Analytica Chimica Acta 387 (1999) 103112

An integrated three-electrode system with a micromachined

liquid-junction Ag/AgCl reference electrode
Hiroaki Suzukia,*, Taishi Hirakawaa, Satoshi Sasakib, Isao Karubeb
a
Institute of Materials Science, University of Tsukuba, 1-1-1 Tennodai, Tsukuba Science City, 305-8573 Japan
b
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904 Japan

Received 17 July 1998; received in revised form 4 January 1999; accepted 9 January 1999

Abstract

A micromachined liquid-junction Ag/AgCl reference electrode was employed to fabricate an integrated three-electrode
system. The reference electrode features a durable thin-lm Ag/AgCl element whose entire surface was coated with a
hydrophobic membrane and AgCl layer grown from its periphery. On the other side of the chip were formed a platinum
working electrode and a platinum counter electrode. The cleanliness of the thin-lm platinum working electrode was
satisfactory to conduct electrochemical analyses including cyclic voltammetry. No substantial difference was observed in the
cyclic voltammograms between the integrated system and the macroscopic three-electrode system in terms of their peak
current and half-wave potential. The integrated electrode system was also applied to fabricate one-chip bio/chemical sensors.
The miniature glucose sensor showed a distinct response against step changes of glucose concentration. The one-chip
hydrogen electrode was used to conduct potentiometric titration. A good coincidence was again conrmed between the
integrated system and the macroscopic system. # 1999 Elsevier Science B.V. All rights reserved.

Keywords: Ag/AgCl; Liquid junction; Platinum; Cyclic voltammogram; Glucose; Potentiometric titration

1. Introduction [1] and was applied to a micromachined liquid-junc-

One of the most troublesome problems in the
development of micro electrochemical sensors has
been the unavailability of a miniature reference elec-
trode with a liquid junction. In realizing such a
reference electrode the foremost issue has been the
development of a durable thin-lm Ag/AgCl element
which functions even in a solution saturated with KCl.
In our previous study a novel thin-lm Ag/AgCl
element with satisfactory durability was developed
*Corresponding author. Fax: +81-298-55-7440; e-mail:
hsuzuki@ims.tsukuba.ac.jp
tion Ag/AgCl reference electrode [2]. Although there
is still room for its performance to be improved
further, its realization made the integration of all
the electrochemical system possible. In this paper
we will show a device on which the working, counter,
and reference electrodes are integrated.
There are some potential applications for the
device. Chemical analysis on samples of minute
quantities is attracting attention in medicine and biol-
ogy. Electrochemistry is one of the most powerful
techniques for selective and sensitive detection in such
an environment. In basic researches the cost and/or
difculty in obtaining puried analyte sometimes pose

