Journal of Automatic Chemistry, Vol. 12, No. 3 (May-June 1990), pp.

116-128

Evaluation of the Jokoo-ION 150 AC:

Guidelines for the evaluation of analysers by
ion-selective electrodes

-
Joan Farr6, Carmen Biosca and Romin Galimany the selective membrane by an electrolytically simple
Servei d’Anitlisis Cliniques, Hospital de Badalona ’Germans Trias Pujol’, solution of the ion to be evaluated. The difference of
Apartado de Correos, 72, Can Ruti., 08916 Badalona, Barcelona, Spain potential measured at the extreme points of the system is
derived from the equation of Nernst:
The Jokoo-ION 150 A C is an automatic sodium, potassium and
chloride analyser which uses ion-selective electrodes. RT
E E0 + Ep + 2"3 logA [1]
The sample mode can be whole-blood, serum or urine. To evaluate a
urine sample, a previous dilution (1:6) with the standard 1 E0 is the sum of the fixed potentials depending on the
solution is required. For three concentrations of control materials, measuring and reference electrodes structure, E/ is the
the total precision (CV) ranged from 0"17 to 1"22% for sodium, sum of the variable potentials, ’parasites’, that have to be
0"22 to 1"69% for potassium and 0"16 to 0"74% for chloride. annulled, minimized or at least stabilized, R is the perfect
The system demonstrated acceptable performance in detection limit, gases constant, Tis absolute temperature, F is Faraday’s
linearity, drift and carry-over. Patients’ resultsfrom ION 150 A C number, and n is valence.
correlated well with those from a SMA C H and an IL 943. In a A=yxC
study on potential interferences, a slightly negative interference with logy 0"509 n 2 X/I
potassium was found with increases in lithium; and only high I 1/2 Z(Cini) [2]
ammonium ion concentrations caused a positive interference on the
potassium ion. A slightly positive interference of bromide on the where Ci is concentration, ni is valence (of each ion
chloride ion was also found. present in the solution),A is the activity, y is the activity
coefficient, C is the molar concentration, and I is the ionic
The electrode slope, electrode response, sample tempera- force.
ture and pH effect, effects of high concentrations of
proteins or lipids, and haematocrit influence on the At very dilute concentrations (10 -7 M), the activity
sodium, potassium and chloride ion concentration were coefficient is very near to unity but at higher concen-
also evaluated. The strategy adopted in this study trations it diminishes [3 and 14]. Thus, the calibrator
provides an ideal framework for future evaluations of ion- must have an activity coefficient similar to that of the
selective electrode analysers. sample.
Ion-selective electrodes have a big international market
Introduction due to their simplicity in use and their inclusion in
automatic analysers.
The device which is commonly called an ’ion-selective
electrode’ (’ISE’) determines the activity (A) of an ion in The unequal quality of the sensors currently available,
solution (in clinical biochemistry it usually refers to together with the sparse experience that analysis labora-
cations). The essential part of this device is a membrane tories have in the field of electrometry and the limited
with properties related to the ion being evaluated: it is quality control materials at present available from
specifically, or at least selectively, permeable to this ion. manufacturers, means that these instruments are often
Therefore, a potential difference is established between difficult to work with.
both sides of the.membrane, which is proportional to the
ion activity difference. The technology behind electro- The present paper presents an evaluation of an ion-
metric instruments aims to make the difference in selective electrode analyser (the Jokoo-ION 150 AC) and
potential as small as possible. suggests guidelines for studying, controlling and evaluat-
ing ion-selective electrode analysers.
As a result, a measuring chain is used where a series of
different devices are capable of exploiting this small
Materials and methods
potential. These complementary devices are called refer-
ence electrodes, and are characterized by the stability of
their own potential. There are two classes of reference
Automatic analyser SMA C H (Technicon Instruments
Corporation, Tarrytown, New York 10591, USA)
electrode; silver chloride and mercury chloride (calomel).
The chain of measurement is composed of a sequence SMAC II is a continuous flow analyser, which evaluates
including a reference electrode (called ’external’) and a sodium and potassium ions by indirect potentiometry.
measuring electrode- the latter being a reference For sodium the total dilution is 16"2, and for potassium
electrode designated ’internal’, which is separated from 1:18"8.

