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Inductively Coupled Plasma Mass
Spectrometry for Direct Multi-element
Analysis of Diluted Human Blood and
Serum†
EBBA BARANYa, INGVAR A. BERGDAHL*b , ANDREJS SCHÜTZb ,
STAFFAN SKERFVINGb AND AGNETA OSKARSSONa
aDepartment of Food Hygiene, Swedish University of Agricultural Sciences, S-750 07 Uppsala, Sweden
bDepartment of Occupational and Environmental Medicine, L und University, S-221 85 L und, Sweden
A method for the inductively coupled plasma mass
spectrometry (ICP-MS ) multi-element analysis of diluted
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human blood and serum was used for the following elements:
Co, Ni, Cu, Zn, Ga, Se, Rb, Mo, Rh, Pd, Cd, Sn, Sb, W, Pt,
Hg, Tl and Pb. Sample pretreatment was a simple dilution
(ten times for blood and five times for serum) with a solution
containing 5 g l−1 of 25% ammonia, 0.5 g l−1 Triton X-100,
and 0.5 g l−1 EDTA in Millipore water. In and Sc were used
as internal standards. For sample introduction a flow-injection-
type technique (based on time instead of volume) was used.
The determinations were carried out first in a peak-jumping
mode for selected masses, and then in a scanning mode. Each
determination of a preparation took 75 s. The results for
reference samples agreed with recommended or certified values
for Co, Cu, Zn, Rb, Cd, Tl and Pb in blood, and for Rb, Mo
and Cd in serum. For Ni and Hg in blood, and Cu and Zn in
serum, the results agreed with one of two reference samples.
The detection limits for all these elements (except for Tl ) were
sucient for analysis of samples from the general population.
On the other hand, the results for Se in blood, and for Co, Ni,
Se, Sn and Hg in serum did not agree with recommended or
certified values. No reference samples are available for Ga,
Mo, Rh, Pd, Sn, Sb, W, or Pt in blood, or for Ga, Rh, Pd,
W, Pt, or Pb in serum. Generally, the limits of detection for
the elements in the latter group (below 0.15 mg l−1) are close
to or above the levels present in the general population.
Keywords: Inductively coupled plasma mass spectrometry;
blood; serum; blood plasma; multi-element; flow injection
The concentration of an element in human blood (in this text
‘blood’ will refer to whole blood) or serum can often be used
as a biological indicator of the absorbed dose of the element.1
For example, Pb, Hg and Cd are often determined in blood
from people occupationally or environmentally exposed to
these toxic metals. Lately, certain ‘rare’ elements, e.g., Ga and
Pt, have found increasing industrial use, but the levels of these
elements in blood and serum are still not well known.
Inductively coupled plasma mass spectrometry (ICP-MS) is
one of the most sensitive trace element techniques available,
and several ICP-MS methods for the determination of di
erent
elements in serum or blood have been published.2–10 It is also
a rapid technique with the capability of multi-element determi-
nation, thereby making it ideal for screening of populations to
gain knowledge of which concentrations are ‘normal’ and to
perform epidemiological studies. The technique may also be
of use for clinical purposes and in forensic medicine, since
† Presented at the 1997 European Winter Conference on Plasma
Spectrochemistry, Gent, Belgium, January 12–17, 1997.
Journal of Analytical Atomic Spectrometry, September 1997, Vol. 12 (1005–1009)
acute poisoning by di
erent elements may give similar
symptoms.
We have begun working on a study where a large number
of blood and serum samples from ‘normal’ teenagers will be
analysed. Few of the methods published so far have, however,
been concerned with the application of ICP-MS to large-scale
multi-element determinations, where issues such as ease of
sample preparation and sample through-put have to be
considered.
The sample preparations generally used may be divided into
two categories, sample dilution or digestion. Simple dilutions
are mainly used for serum analysis,3,4 while blood samples
may be digested5 or deproteinised by addition of acid.6–8
Concern about blocking of the nebulizer or the torch seems
to have prevented development of analytical methods for
undigested blood.9 Digestions are often carried out in nitric
acid in pressurized vessels in a microwave oven. However,
microwave ovens for digestion are expensive, the sample
preparation is time consuming, and there is a risk of carry-
over e
ects in the digestion vessels. Moreover, the extensive
handling of the sample increases the risk of contamination.
Furthermore, the digested samples have to be diluted to
decrease the acid concentration prior to ICP-MS analysis.
Typically, an original sample mass of 0.5–1 g is after digestion
diluted to 50 ml, giving a dilution factor of 50–100. Thus, the
principle of direct analysis after a simple dilution is attractive
in its simplicity.
Based on previous work10,11 Lutz et al.2 presented a sample
preparation method for blood analysis using ICP-MS after a
simple dilution with an alkaline solution. This method has
previously been modified and used for determination of lead
in plasma and blood.12,13 In the present work the suitability
of the method for multi-element determination was
investigated.
EXPERIMENTAL
Sample Preparation
Human blood and serum were diluted ten times for whole
blood and five times for serum, with a solution containing
5 g l−1 of 25% ammonia (ARISTAR, Merck, Poole, UK),
0.5 g l−1 Triton X-100 (Sigma, St Louis, MO, USA), and
0.5 g l−1 ethylenediaminetetraacetic acid disodium salt dihy-
drate (EDTA) ( pro analysi, Janssen Chimica, Geel, Belgium)
in Millipore water. The dilution was carried out in screw-
capped 12 ml polypropylene test tubes (Sarstedt, Nümbrecht,
Germany) rinsed with de-ionized water. Internal standards, In
and Sc in 0.14 mol l−1 nitric acid (ARISTAR, Merck), were
added to a concentration of 10 mg l−1 in the final sample
dilution. All samples were prepared in duplicate.
More diluted blood (20 times instead of 10) and serum (10
1005

