Ann Clin Biochem 1985; 22: 46D-488

Review Article

Electrodes In clinical chemistry*

A D HI R S T t and J F S T EVE N S :j:
From the i Department of ChemicaL PathoLogy, Kings College HospitaL, London SE5 and
i Department of ChemicaL PathoLogy, St Stephen's HospitaL, London SWlO, UK

It has been possible to detect salts and bases in one of its salts, some metal ions enter solution,
solution by electrochemical means for many leaving electrons behind on the metal strip (Fig.
years. This technique, which has proved to be 1). The result of this dissociation is a small
particularly useful for the analysis of simple negative charge on the metal (b-) and a similar
aqueous solutions, was found to have limita- small positive charge in the solution. This in
tions in the analysis of biological fluids. The turn causes metal ions to tend to return to the
most serious limitations were the lack of sensi- metal strip; eventually a dynamic equilibrium is
tivity and specificity of detectors, and a varying established.
relationship between calibrants and samples. The potential between the metal strip and the
Other problems associated with electrodes were solution, which is produced by the small charge
related to the non-linear response and the time differences is known as the electrode potential.
taken to respond to a change in concentration It is dependent on the nature of the metal and
of an analyte. the concentration of the salt. No current can be
Considerable progress has been made in the drawn from this system alone-it forms a half
improvement of specifity, or selectivity, of cell. If a similar half cell with a different
electrodes. Modern electronic techniques and concentration of salt is connected to the first,
microprocessors have reduced the problems of with an electrical conductor between the two
sensitivity and non-linear response. Such im- solutions (eg a 'salt bridge') and the two metals,
provements have led to the introduction of the difference in electrode potential between
analytical techniques which have certain advan- the two half cells can be measured with a
tages over conventional techniques. The prob- suitable voltmeter. The potential difference or
lem of calibration has not yet been solved, and voltage of the cell (two electrically connected
there is confusion caused by an apparent half cells), when a minimal current is flowing,
inaccuracy of results produced by these techni- relates to the difference in concentration of the
ques, when compared to results using conven- solutions. The relationship is described by the
tional photometric analysers. It is possible that Nernst equation:
electrochemical methods will gradually replace
conventional techniques because they offer the RT al
E=-log., -
advantage of speed and sensitivity, and because nF a2
they do not require optically-clear solutions.
The following is an account of the principles
of electrode action and the possible applications 8-
<,
of electrodes in clinical chemistry. Zn
8+
Electrochemical ceUs -,
ELECTRODE POTENTIALS Zn 2 +
--~
Jr
- - -
When a strip of metal is placed in a solution of
- - "-'- Zn 2 +
- - - +2e-
- SO 42- - - - - - -
"This review was prepared at the invitation of the - - - - - - -
Scientific Committee of the Association of Clinical - - - - - - -
Hi. chemists (Instrumentation and Data processing
sub-committee'Lbut does not necessarily reflect the
views of the Scientific Committee. FIG. 1. A zinc half cell.

460

where E is the potential difference of the cell
for given chemical activities, at and a2, of ions
in the two solutions. The standardisation of one
of these electrodes would enable a practical use
of this system for estimating the chemical
activity of unknown solutions. To this end, the
hydrogen half cell has been adopted as the
primary reference electrode.
THE HYDROGEN HALF CELLI
This electrode is, in principle, the same as the
zinc model discussed earlier, or any other metal
electrode, with hydrogen replacing the zinc and
hydrogen ions in solution replacing the zinc
ions. Hydrogen, being a gas, cannot be used
directly as an electrode. The electrode consists
of platinum, which is inert as an electrode,
coated with finely divided platinum (platinum
black), which is in contact with hydrogen gas.
In Figure 2, which shows two hydrogen half
cells connected by a salt bridge, the left-hand
side cell is' accepted as a reference, against
which the electrode potentials of other cells are
measured u'nder all conditions. The reference
electrode consists of platinum-platinum black
in contact with hydrogen gas (bubbled past) at a
pressure of one atmosphere, in a solution of
1 M hydrogen ion activity. The right-hand cell
contains a 0·1 M H+ solution which is being
compared with the reference.
In the solution, hydrogen molecules are
adsorbed onto the platinum black and disso-
ciate into atoms which may then ionise. The
following equilibrium is established:
Pt
H2 _::
.. _ -
I atmosphere
FIG. 2. Two hydrogen half cells.
Electrodes in clinical chemistry 461
H2 ~ 2H ~ 2e- + 2H+
hydrogen platinum surface solution
gas (negative) (positive)
At equilibrium, the rate of ionisation of hy-
drogen atoms is proportional to the hydrogen
molecules adsorbed onto the platinum, and
therefore to the pressure of hydrogen gas in
contact with the electrode. Stability is main-
tained by bubbling hydrogen gas at constant
pressure around the electrode, replenishing the
hydrogen consumed by adsorption and ionisa-
tion.
While hydrogen electrodes are said to be not
too difficult to construct.i they are not suitable
for routine measurements of pH, or the concen-
tration of any other ion. The use of the
hydrogen electrode is reserved as a reference
standard for the measurement of relative elec-
trode potentials of other half cells. A suitable
replacement would be an electrode with a
known potential relative to the hydrogen elec-
trode which is simple to use and stable over
long time periods. This type of half cell is called
a reference electrode, as opposed to the hy-
drogen electrode which is called 'the reference
electrode' .
REFERENCE ELECTRODES
For electrodes to be used as sensors, two
connected electrodes (or half cells) are required
to enable electrochemical measurement to be
made. The most practical arrangement is to
have one half cell which has a constant potential
00
o
o 0
:.0 II"
H+
462 Hirst and Stevens
(a reference electrode), against which the
potential of a second, variable half cell (a
measuring electrode), is compared.
The most common type of reference elec-
trodes in use are two chloride electrodes:
Ag;AgCIJ0·1 M HCI (silver-silver chloride),
and
Hg;Hg2Cl2/3·5 M KCI (mercury-mercurous
chloride-calomel) .
Both silver chloride and mercurous chloride are
highly insoluble in aqueous media.
For the calomel electrode at 25°C, variation
in the electrolyte concentration would, as ex-
pected, produce a variation in potential. The
standard potentials, relative to the normal
hydrogen electrode are.:'
+0·3376 V for 0·1 M KCI,
+0·2848 V for 1 M KCI and
+0·2458 V for saturated KCI.
A CELL
A cell consists of two half cells which are
electrically connected to enable electrochemical
measurements to be made. Each half cell has an
electrode potential (but cannot produce a cur-
rent); it is the potential difference between the
two, when connected together, which we are
able to measure with a millivoltmeter.
For the measurement of a potential differ-
ence, electrical continuity is required within the
cell. To satisfy the conditions of Ohm's law, the
measured voltage, or potential of a cell is
determined as the product of the current and
resistance of the circuit (E=IR). If there was a
lack of electrical continuity, the resistance
would be "infinite and the current would be
zero. The resulting potential would be indeter-
minate. For a cell potential to be measured, a
small, finite current must flow around the
circuit. In practice the current may be as low as
10- 10 amps.
A salt bridge is usually chosen for electrical
contact in the model shown in Figure 2. A
direct contact between the two solutions with
flow restrained by a ceramic plug will also
suffice. This type of contact, which will be
described later, is widely used.
Under ideal conditions, the potential differ-
ence of the cell, which should be measured at
minimal current, should be equal to the differ-
ence in electrode potentials of the two cells. In
practice, a third potential created at the june-
tiO:1S between the salt bridge and the two
solutions also contributes to the total potential
difference of the cell. This potential, called 'the
junction potential', will be discussed later.
THE NERNST EQUATION
Assuming that the junction potential is negli-
gible, the hydrogen cell may be used for the
estimation of hydrogen ion activity (or pH) in
unknown solutions. The relationship between
the potential differences obtained and activity
of hy.dro§en ions is defined by the Nernst
equation."
EMF=E reference-E unknown
RT (H+reference)
= - log,
nF (H+unknown)
R=the gas constant (8,315 J/molJdegree);
T=absolute temperature;
n=valency charge;
F=Faraday (96 500 coulombs).
If the reference and second half cell solutions
are 1 M and 0·1 M for hydrogen ions respec-
tively, then at 25°C:
8·315xT (H+reference)
EMF- ·2·303·log lll -----:----
1x96500 (H+unknown)
=0·OOO1983·T·(logI01-logI01O- 1)
=0·0591 (0+ 1)
=59·1 mV
ie a lO-fold (decade) difference in activity for a
monovalent ion at 25°C will produce a potential
difference of 59·1 mV for a perfect electrode.
This relationship applies at all concentrations at
which the Nernst equation can be applied to the
response of the electrode. The relationship
applies equally well to metal electrodes such as
the zinc model shown earlier (though changes
in activity rather than concentration will be
reflected by this relationship).
The Nernst equation shown here is only
applicable to an ideal cell, in which there are
only two sources of potential. There are other
potentials which must be incorporated into the
equation, one of which-the junction potential--
is caused by the introduction of the salt bridge.
JUNCTION POTENTIALS
The junction potential arises at the junction of
two dissimilar solutions when the mobilities of
some of the ions at the interface differ, resulting
 Electrodes in clinical chemistry 463
I
I En
M+
I
(. I
I
, M+
I Relative error=---, where n is the valency.
0·2568
I x-I-' x-
I I I There are three approaches used to reduce
M+ • I I M+
I M+ x- the error due to the junction potential.
I x-~ x- To change the nature of the electrode. One
I'. , M+
M+ I I
alternative to using the calomel electrode is to
I X-~ X- use a second measuring electrode as depicted in
I I Figure 4, immersed in a solution similar to the
-----
Charge separation,
giving rise to
one being analysed. An attribute of reference
electrodes such as calomel or silver-silver chlor-
junction potential (Ej) ide is their low internal resistance (impedance).
The impedance of an electrode is in parallel
FIG. 3. Representation of a liquid junction of a with the input impedance of the voltmeter used
solution of M+ X- with water. If M+ diffuses more
rapidly than X-, a potential will be established at the for detection. The two impedances should be
junction (Ej). matched," otherwise one will short the other (to
ground). Standard pH meters, used as millivolt-
meters for most ion-selective electrodes, have a
in a charge separation at the interface, as high-impedance input for a high-impedance
depicted in Figure 3. The potential of the cell glass electrode, and a low-impedance input for
described will now be: a reference electrode.
If a high-impedance (eg glass) electrode is
E=Em-Er-Ej used in place of a calomel or silver-silver
chloride reference electrode, a special volt-
where Em is the measuring electrode potential, meter with two high-impedancevinputs would
Er is the reference electrode potential and Ej is be required.P '
the junction potential. To change the physical shape of the electrode.
The junction potential may not only be large, The conventional reference electrode, which is
but may also fluctuate by several millivolts. a vertical calomel electrode with a ceramic plug
Since, for a monovalent ion, a change in at the bottom forming a junction, is far from
potential of 59·1 mV represents a decade (10- ideal. This electrode, which owes its popularity
fold) change in concentration, an error of 1 mV to hardiness and low KCl leakage (approxi-"
would produce an error in concentration terms mately 50 IJ.Uday), may be acceptable for the
of approximately 4%4 measurement of large changes of an ion, eg an