116
0142-0453/90 $3.00 () 1990 Taylor & Francis Ltd.
J. Farr et al. Evaluation of the Jokoo-ION 150 AC
The sodium ion utilizes a sodium-aluminium glass liquid) (Ag/AgC1 in saturated KC1 solution), which
electrode where sodium selectivity against potassium provides a stable potential that can be used as a reference.
amounts to 1000" 1; the serum sample is buffered at The sodium electrode is a glass membrane electrode
pH 8"0 to eliminate any interference by the hydrogen ion (sodium aluminium silicate). The potassium electrode is
[4]. a liquid membrane electrode, which is based on a neutral
carrier (Valynomicine-PVC, polyvinyl chloride as a
The Valynomicine electrode used for the potassium ion support). Finally, the chloride electrode is a liquid
has a selectivity of nearly 4000:1 against the sodium ion. membrane electrode, which is based on ionic exchange
Valynomicine is an antibiotic which is insoluble in water, using quaternary ammonium salts as an ion-exchanger in
but is soluble in organic solvents, such as ether or polymeric solvent..
acetone. It is suspended at the end of the electrode in a
porous membrane. The inner solution, formed by potas- Also used in the evaluation were a tester (Kaiser model
sium chloride, is in contact with the Ag-C1Ag inner SK-6300); a voltimeter of 1014 Ohms and a sensitivity of
medium from the reference electrode [5]. 0"1 milivolt (mV); and an analogous registrator (Linear
Instruments Corporation Model 1201; scale mV-5 V).
The evaluation of the chloride ion uses a mercuric
thiocyanate colorimetric method based on Zall et al.’s Calibrating solutions
manual method, automated by Skeggs and later modified For the SMAC II calibration two seric multiple consti-
by Morgenstern et al. The absorbance reaction is tuent calibrators at two different concentrations were
measured at 480 nm [6]. used: 144 mmol/1 sodium, 6" mmol/1 potassium and 112
rnmol/1 chloride (calibrator 1); 100 mmol/1 sodium, 3
IL 943 flame photometer (Instrumentation Laboratory, Paderno, mmol/1 potassium and 85 mmol/1 chloride (calibrator 2)
Dugnano, Italy) (Technicon Instruments Corporation).
The IL 943 has a dilutor incorporated, providing
For the flame photometer calibrations a water calibrator
automatic calibration and sampling. The calibrator and
was used, which was common to both ions, sodium and
sample dilution is 1:100. It is made with a caesium potassium, in concentrations of 140 mmol/1 sodium and 5
chloride solution, which has final concentration of mmol/1 potassium, for serum samples (I1 catalogue
1.5 mmol/1. The instrument’s zero is regulated with
number 97.517-50). For urine samples, the calibrator
distilled water at 100 with the inner standard (caesium
concentration runs up to 100 mmol/l of sodiun and 100
chloride solution [7]. The gas used is 99% pure propane mmol/1 of potassium. (I1 catalogue number 35.100);
at 3"6 psi.
whilst, for the lithium ion calibration, the concentration
runs up to 140 mmol/1 sodium, 5 mmol/1 potassium and
Jokoo-ION 150 A C analyser mmol/1 lithium (I1 catalogue number 33.313-50). To
calibrate the Jokoo-ION 150 AC analyser, two solutions
This analyser measures sodium, potassium and chloride
were used: C1 lot 0625397, with 140 mmol/1 sodium, 4
ion concentrations simultaneously for every sample
mmol/1 potassium, 100 mmol/1 chloride; and, C2 lot
through selective electrodes by direct potentiometry.
0918407, with 160 mmol/1 sodium, 6 mmol/1 potassium
and 120 mmol/1 chloride.
The sample volume needed is 100 1, plus 50 tl residual
volume. This sample can be whole-blood, plasma or
Controls
urine. To evaluate a urine sample, a previous dilution
(1:6) with standard solution is required. The through- Three commercial lyophilized controls ofhuman origin at
put is 150 samples/hour. three concentrations (low, medium and high) were used.
These were selected to represent common clinical
When the system is ready, a complete calibration is decisions. The manufacturer’s recommendations were
realized, being recalibrated automatically at two points followed to reconstitute the lyophilized control. A pool
every 30 min (this interval is optional). was prepared and stored in aliquot parts in the freezer
(-20 C). As urine control, a pool of urine was used to
The automatic sampler holds 30 sample cuvettes in a study the within-run imprecision.
gyratory disk, allowing positional identification of the
sample. This unit can use manual sampling through Reagents
syringes or capillaries, and stat sampling. It has an
automatic sample detector. Results are obtained in visual (1) Kit for determination of ammonium in blood
digits and are also printed-out. (SIGMA Diagnostics, St. Louis, USA). This
method is based on the ammination of 2-
The sample is aspirated through a capillary and is then oxoglutarate by means of glutamate dehydrogenase
transported directly to the electrode unit by a peristaltic (GLDH) and reduced nicotinamide adenine dinuc-
leotide (NADH). The absorbance decrease, mea-
pump which is placed after the measuring unit. The sured at 340 nm, as a result of the NADH oxidation,
capillary is rinsed automatically with standard solution 1. is proportional to the concentration of plasma.
There are four electrodes in the measuring unit: sodium, (2) Buffer solution Tris-hydroxymethyl aminomethane
potassium, chloride and the reference electrode. The orthophosphoric acid pH 7’35 (Tris 0"2 M, ortho-
reference electrode system is a double union type (liquid- phosphoric acid 0" 15 N).

117
j. Farr et al. Evaluation of the Jokoo-ION 150 AC

(3) Human albumin Hubber at 20% (ref. 70281 1). Table 2. Day-to-day imprecision study.
(4) Lipaemic solution at 10% Tutolipid (ref. 963769). Sodium Potassium Chloride
(5) Lithium heparin salt (vacutainer tubes).
Serum 7 CV 7 CV 7 CV
(6) Sodium chloride (Merck art. 6404, 99"5% purity) N=20 (mmol/1) (%)(mmol/1)(%)(mmol/1)(%)
(7) Potassium chloride (Merck art. 4937, 99"5% Low 121’39 1"22 3"12 0’69 96"21 0"62
purity). Medium 139.14 0"84 6"31 1"02 116"88 0"47
(8) Ammonium chloride (Merck art. 1145, 99"8% High 153’45 0"64 7"19 1"69 125"14 0"53
purity).
(9) Brij-35, 30% (Technicon product No. T-21-0110).
two working days. The mean, standard deviation (SD)
(10) Bidistilled and deionized water. and the coefficient of variation (CV) on each concen-
(11) Potassium bromide (Merck art. 4905, 99"5% tration level were calculated.
purity).
The urine pool was prepared in the laboratory; it was
(12) TRIS (Merck art. 8382, 99’5% purity). diluted at 1" 6 in standard solution 1, and was evaluated
(13) Orthophosphoric acid (Merck art. 565, 98% 30 times within the same run/test. The mean, SD and CV
purity). were also calculated.

