janata and Jan Langmaier
Department of Materials Science and Engineering, University of Utah, Salt Lake City, UT 841 12, USA
The key issue in the development of chemical sensors is the
design of new chemically selective layers. Organic semiconduc-
tors have attracted considerable attention as the materials for
such diverse applications as power sources, non-linear optics,
superconductors and molecular electronics. Not surprisingly,
they are also being investigated as possible sensor materials in a
wide range of applications. The parameters that are modulated
by the chemical interactions include mass, optical absorptivity,
conductance, Galvani potential and work function (WF). This
paper will be restricted to a discussion of the interactions of
organic semiconductors with the gaseous phase and will focus
particularly on the chemical modulation of the WF.
The electron WF is one of the fundamental material
constants. It plays a key role in the distribution of electrons in
solid-state structures, affects the catalytic properties of solid
materials and determines the rate of corrosion and chemical
resistance of materials. It is defined as the work that has to be
used in order to extract an electron from the interior of a phase
and place it outside the reach of image forces, in the so-called
vacuum reference level. There are two components of the WF:
the bulk contribution, which is related to the chemical potential
of an electron in the phase and represents the affinity of the
electron for the matrix; and the surface contribution, which is
related to the electric field resulting from the surface dipole
layer.
Measurement of the WF provides information about the
electronic properties of a material. For example, organic
semiconductors prepared by electro-oxidation under different
conditions have a different WF, which is further affected by the
presence of other electron donors/acceptors. This phenom-
enon is the basis for a new class of electrochemical sensors,
particularly for the sensing of gases. To date, the WF has been
used analytically only rarely. The best known device which
measures the WF is the vibrating capacitor (Kelvin probe).
Because of its size it is impractical as a sensor, although it was
proposed as a gas chromatographic detector almost 40 years
ago. The operating characteristics of solid-state devices based
on metal-insulator-semiconductor junctions, such as insulated
gate field-effect transistors or metal-insulator-semiconductor
diodes, depend directly on the WF of the gate metal. This fact
has been exploited in the design and development of well-
known potentiometric sensors for hydrogen, which use palla-
dium as the gate metal, and in transistors using organic
semiconductors which have been shown to respond to a variety
of organic vapours. In this paper the fundamental aspects of
the WF will be reviewed, its potential for chemical analysis of
gases assessed and examples of general solid-state chemical
sensors based on its chemical modulation will be given.
View Online
ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
References
1 Krull, U. J., and Thompson, M., ZEEE Electron Devices, 1985,
32, 1180.
2 Sugawara, M., Kojima, K., Sazawa, H . , and Umezawa, Y . ,
Anal. Chem., 1987, 59, 2842.
3 Moody, G. J., Sanghera, G. S., and Thomas, J. D. R., Analyst,
1986, 111, 1235.
4 Ghaemmaghami, V., Kallury, K. M., Krull, U. J., Thompson,
M., and Davies, M. C., Anal. Chim. Acta, 1989, 225, 369.
It is necessary to realize that organic semiconductors can be
deposited in different ways, e.g., by sublimation, solvent
casting or electrochemical deposition. The choice of the
deposition technique used is determined by the constraints
given by the sensor itself. In WF sensors it is a necessity that at
least one interface of the selective layer is capacitively coupled
to the remainder of the sensor structure.' This condition is
satisfied in a suspended gate field-effect transistor (SGFET)2
and in its macroscopic counterpart the vibrating capacitor. The
latter has been used in this work to develop and characterize a
selective layer for hydrogen cyanide based on electrochemi-
cally prepared polyaniline (PANI) .
Aniline can be easily polymerized from acidic media3 on the
Pt plate of the vibrating capacitor and its WF can be measured
against a suitable reference plate, e.g., stainless steel. A
considerable advantage of this approach is that the film on the
working plate can be examined by a variety of auxiliary
spectroscopic and microscopic techniques.
The WF of PANI itself is not affected by HCN. However,
when the Ag-AgCN system is incorporated in PANI the layer
becomes sensitive to HCN. The kinetics and the polarity of the
change of the WF depend on several factors. When the
PANI*Ag layer is prepared in the reduced state (PANI *Ag is
a heterogeneous system; the asterisk signifies the heterogen-
eity) its WF decreases on exposure to HCN. The response
becomes reversible when the electropolymerization is carried
out from a glycerine-1 mol dm-3 HZS04 mixture. Glycerine
facilitates proton transfer from the silver clusters to the PANI
backbone :
[Pt-PANI-Ag] + HCN -Slow 1.c--
[Pt-PANI-H+,AgCN]
T -
Glycerine
e
~ '
On the other hand when the film is prepared in the oxidized
state and/or when the AgCN complex is present the reaction
proceeds according to the scheme:
[Pt-PANI-Ag-AgCN] + HCN - Fast
- e-
1
[Pt-PANI-H+ ,Ag( CN)2]
?-A
tilycerine
In this instance the HCN molecule dissociates forming a
strong Ag(CN)* complex and the proton is again transferred to

ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
the PANI backbone. The electroneutrality of PANI is main-
tained by the inflow of electrons from the reference plate which
is equivalent to the increase of the WF. This result confirms the
mechanism of charge transfer between the guest molecule and
the m a t r i ~if: ~the matrix is reduced (high value of WF) it acts
as an electron donor and the WF decreases. When the matrix is
oxidized (low WF) it can accept electrons and the WF
increases. For polypyrrole and polythiophene this proton
transfer reaction is absent and the response to HCN is
irreversible.
The response is logarithmic in the range 5-50 ppm and has
square-root-time dependence, indicating predominantly bulk
interaction. The selectivity of this material to a variety of
environmentally important gases remains to be tested.
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
Downloaded by University of Missouri-Columbia on 21 September 2010
Applications of Electropolymerized Films in Electrochemical Sensors
Philip N. Bartlett
Department of Chemistry, University of Warwick, Coventry CV4 7AL
Electrochemical polymerization is a very convenient technique
for the production of modified electrode surfaces. The method
is simple to carry out and the deposition of the film is readily
controlled by control of the electrode potential. The method is
well suited to the deposition of films onto microelectrode
electrode structures formed by photolithography.'
We have investigated the application of electropolymerized
films as a method for producing modified electrode surfaces for
use in chemical sensors and biosensors. Much of this work has
concentrated on the use of conducting polymer films formed
from substituted pyrroles or other heterocyclic monomers but
we have also made use of poly(pheno1ic) films.
Ion-sensitive Polymers
Films of poly(5-carboxyindole) can be grown from acetonitrile
solutions. The resulting conducting polymer has a carboxylic
acid substituent on every monomer unit of the chain. When the
polymer is transferred into aqueous solution the degree of
ionization of these groups varies with the solution pH and this,
in turn, alters the electrochemistry of the film. The films are
stable at pH values less than 5 but dissolve slowly in aqueous
solution above pH 7. The pH dependency of the electro-
chemistry of poly(5-carboxyindole) can be used to make small
pH-sensitive electrodes. These were made by the electropoly-
merization of the monomer from acetonitrile onto 125 pm
diameter platinum wires sealed in heatshrink PTFE. The
coated electrodes were then transferred into buffered aqueous
solution at pH 2 and cycled until a stable response was attained,
and then held at +800 mV versus SCE and the current allowed
to decay to zero to convert the film into its oxidized form. pH
measurements were made using the oxidized electrodes with a
commercial pH meter and calomel reference electrode.
At pH 7 some portion of the carboxylate groups in the
poly(5-carboxyindole) are deprotonated and can be used to
hydrogen bond to protonated lysine residues around the haem
edge in cytochrome c. This leads to the adsorption of
cytochrome c at the polymer surface in the correct orientation
for electron transfer between the polymer and the haem
group .2 This interaction between the carboxylate groups and
the lysine residues mimics the hydrogen bonding interactions
which are believed to play an important role in electron
transfer between cytochrome c and cytochrome oxidase, its
natural redox partner.
In principle, it should be possible to confer ion selectivity on
a conducting polymer film by attaching suitable substituents to
View Online
373
This work was supported by a contract from BWB and by a
grant from the EPA Office of Exploratory Research, Grant
NO. R-816491-01-0.
References
1 Janata, J . , Principles of Chemical Sensors, Plenum Press, New
York, 1989.
2 Josowicz, M., and Janata, J., in Chemical Sensor Technology,
ed. Seiyama, T., Elsevier, Amsterdam, 1988.
3 Zotti, G . , Cattarin, S . , and Comisso, N., J. Electroanal. Chem.,
1987, 235,259.
4 Blackwood, D., and Josowicz, M., J . Phys. Chem., 1991, 95,
493.
the polymer. This might be advantageous because the mixed
ionic and electronic conductivity of the resulting conducting
polymer could then be utilized to make small ion-selective
sensors. We have investigated this approach by synthesizing
pyrrole substituted in the N-position with a benzo-15-crown-5
derivative. The resulting monomer can be electropolymerized,
and the properties of the resulting conducting polymer films
have been in~estigated.~
Immobilization of Glucose Oxidase
The electropolymerization of pyrrole and N-methylpyrrole
from buffered aqueous solutions can be used to immobilize
glucose oxidase and other enzymes at electrode surfaces. In the
resulting enzyme-loaded films the behaviour is determined by
the balance of the diffusion of reactants and products within
the film and the kinetics for the immobilized enzyme. In order
to understand the behaviour of these films it is necessary to
model these processes and to compare the predictions of these
models with the experimental r e s ~ l t s . ~
Studies of glucose oxidase immobilized in poly( N-methyl-
pyrrole) films show that when the natural redox partner,
oxygen, is used as the mediator species the hydrogen peroxide
produced is not oxidized on the polymer but rather must
diffuse to the underlying electrode to be detected.' By using
tritium-labelled glucose oxidase we have determined the
concentration of enzyme entrapped within our electropolymer-
ized films and, by using these data and data from the kinetic
analysis of the response of these films to glucose, we have
estimated the kinetics for the oxidation of glucose by the
immobilized enzyme. It was found that these kinetics are not
significantly different from the values obtained in homo-
geneous solution.
Studies in which oxygen is replaced by ferrocenecarboxylic
acid or hexacyanoferrate(rI1) as an artificial mediator species
show similar behaviour. In each instance the polymer appears
to be electro-inactive towards re-oxidation of the mediator.
Fourier transform infrared studies of the polymer films indicate
that this loss of conductivity of the film is caused by reactions
with hydrogen peroxide generated by the enzyme.
Electrochemically polymerized films of phenols can also be
used to immobilize glucose oxidase at an electrode surface.6
The immobilized enzyme remains active and glucose can be
detected by using electrodes of this type either with oxygen as
the mediator or when using an artificial redox mediator such as
ferrocenecarboxylic acid. Analysis of the responses of such
 View Online