Like electrodes

-: FIG. 4. A system employing similar electrodes

for both sample and reference electrode.

/
Unknown
solution
Liquid junction
\
Constant solution
similar to unknown
464 Hirst and Stevens

H+ change of two decades, with a correspond-
ing change in EMF of approximately 120 mV.
The detection of small changes of, say,
0·2 mmolJL of potassium at a concentration of
5 mmol/L, is not so easy. This is equivalent to
approximately 1 mV EMF compared with a
possible junction potential variability of several
millivolts.
The size and variation of the junction poten-
tial may be reduced by having the KCI below
the test solution, and by the introduction of
cylindrical symmetry. 2 Several alternative
liquid junctions have been tried on a commercial
basis, most of which have a more stable
potential than the ceramic plug. A few of these
are illustrated in Figure 5. The advantage of
these junctions is that they reduce the effects,
mentioned earlier, of gravity and symmetry.
To change the composition of the electrolyte.
It is possible that, by correct choice of electro-
lyte in the reference electrode, the junction
potential may be kept constant or reduced to a
negligible voltage. The following three solu-
tions to this problem have been proposed:
(a) use of a double junction. eg
Pt;H2!S!! NaCI(0'15 M) !!KCI (sat)! Hg 2Cl2;Hg
(S=sample,!! = junctions, ; = phase boundary);
(b) addition of an inert electrolyte to the
sample as used in the Harned cell for the
determination of the pK of a weak acid. eg
Pt; H 2! S, Na2Cl04!! HCl!AgCI; Ag
(S = sample,!! = junctions,; = phase boundary);
(c) use of salt M+X- where M+ and X-
have similar mobilities, eg KCl.
A titrimetric system has been described" in
which both options (a) and (b) were used,
together with similar electrodes (fluoride) for
both sensor and reference,
LaF3 !Fx, KN0 3 (0·1 M)!!KN0 3 (0·1 M) !! KN0 3
(0·1 M), Fs!LaF

Calomel
electrode

Calomel
electrode

KCL
Porous
ceramic
plug Liquid
junction Cellophane
membrane

(a) (b 1

Ag/AgCL
reference
Liquid
junction
~_ISE
Buffer =..;....
....".--~~----.....,r------.=
~
-.=-- Sample

(c 1
Waste

FIG. S.. Ex.mples of alternative reference cell junctions: (a) a conventional dip electrodewithstaticsample and
KCI; (b) a ellophane membrane junction (Instrumentation Laboratories), with stopped flow sample and static
KCI; (c) a.liquid. junction (Technicon Ltd) with flowing sample and reference buffer.

(Fx and Fs are unknown and standard fluoride
solutions).
In this system, the left-hand side of the cell is
kept constant, and the right-hand side is titrated
with fluoride to produce a null reading. Of the
three solutions, the most common by far in
clinical use is the third, in which the reference
half cell consists of a calomel electrode
(Hg;Hg2Cl2 ) in contact with saturated or 3·5 M
KCI. Potassium chloride is chosen because K+
and Cl" have similar (but different) mobilities.
The choice of concentration is influenced by the
finding that for a single junction cell, the
junction potential decreases with increasing
KCl concentration.?
A drawback associated with the calomel
electrode is that it exhibits a temperature
hysteresis effect. While this effect is not very
large within the temperature range encountered
in clinical laboratories (2D-37°C) , the use of this
electrode is contra-indicated if wide variations
in temperature are likely to be encountered.
This electrode is unsuitable for use in chlor-
ide measurement systems. An alternative is
Hg;HgS0 4lK2S04(saturated), though this elec-
trode has larger and more variable junction
potentials." An alternative reference electrode
in common use is silver-silver chloride in
contact with 0·1 M HC!. This electrode has a
stability similar to the calomel electrode, but
also exhibits a temperature hysteresis effect.
Silver-silver chloride electrodes are usually
chosen as direct-contact electrodes in gas-
diffusion electrodes, such as the CO 2 and NH 3
electrodes, and in continuous flow systems (Fig.
5). Silver-silver chloride electrodes also have
photo-electric properties and should be
shielded from strong light.
If large variations in temperature are to be
Gloss
electrode
Unknown solution
Electrodes in clinical chemistry 465
encountered, a suitable reference electrode
may be the thallium-amalgam thallous chloride
electrode. It has been claimed that this elec-
trode does not exhibit a temperature hysteresis
effect. This electrode is commercially available,
but it is not in common use in clinical measure-
ment systems.
To summarise, for the choice of reference
electrode in the detection of small changes in
concentration of an ion, the following points
should be considered for low, constant liquid-
junction potentials:
(a) there should be a good liquid-liquid
junction, constantly replenished with cylindri-
cal symmetry if possible;
(b) the reference electrolyte should be satu-
ratedl3·5 M KCl if a calomel electrode is
chosen with a static system or a solution similar
in composition to the test solution if a silver-
silver chloride electrode is chosen with a
continuous flow system;
(c) the entire measurement system should be
isothermal."
It should be noted that if the choice of
reference electrode is inappropriate, this elec-
trode is as likely to be the cause of problems as
is the measuring electrode.
MEMBRANE POTENTIALS
The half cells described up to this point consist
of simple electrodes in direct contact with a salt
solution. These electrodes may be useful for the
analysis of pure aqueous solutions, but are of
no practical value in dealing with the problems
of analysis of biological fluids, or any multi-
component solution. When placed in a multi-
component solution, the electrode will no
longer respond specifically to the ion under
consideration, but will respond with differing
FIG. 6. An electrode with an ion-
selective membrane (glass), used in
conjunction with a reference
electrode.
466 Hirst and Stevens
characteristics to all ions of the appropriate
charge in the solution.
The selectivity of response of an electrode to
an ion is improved by placing an ion-selective
membrane between the multi-component solu-
tion and the electrode. Figure 6 shows an
example of such an electrode. The characteris-
tics of membranes, such as selectivity, will be
described later. In this section, the potentials
arising in different parts of the cell are under
discussion, and the membrane is described here
because it is the source of another potential-
the membrane potential.
When a membrane electrode, permeable to
metal ions M+ (Fig. 7) is dipped into a salt
solution M+ X- , there will be a momentary flux
of M+ ions through the membrane from the
stronger to the weaker salt solution. The
anions, X-, being unable to migrate through
the membrane, will remain in situ, the two
populations separated by the membrane. A
charge separation will occur between the
mobile M+ ions entering the membrane and the
static X- ions remaining in solution. This will
result in a potential in the same way that a
charge separation is responsible for junction
potentials.
Since electrical neutrality must be main-
tained, migration of the M+ ions will quickly
cease, opposed by the potential set up between
the membrane and solution (the membrane
potential). The potential will be related to the
differences in activity of M+ on each side of the
membrane. There will be no further net move-
ment of M+ ions from one side of the mem-
brane to the other.
The equilibrium established at the membrane
M+
I J.l.~t
-
t-
Hydrated layer
----------------
4~
M+
Dry layer
Membrane-salut ian
interfacial
patent ial
EKternal salut ian
surface will be changed if a current is drawn
from the cell. It is for this reason that a high
input impedance voltmeter is required.
The membrane potential (Em) has a Nern-
stian type of response:
RT al
Em=-log.,
nF az
where al and az are the activities of the salt on
either side of the membrane.
This equation is an over-simplification of a
complex situation, in which variables such as
diffusion rates and effects of interfering ions are
not taken into account (to be described later).
The point of. this simple model is to show
that, by introducing a selective membrane, the
potential which will vary with the concentration
of the ion under consideration is no longer at
the electrode surface, but across the selective
membrane. Although the nature of this poten-
tial is not quite identical with the electrode
potential, and may vary with the nature of the
membrane, the response may be described as
Nernstian under some conditions. The variable
component of the potential of conducting mem-
branes is the membrane--external solution in-
terfacial potential.
STREAMING POTENTIAL
Liquids flowing through tubes, as encountered
in continuous flow analysers, may give rise to
streaming potentials." The potential produced
is dependent on several factors including the
flow rate, di-electric constant and length of the
tube, and is also inversely proportional to the
FIG. 7. An expanded representation
of a glass membrane.