The results are shown in table 1" precision for the three
Results ions for all three concentration levels was high as was
precision with the urine pool.
Imprecision
Day-to-day imprecision
Within-run imprecision
One sample of each of the three control concentration
A sample of each of the three controls was analysed 20 levels was evaluated each day over 20 days. The mean,
times in the same run without interruption for calibration SD and CV were computed for each of the three
or recovery of the base-line. This process was repeated for
concentration levels. The results are presented in table 2.
A high precision for the three components in the three
Table 1. Within-run imprecision. studied concentration levels is shown.

First Second Third Inaccuracy

Serum samples N 100 series series series
Because there were no commercial controls available [br
Sodium evaluating the system, primary standards at three
Low level (mmol/1) 8 120 120 concentration levels were prepared by weighing them
CV (%) 0"29 0"47 0"35 into buffer Tris-orthophosphoric acid (Tris 0"2 M, ortho-
Medium level (mmol/1) 140 138 140
phosphoric acid 0"15 N), adjusted to pH 7"35, and
CV (%) 0"2 0"19 0"21
High level (mmol/1) 149 154 155 adding a surfactant (brij 30%) at ml/1. Table 3 shows
CV (%) 0"37 0"27 0"17 the concentration levels which were examined. A dupli-
cate determination was carried out and the theoretical
Potassium value was checked by flame photometry. To calculate the
Low level (mmol/1) 3" 3" 3"
CV (%) 0"23 0"63 0.47 inaccuracy percentage, the following formula was used:
Medium level (mmol/1) 6" 5 6"4 64
CV (%) 0"9 0" 0" Table 3. Inaccuracy study.
High level (retool/l) 7’2 7"3 7.3
CV (%) 0’7 0" 0" Primary standard Measured Theoretical Inaccuracy
Chloride N 20 (mmol/1) (mmol/1) (%)
Low level Y (mmol/1) 96 95 95

-
CV (%) 0’5 0’7 0’6 Sodium
Medium level (mmol/1) 119 116 116 Low 118.16 120 1.53
CV (%) 0.29 0"23 0.19 Medium 135"06 140 3"53
High level (mmol/1) 127 125 126 High 154"65 160 -3’60
CV (%) 0’47 0"6 0’74 Potassium
Low 2’41 2"5 -3’6
Medium 3"84 4 -4
Sodium Potassium Chloride High 5’82 6 -3
Urine CV 7 CV 7 CV Chloride
N= 30 (mmol/1) (%)(mmol/1)(%)(mmol/1)(%) Low 110"67 122"5 -9"66
Medium 131.01 144 -9"02
65 2 26"1 0.47 77 1’85 High 151.86 166 -8"52

118
j. Farr6 et al. Evaluation of the Jokoo-ION 150 AC

Table 4. Relalive inaccuracy. Detection limit

(a) Correlation between indirect potentiometry and chloride determination The detection limit was taken to mean the least quantity,
(SMA C lI) and direcl potentiometry (Jokoo-ION 150 A C). or concentration, of a substance that can be distinguished
Correlation Least Deming’s with a given probability of a reaction blank carried out
N 100 coefficient square regression under the same conditions [10]. This ideal reaction blank
is executed by means of the Tris-orthophosphoric acid
Sodium (serum) 0"9604. a 23’514 a 17"76 buffer at pH 7"35 and without the ions to be determined.
b 0"815 b -0"856 The ionic concentrations of sodium, potassium and
Potassium (serum) 0"9914 a- 0"028 a --0"045 chloride were determined in a series of 10 determinations
b 0"971 b 0’987 in 3 days (thus obtaining a total of 30 values per ion). For
Chloride (serum) 0.9643 a 11"121 a 4.515 arisk oc to 5% and large samples (V> 30) the upper limit
b 0"902 b 0’968 of the blank confidence interval was calculated as being 2
+ "97, a value that corresponds to the detection limit for
(b) Correlation between flame photometry (IL 943) and direct this risk. The results are shown in table 5.
polentiometry (Jokoo-ION 150 A C).