374 ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28

electrodes enables the rate constants for the enzyme-catalysed References

reactions to be estimated and the effect of the substituents on
the phenol on the properties of the immobilized enzyme to be
investigated.
This work was supported by the SERC (Grant G R E 67108)
and by MediSense. I thank the following colleagues and
students who have contributed to this work: P. Moore, P.
Tebbutt, A. Benniston, L-Y. Chung, R. Whitaker, Z . Ali, J.
Farrington, D. Dawson, V. Eastwick-Field, D. Caruana, P.
Birkin, V. Rhodes and C. Tyrrell.
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
1 Bartlett, P. N., Gardner, J. W., and Whitaker, R. G., Sens.
Actuators, 1990, A21-A23, 911.
2 Bartlett, P. N., and Farrington, J., J . Electroanal. Chem., 1989,
261, 51.
3 Bartlett, P. N., Benniston, A. C., Chung, L-Y., Dawson, D. H.,
and Moore, P., Electrochim. Acta, 1991, 36, 1377.
4 Bartlett, P. N., and Whitaker, R. G., J . Electroanal. Chem.,
1987, 224, 27.
5 Bartlett, P. N., and Whitaker, R. G., J . Electroanal. Chem.,
1987, 224, 37.
6 Bartlett, P. N., and Whitaker, R. G., Biosensors, 1987/88, 3 ,
359.