conductivity of the solution. Peristaltic pumps
produce fluctuating streaming potentials which
may be detected by the electrode. It is often
necessary to remove these potentials from
continuous flow systems by electrical isolation
and/or shorting from the liquid to the shielding
of electrode.
SUMMARY
The EMF produced by a membrane electrode is
composed of internal electrode potentials, Et,
Er; a junction potential, Ej; and a membrane
potential, Em (Fig. 8). For a test solution and a
cationic electrode, the EMF generated, Ex,
may be described in simple terms by the
equation:
Ex=Et-Er+Ej+Em.
RT
If Em=- loge ax, where ax is the activity
nF
of Ion X:
RT
Ex=(Et-Er+Ej)+- log, ax
nF
RT
Ex=E u + - log, ax or, for an anionic
nF
responsive electrode:
RT
Ex=Eo - - loge ax.
nF
EO is a function of the internal potentials and
junction potential, and is only constant if
temperature and junction potentials are con-
stant.
Ion selective electrodes
Electrodes have been constructed which re-
spond in a relatively specific manner to a
chosen ion. By varying the composition of the
membrane, a large variety of electrodes have
been produced with a selectivity which gives
them a useful role in clinical areas.
There are three basic types of membrane: the
solid state, the liquid ion exchanger, and the
neutral carrier membranes. The mechanism of
action of these three types differ, particularly
with respect to selectivity and establishment of
membrane potentials. The end result, however,
which is important to clinical chemists, is that
Electrodes In clinical chemistry 467
Test electrode Reference (calomel)
electrode
~ ,
t- 1-
d
~
Er ~
Et
Em Ej
FIG. 8. A typical membrane electrode-reference
electrode, with an indication of the potentials in-
volved.
they have a Nernst type of response over
a limited range of concentration, typically
10- 5_10- 1 M.
SELECTIVITY
Electrode response
All electrodes in current use respond in varying
degrees to more than one ion. It is sometimes
possible to suppress the contribution of inter-
ferents by modifying measurement conditions
(eg pH). For clinical applications, it is often
difficult or inappropriate to change conditions,
or remove interferents. It is important, there-
fore, to be aware of the response characteristics
of an electrode.
The membrane potential of an electrode
includes contributions from all the ions to which
the membrane is responsive, each weighted by
the selectivity of the membrane to that ion. In a
simple form, the Pungor generalisation of the
Nicolsky-Eisenrnan" equation may be written
as:
RT
EMF=Eo+-loge(Ai+Sijl·Ajl+Sij2·Aj2 .... )
nF
where Ai is the activity of the major ion being
measured, Aj 1,Aj2, etc are activities of in-
terfering ions of the same charge, Sijl , Sij2, etc
468 Hirst and Stevens

are the selectivity constants of the membrane ion exchange preference (Kij) for potassium
towards intefering ions. over sodium is approximately lOO-fold, but this
The contribution of each ion to the mem- is counteracted by a membrane mobility of
brane potential is the product of the activity of sodium ions which is 10 times greater than
the ion and the selectivity constant for that ion. potassium ions. The result is a lO-fold selectiv-
Selectivity constants are relative to the major ity for potassium over sodium.
ion (Ai), the selectivity constant of the major Liquid ion exchange membranes have a
ion being unity. rather more complex mechanism for the deter-
This relationship is an approximation, and mination of selectivity, while for neutral carrier
may vary with the type of membrane and membranes the selectivity constant is simply
composition of the electrode internal solution. related to the equilibrium state."

Selectivity constant SOLID ION EXCHANGE MEMBRANES

The selectivity constant may be defined and Glass membranes'"
estimated in several ways, depending on the The most common electrode of this type is the
type of membrane. For a solid ion exchange glass electrode, used as a sensor for H+ or Na+
membrane (eg glass), the selectivity for the in biological solutions. Glass membranes con-
membrane between two ions 1+ and J+ is sist of a three-dimensional matrix of a varying
dependent on the mobility of both ions in the mixture of oxides of elements such as silicon
membrane, and on the equilibrium constant and aluminium (oxidation state III), with
(Kij) of the exchange of ions between the oxides of elements such as sodium or calcium
membrane and aqueous phase.? (oxidation state I or II). The most common type
of glass used is sodium aluminium silicate. The
V'
.. ] K"IJ
SIJ=-X membrane acts as a cation exchanger; monova-
Vi lent cations (eg H+, Na+) being the most
mobile charged species in the matrix.
where Sij is the selectivity constant, Vi and Uj
are the mobilities of ions in the membrane, and
The hydrogen electrode
Kij is the equilibrium constant of the reaction,
A thin glass membrane consisting of three
Kij layers separates two solutions of different hy-
J+ (membranej--I" (aqueous) drogen ion activity (Fig. 9). After a conditioning
:;:=:J+ (aqueousj--I" (membrane). period the two outer layers become hydrated,
while the middle layer remains dry. Hydrogen
In the case of the glass potassium electrode, the ions from the internal and external solutions

Hyd rated lcyers

'" '"
'"
/ \ I
I
" I
"
"" ""
....
I
I
....
..... I
Glass electrode ..... H+ H+ I
I
H+ H+
in standard .....
.... I
I
or unknown .... I I
solution
Glass membrane
(expanded)

~IG. 9. A glass electrode responsive to hydrogen ions, showing the differing potentials on either side of the dry
layer.

exchange with hydrogen ions in the two hy-
drated layers, giving rise to a membrane poten-
tial.
The membrane p, tential is related to the
difference in hydrogen ion activity on either
side of the membrane:
where a1 and a2 are the activities of H+ on
either side of the membrane.
If the hydrogen ion activity of the internal
solution is constant, then
RT
E=const+- loge a1
F
Although there may be a small, but finite,
current when used in conjunction with a refer-
ence electrode, hydrogen ions do not travel
across the membrane. There is movement of
ions through the hydrated layer, but the current
in the dry layer is thought to be mediated by
charge transference, eg from one atom to an
adjacent one, with a small amount of move-
ment.
Glass electrodes have a very high resistance
(600-900 Mohm), and require measuring de-
vices with a high input impedance.
The sodium glass electrode
At a low H+ concentration relative to sodium,
the sodium component of the membrane poten-
tial of a hydrogen ion responsive glass electrode
will become significant. 10 This effect is known
as the 'alkali error'. II The alkali error may be
exaggerated by modifying the composition of
the glass matrix to such an extent that under
non-acid conditions, the electrode is highly
selective towards sodium.
Other glass electrodes
Several other glass electrodes have been pro-
duced, but low specificity, sensitivity or stability
limits their suitability for clinical use. Silver and
lithium electrodes are claimed to be among the
better group. III
Solid state crystal membranes
It is possible to extend the range of ions which
can be estimated by substitution of the glass
membrane with a crystal lattice. A few crystals
share the properties required for use as a
Electrodes in clinical chemistry 469
membrane. To be suitable for use as a
membrane.F the crystal should be:
(a) chemically inert;
(b) highly insoluble;
(c) non-porous;
(d) mechanically strong in thin sections.
The crystal should also be capable of watertight
sealing to an electrode body and have a rapid
Nernstian response.
The mode of action of these membranes is
not quite the same as the ion-exchange phe-
nomenon of glass. Conduction through the
crystal occurs by the movement of mobile ions
into adjacent defects in the crystal lattice. The
size of crystal defects restricts the movement to
one particular species of ion. Ions which are the
wrong size or charge cannot move in this way,
and hence cannot contribute to the conduction.
Membranes constructed from crystals with a
very low solubility product tend to be highly
selective. The smaller of the favoured ions tend
to be the most mobile. This has a bearing on the
mechanism of action of the electrode which will
be discussed in the section on halide electrodes.
Crystals of lanthanum fluoride and silver
sulphide have the necessary properties for use
as membranes. Silver halides alone, although
highly insoluble, are susceptible to photo-
electric effects. Silver chloride membranes
have, additionally, a relatively high electrical
resistance which may present voltage measure-
ment problems.
The lanthanum fluoride electrode
This electrode.':' (Fig. 10) may be used as a
sensor for lanthanum or fluoride, and is the
~ Internal electrode
PTFE
Internal solt solution
0·1 M F-
0') M CC
1
LoF 3 crystal
·r
FIG. 10. A crystal membrane electrudr .(LaF x)·
470 Hirst and Stevens

basis of a method for the estimation of F- in acts as an inert matrix for the silver halide
blood and urine.!" The membrane consists of a which is in equilibrium with the sample solution
thin crystal of LaF] , sometimes doped with at the membrane-liquid interface. The silver
traces of europium (Eu3+) to increase the ions, being smaller than halide ions, are the
conductivity of the crystal. mobile species within the membrane.
Crystal lattice membranes suffer interference At the membrane surface, the activity of
from ions of like size and charge to the one silver in solution (aAg ") is given by:
being measured. In the case of fluoride, the
greatest interference is from OH-. This in- K (AgX)
terference is suppressed by buffering the a Ag"
sample at a pH 5-5·5 using total ionic strength
adjusting buffer (TISAB).15
where aX is the activity of halide, and K (AgX)
is the solubility product. The EMF of a halide
Silver sulphide-silver halide membranes
electrode is:
Silver sulphide is well suited to the re~uire­
ments of a crystal lattice membrane. 2 In
RT
addition to suitable physical properties, silver E=EO+- loge a(Ag+)
sulphide is not readily oxidised or reduced, and F
is extremely insoluble. An electrode with a
RT K (AgX)
silver sulphide membrane is capable of detec- =El)+ _ log
tion of Ag+ or S2- down to a concentration of F e aX-
the order of lO-H M. l3
RT
The use of pure silver halides for the = E lI ' - - log aX
construction of halide selective membranes is F e

associated with problems, but when silver

halides are incorporated with a silver sulphide
ie the electrode responds to the halide activity.
matrix, a series of stable and relatively selective
There is a clinical use for the chloride electrode
halide electrodes may be produced, as shown in
in the measurement of plasma and sweat'" 17
Fig. 11. In these membranes, the silver sulphide
chloride. Little use has been made of this
electrode for plasma estimations, possibly due
to the effect of proteins. The membrane should
be protected from direct contact with protein
with a semi-permeable membrane, such as
cellophane, to prevent precipitation or adsorp-
tion of protein on the membrane surface.
Internal
electrode
Careful consideration should be given to the
choice of reference electrodes when estimating
chloride, since a saturated KCL salt bridge may
contaminate the sample. A double junction
Internal reference electrode is often used, with, eg, a
0+- f-
elec t ro Iyte potassium-nitrate bridge in contact with the
specimen.