N Lcast Lineariy limits

100.(serum) Correlation Deming’s
N 90 (urine) coefficient square regression Linearity is expressed by the equation
Sodium (serum) 0’9752 a 17"568 a 13’052 y= a + bx [10].
b 0" 866 b 0"899
Potassium (serum) 0"9827 a -0" 15 a -0"312 Because it was impossible to obtain a standard containing
b 1"04 b 1"076 the pure ion to be determined, sodium chloride and
Sodium (urine) 0.9789 a 23"994. a 22’493 potassium chloride were used. On the basis of a stock
b 0"801 b 0"820 standard prepared by weighing in Tris-orthophosphoric
Potassium (urine) 0"9933 a 2"222 a 2’095 acid buffer at pH 7"35, a series in triplicate of decreasing
b 0"829 b 0"832 dilutions was pertbrmed with the same buffer up to a total
of 12 points, which were evaluated on three different days
with a total of nine determinations per point. Then for
Inaccuracy (%) each point the mean value and the standard deviation
were calculated. The mean of the observed values
measured values theoretical value
x 100 [] corresponds to the ordinate axis, and the theoretical
theoretical value values to the abscissa axis. The bisector (y bx, b 1) is
This process was repeated for nine days, and the drawn and the linearity limits correspond to the maxi-
mum and minimum points ot’ which the interval of
inaccuracy was calculated taking the mean of the
measured values. Good accuracy was observed for the confidence cut the bisector. The interval was calculated
sodium and the potassium ions, although the system for a risk oc of ,5% and 8 degrees of freedom by X +_
tended to give values which were inferior to the 2.306 SD.
theoretical ones. The results obtained for chloride ions
were substantially below the theoretical ones. The least squares line and the correlation coefficient
corresponding to the linearity interval was obtained. The
The correlation between direct potentiometry and flame results are shown in table 6.
photometry and the indirect potentiometry (SMAC II)
was also studied. One hundred serum samples of patients Estimation qf the total error
with concentrations varying from the studied components The errors corresponding to the inaccuracy and impre-
were assessed, in duplicate and by the three methods. The cision of a technique, which can sum up or compensate
duplicates were distributed throughout the series, includ- each other, can be calculated as follows:
ing lipaemic, icteric and haemolitic samples. In addition,
90 urine samples were processed on the same day, by both A-I x SD and A + x SD (10)
the Jokoo analyser and by flame photometry. where A inaccuracy, SD standard deviation of the
day-to-day imprecision, and student value according
The correlation coefficient was calculated, the least to the risk oc and the degrees of freedom. Table 7 shows
squares line and the least squares regression according to the results.
Deming [9]. The results are shown in the table 4.
Carry-over
Table 5. Detection limil calculation for a oc 5% risk. The evaluations of carry-over used three protocols [1 1-
13] -each author reports on different aspects, all of which
Detection can affect the contamination between samples evaluated
N 30 mmol/l ;V SD limit
in series. For clarity, this report describes each of the
Sodium 9" 16 "66 12"48 three protocols, and results are given for each method.
Potassium 0"15 0"02 0"19
Chloride 12"7 1"03 14"76 Primary standards were prepared in the same way as
those used in the linearity and inaccuracy study.

119
j. Farr et al. Evaluation of the Jokoo-ION 150 AC

Table 6. Linearity limits calculated assuming oc 5% risk and 8 freedom. Linearity limits are indicated by asterisks. (N 9.)
Sodium (mmol/1)
Theoretical 1000 800* 500 250 125 62"5 31"2" 15"6 7"8 3"9
Observed 809 494 241 123 64.4 34 24"3 16.1 12
SD 45"5 21 5"4 2"63 2"18 1.74 2"48 1.93 2’1
Confidence limits 704"2 445"7 228"6 117"1 59"4 30 18"6 11’6 7’15
914"2 542"5 253’4 129"3 69"4 38 30 20"6 16’8

Potassium (retool/l)

Theoretical 64 40* 20 10 5 2"5 1"25 0"65 0"31" 0"15

Observed 50"34 39’26 19"3 9’46 4"84 2"37 1’26 0"68 0"38 0"28
SD 0"44 0"35 0"31 0"25 0"15 0"11 0"09 0’08 0"045 0.04
Confidence limits 49"3 38"4 15"58 8"88 4"49 2’12 1’05 0"5 0"28 0"19
51"3 40"1 20"01 10"04 5"18 2"62 1"46 0’86 0-48 0"37

Chloride (mmol/1)
Theoretical 1080 864 540* 270 135 67’5 33"75* 16"87 8"45 4"21
Observed 866 698 509 258 127’8 65"4 35"4 23"5 17"8 4"6
SD 30"2 24"9 13"6 5’62 3"31 2"66 1"52 1"68 1"35 0"75
Confidence limits 796"4 640"4 478 245"2 120"2 59"3 31’9 19"6 14"7 12"9
935 755"5 540"8 271"2 135’4 71"5 38’9 27’4 20"9 16"3

Table 7. Total error estimation for a risk of oc 5% and 19 freedom.

Imprecision Total error
N 20 mmol/1 Theoretical Observed A (SD) variation

Sodium 140 135"06 -4"94 1" 17 7’ 39 to 2"49

Potassium 4 3"84 -0’ 16 0"021 -0"2 to -0" 116
Chloride 122"5 110"67 11"83 0"77 13"44 to 10"22

Table 8. Carry-over studies according to Young-Gouchman’s Interaction: Interaction was evaluated according to Young
(interaction) and Bennet’s protocols. and Gouchman [11]. A series of samples were analysed
following the sequence below:
Sequence Inter- Carry- Sequence Carry-
H-- L actions over L-- H over H1-H2- Ha- L1 -L2- La- H1- H2-Ha
(mmol/1) % % (mmol/1) % H being the high sample concentration, and L the low
Sodium 196 49 0"55 0’82 49 141 -0"14
sample concentration. Each was evaluated in triplicate
178 99 0 0 141 178 -0"17 by applying the following formula:
159 81 0 -0"11 99 159 -0"37
% Interaction
L1La x 100
121 49 1"78 0"65 81 141 -0"2
196 141 0"2 0"20 49 196 -0"39 H La
Potassium 16 0"15 0 4 -0"85
where L1 is the contaminated sample, and Ha is the
8 2 0"21 0"51 4 16 -1"69 contaminating agent.
6 1"5 0"56 1"77 4 8 0"15
16 4 0"38 1"6 1"5 6 -1"1 Carry-over, according to Annette Bennet’s protocol [12],
8 4 0 0"2 16 -0"35 uses the same sequence of samples as in the previous
report. The following formula is applied:
Chloride 190 60 -0"39 -0"66 135 190 -0"58
170 100 0 -0"2 60 140 -0"28 % Carry-over
L1 L x 100
160 80 -0"15 0 140 170 0"89
120 60 0"49 -0"49 60 190 -1
190 140 -1’77 -0"29 80 140 -0"42 If the sample, L, is assumed to have been contaminated
by the sample H.
120
J. Farr et al. Evaluation of the Jokoo-ION 150 AC