0003-2670/99/$ see front matter # 1999 Elsevier Science B.V. All rights reserved.
PII: S0003-2670(99)00063-X
104 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112
a serious problem and the analysis in a small sample
volume is imperative. An extreme case is the chemical
analysis of a single cell [3,4]. In order to achieve such
an electroanalysis in microvolume, several micro
systems have been proposed [3,511], some of which
have been fabricated using thin-lm technologies and
micromachining [3,911]. By using these systems not
only can the dilution of the analyte due to the diffusion
to the bulk solution be alleviated to maintain the low
analyte levels (in the order of femtomolezeptomole)
present in the single cell, but also a fast response is
obtained because the diffusion length (or the distance
between the cell and the working electrode) is reduced
[3]. With the recent advent of mTAS (micro total
analysis system) technology the integration of the
three-electrode system in a small space is becoming
more and more important. Once a reliable miniature
integrated system is realized, it would not be difcult
to modify its design and use it as a detector in other
microanalytical devices such as a microfabricated
capillary electrophoresis system [12].
Another important application of the integrated
device is in vivo, in situ measurements. Hundreds
of papers have been published which deal with micro
electrochemical sensors including ion sensitive eld
effect transistors (ISFETs). However, in almost all
cases the miniaturization is limited to the working or
indicator electrode and a macroscopic reference elec-
trode (and a macroscopic counter electrode for
amperometric sensors) are used. It might be enough
for a basic research but it is actually the cause which
hinders further practical application of these miniature
sensors to many potential elds. For in vivo and in situ
measurements, all electrode elements must be inte-
grated on a chip. In order to show a possibility of
application of this integrated device to one-chip
microsensors, a glucose sensor and a hydrogen elec-
trode were fabricated. The performance of the inte-
grated electrode system and the one-chip sensors will
be presented in this report.
2. Experimental
2.1. Materials
Glass wafers (No. 7740, 3 in., 500 mm thick) were
purchased from Corning. (1 0 0)-oriented silicon
wafers (3 in., 400 mm thick) were purchased from
Japan Silicon. A photo-patternable polyimide prepo-
lymer (Photoneece 3100) was a gift from Toray. A
positive photoresist (S1400-31) used in fabrication
was purchased from Shipley Far East, Tokyo. A
negative photoresist, OMR-83, used in fabrication
and the electrode structure was purchased from Tokyo
Ohka Kogyo, Kawasaki, Japan. A photocurable adhe-
sive (BENEFIX VL) was purchased from Adell,
Tokyo. The reagents used in fabricating the electrode
were of semiconductor grade and purchased from
Kanto Chemicals, Tokyo, and Wako Pure Chemicals,
Osaka. All reagents used to examine the electrode
performance were purchased from Wako Pure Che-
micals. They were of analytical reagent grade and
used without further purication. Glucose oxidase
(type X-S, from Aspergillus niger) and bovine serum
albumin (BSA) were used as supplied from Sigma.
Distilled water was used throughout the experiments.
2.2. Structure and fabrication of the integrated
three-electrode system
The integrated three-electrode system was fabri-
cated on both sides of a glass substrate (Fig. 1).
Ordinary semiconductor and micromachining pro-
cesses were followed, which will not be described
in detail here. Metals were deposited by sputtering.
The dimensions of the chip were approximately
1.5 mm wide, 13 mm long and 0.9 mm thick. On
one side were formed a working electrode and a
counter electrode both of which were made from
300-nm-thick platinum (Fig. 1(a)). A 40-nm-thick
chromium layer was used as an adhesive layer. In
order to delineate the electrode areas and insulate the
lead patterns, a 600-nm-thick polyimide layer was
formed. The exposed areas on the working and counter
electrodes were 0.5 mm0.5 mm and 0.5 mm
6.4 mm, respectively. In curing the polyimide patterns
the devices were subjected to bakings for 15 min at
1508C, 15 min at 2008C, and 30 min at 3008C.
The miniature liquid-junction Ag/AgCl reference
electrode was formed on the other side of the glass
substrate (Fig. 1(b)), the structure and fabrication of
which were the same as reported elsewhere [1,2]. In
brief, the thin-lm Ag/AgCl element used in this
device consisted of a silver layer (300 nm thick)
formed on a gold backbone layer (200 nm thick)
which made partial contact with the silver layer. A
 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112 105

Fig. 1. Structure of the integrated three-electrode system: (a) glass substrate on which the working and counter electrodes are formed; (b) the
other side of (a) on which the novel thin-film Ag/AgCl element is formed; (c) silicon substrate on which the container for the electrolyte
solution and the liquid-junction are formed. The surfaces of (b) and (c) are bonded to make (d); (d) completed liquid-junction reference
electrode. The other side of the silicon substrate shown in (c) can be seen; (e) cross section of the currently-used thin-film Ag/AgCl element;
(f) cross section of the thin-film Ag/AgCl element shown in (a) along the line xx0 .