times instead of 5) were tested with respect to precision and
detection limits.
Screening of all standard solutions used for calibration was
carried out using ICP-MS before multi-element stock solutions
were made, which showed only negligible contaminations with
other elements.
Calibration curves were obtained using outdated blood or
plasma from blood donors (serum was not available). Blood
and plasma aliquots of 50 ml were spiked with multi-element
stock solutions to produce calibration curves in physiologically
relevant ranges. For elements with high natural concentrations
(e.g., Cu, Zn, Se and Rb) the highest calibration standard’s
concentration was approximately twice the original concen-
tration. Elements with low concentrations (e.g., Cd, W, Pt and
Hg) were calibrated against blood or plasma spiked with 1, 2
and 5 mg l−1. Di
erent stock solutions were prepared for
standards which were delivered in nitric acid and for those
delivered in hydrochloric acid.
The blood and plasma standard with the highest concen-
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trations were used for assessment of method precision.
Polyatomic Interferences
To study the e
ect of ethanol addition, 1, 3 and 6% (v/v)
ethanol solutions were added to prepared blood samples. To
study the e
ect of S and Zn polyatomics on 82Se, additions
with S and Zn were made to approximate a duplication of the
concentrations in the sample. Thus the addition of S [a solution
prepared from sodium sulfate (p. a., Merck, Darmstadt,
Germany)] corresponded to 1200 mg l−1 in blood, and the
addition of Zn (prepared from Merck standard) corresponded
to 5 mg l−1.
Instrumentation
The ICP-MS instrument was a PQ2+ from Fisons Elemental
(Winsford, Cheshire, UK) with a Gilson 222 autosampler
(Gilson, Villiers, France). A PTFE V-groove nebulizer, a Ni
sampler cone with a 6 mm Pt tip and a Ni skimmer cone were
used (Fisons Elemental ). The gas flows were 13.5 l min−1 for
the coolant gas, 1.0 l min−1 for the auxiliary gas and 1.0 l min−1
for the nebulizer gas. The forward power was 1350 W, and
reflected power <5 W. The sample flow was 1.0 ml min−1.
The sample was introduced in a segmented-flow mode,
which is similar to flow injection, but with no flow-injection
valve. The autosampler was programmed to take up sample
for 20 s and then return to rinse position where it remained
for 55 s. The same solution that was used for dilution of the
samples was continuously pumped into the rinse. The solution
from the rinse then acted as a carrier flow for the sample. The
instrument was programmed to start acquiring data just before
the sample reached the plasma, and continue for 30 s, thereby
covering the whole segmented-flow peak. The programming is
easily done with the ICP-MS software. The total time for
analysis of a sample preparation was 75 s.
All samples were first run in the peak-jumping mode for
45Sc, 59Co, 60Ni, 114Cd, 115In, 118Sn, 202Hg, 205Tl and, in serum
also, for 208Pb (3 points per peak, 10 ms dwell time for internal
standards and Sn, and 50 ms for the rest of the analytes). After
that all samples were run in the scan mode for determination
of the other elements (m/z: 5–10, 42–50, 58–78, and 82–239;
320 ms dwell time, 19 channels per u). In both peak-jumping
and scan mode three repeats were made on each preparation.
Calculations and Interference Corrections
The calibration procedure in the ICP-MS software was used.
For all samples, the results for a reagent blank were subtracted.
Interference corrections were made for 114Cd (corrected for