% Carry-over
H1 H2 x 100 V
H
if the sample H is supposed to have been contaminated by
the sample L.

Table 8 shows the results obtained for each of the

concentration levels of the studied components, applying
the two previous protocols.

For the majority of the studied concentration levels the

contamination can be ignored because it was always less 100-
than 2%.

Carry-over, according to Broughton’s protocol [13], uses

the following sample sequence:
150-
Hi H2- L L2-...- L L
for 30 samples, H being the high concentration level and
L the low. The following formula was used:
K=
L1 Lx 100 200"
H2 L
considering that, if K is smaller than 2, there is no
’DECADE’
contamination.

The following results were obtained: K 0"038% for -250 IIII IIIII ’1 III1’
sodium; K 0% for potassium; K 1"47% for chloride. 10 100 1000
Contamination, then, is practically negligible for the Log. concentration
three studied components.
Figure 1. Sodium electrode slope study.

Drift
A commercial control series and having realized various calibrations. This was
was used- the level of which
coincided with the physiological values for each compo- performed over six days.
nent. Controls were evaluated at the beginning of a series
of samples (after the first calibration), at the end of this Therefore, the drift has been studied both in a series and
series of samples (prior to the recovery of the base-line also during the day’s work. The values obtained at the
and to the second calibration) and, finally, at the end of beginning of the series were compared to those at the end
the day’s work after having processed various sample of the series. In addition, those values obtained at the
beginning of the series were compared to those from the
end of the day’s work.

Table 9 shows the results obtained from both studies- the

drift was negligible.
Table 9. Drift study. (N 6.)
Calibration drift
Electrode slope study
Initial Final Drift For the electrode slope study, the same stock standards
mmol/1 mmol/1 % and solutions with decreasing concentrations in the
Sodium 138"2 138"2 0 linearity study were used. The negative pole of the
Potassium 3"63 3"61 -0"55 voltimeter was connected to the external reference
Chloride 110"97 112"28 1" 18 electrode, and the positive pole to the electrode under
evaluation. The results were the concentrations and the
corresponding responses in mV from the voltimeter (see
Day’s work drift figures 1-3). The abscissa axis is the concentration
Initial Final Drift
logarithm and the ordinate axis the corresponding mV-
mmol/1 mmol/1 % value; the slope of every electrode was obtained by giving
it the mV-values corresponding to 10-times increase in
Sodium 140" 140.4 0"21 the concentration of the ion (a ’decade’) [14].
Potassium 3"095 3"077 -0"58 Considering the theoretical value of the slope of 59 mV as
Chloride 118’ 119" 15 0"89 ideal for monovalent ions, acceptable results are observed
for the three electrodes.

121
J. Farr6 el al. Evaluation of the Jokoo-ION 150 AC

mV mV
80-

70-

60

150-

-40
40-
46.75 mV
3O

-60.-

10-
-80
’DECADE’ 0
’DECADE’

-I0
-I00 L_2__2.,,,,, ,,, TT-
10
f-----T--T--F-TTTT-I
100 1000
0.1 10 100
Log. concentration
Log. concentration
Figure 3. Chloride electrode slope study.
F{_ure 2. Potassium electrode slope sludy.
Electrode response study series of decreasing dilutions was then executed, with a
constant pH being maintained over the series. The
The response of each electrode was studied separately, concentrations were evaluated in duplicate; this value
using a serum pool and concentrating on the response was used on the ordinate axis and the corresponding
time and the stability of the steady state obtained [15]. theoretical value on the abscissa axis. By joining the
For that purpose an analogue registrator was connected intersection points of each series with identical pHs, the
to the outlet of every electrode. The stability of the steady deviations suffered by the ion values as a consequence of
state was good for all three electrodes, with a very short pH variation was plotted- see tigures 7, 8 and 9.
response time- less than s- in achieving the plateaus
and with a total duration of less than 22 s. In a similar way the pH levels of interest to clinical
practice were measured, and a minimm influence within
The results are shown in figures 4, 5 and 6. lhe pH-margins were observed -see table 11.
Sample temperature effect
Three commercial controls were evaluated on being Interference studies
removed from the refrigerator (-4 C), and after being
stored at room temperature (26 C). The controls were Influence of the ,;odium ion on the potassium ion electrode. A series
all evaluated after 5, 15 and 30 min, and always within of solutions were prepared with a constant concentration
the same analytic series. of 4 retool/1 of potassium and decreasing sodium con-
centrations of 200, 100, 50 and 8 mmol/1, in
The percentage of variation for each concentration level Tris-orthophosphoric acid buffer with ml/1 of a surfac-
was calculated, with reference to the concentration which tant. Each solution was evaluated five times, the mean
was evaluated initially (--4C). Table 10 shows the value taken and the effect of the sodium ion concentration
over the constant value of the potassium ion calculated.
percentages obtained in every measurement with refer-
ence to the initial values. The concentrations of this series were also evaluated by
flame photometry, in order to check the accuracy of the
concentration of each ion.
Effect of pH on electrode response
Three stock standards of identical ionic concentration In figure 10 the concentrations of the sodium ion are
were prepared. The standards all had different pHs: 8’48, shown on the abscissa and those of the potassium ion on
5" 17 and 7"37 with Tris-orthophosphoric acid buffer. A the ordinate- thus the evolution of the potassium ion