hydrophobic negative photoresist layer (2.7 mm thick)
covers the entire silver layer. AgCl layers were formed
not from the surface but from the periphery of the
silver pattern as shown in Fig. 1(b) and (f). The silver
layer was transformed to AgCl by immersing the chip
in 1.0 M FeCl3 for 50 min at 308C. The microcavity
for the internal electrolyte solution was formed by
anisotropically etching a silicon substrate in a KOH
solution at 808C (Fig. 1(c) and (d)). The glass and
silicon substrates were bonded with a photocurable
adhesive and the end of the liquid junction was closed
with cellulose acetate as described elsewhere [2]. In
106 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112
performing the work, the working and counter elec-
trodes and their leads were protected with a poly-
(vinylalcohol) (PVA) coating to avoid mechanical
damage, which was removed just before the introduc-
tion of the electrolyte solution. The internal electrolyte
solution for the reference electrode was a solution
saturated with KCl and AgCl, which was introduced
into the microcavity by immersing the chip in the
electrolyte solution in a beaker, placing it in a cham-
ber, and evacuating it [2,13]. Air bubbles often trapped
at the liquid junction were removed thoroughly by
centrifuging the chip. Ninety devices were batch-
fabricated on a 3 in. wafer and were diced into chips.
In fabricating the one-chip glucose sensor, the
working electrode area of the chip was immersed in
10% 3-aminopropyltriethoxysilane for 1 h, rinsed
with distilled water, and dried. One mg of glucose
oxidase was dissolved in 25 ml of 2% BSA solution,
which was subsequently mixed with 15 ml of 10%
glutaraldehyde solution with a small rod. The working
electrode area of the chip was immediately immersed
in the mixture to coat it, which was allowed to react for
about 10 min, then the chip was rinsed with distilled
water and was immersed in 0.1 M glycine solution for
30 min. The whole immobilization procedure was
carried out at room temperature.
In forming the hydrogen electrode, the platinum
working electrode site and approximately half of the
platinum counter electrode site were platinized in a
solution containing 66 mM H2PtCl6 and 0.88 mM
(CH3COO)2Pb at 30 mA/cm2 for 60 min. Current of
the opposite polarity was own for a brief period every
10 min. The reference electrode site was protected
with the positive photoresist during the platinization.
The platinum working and counter electrode sites and
the obtained platinum black were activated before and
after the procedure in a 0.1 M H2SO4 solution by
polarizing it repeatedly between 0.5 and 0.5 V
(vs. Ag/AgCl). The reference electrode was formed
and assembled after the procedure.
2.3. Procedure
In order to evaluate some electrochemical charac-
teristics two sets of three-electrode system were exam-
ined in combination with a Hokuto-Denko HA-151
potentiostat/galvanostat. The rst type uses the plati-
num working electrode on the chip, a macroscopic Ag/
AgCl reference electrode (Horiba 2080A-06T with a
ceramic-plug junction), and a macroscopic platinum
plate counter electrode. The internal lling solution
for the macroscopic reference electrode was saturated
with KCl and AgCl. The second type used the work-
ing, reference, and counter electrodes on the chip.
Most of the experiments were conducted in a 100 ml
solution lled in a 100 ml beaker. The latter type was
also used to take cyclic voltammograms in a 100 ml
solution lled in a plastic tip of a 100 ml micropipette
whose end was closed by silicone rubber. In conduct-
ing these experiments the platinum working electrode
site was cleaned by repeatedly cycling its potential
between 0.2 and 1.3 V (vs. Ag/AgCl) at 1 V/s. The
sample solutions to be analyzed were bubbled with
pure nitrogen beforehand.
The performance of the glucose sensor was eval-
uated in a 20 mM KH2PO4-NaOH buffer solution (pH
7.0) containing 0.1 M KCl. Applied potential to the
working electrode was 0.7 V (vs. Ag/AgCl). 1.0 M
glucose sample solution was prepared and a prede-
termined amount of the solution was injected into the
buffer solution to check the sensor response.
In conducting the potentiometric titration the inte-
grated electrode chip was immersed in 100 ml solution
of 20 mM HCl and pure hydrogen gas (1 atm) was
bubbled into the solution. After the response of the
hydrogen electrode was allowed to stabilize, 2.0 M
NaOH solution was added progressively to conduct
the titration. In one experiment the site of the platinum
black and the miniature reference electrode on the
chip were used. The same experiment was repeated