1006 Journal of Analytical Atomic Spectrometry, September 1997, Vol. 12
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spectral overlap from Sn, measured at m/z 118), 55Mn (overlap
from ClO, m/z 51), 87Rb (overlap from Sr, m/z 88) and 108Pd
(overlap from Cd; the corrected measure at m/z 114 was used).
Since the isotopic composition of Pb may vary, the sum of the
three isotopes at 206, 207 and 208 was used in scan mode (i.e.,
for blood analysis, but not for serum analysis). Moreover, to
improve precision for some elements, the signal from two
isotopes were summarized: Pd (m/z 106 and 108), Sb (121,
123), W (182, 184) and Pt (194, 195).
Reference Materials
The reference materials used were Seronorm Trace Element
Human Whole Blood ( batches 205052 and 203056) and Serum
( batch 311089; Nycomed AS, Oslo, Norway), A-13 Freeze
Dried Animal Blood (International Atomic Energy Agency,
Vienna, Austria), and a ‘second generation’ Human Serum
from Gent University, Belgium.14,15
RESULTS AND DISCUSSION
Sample Dilution
At the dilution used the signal of the internal standards added
to blood and serum samples was approximately 60% of that
for internal standards added to the dilution reagent. That is a
considerable signal supression, caused by the high content of
dissolved inorganic and organic material. An increase of the
dilution by a factor of two (20 times for blood and 10 times
for plasma) decreased the signal supression, giving a signal of
the internal standards of approximately 80%. However, the
precision was not improved by the further dilution and, as a
consequence, the detection limits (in the original samples)
increased. Given a choice between less signal supression
or better detection limits, we chose to give priority to the
detection limits.
The sample-to-sample variability in the matrices of blood,
plasma, or serum is very small, as compared with e.g., urine
or freshwater where the concentrations of dissolved solids may
vary by orders of magnitude. Therefore, the considerable signal
supression in blood, plasma and serum samples should not
present any major problems as long as internal standards are
used and the calibration is made by using spiked blood
or plasma.
Sample Introduction
Continous injection of the sample would make analysis of a
large series impossible, since denatured proteins would block
the torch. Therefore, a segmented-flow technique was used.
Single injections of 220 sample preparations could be made
without blocking the torch. This corresponded to 80 blood
samples in duplicate, plus samples for calibration and quality
control. One such run took five hour. Denatured proteins did
however build up a ring inside the sample injector of the
plasma torch, approximately 15 mm from the tip, making
cleaning of the torch necessary after the five-hour run.
Conventional flow injection can also be used but the present
segmented-flow technique has some advantages compared with
conventional flow injection. The main advantage is that no
valve is required, hence, instrument costs cleaning procedures
and in most cases also analysis time are minimized. Moreover,
as the autosampler moves between the rinse position and
sample positions with the sample introduction pump running,
a small air bubble is introduced between the carrier solution
and the sample. Introduction of gas in the flow is sometimes
used in advanced flow-injection systems to reduce dispersion
of the sample, giving a sharper flow-injection peak.16 On the
other hand, a conventional flow-injection system may utilize
further automation, e.g., by on-line dilution of the samples.