Silver sulphide-metal sulphide membranes

. =f 0·1 mm
Replacement of the silver halide with a metal
.. sulphide may produce a membrane which is
responsive to the metal in question.V Elec-
0'5-1 cm
trodes have been produced using this principle
I which are responsive to copper, lead and
cadmium. These electrodes have a limited
clinical application because, even if they had
Compressed disc of a mixture the required sensitivity for blood concentra-
of Ag 2S and AgCL tions, the specimens would require pre-
FIG.11. A halide. electrode with a mixed Ag 2S/ treatment to release the metals from the proteins
AgCl membrane. to which they are bound, or from erythrocytes.
 Electrodes In clinical chemistry 471

LIQUID ION EXCHANGE ELECTRODES

Liquid ion exchange membranes
A liquid ion-exchange mernbrane'" consists of a
water-immiscible solvent which contains an
ionisable species preferentially soluble in the
solvent phase, eg, a fatty acid. This membrane
separates the inner electrode solution from the PTFE body
outer sample solution. For physical stability,
the membrane is often supported by an inert
porous matrix such as cellulose acetate. This
~embrane .differs from the solid ion-exchanger
In that the IOn exchange sites are mobile within
the membrane.
If the membrane separates two aqueous
solutions of a salt M+ X-, as illustrated in
Figure 12, the cation exchanger, S-, will be
Exchangeable
capable of selectively binding and transporting
end-cap
the M+ ions across the hydrophobic membrane. Ag/AgCl
The X- ions on either side of the membrane --.Q<'"
internal Internal
electrode solution
will be excluded and will remain in situ. In a CaCl 2 (10- 3 hi)
~imilar wa~, the cation exchanger, ideally being
Insoluble In water, will remain trapped within
the membrane. Membrane
FIG. 13. A calcium electrode with a liquid ion
Selectivity exchanger membrane.
The behaviour of the membrane in determining
the selectivity of the electrode is complex and
The calcium selective membrane consists of the
varies with the solvent. A solvent with a high
calcium salt of an alkyl phosphate dissolved in
di-electric constant will contain a dissociated
di-n-octylphenyl phosphonate, supported in an
ion exchanger. In this case the selectivity of the
inert matrix such as PVc. 19
membrane is dependent only on the solvent. As
The calcium electrode has sufficient selectiv-
the di-electric constant of the solvent is de-
ity over magnesium, sodium and potassium to
creased, the association of ion and ion-
be useful for serum calcium estimation, particu-
exchanger increases, and the relative mobility
larly free (ionised) calcium. These electrodes
of ions and interfering ions (and binding sites)
have a high resistance (approximately 10
become more important, though not to the
Mohm), a detection limit of 10- 5 rnol/L Ca 2+
and a slope of 25-29 mV per decade change i~
same extent as in solid ion exchangers.
concentration. (Calcium is divalent.)
The calcium electrode
The electrode of this type in most common
NEUTRAL CARRIER ELECTRODES
clinical use is the calcium electrode (Fig. 13).
Neutral carrier membranes
Membrane Solution 2
Analysts have long envied the ability of living
Solution I
cell membranes to transport ions and small
compounds, often against opposing concentra-
tion gradients, with a very high selectivity.
Progress was made towards reproducing this
characteristic with the discovery that macro-
x- _ _ S----..J- x- cyclic antibiotics caused an increased ion per-
meability of cell membranes.r'' Membranes
1~ constructed of the antibiotic in phospholipid
bilayers or organic solvents exhibit the same
1 - - - MS - - - - . I
property. The antibiotic, valinomycin, and a
synthetic analogue, dibenzo-18-crown-6 (a
"!'
FIG. 12.. liquid ion exchanger membrane, showing 'crown compound'S")' (Fig. 14), are ring com-
permeability to cation M+. pounds containing several ring oxygen atoms.
472 Hirst and Stevens
valinomycin
dibenzo-18-crown- 6
FIG. 14. Valinomycin and dicyclohexyl-18-crown-
6. 4
The oxygen atoms may act as a substitute for
the hydration shell of some cations, eg, potas-
sium, rubidium and caesium. These cations can
move from aqueous solution to the ring com-
pound and can be selectively transported
through a hydrophobic matrix containing the
compound.
Such complexing agents (unlike the ion-
exchange carriers) are electrically neutral, and
carry the charge of the cation in the complexed
form.
Selectivity
Assuming that the different ion complexes of
the macrocyclic molecule have similar mobili-
ties, selectivity is largely determined by the
relative affinity of cations for the carrier, and is
independent of solvent. The simplicity of this
function of selectivity should result in large
future expansion of the use of this type of
membrane.
The potassium electrode
This is the most common example of a neutral
carrier type of electrodef in clinical use. The
most Common carrier is valinomycin rather than
crown compounds, dissolved in one of several
solvents (usually nitroaromatic) supported in an
inert matrix such as PVC or silicone rubber
(Fig. 15). The potassium electrode has a
measuring range of 10- 5_10- 1 mmoUL and a
response of 57-59 mV/decade. The membrane
has a limited life because valinomycin is slowly
leached out into the aqueous phase. Substitu-
tion of the valinomycin by crown compounds
with hydrophobic substituents should eventu-
ally improve membrane life.
A summary of electrodes suitable for clinical
use is shown in Table 1.
Other electrodes
In addition to the basic ion-selective electrodes
described above, there are other types of other
electrode with a different mode of operation,
which may be direct, or, more often, indirect.
The three further categories are solid state, gas
dialysis and enzyme electrodes.
SOLID STATE ELECTRODES
The EMF generated by a cell consists of several
constant potentials and the membrane poten-
tial, which is variable. Since there is no require-
ment for the internal reference electrode of the
measuring half cell to produce a potential (this
being constant), attempts have been made to
produce direct contact between electrode and
membrane, omitting the internal aqueous
phase. A functional solid state sensor may be
produced if there is electron conduction
through the membrane and reversible electron
exchange to replace reversible ion exchange at
the inner surface of the membrane. The
membrane--external solution interfacial poten-
tial is the variable being measured. The most
r--t;:::::=l---l- electrode
Ag/AgCL
Internal
--+--+-- solution
0·1 M K+
Valinomycin
membrane
FIG. 15. A potassium electrode.
 Electrodes in clinical chemistry 473
TABLE 1. A summary of electrodes suitable for clinical use

Concentration Special
Electrode range (M) pH conditions Application

Ammonia 1~-Hf >11 adjust pH NH3 , serum, urine

(NH 3 ) NH3 , as a reaction
product, eg, urease
Carbon dioxide 1O-5_W--2 <pH 5 for total CO 2 PC0 2 , blood, serum
reaction product
Calcium 1O-5- Hf 6-8 Calcium, blood
serum, urine
Chloride 10-5_10° 2-11 Requires a double Chloride, sweat
junction ref., serum, urine
poisoned by protein
Fluoride 10-7_10° S-8 Sample requires Fluoride, serum, urine
buffering
Iodide 10-7 - 10" 3-12 1-, reaction product
Potassium 1(}-5_100 3-10 Requires a double Potassium, blood
junction ref. serum, urine
Sodium 1~-101l >pNa+3 Sodium, blood
serum, urine

Electrodes with limited clinical use Limitation

K+fNH 4+ 1O-5_Hi1 Low selectivity, interference from Na+, K+ and NH 4+

Lithium 1(}-5_100 Selectivity, interference from Na" (SN.=5·1O- 2 )
Other metals 1~-10" Most metals are protein-bound and would require sample
Pb, Cd pretreatment
Ag, Cu

stable readings are obtained if an electrode
membrane has dimensions (area, thickness)
which produce a DC resistance of 1-100 Mohm.
The input impedance of the measuring device
should be of the order of 1000 X the resistance
of the electrode."
The two most common variations of the solid
state electrode employ membrane contact
either via carbonlgra~hite, eg, the Pungor-
Radelkis electrode/!' 2. (Fig. 16) or directly to
metal. The physical form of direct metal contact
varies from electroplating of glass to coating a
metal wire with a membrane" (Fig. 17). Coated
wire electrodes have been produced for calcium
and potassium estimations, but are not used
routinely.