using the same platinum black site and the macro-
scopic Ag/AgCl reference electrode.
All pH measurements were made using a TOA
Electronics HM-20S pH meter, with a combination
pH glass electrode previously calibrated in precision
buffers of pH 4.01 and 6.86 (TOA Electronics). All
experiments were conducted at a temperature of
25.00.18C.
3. Results and discussion
3.1. Cleanliness of the thin-film platinum working
electrode
Generally speaking, cleanliness of the surface of a
thin-lm electrode is preserved relatively well in
 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112
Fig. 2. Cyclic voltammogram in 100 ml solution of 0.5 M H2SO4
at 258C obtained using the platinum working electrode on the chip.
A macroscopic reference electrode and a macroscopic platinum
plate counter electrode were used. Scan rate: 50 mV/s.
microfabrication processes as can be imagined from
the quality of the cleanliness which the current semi-
conductor process requires. However, depending on
the facility and materials used, residues of various
polymers used in fabrication might remain on the
surface and affect electrochemical characteristics of
the system. It is known that a residue of polyimide
always persists on the surface of a thin-lm metal
pattern. In order to obtain a clean surface which is free
from such a contaminant, a nickelchromium sacri-
cial layer was used and was removed by chemical
etching [10]. Because a photopatternable polyimide
was used to delineate the working and counter elec-
trodes in our device, the cleanliness of the platinum
working electrode was checked.
Fig. 2 shows a cyclic voltammogram using the thin-
lm platinum working electrode in 0.5 M H2SO4. A
well-known shape of the voltammogram was obtained
[14]. The peaks corresponding to the adsorption and
desorption of hydrogen, formation and reduction of
oxide are clearly distinguished, which indicates that
the cleanliness of the surface is preserved well for use
in electroanalysis. Although there were chips which
did not show the cyclic voltammograms as in Fig. 2
possibly due to the polyimide residue on the working
electrode, the clean surface and the clear cyclic vol-
tammograms could be obtained by repeatedly cycling
107
the potential of the working electrode in the 0.5 M
H2SO4 solution. This indicates that the cleaning pro-
cedure is effective even if some organic residues were
left during the fabrication process. During the poten-
tial scan neither dissolution of the chromium layer nor
corresponding waves in the voltammogram were
observed judging from Fig. 2 and visual inspection.
3.2. Application to electrochemical analysis
In examining the applicability of the integrated
micro system to electroanalysis, cyclic voltammetry
is a good test. Fig. 3 shows cyclic voltammograms of
3 =4
FeCN6 redox couple. Curve (a) is a voltammo-
gram taken in a plastic tip of a 100 ml pipette using the
integrated working, reference, and counter electrodes
on the chip. Curve (b) was taken in a 100 ml beaker
using the same working electrode on the chip and the
macroscopic Ag/AgCl reference electrode and the
macroscopic platinum counter electrode. The shape
and the positions of the peaks of the voltammogram
obtained in the plastic tip are identical with those
obtained in the beaker and coincide with the fre-
quently described curves in related literature [15].
Although a liquid-junction potential of several mV
had been measured for the present miniature liquid-
junction reference electrode [2], the difference was not
clear on the scale shown in the gure. The peak-
shaped curves indicate that planar diffusion perpen-
dicular to the working electrode dominates.
The difference between the two cases was examined
in more detail (Table 1). Overall, a good coincidence
between the integrated system and the macroscopic
system was conrmed. The difference in the cathodic
peak current between the integrated system and the
macroscopic system or among chips is possibly due to
the slight difference in cleanliness of the working
electrode surface. Although a couple of E1/2 values
deviate from the average, this is due to the unsatis-
factory function of the cellulose acetate liquid
junction. Furthermore, the dependence of the peak
currents on the square root of the scan rate was also
conrmed in both systems. Note that the overall
coincidence of the results could be achieved only
by the liquid-junction reference electrode. In this
sense, our study is essentially different from the other
related ones using pseudo-reference electrodes. The
results obtained in the experiments show that not only
108 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112

