Journal of Analytical Toxicology 2013;37:222 –226
doi:10.1093/jat/bkt015 Advance Access publication March 6, 2013
Application of ICP-OES to the Determination of Barium in Blood and Urine in Clinical
and Forensic Analysis
Teresa Lech1,2*
1
Institute of Forensic Research, Westerplatte 9, 31-033 Krakow, Poland and 2Department of Toxicology, Faculty of Pharmacy,
Collegium Medicum, Jagiellonian University, Medyczna 9, 30-688 Krakow, Poland
*Author to whom correspondence should be addressed. Email: tlech@ies.krakow.pl
Exposure to barium (Ba) mostly occurs in the workplace or from
drinking water, but it may sometimes be due to accidental or inten-
tional intoxication. This paper presents a reliable, sensitive method
for the determination of Ba in blood and urine: inductively coupled
plasma optical emission spectrometry (ICP-OES) after microwave
digestion of samples. The overall procedure was checked using
Seronorm Whole Blood L-2, Trace Elements Urine and spiked blood
and urine samples (0.5 –10 mg/mL of Ba). The accuracy of the
whole procedure (relative error) was 4% (blood) and 7% (urine);
the recovery was 76 –104% (blood) and 85– 101% (urine). The
limits of detection and quantification (Ba l 5 455.403 nm) were
0.11 and 0.4 mg/L of Ba, respectively; precision (relative standard
deviation) was below 6% at the level of 15 mg/L of Ba for blood.
This method was applied to a case of the poisoning of a man who
had been exposed at the workplace for over two years to powdered
BaCO3, and who suffered from paralysis and heart disorders. The
concentrations of Ba, in mg/L, were 160 (blood), 460 (serum) and
1,458 (urine) upon his admission to the hospital, and 6.1 (blood)
and 4.9 (urine) after 11 months (reference values: 3.34 + 2.20 mg/L
of Ba for blood and 4.43 + 4.60 mg/L of Ba for urine).
Introduction
Barium (Ba) compounds are widely applied in many fields,
such as in the oil and gas industries; in the production of lubri-
cating oil additives (dinonylnaphthalene sulfonate); in making
paints (sulfate and chloride), bricks, ceramics, glass (carbon-
ate), rubber, vinyl stabilizers and steel hardening (chloride)
(1 –3), fireworks or propellants (carbonate, nitrate and styph-
nate) (4 –5), rodenticides (carbonate and nitrate) (6 –11) and
cosmetics (sulfide) (12). Exposure to this element mostly
occurs in the workplace (13 –15) or from drinking water (2 –3,
16). There may also be accidental exposure from other sources
(e.g., from contaminated flour) (10) or from intentional intoxi-
cation (forensic cases) (7, 11, 12, 14, 17 –20). The health
effects of the different barium compounds depend on how
well the given compound dissolves in water. Only barium
sulfate is considered to be an innocuous, or at worst, a minim-
ally harmful compound (1 –3, 12); however, cases of non-fatal
or fatal poisonings after oral administration of barium sulfate
for contrast radiography have been described (21 –22). The
barium cation is extremely toxic and causes characteristic
gastrointestinal symptoms, periorbital and extremity paresthe-
sia, hypertension and progressive flaccid muscular paralysis
(1 –3, 12) that can result in death, a condition referred to in
the past as Pa Ping (16). Rarely, rhabdomyolysis, respiratory
failure and hypophosphatemia may develop (7). Profound
hypokalemia can also be induced (1– 3, 12).
# The Author [2013]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com
Article
Concentrations of Ba in blood and urine that affect human
health can be rather low. The concentrations of Ba in biological
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specimens vary broadly from approximately 1 mg/L or less in
blood and plasma/serum, below10 mg/L in urine to a few mg/g
in bone, hair, nails and teeth (23). Therefore, reliable analytical
tools for the clinical and forensic analysis of biological material
for barium need to be applied, because some basic methods,
such as flame atomic absorption spectrometry (FAAS), are not
adequately sensitive [limit of detection (LOD) of approximately
10 mg/L in aqueous solutions, but several times worse for real
samples]. To determine the normal concentrations of Ba in
body fluids, inductively coupled plasma mass spectrometry
(ICP-MS) or electrothermal atomic absorption spectrometry
(ET-AAS) can be used, because they offer excellent detection
capabilities (LOD of 1.2 ng/L) (1 –2, 13, 17, 23). In cases of
chronic or acute poisonings, however, other methods may be
applied (1, 23). In ET-AAS, the atomization efficiency of bio-
logical samples is rather low due to the formation of numerous
side products such as BaC2, BaCN, BaO, Ba(OH)2, BaS, BaCl and
BaCl2 (23). In the inductively coupled plasma optical emission
spectrometry (ICP-OES) technique, only boric acid or sodium
borate were reported to interfere with the line emission
spectra of barium at 455.403 nm (2). Neutron activation ana-
lysis (NAA) used to be applied to different kinds of samples
(hair and bone biopsies with LOD of 0.2 mg/g), but it is no
longer used (23).
In this paper, a simple, sensitive and reliable method involv-
ing moderate costs and fairly rapid analysis time has been pro-
posed for the determination of Ba in biological samples (blood
and urine) by ICP-OES after microwave digestion with nitric
acid and hydrogen peroxide. The aim of the study was to evalu-
ate whether the ICP-OES method is suitable for the determin-
ation of Ba in body fluids (blood, serum and urine) in cases of
chronic and acute poisonings, and possibly for the determin-
ation of normal (reference) levels of Ba.
Experimental
Samples
Samples of blood, serum and urine were collected (Sarstedt
Monovette Li-Heparin LH/2.6 mL tubes for blood, Eppendorf test
tubes for serum, 100 mL plastic vessels for urine) from a patient
of a hospital in Poland who had been exposed to a barium com-
pound at his workplace (a ceramic factory) for over two years.
The patient probably inhaled barium carbonate powder.
Samples of blood (n ¼ 24) and urine (n ¼ 25) were obtained
from non-exposed people ( patients of a hospital in Krakow,
Poland) to validate the procedure and to estimate reference
levels.
Table I.
Sample Preparation