11-4-tt-- Ag
Interna I
t--H-- electrode
Glass

___ Graphite
Membrane
coating

- - - - Ag/AgCL
- Membrane
FIG. 16. A solid state electrode using a graphite FIG. 17. A coated wire electrode, consisting of
contact. anodised silver coated with a selective membrane.
474 Hirst and Stevens
ION SELECTIVE FIELD EFFECT
TRANSISTORS (ISFET)
Bipolar (normal) transistors are current-driven
semiconductors, ie, they amplify a current.
They have a low input impedance, and are not
suitable for the first (input) stage of an elec-
trode voltmeter. Field-effect transistors (FET)
are voltage-driven semiconductors; amplifica-
tion is related to the voltage applied to the
device which produces an electrostatic field.
FETs do not consume current, operating only
on voltage. They have, therefore, a high input
impedance, and are widely used for the first
stage of an electrode voltmeterv: 26 (Fig. 18).
The ion selective component of this sensor is
similar to membranes described earlier.
The elimination of intermediate processes is
taken one step further than with the coated wire
electrode by connecting the first stage of the
millivoltmeter (a field-effect transistor) directly
with the ion-selective' membrane. The advan-
tage of this arrangement is that the high
impedance of the electrode is transformed
directly to the low impedance output of the
transistor, eliminating the need for heavy
shielding.
The attraction of this approach is that with
current micro-electronic technology, it should
be possible to construct a cluster of electrodes
on one chip, for possible in vivo applications.f"
ISFETs have been developed for Na+, K+,
Ca2+, H+, and Nat and have been used in
vivo. Unfortunately development of this sensor
has been retarded by problems of electrical
insulation of the chip from the solution. The
lon-selective membrane
n!2!:i:~~ni4---Gate
FIG. 18. An ion-selective field-effect transistor. Two
areas of silicone with 'negative holes' (neg Si) are
separauI by one with 'positive holes' (pos Si).
Application of a potential to the gate will induce a
current to flow through the pos Si area.
present ISFETs have a limited active life (1G-20
days). Mass production of stable ISFETs would
provide inexpensive assays for many analytes.
GAS DIALYSIS ELECTRODES (NH3 AND COz)
In this type of electrode ,2M. 29 the gas to be
determined diffuses across a membrane, fol-
lowed by detection of the gas on the sensor side
of the membrane. In two cases (carbon dioxide
and ammonia) the gas changes the pH of a
buffer in contact with a pH electrode (Fig. 19).
Since the gas is neutral, movement across the
gas membrane is 'halted not for preservation of
electrical neutrality, but upon achievement of
equilibrium of the buffer reaction. For the CO2
electrode the reaction is:
The production of carbonic acid (first step) is
the rate-limiting step, and this, rather than
diffusion through the membrane, is the reason
for the slow response of CO 2 electrodes. The
rate of reaction may be increased by the
addition of carbonic anhydrase (eg, lysed red
cells), though this is not feasible for everyday
use.
The membrane is a gas-permeable hydropho-
bic material such as polypropylene, teflon, or,
more recently, silicone rubber. The ammonia
electrode incorporates nonactin, a macro cyclic
neutral carrier, in a polytetrafluoroethylene
membrane.'?
For clinical use, selectivity between CO 2 and
NH 3 is based on the pH of the external
solution, the ammonia electrode requiring an
Outer ----..; ~
body
pH electrode
Pt internal
electrode Ag/AgCL
l--- reference
I-
electrode
Buffer
solution -
~
Rubber a-ring ~ ~
f
Gas dialysis
membrane
FIG. 19. A carbon dioxide-ammonia electrode.

alkaline solution to convert NHt to the diffu-
sible NH 3 molecules. Both electrodes have a
useful clinical function. The CO 2 electrode may
be used for the estimation of PC02 and indi-
rectly, total CO 2 in a large variety of fluida."
The ammonia electrode may be used for the
estimation of plasma NH 3 , 32 and for the detec-
tion of NH 3 as the product of a primary
reaction. The plasma or reaction solution must
be made alkaline prior to ammonia estimation.
The two gas electrodes described respond to
the partial pressure of gas. The response over
small changes of concentration is almost Nern-
stian in that the internal pH electrode has
Nernstian response to changes in the H+
activity of the buffer, but deviates according to
the characteristics of the buffering system.
These gas dialysis electrodes are indirect, or
double membrane electrodes, in that the poten-
tial arises not at the gas-permeable membrane,
but at the glass membrane of the pH electrode.
In this sense, they are comparable with 'enzyme
electrodes', (see Enzyme and other indirect
electrodes) .
The oxygen electrode shares the characteris-
tics of the gas-permeable membrane, but has a
different detection system (polarography).
POLAROGRAPHIC ELECTRODES
Polarographic, or amperometric, analysis in-
volves the measurement of current produced in
electrochemical reactions. The electrodes dis-
cussed up to this point have been voltametric;
voltage is measured under equilibrium condi-
tions at negligible current. Polarographic
systems are designed to measure a current
produced by an electrochemical reaction at an
electrode surface. A polarographic electrode
Current (/Lamps)
+ The Clarke oxygen electrode in common clinical
Valtage (V)
FIG. 20. The relationship of current to voltage for
solutions with and without electroactive compounds.
Electrodes in clinical chemistry 475
will produce a current at constant potential, as
opposed to a potential at zero current. The
response of a polarographic electrode tends to
be linear, rather than Nernstian.
The relationship between voltage and current
which is seen in oxygen-free solutions at low
voltage becomes distorted in the presence of
oxygen and other electroactive compounds such
as paracetamol and catecholamines. Plateaux,
such as those shown in Figure 20, are found.
Reduction reactions occur at negative vol-
tages, and the current passing shows a plateau
at low voltage which corresponds to energy
consumption by the following reactions:
-0·7 V
02+2H20+2e----+H 202+20H-
+0·7 V
H 202 ---+ 2H+ +02+2e-
It is possible to estimate dissolved oxygen in a
solution by measurement of the current at a
constant potential of -0·6 to -0·7 V. The
current will be directly proportional to the
oxygen consumed which is itself proportional to
the P0 2 of the test solution. Peroxide may be
similarly estimated if a potential of +0·7 V is
applied. The earliest form of polarographic
electrode, the dropping mercury electrode,
suffers from many disadvantages, such as pro-
tein poisoning and lack of specificity, and is not
suitable for routine laboratory use. Alternative
electrodes which do not suffer from these
disadvantages are now widely used in clinical
laboratories.
A large number of electroactive compounds,
such as those shown in Figure 21/1 are detect-
able by this technique. Oxidation (positive
voltage) or reduction (negative voltage) condi-
tions, or a combination of the two, may be used.
If 100% of the analyte is oxidised (or reduced),
the technique is referred to as 'coulometric
analysis'.
THE OXYGEN ELECTRODE
use is an amperometric electrode based on
polarographic detection.'! Solid metal elec-
trodes are protected from protein poisoning by
a gas-permeable membrane (eg, polypropylene).
The dimensions of the Clarke electrode (Fig.
22) are critical. The diameter of the cathode tip
must be such that there is not excessive con-
sumption of oxygen. If oxygen consumption is
too high, diffusion nither than partial pressure
476 Hirst and Stevens

becomes rate-limiting. As gaseous oxygen dif- be difficult to measure. As a compromise, the

fuses more rapidly than dissolved oxygen, a gas
and a liquid with identical P0 2 will yield
differing currents. On the other hand, a very
small cathode will yield a low current which will
Reduction Oxidation
platinum tip will be 1-100 urn in diameter, and
will yield a current of the order of 10 uamps for
a normal blood Po 2 •
The oxygen electrode in this form is in
widespread use for P0 2 estimation of blood and
other aqueous solutions, and is used as an
oxygen detector in indirect systems (see En-
zyme and other indirect electrodes).

-Olefins

Esters
Hydrocarbons
4

Azinesl
triazines
•
The selectivity of the electrode for oxygen is
derived from both the membrane and the
polarographic detection. Some foreign volatile
• compounds, eg, halothane.P' have been re-
Aminesl
Ketones amides ported as interferents, but normal blood consti-
tuents do not affect the estimation.
Aldehydes Phenathiazines
• OTHER POLAROGRAPHIC ELECTRODES
Olefinic Peroxide
esters Phenols Amperometric detection of peroxide is the
basis of an indirect enzyme electrode for glucose

-
Organometalics Aromatic OH estimation which is in widespread use. (See
Enzyme and other indirect electrodes).

-Diazo

Nitro
Quinoli nes

Imines
Electrochemical detectors for HPLC
Polarographic, or 'electrochemical', detection
has been successfully used in HPLC for com-
pounds which would otherwise re~uire deriva-
Halogens Halogens
... .........-..... tisation for optical detection.P: 33, 5 In the case
I
I
of catecholamines, the reaction at the electrode
,
I is:
-2 -I 0 2
HO~R +O.5V O~R
(Volts vs calomel electrode)
FIG. 21. Electroactivity of organic compounds. HoN ----- oN + 2H+ + 2e-

Gold, and gold amalgam electrodes have

been commonly used for (oxidative) polaro-
graphy; gold amalgam being particularly useful
in detecting thiols. There is considerable interest
in clinical chemistry in the development of a
+lfH--Ag anod e carbon electrode with a highly polished, hard
surface (glassy carbon). Typical working ranges
of the three electrodes are shown in Figure 23.
Electrolyte Voltage stabilisation is necessary because when
a current flows there is a potential drop
between the two electrodes, which will vary
with the composition and flow rate of the
solution. A constant voltage is maintained by
the introduction of a third 'counter' electrode
t4t-f5I-:-- Pt cat hod e
into the measurement cell, with a potential
Rubber O-ring which is controlled from the reference electrode
via a feedback circuit. An example of such a
Gas permeable circuit is shown in Figure 24. Continuous use of
Site ",f reduction membrane a polarographic cell may result in the accumula-
(inside 'hemembrline)
tion and adsorption of reaction products on to
FIG. 22. A C!lirkeoxygen electrode. the electrode surface. The electrode will then
 Electrodes in clinical chemistry 477

become inactive. A further electronic refine- the HPLC solvent, and continuously purging it
ment to maintain the activity of the electrode is with nitrogen or helium.
to apply alternating pulses of a polarising
ENZYME AND OTHER INDIRECT
potential and a lower 'cleaning' potential at 36
E LECTROD ES
which the electrode reaction is reversed.
This section is concerned with electrodes with a
In practical terms there are two further
membrane in addition to that of the sensor. The
effects which should be considered for HPLC
gas-permeable membranes were described
applications. One is pulsation of the liquid flow,
separately because they allow gases to diffuse
producing streaming potentials which will be
unchanged through them. The membranes to
detected by the electrode. A pulse-free pump is
be described in this section allow the selective
essential for low background noise. The second
conversion of substances to products which are
effect is produced by the presence of dissolved
readily detectable by electrochemical means. A
oxygen. In the reduction mode, dissolved
prime requirement of this system is that either
oxygen will produce a high background current
the membrane or the detector must be selec-
(Figure 20). In the oxidation mode (positive
tive.
voltage), high noise signals are often found,
possibly due to the transient production of The urea electrode
electroactive species. Both the background One of the earliest enzyme electrodes, the urea
current and noise can be reduced by de-gassing electrode" consisted of an ammonium (NHt)
electrode enveloped by urease, as illustrated in
Figure 25. This electrode was unsuitable for
Glassy
car ban

u:;
c.
E
c
:L
....~ NHl glass
Illet radl

Cillophoni

0·1 0'8 1·6 Ureon

Valts
FIG. 23. Working ranges for three electrochemical
electrodes: response to thiols . FIG. 25. A urea electrode.