122
J. Farrd e! al. Evaluation of the Jokoo-lON 150 AC

Baseline

Sample (sodium 160 mmol/L) Air and reference Sample (potassium 6 mmol/L) Air and reference
solution solution

--I
-5 0 5 10 15 20 25 30 -5 0 5 10 15 20 25 30
Time (seconds) Time (seconds)
Figure 4. Sodium electrode response study. Figure 5. Potassium electrode response study.

values against the variations of the sodium ion concen- since the volume used is very small. Thus a lithium
tration, can be observed. Between the margins corre- concentration gradient was obtained. Seven dilutions
sponding to 50 mrnol/1 and 200 mmol/1 of sodium, the were obtained and each was evaluated for concen-
effect on the potassium ion has no significance. tration of sodium ions, potassium ions, chloride ions and
lithium.
The same sample series was processed by the ion
analyser, tbllowing the protocol fbr urine samples, and no Figure 12 gives the concentration percentage variations of
effect was found on the series. the ions on its ordinate axis, and the lithium ion
concentration on its abscissa. A reduction in potassium
ion on the sodium ion electrode. For a
Effect of the polassium ion concentration can be observed with an increase in
constant concentration of 125 mmol/1 sodium, potassium lithium ions.
concentrations of 250, 125, 62, 30, 15, 7"5 and 3"2 retool/1
in Tris-orthophosphoric acid buffer were prepared. Each Influence of the ammonium ion. A human serum pool was
solution was evaluated five times, and the mean value was prepared. Ammonium chloride was dissolved to give a
determined later. final concentration of 100 mmol/1, and by means of
another pool fraction, eight solutions of decreasing
The potassium ion concentration effect on a constant concentration of ammonium chloride were prepared. The
sodium ion concentration was also calculated. The ammonium ion concentration in each solution was
accuracy of the concentrations of both ions in this series evaluated in line with the previous method. At the same
was checked by flame photometry. Figure 11 shows the time, the sodium and the potassium ion concentrations
potassium ion values on its abscissa and the sodium ion were evaluated in the original pool and in the ammonium
values, found as a result of the different potassium ion ion solutions.
concentrations, on ts ordinate axis. The influence of the
potassium ion on the electrode of the sodium ion is Figure 13 gives the sodium and potassium concentrations
insignificant. on its ordinate axis, against ammonium ion concen-
trations. Neither ion is subject to any interference from
lithium ion as a component of the anticoagulant
Influence oj the ammonium ions at physiopathological concentrations.
agent. The human serum pool was used for serial dilutions Significant interference was seen with experimental
of the samples in tubes with lithium heparin. The lithium concentrations of ammonium higher than 10 mmol/1
heparin dilutor effect can be considered insignificant, (1"38%) and 100 mmol/1 (28.7%), which might affect the
123
J. FartS. et al. Evaluation of the Jokoo-ION 150 AC
is a geometrically proportional influence of bromide on
chloride, which, in spite of the fact it is small at lower
concentrations, should be taken into account for patients
who are receiving bromide and other halogens in
substantial amounts.
mV
Influence oflipid and protein concentration. The protein
concentration effect of direct potentiometry, indirect
potentiometry and flame photometry on sodium and
potassium ions has also been studied.
A human albumin solution at 20% was used which was
poor in ions. A lipaemic solution which was 10% free of
ions was also used. Decreasing dilutions were prepared,
in duplicate, maintaining a fixed ion concentration and
processing each of them following three survey methods.
Tables 12 and 13 show the variation of direct potentio-
metry in relation to indirect potentiometry and flame
photometry.
Due to the effect of the sample content of plasmatic water
[16], some sodium and potassium ion values could be
observed to be inversely proportional to the protein
Sample (chloride 120 mmollL) Air and reference concentration in those methods that employ dilution
solution relating to direct potentiometry. The same effect, but to a
minor degree, could be seen in the 10% lipid series (long-
chain triglyceride).
-5 0 5 10 15 20 25 30
Time (eecond$) Whole blood samples
Figure 6. Chloride electrode response study. Haematocrit influence. The eventual haematocrit influence
on the sodium and potassium and chloride ion concen-
potassium evaluations for urine samples with high tration evaluation were checked. Starting from blood
ammonium ion concentrations. samples obtained with lithium heparin anticoagulant of
the same patient, a series of whole blood samples was
Effect of halogenson the chloride electrode. For a constant prepared with a haematocrit varying from 4 to 70%. This
concentration of 125 mmol/1 chloride, decreasing bro- preparation was made by adding or subtracting plasma,
mide concentrations of 250, 125, 62, 30, 15, 7"5 and 3"2 after the corresponding phases of centrifugation, until the
mmol/1 in Tris-orthophosphoric acid buffer were desired haematocrits were achieved.
prepared. Each solution was evaluated five times and the
mean and the bromide ion concentration impact on a In this assay, very small differences in the chloride ion,
constant chloride concentration were calculated. potassium ion and sodium ion evaluations were observed,
except for the samples with a haematocrit higher than
Bromide values are shown on the abscissa axis.of figure 65%, where the potassium ion concentration value was
14, and the ordinate axis gives the chloride values found subject to a strong increase. This effect could result from
as a result of the different bromide concentrations. There the way the sample flows through the capillary which

Table 10. Study of the influence of temperature on concentration (mmol/1).