Material Volume of sample Microwave digestion Final volume Dilution with water before measurement
(mL) (mL)
HNO3 H2O2
Blood: Seronorm Whole Blood L-2 2 3 1 10 1:10, 1:5 or 1:2 (similar results obtained)
2 3 1 10
Serum 0.5 3 1 10 1:10, 1:5 or 1:2 (similar results obtained)
Urine: Seronorm Trace Elements Urine 10 3 1 20 1:10 or 1:5 (similar results); 1:2 (lower results obtained)
5 3 1 10

Before determination, samples of investigated material (2 mL Table II.

of blood, 0.5 mL of serum or 20 mL of urine) were digested by Specifications and Working Conditions of the ICP-OES Device

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nitric acid (3 mL) and hydrogen peroxide (1 mL) in an Ethos 1
Monochromator Echelle type
microwave digestion system (Milestone, Italy). Details are given
in Table I. Detector CID
Radio frequency generator power 1.150 kW
Radio frequency 27.12 MHz
Plasma observation Radial/axial (auto view)
Instrumentation Pump rate 50 rpm
The analysis was conducted by using an iCAP 6300 duo plasma Integration time (low/high wavelength) 15 s/15 s
emission simultaneous spectrometer (Thermo Electron, Waltham, Auxiliary gas flow 0.5 L/min
Coolant gas flow 20 L/min
MA), allowing the recording of the full emission spectrum of the Spray chamber Cyclonic with concentric nebulizer (Meinhard)
sample in the range from 166.250 to 847.000 nm with the help Nebulizer gas pressure 0.14 MPa
of a charge-injection device (CID). The fundamental features of
the spectrometer and measuring conditions applied in the ana-
lysis are presented in Table II. The instrument was calibrated
against multi-element standards. Linear regression analysis gave Table III.
a regression coefficient (R 2) of 0.999 for an emission line of Analysis of Certified Materials
l ¼ 455.403 nm in the concentration range up to 9.0 mg/mL of Sample Certified value Found Accuracy*
Ba, and for l ¼ 233.527 and 230.527 nm up to 50.0 mg/mL of Ba (mg/L of Ba) (mg/L of Ba) (%)
(R 2 . 0.999). Seronorm Whole Blood L-2 (n ¼ 5) 66 + 4 69 + 4 4.0
Seronorm Trace Elements Urine (n ¼ 5) 53 + 5 57 + 5 7.0

*Relative error in percentage.