Disposable Single-use Sensors

Downloaded by University of Missouri-Columbia on 21 September 2010

Monika J. Green and Paul 1. Hilditch

MediSense Inc., Units 3 & 4, 14/15 Eyston Way, Abingdon, Oxfordshire OX14 ITR

For a medical diagnostic product, the advantages of a single- Conductive Working

use disposable sensor are clear. Most importantly, problems
with contamination or carryover are eliminated and steriliz-
ation by the user is unnecessary. Further, with a re-usable
sensor there might be concerns with drift and the need for
recalibration, which are not an issue with a disposable device.
In addition, the low cost of disposable sensors reduces the fear
of expensive damage associated with a re-usable device.
The principal criterion for successfully producing or adapt-
ing a diagnostic test in disposable form is that a means must be
found of constructing a device containing all the necessary
components for the test, avoiding the need to add reagents at
the time of use. The device should have a stable shelf-life of at
least 12 months, and the whole should be capable of
manufacture in large numbers relatively cheaply.
Where the addition of reagents at the time of the test seems
unavoidable, the usual solution has been to employ one of the
large automated analysers to perform the test. This type of
instrument, however, has little part to play in the rapidly
expanding market of decentralized testing, which offers much
for improved efficiency and quality of patient care.
Glucose Monitoring
The basic concept of a biosensor for personal glucose
monitoring is simple: reagents and biochemicals necessary for
detection are deposited on a disposable sensor device; this can
be interfaced with an electronic meter system which monitors
(and may control) the progress of the reaction, and interprets
and displays the results.
The detection chemistry which has been most successful in
this area is that based on the interaction between glucose
oxidase (GOD) and derivatives of ferrocene [bis(cyclopentadi-
.’
enyl)iron] The oxidized forms of ferrocenes (ferricinium
ions) are capable of accepting electrons from GOD which has
been reduced by the reaction with glucose. It is therefore
possible to set up a system whereby ferrocene, oxidized at an
electrode to form ferricinium, is reduced by GOD in the
presence of glucose, and re-oxidized at the electrode producing
PVC substrate silver track elec!rode
\- ,Contacts I I
I I
I I
Conductive
carbon track Dielectric Ag-AgCI reference
layer electrode
Fig. 1 Schematic representation of a disposable glucose sensor strip
two electrodes (working and reference) necessary for the
electrochemical reaction, and transmit current and potential
information to the meter during the measurement, and one of
which contains enzyme and mediator in a labile matrix.
A ‘second-generation’ sensor, which is also available in desk-
top format, has several improved features including automatic
detection of blood application (no button pressing), tempera-
ture com ensation and almost complete freedom from inter-
ferences.’ The last of these has been achieved by the
incorporation on the sensor strip of a third electrode, which
contains all the components of the normal working electrode
except for the enzyme. Any redox-active interferents present
in the sample will affect the response of this ‘dummy’ electrode
in the same way as the working electrode, and a correction is
performed by the meter to give the glucose value. The linearity
and accuracy of this system are illustrated in Fig. 2.
Acetaminophen
Known in the UK as ‘paracetamol’ and in the US as ‘tylenol’,
acetaminophen is a widely used analgesic. Its ready availability
and toxic effect in overdose make this a substance which is
frequently suspected in emergency room admissions. In an
acetaminophen ~ e n s o rthe
, ~ enzyme aryl acylamidase catalyses
the deacylation of acetaminophen to yield p-aminophenol,

-
which can be oxidized at an electrode to yield quinoneimine
an electrical current dependent on the glucose concentration: with consequent measurable current:
Glucose + GOD,, gluconolactone + GODred (1)
G0Dp-d + Fe(cp)2+ + GOD,, + Fe(cp)2
) ~Fe(cp)2f + e-
F e ( ~ p+
This chemistry is produced commercially in the form of a
(2)
(3)

disposable sensor .2 A poly(viny1 chloride) (PVC) substrate is

printed with several layers (Fig. l),some of which provide the
0 -@
NHCOCH3
I

OH
+ H20
NH2
I

OH
+ CH3COOH
(4)

ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
+ &i
++ + Economical (<$1per test)
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
Downloaded by University of Missouri-Columbia on 21 September 2010
0 3 6 9 12 15 18 21 24 27 30 33
Mean YSI venous reading/mmol I-'
Fig. 2 Clinical response of the Satellite G glucose sensor compared
with a laboratory reference analyser [Yellow Springs Instruments
(YSI)]. The line on the graph is the line of identity
OH Q for theophylline exploits the interaction of theophylline with
A potential interferent is N-acetylcysteine, which is adminis-
tered as the antidote for acetaminophen poisoning; this (and
other thiol reagents) can react with the quinoneimine product
of the first reactions, forming a substituted aminophenol which
can be reduced at the electrode, augmenting the current
already observed:
NH NH,
NH2 NH
In order to eliminate this possible interference, the sensor strip
described includes an absorbent overlay containing gluta-
thione. This reagent ensures that the thiol augmentation of
response is fully saturated irrespective of the thiol content of
the blood.
Salicylate
N-Acetylsalicylate (aspirin) is another widely used analgesic,
and is monitored in emergency situations for the same reasons
as acetaminophen. Aspirin is also used as an anti-inflammatory
agent in the management of conditions such as arthritis, and
because of the toxicity of salicylate, monitoring of drug
concentrations is desirable, N-Acetylsalicylate is rapidly con-
verted by hepatic esterases into salicylic acid, which is the
therapeutically active form of the drug, and it is this which is
measured. The principle of the assag is similar to that for
acetaminophen: salicylate is converted into catechol by the
View Online
375
Table 1 Design specifications for the bacterial assay system
Alternative to 'standard plate count'
Capable of producing results rapidly (<1 h)
Moderate sensitivity (10s cfu ml-' or better)
enzyme salicylate hydroxylase. Catechol is oxidized to o-
benzoquinone by the electrode and the current measured. The
cofactor NADH is required by the enzyme, and must be
provided in the sensor. Linearity of response is obtained by the
inclusion of sodium benzoate in the reaction chemistry. This
acts as a pseudo-substrate for the enzyme, effectively modify-
ing its kinetics to give a near-linear response in the clinical
concentration range.
Theophylline
There is a considerable market for sensors capable of
monitoring therapeutic drugs present in blood at micromolar
concentrations. These substances will usually not be directly
detectable in the blood matrix, leaving as options an immuno-
assay test or some other format such as enzyme inhibition. For
theophylline, the latter approach has been p u r ~ u e d . ~
Theophylline, a member of the xanthine family of com-
pounds, is prescribed as a bronchodilator in cases of asthma, on
r
a proph lactic basis. Above its therapeutic range of 55-110
ymol 1- , theophylline rapidly becomes toxic and so effective
monitoring of concentration is desirable. A disposable sensor
the enzyme alkaline phosphatase: the drug behaves as an
uncompetitive inhibitor, binding to the enzyme-substrate
complex and preventing its further reaction.
Alkaline phosphatase will hydrolyse a wide range of
substrates, and that which is of most interest electrochemically
is p-aminophenol phosphate. The parent compound is readily
distinguishable electrochemically from the product, p-amino-
phenol, and so the activity of a sample of alkaline phosphatase
can be measured by providing p-aminophenol phosphate and
measuring electrochemically the amount of p-aminophenol
produced. Equally, providing a known amount of enzyme and
substrate allows the concentration of inhibitor (theophylline)
to be assessed.
Bacterial Assay System
A final example of a disposable sensor system is that developed
for the detection and measurement of micro-organisms. The
opportunities for this area of testing are almost endless,
encompassing clinical, food and environmental applications.
The objectives set out for the system described by Hilditch
et ~ 1 are. summarized
~ in Table 1. The approach taken was the
electrochemical interrogation of bacterial respiration using a
mediator compound; the assay therefore measures viable
biomass.
The mediator used in the assay is p-benzoquinone, which
resembles the quinones which occupy a pivotal position in the
electron transport chain of bacteria and may compete with
these for respiratory electrons, although the exact site of
mediation is unclear. A wide range of quinones and related
compounds have been screened and the simplest, p-benzo-
quinone, has been found to be the most satisfactory. Bacteria
respiring in the presence of p-benzoquinone cause it to become
reduced to hydroquinone. This compound can then be re-
oxidized at an electrode at the appropriate potential; the
amount of hydroquinone formed in a given time is a measure of
the rate of bacterial respiration in the sample and hence of the
cell numbers.
A simple assay of this type has certain limitations, in
particular that its sensitivity is limited to about lo6cfu ml-' (cfu
= colony forming units) by the rate of bacterial respiration,
 View Online