Polyatomic Interferences for Se
Selenium is dicult to determine owing to its low degree of
ionization in the plasma and the polyatomic interferences. In
order to be able to correct for polyatomic interferences on m/z
82 the samples were spiked with elements which may cause
polyatomic interferences at that mass.
Possible interferences on 82Se were SO 17 or ZnO, but
3
neither addition of S nor Zn increased the signal on m/z 82 in
blood or reagent blank. We were therefore not able to identify
the nature of any polyatomic interferences on m/z 82. The
additions of S and Zn were confirmed by increases of the
signals for SO and Zn, respectively.
It has been shown that addition of ethanol to serum and
urine diluted eight-fold with 1% nitric acid increases the signal
for Se and makes it possible to decrease polyatomic inter-
ferences by optimizing the nebulizer gas flow.18 In the present
work, however, ethanol addition to blood increased the Se
signal by only 10–20%. Attempts to optimize the ratio between
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Se and interfering polyatomics by adjusting the nebulizer gas
flow between 0.8–1.2 l min−1 did not noticebly improve the
performance. A likely explanation for the lack of a more
pronounced e
ect by ethanol addition on undigested blood is
that the e
ect of ethanol may depend on the carbon content.
It has previously been shown that addition of methane
increases the Se signal,19 and recent investigations indicate
that any organic material may increase the ionization of Se
through interaction with ionized carbon.20 Perhaps the solution
of 10% blood contains so much carbon that the influence
from addition of 1–6% ethanol becomes small. An altern-
ative explanation may be that the interferences di
er between
di
erent ICP-MS instruments.
Polyatomic Interferences for Cu, Zn, Mn and Cr
Spiking of blood with sulfur to approximately double the
concentration did not give any statistically significant increase
in the signals for 65Cu or 66Zn. The tendency was actually
towards lower signals. In the reagent blanks, increases were
seen (approximately 10 and 50 mg l−1, respectively). There was,
however, an increase in the signal for 68Zn with 25 mg l−1,
indicating some Zn contamination of the sulfuric acid used
for spiking.
The method might lead to overestimation of Cu and Zn
concentrations due to interferences from sulfur. This inter-
ference should contribute less than 10 mg l−1 for Cu, and less
than 25 mg l−1 for Zn. At the concentrations in the blood
reference samples this would correspond to 0.5–2.5%.
As the 55Mn peak was a
ected by significant overlap from
Fe and ArO of m/z 56, no attempt was made to determine Mn
in blood. In serum the Fe concentration is three orders of
magnitude lower, but the peaks were still not resolved.
The determination of Cr on a quadrupole ICP-MS requires
digestion of the samples to decrease the carbon content5 and
in the present work no attempt was made to determine Cr.
Peak Jump or Scan Mode
The scan mode is the obvious choice for a multi-element
method but for ultimate detection limits the peak-jumping
mode has to be used. It is then necessary to restrict the number
of m/z values monitored, otherwise the gain from increased
measurement time on each peak will be lost. We chose to use
the peak-jumping mode for a few elements (Co, Ni, Cd, Sn,
Hg, Tl, and in serum also for Pb) which may be of interest for
future epidemiologic studies and for which the signal was low.
After the run in peak-jumping mode all the samples were run
again in scan mode to determine other elements.
Journal of Analytical Atomic Spectrometry, September 1997, Vol. 12
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Whole Blood Reference Samples
For whole blood, the results for Co, Cu, Zn, Rb, Cd, Tl and
Pb agreed with the recommended or certified values. With the
exception of Tl, the detection limits for these elements seem to
be sucient, although those for Co and Cd are close to the
levels that may be expected in the general population (Table 1).
Thus, the Co and Cd results in individual samples will have
to be interpreted with caution. If these elements were to be
determined by peak-jumping with longer measurement time
this should improve the detection limits slightly.
For Ni and Hg the results agreed with the recommended
values for one of the reference samples, and disagreed for
another. There was a surprisingly large deviation for Ni in the
IAEA A-13 sample. A Ni sampler cone with Pt tip and a Ni
skimmer cone were used, but this does not seem to provide a
reasonable explanation of why falsely low results for Ni in
that sample would appear. The explanation might be that Ni
in the Freeze-Dried Blood was not dissolved in the dilution
reagent. The reference sample is produced for use in digestion
methods. The recommended value (95 mg l−1) seems to be
extremely high for a blood sample; the concentration in human
blood from unexposed adults is reported to be below 1 mg l−1.5
Thus, the Ni may come from use of stainless steel in the
preparation of the sample and might be in a form that is not
soluble using the present method.
For Hg the memory e
ects arising from adsorption on
tubings and/or glass surfaces may a
ect the results. It is thus
important to avoid use of standards with too high concen-
trations, and to make sure that no results are falsely high due
to a preceeding sample with a high Hg concentration. Errors
due to memory e
ects from a preceeding sample can be avoided
by quality control procedures if the duplicate samples are run
in di
erent series with di
erent sample sequences, e.g.,
reversed order.
The Se results did not agree with the recommended values.
We were unable to identify any polyatomic interferences on
m/z 82, but cannot rule out that as a reason. It can also be
speculated that variations in the ionization of Se may be
involved.
As no reference materials are available for Ga, Mo, Rh, Pd,
Sn, Sb, W or Pt in blood, the accuracy of the determination
of these elements cannot be evaluated in this single study.
Spike-recovery experiments could have been made but would
not be of particular value because the calibration was already
made against spiked blood and plasma. The detection limits
for some of the elements are below the concentrations in
normal populations, but the results may be used to establish
a level which is not exceeded in the general population. This
is true even though Rh and Pd have been shown to have
spectral interferences,7 since these will lead to overestimations.
Serum Reference Samples
For serum, the results for Rb, Mo and Cd agreed with the
recommended or certified values, while the results for Zn and
Cu deviated slightly (Table 2). For these five elements the
limits of detection seems to be sucient.
The results did not agree with the recommended or certified
values for Mn, Co, Ni, Se, Sn and Hg. For Se and Hg the
problems are probably the same as for blood. For Ni the lower
dilution, compared with blood, may cause a more significant
contribution from the cones. For Co and Sn we have no
explanation to the discrepance between our results and the
recommended values.
No reference materials are available for Ga, Rh, Pd, W, Pt
or Pb in serum or plasma, so the accuracy for these elements
could not be evaluated. Especially for Pb, an important
1007
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Table 1 Analysis of whole blood reference materials (all results in mg l−1)