.....----.,-----{ A)-----,
Warking
electrade

Reference Pawer
supply FIG. 24. Circuit diagram for pro-
viding a constant potential for a
Caunter 3-electrode cell.
478 Hirst and Stevens

biological fluids, due to the poor selectivity of

the NHt electrode. More specific systems have
been described" using the more selective
ammonia electrode.
An instrument which is widely used (Beck-
man Instruments), for electrochemical urea N
determination is based upon the change in o
It
conductivity of a sample following the addition
of urease, due to the reaction:
NHz CONH z + 2H zD---+2NH 4+ -cos
The glucose electrode
The electrochemical detection of glucose is
based upon the action of glucose oxidase which 30 60
catalyses the reaction: Time (s)

glucose--Oj-e HzD---+gluconic acid+ HzO z FIG. 27. The change in Po; with time in a glucose
electrode exposed to air.
There are two types of glucose electrode in
common use, both of which use polarographic
The glucose-peroxide electrode. The
detectors. The first, and earliest, records the
electrode." illustrated in Figure 28 forms the
consumption of oxygen, or decrease in POz,
detector of a widely used glucose analyser
while the second records the production of
(Yellow Springs Instruments Ltd). The mem-
peroxide.
brane contains immobilised glucose oxidase at
The glucose-Paz electrode. The glucose
which site hydrogen peroxide is formed. The
electrode" as illustrated in Figure 26 may take
HzO z diffuses through the membrane to the
many forms. The glucose oxidase may be added
polarographic cell where it is detected at a
as a wet reagent, or may be immobilised eg on
potential of +0·7 V (see section on polaro-
to the stirrer. The drawback which applies to all
graphy).
these systems is that if the system is exposed to
This electrode does not suffer from the
~ir the oxygen consumed will be replenished,
problem of re-equilibration with atmospheric
ie, the POz will fall then rise until it is again in
oxygen, enabling the total reaction to be
equilibrium with air, as seen in Figure 27.
followed. The membrane is not, however,
Glucose is estimated by using relatively high
entirely selective for peroxide. The system may
glucose oxidase activity and detecting either the
respond to foreign substances such as
peak activity (point P), or the peak rate of
paracetamol. 3Y
reaction. Both approaches require a fast re-
Other enzymes. The use of the polarographic
sponding POz electrode, and may suffer poor
and the potentiometric type of enzyme elec-
precision.
trode. is ~ot restricted to glucose estimation. By
substituting the glucose oxidase with other

Reaction
chamber

Polarographic
......,;...q._ Reactian cell
chamber

Stirring bar P t - - - -....

~ -Membrane

/1 l~--- Oz detector
Ag-----iI

I----t
-
Double layer membrane
Pt cathode Annular silver with glucose oxidase
anode trapped between

FIe. 26. A glucose-Po, electrode. FIG. 28. A glucose-peroxide electrode.


enzymes, electrodes have been produced for
urate, cholesterol and ethanol.r"
The lactate electrode
This electrode'" 40 uses a yeast lactate dehy-
drogenase which catalyses the reaction:
lactate + 2Fe( CN)~ - ~pyruvate + 2Fe( CN)~- + 2H +
PI
2Fe(CN)~--->2.Fe(CN)~- -ze
The ferricyanide is detected by polarography
with an 80 mV polarising voltage.
Other enzyme/redox systems
Although many other enzyme electrodes have
been described, such as deaminating enzymes!
ammonia probes, few are suitable for routine
clinical use. Several amperometric techniques
for the NADH-NAD+ system have been des-
cribed, which could have wide clinical applica-
tions. Unfortunately the reduction of NAD+ at
the electrode surface is complicated by absorp-
tion and dimer formation. As a result, these
electrodes are not suitable for continuous use,
and are at present of academic interest only to
clinical chemists.
Future developments
(a) ISFETs. The range of ISFETs is expanding
and now includes enzyme and immunochernical
sensors. 26
(b) lmmunochemical sensors. If an
antibody-antigen reaction causes a change in
charge density, immobilisation of an antibody
could be used to produce a potentiometric
membrane. Attractive though this concept may
sound, practical systems have not yet been very
successful.
(c) Bipolar pulse conductance/" This techni-
que involves the excitation of an ion-selective
electrode using two equal and opposite 3 V
pulses lasting for 100 us. At the end of the
second pulse the current is recorded, this being
related to the interfacial charge transfer and
conductance. The advantages of this technique
are speed (several readings may be taken over a
short time) and elimination of a reference half
cell. A metal counter-electrode may be used,
eliminating problems of liquid-junction poten-
tials. A disadvantage is that, at present, the
technique cannot be used for enzyme elec-
trodes.
Factors affecting ISE measurements
There are several factors which cause elec-
Electrodes in clinical chemistry 479
rodes to yield results that may be significantly
different from those obtained by conventional
photometric methods. Some factors such as
interfering ions and junction potentials have
already been discussed. Among these factors
are:
A. Effects on the activity detected by the
electrode
(i) variations in ionic strength and activity
coefficient: both affect the activity of a given
molar concentration of an ion.
(ii) chemical interference.
(iii) space occupying effects of protein and
lipid in the sample.
B. Factors affecting the total potential of the
system
(i) selectivity of the membrane.
(ii) junction potentials.
(iii) rate of response.
(iv) temperature.
C. Calibration
(i) the nature of the cali brant.
(ii) the algorithm used for calibration.
Of these factors, the activity coefficient is the
most important and is influenced by ionic
strength, particularly if direct measurements
are required.
ACTIVITY
Activity coefficient
As discussed earlier, the EMF of a poten-
tiometric electrode is related to the activity of
the analyte in solution which is related to the
concentration of analyte by the equation:
Ai=yi'Ci
where Ai is activity, yi is the activity coefficient
and Ci is concentration. Activity coefficients in
aqueous solutions or blood/plasma are not
known exactly, though an approximate value
can be derived for dilute solutions using the
Debye-Huckel equationY
A·z 2 vT
logy=
l+BaYI
where A and B are constants, Z is the ion
charge, a is the ion size parameter (nrn), and I is
the total ionic strength (mmol/L). This equation
should only apply when ! is less than 0·1
mrnol/L and therefore is no more than 'an
480 Hirst and Stevens
approximation for plasma. (Appendix A shows
an example of an ionic strength calculation.)
Plasma and urine present two separate prob-
lems in estimation of ionic strength.
The ionic strength of blood/plasma is gener-
ally assumed to be 166-168 mmoVL,43 and is
affected principally by variations in plasma
sodium concentration. A change in ionic
strength of 15 mrnol/L, which is frequently
encountered, would result in an error of 2-3
mrnol/L in a sodium estimation.
The composition and ionic strength of urine
varies so widely that direct analysis by electrode
should be avoided. Unlike plasma, a 'sodium
correction' may not provide adequate com-
pensation. An indirect approach is preferable,
in which the ionic strength of urine is adjusted
by dilution with a strong buffer. The rela-
tionship of activity coefficient to ionic strength
is depicted in Figure 29. Since the ionic strength
of plasma and urine is unknown and variable,
the activity coefficients of the constituents is
similarly unknown and variable. As with ionic
strength, estimates of the activity coefficient are
widely used.
There is a large variation in activity coeffi-
cients of different blood constituents, which are
approximately 0·71 for potassium, 0·75 for
sodium and 0·55 for calcium, though widely
differing values have been reported.P The
combination of the low activity coefficient for
calcium, and the electrical charge results in a
sensitivity of approximately 14 mV per decade
change in concentration. A reading error of
1 mV will be equivalent to a 15% error for free
calcium, compared with 4% for potassium.
While there are several schools of thought on
calibration of electrodes for clinical use, com-
parison of the ionic strength of the calibrant with
that of the test solution must be considered.
Monovalent
1'0
~
.
co 0·75
.
.."
a
u
0'5
'"'
> 0·25
£Cl:
10- 7 10- 6 10- 5 10- 4 10- 3 10- 2 10- 1
Ionic strength (mmot/L)
FIG. :l9. The variation- of activity coefficient with
ionicstrength foro monovalent and a divalentcation.
Interferents (chemical)
The effect of chemical interference is to remove
ions by binding so that they are not sensed by
the electrode as 'active'.
In vivo binding compounds. The classic case
is that of calcium in plasma, less than half of
which is free, or 'ionised'. The remainder of the
calcium is bound to proteins (mainly albumin)
or other organic anions. The binding of plasma
calcium to protein is reversible and pH-
dependent." the calcium activity falling as pH
increases. In the case of calcium, where estima-
tion of the ionised species is of clinical interest,
blood specimens must be handled anaerobically
or re-adjusted by titration to the patient's pH or
a standard pH.
Several other compounds in blood are bound
to proteins or cell membranes, including most
of the heavier metals-Cu, Zn, Pb, etc. In these
cases, and for substances such as salicylate,
barbiturates, etc, the total amount rather than
the free ions is clinically important. Use of ISEs
to estimate these ions and compounds, even if
there were suitable ones, would require prior
liberation of the ions from protein.
Bicarbonate has been reported to bind
sodiurn.f' though there is some doubt over the
nature of this effect.
Foreign compounds. Calcium is subject to
further chemical interference, this time from
heparin.t" Two attractions of potentiometric
measurements is that they do not require
optically clear solutions and should operate on
whole blood. While blood is used for acid-base
studies, usually anticoagulated with lithium or
sodium heparin, other popular anticoagulants
such as citrate and EDTA act by removing
calcium, and are unsuitable for calcium estima-
tion. Heparin has a different mode of action,
but nevertheless binds calcium to a small but
significant degree. The two alternatives to using
serum are: (a) to control the amount of
heparin; and (b) to render the heparin non-
binding by presaturation with calcium.
Protein-lipid dilution effect
Colorimetric and potentiometric analyses often
differ in the way in which the sample is treated.
In colorimetric analysis the sample is usually
diluted. Electrode analysis is often performed
10 directly on whole blood or plasma, since cells,
proteins, lipids, etc, should not affect the
reading of the electrode-this being related to
the concentration of analyte in the water
compartment of the plasma (Fig. 30). Since
 Electrodes in clinical chemistry 481