3 =4
Fig. 3. Cyclic voltammograms of FeCN6 couple. The experiments were conducted in a solution containing 5 mM K3Fe(CN)6 and 0.2 M
Na2SO4. (a) The working, counter, and reference electrodes on the chip were used and the cyclic voltammogram was taken in a plastic tip of a
100 ml micropipette. (b) The working electrode on the chip and macroscopic reference and counter electrodes were used in the solution filled
in a 100 ml beaker. Scan rate: 50 mV/s. Insets show the image of each three-electrode system.

Table 1
Distribution of the cathodic peak current ipc and the half-wave potential E1/2 obtained from the cyclic voltammograms of 10 mM K3Fe(CN)6 in
0.2 M Na2SO4

Chip no. Micro Macro

ipc (mA) E1/2 (V) ipc (mA) E1/2 (V)

1 3.98 0.24 3.25 0.27

2 3.86 0.30 3.45 0.26
3 3.63 0.27 3.85 0.26
4 3.56 0.21 3.30 0.26
5 3.49 0.30 3.58 0.28
6 3.44 0.28 3.81 0.28
7 3.33 0.24 3.53 0.27
8 3.76 0.27 3.87 0.27

Average (SD) 3.63 (0.22) 0.26 (0.03) 3.58 (0.24) 0.27 (0.01)
The columns noted as ``Micro'' are the results obtained with the working, reference, and counter electrodes on the chip in the 100 ml solution.
The columns noted as ``Macro'' are the results obtained with the working electrode on the chip, the macroscopic reference electrode, and the
macroscopic counter electrode in the 100 ml solution. Scan rate: 50 mV/s.