Seronorm Seronorm IAEA Animal Blood

Batch No. 205052 Batch No. 203056 A-13 Detection
Element/ limit Precision*
nuclide Found (s) Target (range) Found (s) Target (range) Found (s) Target (range) (3s, n=10) (% RSD, n=10)
Elements with agreement between found and recommended concentrations:
59Co† 0.2 (0.04) <1 6.2 (0.1) 6 0.1 (0.01) 0.02 1
65Cu 820 (20) 880 (30) 410 (20) 410‡ (352–457) 3 2
66Zn 6590 (160) 6320 (280) 1330 (90) 1240‡ (1140–1330) 4 2
87Rb 1370 (30) 2060 (70) 250 (6) 219‡ (162–295) 0.5 9
114Cd† 0.7 (0.1) 0.9 (0.8–1.0) 6.0 (0.2) 6.4 (5.9–6.8) 0.2 (0.02) 0.03 2
205Tl† n.d. 5.4 (0.1) +5§ n.d. 0.05 2
206+207+208Pb 34 (2) 35 (31–41) 380 (10) 383 (361–396) 18 (1) 18(14–29) 0.1 6
Elements with both agreement and disagreement:
60Ni† 3.3 (0.5) 3 8.7 (0.8) 7 1.7 (0.3) 95(57–130) 0.2 4
202Hg† 3.2 (0.1) 3 (2–4) 12 (0.4) 9 (8–9) 0.4 (0.04) 0.1 3
Element with disagreement between found and recommended concentrations:
82Se 120 (10) 83 (79–87) 140 (20) 93 (89–97) 32 (8) 22.9‡ (14.3–29.5) 10 11
Elements for which the accuracy can not be evaluated:
71Ga 2.4 (0.2) 2.4 (0.4) 1 (0.1) 0.2 6
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98Mo 1.4 (0.4) 8.5 (0.6) +5§ 1.5 (0.2) 0.4 7

103Rh n.d. n.d. n.d. 0.1 5
106+108Pd 0.2 (0.1) 0.4 (0.1) n.d. 0.08 8
118Sn† 1.7 (0.1) 2.7 (0.5) 2.3 (0.1) 0.6 4
121+123Sb 0.6 (0.2) 33 (0.6) +25§ n.d. 0.1 4
184W 0.4 (0.2) 0.5 (0.06) 0.2 (0.04) 0.1 9
194+195Pt 0.3 (0.4) n.d n.d. 0.2 9

* The precision was assessed from repeated runs of one of the calibration standards, i.e., a spiked blood sample with approximately a doubled
concentration of major elements and 5 mg l−1 of elements with low original concentrations. † Peak jumping mode. ‡ Certified value. § Added amount.