Taken out Time (min) at room temperature (26 C)
of fridge
(4 C) %A 15 %A 30 %A

Sodium 121.3 121.3 0 121.9 0.49 122"2 0.74

138.9 139.4 0.36 140 0.79 141.3 1.73
153" 154.1 0.65 155.3 1.44 155.8 1.76
Potassium 3" 18 3" 16 -0"63 3" 15 -0"94 3" 14 1.26
6’26 6’24 -0"32 6"24 -0"32 6"25 -0" 16
7’03 7"03 0 7"05 0"28 7"06 0"43
Chloride 96"8 97" 0"31 97"2 0.41 97" 0"31
117.3 117.1 -0.17 117.1 -0.17 117.1 -0.17
124.9 125"2 0"24 125.5 0.48 124"6 0’24

124
j. Farr et al. Evaluation of the Jokoo-ION 150 AC

Observed (mmol/L) Observed (mmol/L)

800
1
500

400
00

BOO

200

100

0
0 100 200 300 400 500 600 0 100 200 300 400 500 600
Theoretical (mmol/L) Theoretical (rnmol/L)

, ,
Figure 7. Sample pH influence on sodium electrode. [], pH 8"48;
pH 7"37; pH 5"17. ,
Figure. 9. Sample pH influence on chloride electrode. V], pH 8"48;
pH 7"37; pH 5"17.

connects with the measuring chamber, where the haema-

tites, owing to their number, could suffer lysis. Thus,
releasing the intraeritrocitary potassium ion would
Observed (mmol/L) increase the total potassium ion concentration (see table
20- 14).

Conclusions

The evaluation described here includes guidelines for ion-

selective electrode analyser evaluations. The following
results were found from the assays reported:

Table 11. Dependence of ionic concentration (mmol/l) variation on

physioj)athological pH values.
Observed values A
Theor- pH 6.9-
etical pH 6"9 pH 7.4 pH 7’8 7"4
Sodium 120 122"6 118.8 116"9 5’7
140 140.3 136"6 134"6 5" 7
160 159"9 156 153"3 6"6
Potassium 2’5 2"45 2"45 2’44 0.01
4 3"85 3’85 3’83 0"02
6 5"86 5.85 5"81 0’05
Theoretical (mmol/L) Chloride 122’5 109"2 109’8 110.3 1.1

8"48; ,
Figure 8. Sample pH influence on potassium electrode. D, pH
pH 7.37; pH 5.17.
144
166
130.4
154’2
131
155.4
131.7
155"7
1’3
1"5

125
 J. Farr6 et al. Evaluation of the Jokoo-ION 150 AC
Potassium (mmol/L) (1) The within-run imprecision for the Jokoo-ION 150
5- AC was smaller than 1%, and. the day-to-day
imprecision less than 2%.
(2) The inaccuracy relating to the primary standard
values was good for the sodium and potassium ion,
and acceptable tbr the chloride ion.
(3) The detection limit tbr the sodium, ion is 12’48
mmol/1, for the potassium ion 0’ 19 mmol/1 and for
the chloride ion 14"76 mmol/1.
(4) The linearity limits are tfigher than 500 mmol/1 for
the sodium ion and chloride ion, and more than 45
mmol/1 for the potassium ion.
(5) Carry-over and drift are negligible (smaller than
2%). The results were not significantly affected by
sample temperature.
(6) In. the study of the electrode slope, some values close
to 59 mV were obtained, which is considered to be
the optimal value for monovalent ions. The re-
sponse time is also very short (less than s), and
stability is Inaintained for 22 s.
(7) Significant variations due to.pH were observed on
both ions (sodium and potassium).
(8) The interference study showed a slightly negative
0 50 100 150 200 250 interference on the potassium ion due to the
Sodium (mmol/L) increment of the lithium ion. Only high ammonium
ion concentrations cause a positive interference on
the potassium ion. It was also observed that there

Figure 10. Influence of the sodium ion on the potassium electrode.

Sodium (mmol/L)

134
Variation (%)
I-

126

124
.’ .’ , "’,. ’ ." ’,.
:’
2 ,(;:
122

-2.5
120 0 2 4 6
0 100 200 300 400 500

Figure 11.
Potassium (mrnol/L)
Influence of potassium on the sodium electrode.
Lithium (mmol/L)
Figure 12. Influence of lithium. V], Sodium 139"6 mmol/1;
potassium 4"4 mmol/1; chloride 109 mmol/1.
,
126
j. Farr et al. Evaluation of the Jokoo-ION 150 AC