Aqueous Normol serum Pathologicol

stondord serum

r r H2O

r
H2O
H2O II 140 mmol

1r
II
140 mmol No+
II
Abnormal protein

L
140 mmol

No+ L Na+ (ag myelomo) or

1-1Ipid(chylomicrons)

r
x x

\
Normal protein
\
Normal protein
and iipid and lipid
(7 % of serum volume)

FIG. 30. The effect of protein and lipid in plasma on the molal and molar concentration of sodium. In each of
the three cases, a direct ISE measurement will detect a constant molal concentration of sodium (140 mrnol/kg
H 20), whereas indirect methods will detect a variable molar concentration (mmoVtotal volume).

normal plasma contains approximately 7%
protein-lipid and 93% water by volume,42 one
would expect results from direct potentiometric
analysis to be higher than techniques involving
dilution by a similar degree. The differences
found are usually smaller than 7% (2-5%)
because this effect is counteracted by activity
coefficient, junction potential effects and the
nature of the calibrant used.
The results obtained by indirect potentio-
metry, when the sample is diluted prior to
analysis, usually compare well with alternative
techniques. The results obtained by direct
potentiometry may be clinically more useful,"
in the cases of plasma sodium and calcium and
particularly when plasma proteins-lipids are
grossly elevated. An alternative to using an
adjusting factor which has been recommended'"
to achieve agreement, eg, with flame photo-
metry, would be to make it plain that the
estimation is not the same as 'plasma sodium'.
FACTORS AFFECTING THE TOTAL
POTENTIAL OF THE SYSTEM
Selectivity
Ionic interference has been described in the
section on Electrochemical cells. The majority
of electrodes in clinical use have been chosen
for their suitable selectivity towards other ions
in biological fluids, particularly blood/serum.
Care should be exercised, however, in using
electrodes for urine analysis, in which the
concentration of potassium, ammonium, phos-
phate and many other ions is much higher than
in serum. Potassium estimation may be affected
by caesium which is occasionally added to
quality control materials. Measurements may
require buffering to a fixed pH as H+ (sodium)
or OH- (fluoride) may interfere.
Junction potentials
The effects of junction potentials were de-
scribed in the section on Electrochemical
potentials. Aqueous calibrants may differ from
biological fluids not only by the activity coeffi-
cients of their constituent ions, but also by the
junction potential at the reference electrode
interface. Although 3 mol/L or > 3·5 rnol/L
KCI is usually chosen to minimise junction
potentials, variation of the potential still occurs
and tends to have the opposite effect of the
plasma protein-lipid dilution effect. The varia-
tion of junction potentials between specimens is
sometimes referred to as the 'residual junction
potential' .
In whole blood, erythrocytes produce a
further difference in junction potential with a
'sedimentation effect'-a form of streaming
potential. It has been clairned'" that the
sedimentation effect is reduced by using a
double junction reference electrode with a
sodium formate salt bridge.
For a situation where the calibrant and
sample solution have differing activity coeffi-
cients and junction potentials, the concentra-
tion of an analyte is given bj the equation.v'
482 Hirst and Stevens

Cs=Cc'explO ( AEC) yc
- - . -·explO (AEj)
--
slope ys slope

where Cs and Cc are concentrations of sample

and calibrant, ys and yc are activity coefficients 101
of sample and calibrant , AEc is the difference
in EMF between sample and calibrant, and AEj
u,
is the junction potential difference between :::;;;: Concentration
sample and calibrant. w change
The last two terms in the expression are error
terms. One approach'! to reduce the difference
in results produced by electrometric and other -10 1
methods is to design the reference electrode
such that the product of the error terms
(derived from the previous equation):

- AEj ) ys
explO ( - - - is unity.
slope yc
Response time
The response of an ISE to a change in analyte
concentration is slow compared with photo-
metry, and may be variable. The rate of
response of an electrode is affected by several
factors such as the thickness, dimension and
nature (ion exchange rates) of the membrane,
change in activity, temperature and stirring
rate. The relationship between EMF and time is
far from simple,49 being based on 3 or 4 time
constants, and can be represented by a series of
exponentials only in an ideal electrode. For the
commercially available electrodes in clinical
use, the two most important variables affecting
response are change in concentration and
stirring rate.
Change in concentration. Electrodes respond
more rapidly to large changes in concentration
than small ones, and more rapidly to increases
than to decreases, though this does not always
apply. Theoretical response times are depicted
in Figure 31. An example of the effect of
concentration can be seen by comparing the
apparently instant response of a pH electrode
over a range of 2 pH units (2 decades change in
concentration) with the sluggish response
(5 min) of a fluoride electrode to changes in
urine fluoride concentration of 0·1 umol/L
(10-,-7 M). Since response time is so variable, it is
not good practice to take a reading at a fixed
time, unless the time interval is large compared
with the electrode response. The best practical
approach is to wait until the rate of change of
signal is negligible before taking a reading.
Stirring (flow) rate. The second variable
which can affect response time is the stirring
Time (s )
FIG. 31. The variation in response rate to positive
and negative changes in concentration.
rate or flow rate of a solution. Figure 32
illustrates the change in response of an elec-
trode with increasing stirring rate. Continuous
flow systems may benefit in a similar way by
using a faster flow rate, provided that this does
not produce a streaming potential.
Temperature
Sensitivity. The EMF of an electrode is
temperature-dependent, the relationship being
shown in the Nernst equation:
RT
E=Eo+- log, a
nF
where T is the absolute temperature. An
increase in temperature of lOoC will produce an
(I)
.L.:.---- (2)
u, '".-t:.-------- (3)
:::;;;:
w
<J
Time
FIG. 32. The relationship of response rate to stirring
rate. As the stirring rate increases from 1 to 3 the rate
of response increases, but the final response de-
creases.