the miniature liquid-junction reference electrode but
also our integrated device as a whole can be used like
the currently-used macroscopic three-electrode sys-
tem.
Although the curve (a) was taken in a tip of a 100 ml
pipette, there was still enough space for the integrated
chip and even 50 ml of the sample solution would have
been enough. A possible application of this study
would be a micro analytical system incorporated in
a micropipette, which would actually be fabricated by
some technical improvements. The micropipette sys-
tem makes transfer of the sample solution to a con-
ventional analysis system (e.g., ow injection analysis
(FIA)) unnecessary. Sensing or electrochemical ana-
lysis can be nished on the spot following suction of
the sample solution. Otherwise, the integrated system
could be incorporated in a small ow cell, which will
contribute to the miniaturization of the existing ana-
 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112
lytical equipment. Although the present integrated
device is a needle-type one, the design can be modied
to a planar-type and the miniaturized electrode system
can be incorporated in a microvial as reported else-
where [3,10]. Furthermore, the present planar working
electrode pattern can be modied to a microelectrode
conguration for more suitable analytical measure-
ments.
As the size of the electrode system is reduced,
electrochemical reaction or equilibrium becomes
adversely inuenced by reaction products from other
electrodes located in the vicinity [16]. This electro-
chemical crosstalk is not specic to the microsystem
but has also been observed with conventional rotating
ring-disk electrodes. Electrodes from an interdigitated
electrode array studied by several groups use this
crosstalk advantageously [17,18]. Given this fact, it
is highly possible, in extremely miniaturized systems,
that the reaction products from the counter electrode
diffuse to the working and reference electrodes (or
from the working electrode to the reference electrode)
located in the vicinity, which either generate a residual
current or might shift the reference electrode potential.
Therefore, each electrode must be separated as far as
possible in a limited microspace. A solution will be to
store each electrode in isolated compartments and
connect them with a liquid junction [13]. Fabrication
of such a device is easy, which will be reported in the
future.
3.3. Effect of using a pseudo-reference electrode
As mentioned before, some microfabricated elec-
trochemical cells used a thin-lm pseudo-reference
electrode which is in direct contact with the test
solution [3,9,10]. Otherwise a microcapillary type
liquid-junction reference electrode was used [11].
These are a compromise due to the unavailability of
a reliable microfabricated liquid-junction reference
electrode. Needless to say, the potential of the former
pseudo-reference electrode varies depending upon the
activity of the primary ion which contributes most to
the equilibrium potential. Unlike macroscopic elec-
trodes a thin-lm reference electrode often behaves
unexpectedly and utmost care must be paid in using it
in real measurements. The latter microcapillary refer-
ence electrode requires some skilled techniques and is
not easy for unskilled persons to handle.
109
The performance of the integrated systems has been
evaluated mainly by cyclic voltammograms, some
of which have been described with the potential
measured against the pseudo-reference electrode.
Although it would be satisfactory as far as the shape
and/or the peak height of the voltammograms are
concerned, reliable information could not be provided
in terms of its potentials. The potential of the pseudo-
reference electrode can be determined with consider-
able accuracy if the activity of the related primary
ion is given. However, such a case will be rare in
real electroanalysis and the sample solution to be
analyzed might contain some redox compounds which
adversely affect the pseudo-reference electrode.
Therefore, if reliability is required, a liquid-junction
reference electrode will be indispensable as in macro-
scopic electrochemical experiments.
An inevitable problem in using a pseudo-Ag/AgCl
reference electrode is its poorly xed potential which
varies according to Cl ion activity, which manifests
itself as a shift of the cyclic voltammogram along
abscissa. The tendency is well described in Fig. 4
which shows the cyclic voltammograms taken against
the pseudo-Ag/AgCl reference electrode on the chip
(i.e., the Ag/AgCl element directly exposed to the
solution without the microcontainer). Five solutions
were used each containing 5 mM K3Fe(CN)6, 0.1 M
KNO3, and various concentration of KCl. The Ag/
AgCl electrode can be regarded as a Cl ion sensitive
electrode. Its potential varies by approximately 59 mV
at 258C as predicted by the Nernst equation, although
it becomes insensitive to Cl activity near the lower
detection limit [19]. The effect is clearly shown as the
shift of the voltammogram. The same experiment was
conducted in solutions using 0.2 M Na2SO4 as a
supporting electrolyte instead of KNO3. However,
the cyclic voltammograms for 10 mM and 0.1 M
KCl, settled virtually the same position. The discre-
pancy between the two kinds of solution with different
supporting electrolyte indicates that the linear range of
the calibration curve of the Ag/AgCl element regarded
as the Cl ion electrode depends on the ionic envir-
onment.
In Fig. 4 the shift of the potential is approximately
100 mV for KCl concentrations between 1 mM and
1 M. Signicance of this shift will depend upon the
sample solution to be analyzed and the required
accuracy. If more than two redox species are contained
110 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112

3 =4
Fig. 4. Cyclic voltammograms of FeCN6 couple obtained using the pseudo-Ag/AgCl reference electrode and the working and counter
electrodes on the chip. The experiments were conducted in 100 ml solutions containing 5 mM K3Fe(CN)6, 0.1 M KNO3, and various
concentration of KCl: (a) no KCl, (b) 1 mM, (c) 10 mM, (d) 0.1 M, and (e) 1.0 M KCl. Scan rate: 50 mV/s.