376 ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28

and that redox-active substances in the sample will interfere. typically found to give a linear response with a sensitivity of
The system described overcomes these problems by concen- about lo4 cfu ml-' (Fig. 4).In addition to pure cultures, results
trating bacterial cells on a filter in close proximity to the
measurement electrodes. This allows amplification of the
response as cells from a relatively large volume of sample are
concentrated into a small reaction volume; moreover, the cells
can be washed when on the filter, removing soluble redox
interferents.
Filter and electrodes are contained in a disposable assembly
2.
loo0

2 100 -
i
C
(Fig. 3) formed from PVC; the lower half has printed 0
CL
v)
electrodes while the upper half has a recess to contain a disc of
glass-fibre filter material. The disposable assembly fits into a [r
10 -
housing built into the meter which automatically interfaces
with the fluid handling and electrical contacts. The meter 0
pumps sample, wash buffer and mediator solution as appropri- r = 0.988
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366

1 I I I
ate through the disposable assembly, which is maintained at 1
103 104 105 106 107 108
constant temperature. After loading sample onto the dispos- Se nsit ivit y k f u mI - 1
Downloaded by University of Missouri-Columbia on 21 September 2010

able assembly, and washing with buffer, the assembly is

flooded with mediator solution (which also contains carbon
source for the bacteria) and is incubated for 10 min. During this
time bacterial respiration is converting benzoquinone into
hydroquinone, which is measured for 30 s at the end of the
incubation. The total assay time is therefore of the order of 15
min.
The performance of the assay has been validated with a
considerable range of aerobic bacterial species; it has been
Embossed and
textured P V C A such as bacterial monitoring promises to extend further the
Fig. 4 Response of the bacterial assay system to bacteria (Pseudo-
monas cepacia)
have been obtained with mixed bacterial cultures, yeasts and
food and environmental samples.
Conclusion
Single-use biosensor diagnostics are characterized by con-
venience of use, rapidity, accuracy and freedom from sample
pre-treatment. Application of the technology to other areas
boundaries of analytical chemistry.

References
Green, M. J., and Hill, H. A . O., J . Chem. SOC.,Faruday Trans.,
1986, 82, 1237.
Matthews, D. R., Holman, R . R., Bown, E., Steemson, J.,

Filter disc c-> Watson, A., Hughes, S., and Scott, D., Lancet, 1987, 1, 778.
Matthews, D. R., Burton, S. F., and Smith, E., in Proceedings,
Artijicial Insulin Delivery Systems Pancreas and Islet Transplan-
tation, European Association for the Study of Diabetes,
Amsterdam, 1991.
Jones, A. F., McAleer, J. F., Braithwaite, R. A . , Scott, L. D.,
Brown, S. S., and Vale, J. A., Lancet, 1990, 335, 793.
Frew, J. E., Bayliff, S. W., Gibbs, P. N. B., and Green, M. J.,
Anal. Chim. Actu, 1989, 224, 39.
Foulds, N. C., Wilshere, J. M., and Green, M. J., Anal. Chim.
Actu, 1990, 229, 57.
Hilditch, P. I., Carter, N. F., Barrett, C. B., Sullivan, D. J.,
with electrodes Charman, K. M., Green, M. J . , and Williams, S. C . ,in Advances
Fig. 3 Disposable sensor element for use in a biosensor bacterial in Bioreactor Monitoring, ed. Wang, N. S . , Academic Press,
assay system New York, 1991, in the press.

Applications of Amperometric Biosensors

Anthony P. F. Turner
Biotechnolog y Centre, Cranfield Institute of Technology, Cranfield, Bedford MK43 OAL

Numerous definitions of a biosensor pervade the recent
scientific literature.' Taking a pragmatic view, a biosensor can
be considered as one possible solution to a particular analytical
problem. Whether this technical route should be pursued or
not depends on the relevance of the features offered by
biosensors to the problem. Biosensors generally provide
continuous information about the concentration of a chemical
or group of chemical^.^,^ They are capable of considerable
specificity and sensitivity even when used directly in real
samples, without necessarily incurring the need for complex
and expensive instrumentation. Coupled with modern micro-
electronics they form the basis for a new generation of
'sensitive' computers. This paper will review the requirements
of several areas where amperometric biosensors are likely to
make an impact. A few recent or new examples of biosensor
technology that are likely to prove a commercial success4 will
be illustrated.
Clinical diagnostics have dominated biosensor development
to date and are likely to continue to be the focus for commercial
successes over the next five years .5 Clinical applications are
driven by the need for cost effective patient care. Analytical
instruments can be comprehensive and automated or
decentralized and specialized. Some conditions, for example
diabetes, benefit from self-tests usable in the home.6 Indeed,