Table 2 Analysis of serum reference materials (all results in mg l−1)

Seronorm Serum Batch No. 311089 ‘Second generation’ Human Serum

Element/ Detection limit Precision*
nuclide Found (s) Target (range) Found (s) Target (range) (3s; n=10) (%RSD; n=10)
Elements with agreement between found and recommended concentrations:
87Rb 5.8 (0.4) 160 (2) 168† (138–198) 0.2 5
98Mo 1.0 (0.1) 0.6 (0.1) 0.68† (0.61–0.75) 0.1 6
114Cd‡ 0.2 (0.01) 0.2 (0.01) 0.18† (0.15–0.23) 0.03 1
Elements with agreement for one and disagreement for one reference material:
65Cu 1260 (60) 1260 (1200–1320) 1070 (30) 1010† (972–1045) 2 5
66Zn 1520 (70) 1500 (1370–1640) 970 (20) 873† (855–890) 3 3
Elements with disagreement between found and recommended concentrations:
59Co‡ 1.2 (0.01) 4.8 (4.78–4.88) 0.6 (0.02) 0.33† (0.27–0.38) 0.01 1
60Ni† 5.1 (0.1) 2.5 (1.9–3.1) 3.2 (0.4) 0.23 0.08 5
82Se 100 (7) 86 (79–87) 120 (6) 95 (90–100) 4 4
118Sn‡ 2.9 (0.2) 1.2 (0.2) 0.77 0.2 4
202Hg‡ 2.4 (0.1) 1.8 (0.1) 0.6† (0.56–0.63) 0.07 5
Elements for which the accuracy can not be evaluated:
71Ga 0.5 (0.04) 0.6 (0.07) 0.06 5
103Rh 0.1 (0.07) 0.07 (0.01) 0.03 3
106+108Pd 0.06 (0.03) n.d. 0.03 3
121+123Sb 9.6 (0.5) n.d. 0.1 3
184W 0.3 (0.04) n.d. 0.03 4
194+195Pt n.d. n.d. 0.06 5
205Tl‡ 0.4 (0.03) 0.03 (0.006) 0.01 3
208Pb‡ 0.8 (0.04) 1.8 (0.05) 0.1 3

* The precision was assessed from repeated runs of one of the calibration standards, i.e., a spiked plasma sample with approximately a doubled
concentration of major elements and 5 mg l−1 of elements with low original concentrations. † =Certified value. ‡ =Peak jumping mode.

environmental pollutant which can be determined in blood
plasma by ICP-MS,12,13 such materials would be valuable.
CONCLUSION
The present method may be used for determination of Cd, Co,
Cu, Pb, Rb, Zn, and possibly also Hg and Ni in blood. In
serum it may be used for Cd, Mo, Rb, possibly also Cu, Hg
and Zn. This study could not show whether the method is
useful for determination of the concentrations of Ga, Mo, Pd,
Pt, Rh, Sb, Sn and W in blood, and Ga, Pb, Pd, Pt, Rh and
W in serum. The detection limits for many of these elements
are not low enough to determine the concentrations in
individual samples.
The preparation of the samples and the ICP-MS analysis
are quick, and make it possible to analyse about 80 samples
in duplicate preparations, plus calibration and quality control
samples, per day.
Anna Akantis is thanked for skilful technical assistance and
Görel Svensson for advice on writing. This work was financially
supported by the National Swedish Environment Protection
Agency, the Swedish Work Environment Fund, the Swedish

1008 Journal of Analytical Atomic Spectrometry, September 1997, Vol. 12


Council for Planning and Coordination of Research, and the
Medical Faculty at Lund University.
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Paper 7/00904F
Received February 10, 1997
Accepted July 16, 1997
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