increase in sensitivity of approximately 3·5%
for a univalent ion and half that for a divalent
ion. While there is little to be gained by
moderate increases in temperature, there is a
possibility of inaccuracy if the temperature of
calibrants and samples is not matched.
Response rate. The response rate of some
electrodes is significantly improved with moder-
ate increases in temperature.
Calibration
CALCULATION
There are several techniques for converting the
millivolts produced by an electrode to concen-
tration, most of which are based on the Nernst
equation.
Direct use of the Nernst equation
The Nernst equation required is in the form:
RT
Ecell=Eind-- loge Ax-Eref+Ej
nF
yxu=Ax 1 analyte is simply read off the graph. The slope
=antilog lO Ecell+Eind-Eref+Ej
O·1984·T
Eind is a composite of internal potentials of the
indicator electrode and includes potentials from
the inner reference electrode and the inner
phase boundary. The uncertainty of Eind, Ej
and the activity coefficient make direct use of
the Nernst equation impractical.
Standard addition or subtraction
The effect of adding a known amount of analyte
(or removal by chelation), on the EMF of a cell
can be used to estimate the original concentra-
tion in the sample. For the unknown solution,
RT
E =Eo+- log u·yx+EJ·
1 nF e
After the addition of an amount ~C of analyte,
RT
E:z=Eo+- loge (u+~C).y~+Ej
nF
if the addition does not significantly alter the
ionic strength and volume of the solution, the
activity coefficient and junction potential
should be unaltered. Then:
~E (ex+~C) RT
--=loglO where slope=2·303·-
slope ex nF
Electrodes in clinical chemistry 483
and the original unknown concentration:
u=~C(1O~EJS-1)
This technique can be used for complex and
variable solutions, eg, urine, but it is not
practical for high throughput serum analyses.
Calibration curves
An electrode with a Nernstian response should
show a limited linear relationship between log
(activity) and EMF. Figure 33 shows a typical
electrode response curve and the relationship
between EMF and log activity is mainly linear.
At the lower end the relationship becomes
unlinear as the limit of detection is approached
(or passed, depending on the definition of
'limit'). At the high end, the relationship with
activity remains linear, but there is a divergence
of the relationship with concentration.
An electrode may be calibrated by plotting
the EMF readings from two or more calibrant
solutions on semi-log graph paper, and assum-
ing a linear relationship. The concentration of
of the electrode response is relatively stable.
Most electrodes may be recalibrated with a
single calibrant, and the slope checked at
infrequent (eg, 6 h) intervals. The slope of
many electrodes is lower than the theoretical
59·1 mY/decade and often slowly decreases as
the electrode ages to less than 55 mV/decade.
, 'Calibration of electrodes by graph plotting is
suitable for clinical applications. This method
has had wide use and is still used in the form of
pH meters with logarithmic 'concentration'
scales. Graph plotting has now been largely
superseded by modern direct readout electro-
meters.
Activity
LJ..
::!: Concentration
w-
~
I
Activity/concentration (rrvnol/L)
FIG. 33. A typical electrode response curve,
484 Hirst and Stevens
Electronic signal linearisation
It is possible to linearise a logarithmic signal
with an antilog amplifier. Unfortunately an
antilog amplifier used alone will take the
antilog of the whole cell potential, and will not
necessarily produce an output which is linear.
For the circuit
Ex Eout
RT
Ex=Eo +- log, Ax;
nF
RT
put a=- and E=antilogcEO, and:
Nf
Ex = 10g.,E +10g.,(Ax)
= 10g.,(E. Ax")
After passing through the antilog amplifier:
Eout=E·(Ax)a
The output has an exponential, rather than a
linear, relationship to activity.
It is possible to linearise the relationship by
introducing a pre-amplifier with gain b into the
circuit:
Eout
. ..
By the same argument, the input to the antilog
amplifier is:
b·log.,(E·Ax a )
and the output will be:
Eout= antilog.,(b·log.,E·Ax")
= antilog c(1og.,E. Ax ab)
=E'(Ax)ab
If the gain of the pre-amplifier is adjusted so
that ab=l, ie, b=l/a, then Eout=E·Ax 1=
(antilog.,Eo)·Ax. ie, the output is directly pro-
portional to activity, but has a slope dependent
on £0 which-may vary. The slope variable may
be rdroved·with the. addition of a further
amplifier before Wlift¢r the antilog amplifier.
The electronic approach is not, then, a simple
solution. Software on a microprocessor may
offer a better means of linearising an electrode
response.
Computer algorithms
A number of instruments now available use a
dedicated computer or microprocessor to com-
pute the concentration of an analyte, eg,
plasma sodium or potassium. The algorithm
used by the computer will depend on the
composition of the manufacturer's calibrant
solutions. Information on either the calibrant
composition or algorithm is not always freely
available, though this information should be
requested by prospective purchasers. Examples
of two commercially-used algorithms are given
in Appendix 2.
CALIBRATION
An electrode is calibrated by converting the
potential (mV) generated by a calibrant solution
to a practical quantity such as activity or molar
concentration. The conversion is made by the
application of a variety of algorithms of greater
or lesser complexity (Appendix 2). The choice
of calibrant and definition of its value presents
certain difficulties. To calibrate an electrode
with a primary aqueous calibrant of defined
activity is not possible at present. It would be
necessary first to find a way of defining and
calculating activity as has been evolved over
many years for the pH scale.
A more practical solution to the problem of
calibration is to use a standard calibrant which
is comparable to the solution being analysed.l"
The EMF obtained could then be converted to
concentration units (eg, rnmol/L) and related to
the activity of analyte in the calibrant. Serum or
blood-based materials used for quality control
in blood gas measurements have been found to
be unstable" and would be unsuitable for use
as calibrants. An alternative to serum is an
aqueous calibrant containing physiological
amounts of the major stable ions and buffers,
but omitting the unstable components such as
proteins, in particular haemoglobin. Such a
calibrant may contain Na+, K+, Ca 2+, Mg2+,
Cl", HC03" and inorganic phosphate, with a
pH adjusted to 7·40.
Most manufacturers have adopted a calibrant
similar in composition to the one outlined, with
a few modifications eg, substituting Tris buffer
for the inorganic phosphate and adjusting the
total ionic strength to 160-166 mmoVL. The
rationale for the adjustment of the ionic
 Electrodes in clinical chemistry 485

strength of calibrants is that the ionic strength
of normal serum is around 166 mmol/L. If the
calibrant has a normal composition (eg, sodium
concentration = 140 mmol/L) the activity and
jun,ction potentials for calibrant and normal
sera should be similar. In practice, at least two
calibrants are required, because the slope of the
electrode must be established before an al-
gorithm derived from the Nernst equation can
be used (Appendix 2). The slope is derived
from the EMF difference of two calibrants.
Many manufacturers maintain a constant total
ionic strength for all calibrants (eg, 160 mmol/L)
by addition of a non-interfering salt. The effect of
this adjustment is that the activity coefficients and
junction potentials for a 'low sodium' calibrant
will differ from those found for a serum sample
with a similar molar concentration of sodium."
(Figure 34).
Since calibrants with an adjusted ionic
strength do not resemble sera with a similar
molar concentration of sodium (ie, they do not
have a physiological ionic strength), the slope
(S2) derived from these calibrants will be
incorrect. The effect of using an incorrect slope
in the calibration will be a discrepancy between
the calculated activity/concentration and true
activity/concentration. The discrepancy will in-
crease with the deviation of the serum sodium
concentration from normality. The agreement
between instruments calibrated in this way and
conventional techniques (eg, flame emission)
will only be close at normal sodium concentra-
tions. Results from flame emission and ion
selective electrodes have been shown to cor-
relate well over a wide range of sodium
concentration.V In this system the ionic
strength of all the calibrants was not adjusted to
160 mmol/L,
SUMMARY
On theoretical grounds it would appear prefer-
able to use calibrants which are not adjusted to
a constant ionic strength, whether concentra-
tion or activity is being estimated directly. For
indirect methods, when sample and calibrant
are diluted in an ionic strength adjusting buffer,
there is probably little difference between
adjusted and unadjusted calibrants. In this
case a simple calculation of concentration is
possible.
A simple algorithm may be used for activity

Molar concentration response

{unilinear due to change in yl

Activity slope /
= S2 adjusted "
ionic strength "

Calibratian
paints
\
L; "
/
"
"
...
",,:,y
LL Activity
:::!:
"
•.
w slope = SI

""
<l physiolog ical
I ionic strength
• .....J"
."" .: I
" I
/" I
" I
I
I

Sodium concentration (mmol/Ll

FIG. 34. Relationship of EMF to sodium concentrations at physiological ionic strength (••••.) anddl~;~sted
ionic strength (- - - -).
i'
486 Hirst and Stevens
estimation. The algorithm for direct concentra-
tion estimation is more complex. If concentra-
tion, rather than activity, is required an indirect
method must be used. If activity is required a
direct method must be used.
The quantitative effects of variations in
calibration procedure may be smaller than
variations in junction potential.
Advantages and disadvantages of electrodes
ADVANTAGES
(a) Analysis is rapid.
(b) Instrumentation is often simple, inexpen-
sive and portable.
(c) Optically clear solutions are not required
(whole blood may be used).
(d) Analysis is simple.
(e) Analysis is often non-destructive.
DISADVANTAGES
(a) Primary calibration is not possible.
(b) Junction potentials may vary between
samples in direct systems.
(c) Analysis may be affected by ionic or
chemical interference.
(d) Response is usually non-linear (except
for amperornetric methods).
(e) Response is sometimes slow, particularly
at low concentrations, limiting the use of
electrodes in automated analysis.
(f) Some electrodes have a limited life, or
are easily poisoned.
Acknowledgements
We are indebted to all the instrument manufac-
turers who supplied information on calibrant
composition and instrument design. We also
thank Professor Smith for his help and en-
couragement, Mrs Margaret Coles for typing
the manuscript and the members of the Instru-
mentation and Data-Processing Sub-
Committee, especially Mr R Beetham, for their
help and advice.
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Accepted for publication 12 December 1984
488 Hirst and Stevens

Appendix 1
The ionic strength of a solution is the sum of
contributions from all the ionised species in
solution and is defined as:
I=ljl~Ci'Zi2
where Ci is the molar concentration of Ion i
with charge Zi. For a simple solution of com-
pletely dissociated salts, the ionic strength is not
difficult to calculate:

Molar Concentration
Salt concentration Ions (mmol/L)

NaHCO] 25 Na+ 110+25 135

NaCI 110
KCI 5 cr 110+5+2 117
CaCI 1
K+ 5 1 5
Ca2+ 1 4 4
HCO; 25 1 25
S=286

Appendix 2 This modification of the measured sodium

The 'KNA1' Sodium-Potassium analyser
(Radiometer Ltd) calculates results by software
from measured electrode potentials using the
following algorithm for sodium:"
Erne••- E I52 )
(
CNa=152·1Q S·61·54
where S
61·54 (log 152-log 40)
152 rnmol/L and 40 mmoUL calibrants are used;
E l52 and E 40 are the potentials of the two
calibrants; Erne as is the potential of the un-
known solution; 61·54 is the theoretical (Ner-
stian) slope in mV at 37°c; and S is the slope
correction factor.
The final result, which is displayed, is calcu-
lated as:
CNare.dout=0·92981·CNameas+5·385.
activity is designed to compensate for the
protein-lipid dilution effect on flame photo-
meters. The system is designed to produce the
same results as flame photometry within the
normal range, as recommended by the IFCC. 47
The 'KNA1', being a direct measurement sys-
tem, produces results which are based on
activity, rather than concentration, and which
will not agree closely with flame photometry at
high or low sodium activity.
The Coming 902 Sodium-Potassium analyser
(Coming Instruments Ltd) uses the following
algorithm for the calculation of sodium activity/
concentration.
( Erne••- E c• 1 )
CNa=CcaJ.10 Slope
This algorithm is in essence identical to the
KNA1 algorithm. Coming also have an addi-
tional (optional) algorithm which is designed to
adjust the reading to a flame emission equiva-
lent, by subtraction of 2% of the calculated
activity.