in the sample solution or multipotential electroche-
mical detection is desired [12], more reliable informa-
tion about the potential will be required. It will not be
denied that a liquid-junction reference electrode is
more suitable for that purpose.
3.4. Application to a one-chip biosensor
In order to test the applicability of the integrated
system to chemical sensors, a one-chip miniature
glucose sensor was fabricated and its performance
evaluated. Its detection was based upon the measure-
ment of H2O2 produced by the enzymatic oxidation of
glucose by glucose oxidase. Fig. 5(a) shows its
response curve. Despite small dimensions of the chip,
a very distinct response curve was obtained. The 90%
response time was within 5 s. Fig. 5(b) shows the
calibration curve. A linear relationship was obtained
in the range of glucose concentration less than about
1.2 mM and the slope gradually decreased above this
concentration.
The data showed that the miniature liquid-junction
reference electrode can be successfully used to con-
struct miniature amperometric sensors. In real situa-
tions, however, a pseudo-reference electrode might be
enough depending on the accuracy and the samples to
be treated. In some commercialized disposable glu-
cose sensors, a pseudo-reference electrode without a

Fig. 5. Application to a miniature one-chip glucose sensor. (a) Response curve against step changes of glucose concentration. Two
hundred mM concentration of glucose was added to the buffer solution stepwise at times indicated by arrows. (b) Calibration curve. Applied
voltage: 0.7 V vs. Ag/AgCl.
 H. Suzuki et al. / Analytica Chimica Acta 387 (1999) 103112
liquid-junction or even one which does not based on a
high exchange current density reaction is used. This
design is allowable when the limiting-current region in
the voltammogram is at enough within the accuracy
of the experiment and the shift of the reference
electrode potential does not cause serious variation
in the output current. Actually, repetition of the
experiment, using the pseudo-Ag/AgCl reference
electrode mentioned before, conrmed no noticeable
difference. However, it should be borne in mind that
reliable data cannot be obtained if the residual current
from interfering materials is affected due to the shift of
the reference electrode potential.
3.5. Application to potentiometric sensing
More stringent stability of the reference electrode
potential is required in potentiometric sensing,
because the shift of the potential immediately man-
ifests itself as an error in the measurement. The
applicability of the miniature reference electrode to
one-chip potentiometric sensing was examined using a
hydrogen electrode formed on the back of the liquid-
junction Ag/AgCl reference electrode. (Note that it
Fig. 6. Variation of the potential of the thin-film hydrogen
electrode during the titration of 100 ml solution of 20 mM HCl
with 2 M NaOH. The reference electrode was (&) miniature
liquid-junction reference electrode, and (*) commercialized
macroscopic reference electrode (Horiba 2080A-06T). The same
platinum black on the chip was used for both the cases.
111
does not mean that the hydrogen electrode itself is
applicable to a micro pH sensor. It was just used as an
example of a potentiometric sensor using platinum as
an indicator electrode.) The titration curves for both
cases showed a good coincidence in terms of their
potential values in particular (Fig. 6). Notice that the
coincidence can be obtained only using a liquid-
junction reference electrode, which exemplies the
applicability of the miniature liquid-junction refer-
ence electrode to potentiometric sensing.
4. Conclusions
A miniature liquid-junction reference electrode was
employed to integrate the three-electrode system on a
chip. Evaluation of the system by cyclic voltammetry
and as an amperometric glucose sensor conrmed that
the system can actually be used in real electrochemical
sensing. Our present study showed a possibility that a
miniature disposable three-electrode system with a
clean working electrode can be provided by mass
production. The results given in Figs. 3 and 5 showed
another new possibility. Namely, incorporation of an
electrode system or microsensors in a micropipette.
Measurements could be made immediately on the spot
after suction of the sample solution and no further
procedure such as an injection of the sample solution
into an analytical system is necessary. Also, it was
shown that the miniature liquid-junction reference
electrode was applicable to one-chip potentiometric
sensors. If the lifetime of the reference electrode is
elongated, the sensors will be used for practical use.
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