Variation (%) Table 12. Influence of lipid concentration. Differences are given in
30- sodium concentration between direct potentiometry and flame
photometry and indirect potentiometry for solutions which are free
of /ps.
Tri- Direct Flame Indirect
glycerides potentio- photo- A potentio- A
mmol/1 metry metry mmol/1 metry mmol/1
20-
0 140.7 143"6 141
0"3 140"6 143 0-5 140 0"9
0"6 140"3 142"4 0"8 141 -0"4
1"75 140 1.42 0"9 140 0"3
2"5 140"9 142"3 1.5 1.40 1"2
113- 5 141.8 142"4 2’3 140 2"1
7"5 144"9 142"7 5"1 141 4"2
10 143’1 140"8 5"2 140 4.4
5-

-5 T-’]-I"’II’I 111I

0.1 10
Ammonium (Log. mmol/L) Table 13. Influence of protein. Differences are given in sodium
concentration between direct potentiometry/flame photometry and
indirect potentiometry for solutions free of proteins. The sodium
concentration is notfixed, as the albumin concentrate* has sodium in
Figure 13. In,/tuence of ammonium on the sodium and potassium it.
electrodes. V-l, Sodium 135"9 mmol/1; @, potassium 4"3 mmol/1. Indirect
Direct Flame
Protein potentio- photo- A potentio- A
Chloride (rnmol/L) g/1 metry metry mmol/1 metry mmol/1
lO00
0 134"9 139"6 135
25’6 139"2 142"8 1.1 136 3"3
46"9 144"2 146"7 2"2 142 2"3
65"8 147"6 149"9 2"4 142 5"7
155"2 3"5 146 10’1

80 -.
79’8 154
99"3 160"4 160 5" 147 13"5
127’7 167"9 163’5 9’1 156 12
263* 57’1 48"6 46

600

400

Table 14. Influence of haematocrits.

Sodium Potassium Chloride
200- Haemato-
crit (mmol/1) A % (mmol/1) A % (mmol/1) A %
Plasma 141"4 4"05 108
5% 141"7 0"21 4"09 0"99 108"6 0"55
0"’ "1 111’1 I11 -r-- 18% 141 -0"28 4"07 0"49 107"5 -0"46
35% 143"6 1"5 4"03 -0"49 107 -0’93
10 100 0"99 4"09 0"99 106"2 1"66
45% 142"8
Bromide (mmol/L) 60% 140-9 -0"35 4.11 1"48 105"2 -2"59
70% 143 1.13 4"42 9"14 107’6 -0"37
Figure 14. Influence of bromide on the chloride electrode.
127
j. Farr6 et al. Evaluation of the Jokoo-ION 150 AC

was a slight positive interference of the bromide ion 6. MOIOEINSaEIN, S., RusI-I, R. and LEHMAN, D., Advances in
on the chloride ion. Automated Analysis. 1972. Technicon International Congress Vol. 1
(Mediad, Inc., White Plains, New York, 1973).
(9) The effect of high protein or lipid concentrations 7. Manual de Instrucciones Flame photometer 943 (Instrumentation
produced ionic concentrations proportionally more Laboratory S.p.A., Vialle dell’Industria, 3 Padermo, Italy).
elevated in direct potentiometry, than in indirect 8. Socidt6 Frangaise de Biologie Clinique. Commission ’Vali-
potentiometry and in flame photometry. dation de techniques’. Dictionaire des termes l’usage de la
validation de techniques. Annales de biologie clinique, 44 (1986),
(10) In the evaluation of whole blood samples significant 679.
increases of the potassium values were observed, 9. PAYNE, R. B., Clinical Chemistry, 30 (1984), 807 (letter).
starting from a haematocrit of 60% on. 10. Soci6td Frangaise de Biologie Clinique. Commission ’Vali-
dation de techniques’. Prolocole de validation de techniques.
References Annales de biologie clinique, 44 (1986), 686.
11. YOUNg, D. S. and (OUCHMAN, N., Methods for assuring
1. TIETZ, N. W., Quimica Clinica Moderna (Interamericana, quality data from continuous flow analyzers. Standard
S. A., Mexico, 1972). Methods of Clinical Chemistry, 7 (Academic Press, New York,
2. MARON, S. H. and PRtJraON, C. F., Fundamentos de 1972), 293.
Fisicoquimica (Limisa, S. A., Mexico, 1972). 12. BENNEa’, A., GARTELMANN, D., ASON, J. I. and OWEN, J. A.,
3. OESCH, V., AMMANN, D. and SIMON, W., Clinical Chemistry, Clinica Chimica Acta, 29 (1970), 161.
32 (1986), 1448. 13. BOtOHa’ON, P.,Journal ojAutomatic Chemistry, 6 (1984), 94.
4. RAo, J., PELAVIN, H. M. and MOROENSa’EIN, S., Advances in 14. SACHS, CI-I., Annales de biologie clinique, 43 (1985), 715.
Automated Analysis: Technicon International Congress 1972. Vol. 15. SACI-IS, CH., Donndes pratiques pour l’6ssai automates
1: Clinical Chemistry Methods (Mediad, Inc., New York, ’electrometriques’. I.S.B., 5 (1982), 397.
1973), p. 33. 16. SACHS, CH. and TRUCHAUD, A., Potentiometrie ’Directe’
5. LAVINIA, A., PIODA, R., SIMON, W., BOSSHARD, H. R. and contre Potenyiometrie ’Indirecte’ apropos d’une guerre
Ctma’itrs, H. C., Clinica Chimica Acta, 39 (1970), 289. picrocholine en electrometrie. LS.B., 9 (1983), 103.

128
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