ANALYTICAL PROCEEDINGS, NOVEMBER 1991, VOL 28
the world's most successful biosensor is a pen-shaped blood
glucose monitor which addresses this market. A key issue in
tackling large markets is the requirement for inexpensive and
reproducible mass production of the disposable biosensor
element. Once investment has been made in such a production
facility it is desirable to find further applications for the
technology. Diagnostic tests that make sense to perform in the
home are few and far between. Pregnancy testing is an obvious
example, but the frequency with which these tests are
performed makes an instrumented assay uncompetitive with
the elegant visual assays now marketed. Other promising
analytes that are attracting commercial development of
pocket-sized devices include total cholesterol, ethanol and
lactate.
The printing technology that has been developed for the
Published on 01 January 1991 on http://pubs.rsc.org | doi:10.1039/AP9912800366
mass production of one-shot devices might find application in
reusable or continuous use devices. Recent work in our
Downloaded by University of Missouri-Columbia on 21 September 2010
laboratories has shown that screen-printed enzyme electrodes
for glucose, using tetrathiafulvalene as mediator,' can be used
over 2000 times. This has been achieved by using proprietary
membrane technology acting as a reservoir of the mediator.'
Such elements might be useful in larger machines where the
throughput permitted by biosensors can out-perform conven-
tional systems or where a competitive advantage can be
achieved by broadening the spectrum of analyses offered to
include, for example, glucose, urea, creatinine and lactate.
Equally, simple amperometric devices can be constructed
which are capable of highly sensitive immunoassay of analytes
such as thyroid stimulating hormone, prostatic acid phospha-
tase (a tumour marker) or estradiol.'**" Some speculative
work is underway to perform electrochemical detection of
DNA hybridization, furnishing the ossibility of detecting a
wide range of pathogenic organisms.Y2
A further area of interest is in vivo m ~ n i t o r i n g . Mediated
'~
amperometric enzyme electrodes offer advantages over oxy-
gen-consuming systems in their relative independence from
variations in oxygen tension. Concern has been expressed,
however, about the toxicity of the mediators used. New
evidence suggests that mediators can be less toxic than feared,
for example, the LD50 in mice of tetracyan~quinodimethane'~
and tetrathiaf~lvalene~~' (two recently discovered mediators) is
1225 and 710 mg kg-', respectively.lS While many promising
sensors have been described, lack of biocompatibility remains
a major hurdle to the application of sensors inside the body for
critical care or long-term therapeutic purposes. 16.17A n inter-
esting compromise has recently been announced by an Italian
company; the proposed product consists of a wearable
microdialysis system incorporating a non-mediated enzyme
electrode for continuous monitoring in diabetics.
Much of the technology developed for clinical applications
will find other uses in industry, l8 environmental monitoring"
and defence. Industry is seeking sensors to improve productiv-
ity, quality and compliance with legislation. Amperometric
biosensors have recently become commercially available for
use in situ in fermentation monitoring with broad consequences
for the food and biotechnology industries. Amperometric
detection of microbial contamination2' of milk has reached the
manufacturing prototype sta e in our hands with a detection
limit of lo4 organisms ml-' obtained within 20 min in a
completely 'hands off' format.
Environmental applications of biosensors are characterized
by a desire to protect workers and the public from toxic
View Online
377
material, and compliance with legislation. One interesting new
development with particular relevance to this area is the
discovery of organic phase enzyme electrodes.21722These
instruments are capable of operating in oils or fats23 and can
also offer improved performance in aqueous samples by, for
example, automatically concentrating contaminants present in
drinking water.
As the academic literature on biosensors continues to
diversify, a clear trend within industry critically t o evaluate and
invest in focused areas is emerging. Technical issues concern-
ing the developers include fabrication technologies and a
general drive towards miniaturization. Medicine and food
provide the prime focus for new products, but many smaller
companies are searching for niche markets and might furnish
some fascinating new gadgets, allowing executives to transform
themselves into skilled analytical chemists overnight.
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