Reagents
All reagents were analytical grade: concentrated nitric(V) acid
of special purity (Suprapur) and 30% hydrogen peroxide
mineralization (matrix dilution can affect the accuracy of the
(Merck, Darmstadt, Germany).
method). The limits of detection (LOD) and quantification
The calibrations for Ba and spiked samples were prepared
(LOQ) measured (n ¼ 20) for the replicates of the blank
with 1,000 mg/L of ICP multi-element (Ag, Al, B, Ba, Bi, Ca, Cd,
(taken through the digestion procedure before analysis, diluted
Co, Cr, Cu, Fe, Ga, In, K, Li, Mg, Mn, Na, Ni, Pb, Sr, Tl and Zn)
1:10 for measurements), accepted to be three times the stand-
standard solution IV (Merck).
ard deviation (SD) for the LOD and 10 times the SD for the
The accuracy of the method was assessed on the basis of
LOQ at the most sensitive emission line of Ba (l ¼
two certified materials: blood (Seronorm Whole Blood L-2) and
455.403 nm), were 0.11 and 0.4 mg/L of Ba, respectively. The
urine (Seronorm Trace Elements Urine) (SERO AS, Billingstad,
precision [relative standard deviation (RSD)], determined on
Norway).
the basis of results obtained for three different samples of
Deionized water obtained from NANOpure Diamond appar-
digested blood, each tested 10 times for Ba, was below 6% at
atus produced by Barnstead (Dubuque, IA) was used to prepare
the level of 15 mg/L of Ba for blood.
working standard solutions and to dilute samples.
The results of the analysis of blood, serum and urine samples in
the case of a man suspected to have been poisoned by barium
carbonate, probably by inhalation at his workplace, are presented
Results
in Table IV. The concentrations of Ba in blood (n ¼ 24) and urine
The overall procedure was assessed by using standard reference (n ¼ 25) in people not exposed to Ba obtained by the author
materials (Seronorm Whole Blood L-2 and Seronorm Trace (the lowest value of the range: LOQ/2) and those found or cited
Elements Urine) and different spiked blood and urine samples by other analysts are summarized in Table V.
(in the range of 0.5 –10 mg/mL of Ba). The accuracy of the
whole procedure—digestion in a microwave system and deter-
mination by ICP-OES—expressed as a relative error was 4%
(blood) and 7% (urine) (Table III). The recovery of the method Discussion
using spiked samples was 76 –104% (blood) and 85 –101% Very little has been published on the analysis of biological
(urine), depending on the dilution of the solution after samples for Ba by AAS. FAAS lacks sensitivity, although it is

Application of ICP-OES to the Determination of Barium in Blood and Urine in Clinical and Forensic Analysis 223
possible to use it only for acute poisonings, and only after ex-
traction, for example by tetrasodium versenate in alkaline solu-
tion, with an LOD of 30 mg/L (32). Most ET-AAS applications
have been used in the analysis of drinking water within the
range of a few mg/L to several hundred mg/L, and in the analysis
of the very challenging matrix of seawater (23); however, some
authors have described its application to the evaluation of
barium concentrations, e.g., in a case of acute poisoning (13). It
seems that emission techniques (ICP-OES or ICP-MS) may be
current methods of choice for the determination of barium in
biological material, not only to evaluate levels in chronic or
acute poisonings, but also to estimate reference levels. Although
ICP-MS offers excellent detection capabilities in the analysis of
and urine with demineralized water and then analyzed them.
The detection limits achieved were 0.06 mg/L of Ba for water,
0.25 mg/L of Ba for urine and 0.6 mg/L of Ba for blood. Schramel
et al. (25) analyzed urine samples for Ba, Sr, Ti and other ele-
ments (B, Ca, Cu, Fe, Mg, P and Zn) by the ICP-OES technique
after acidification of a sample (10 mL of concentrated nitric acid
of high quality/100 mL of urine).
Physiological reference values reported for Ba, which are of
particular interest in the field of occupational medicine due to
relatively common exposure at the workplace, are scarce.
According to Hamilton et al. (34), the normal values for barium
are approximately 4 mg/mL in urine and below 1 mg/mL in
blood. Schramel et al. (25) obtained, for 25 samples of urine,
mean values of 4.5 + 4.2 mg/mL of Ba, as the reference values

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bone, erythrocytes, plasma and other biological tissues (23), it is
one of the most expensive techniques. Because of this, the pro-
posed method using ICP-OES may be very useful in the routine
analysis of clinical and forensic samples for barium, and simul-
taneously for other elements if necessary.
This technique has previously been applied to the multi-
element (including Ba) analysis, of urine, serum, blood (ante-
mortem samples) and bone or other postmortem samples
(2, 27, 33). Some problems, however, concern the pre-treatment
of biological samples before determination by different proce-
dures. Most sample decomposition procedures should be applic-
able, except for those employing sulfuric acid-containing
mixtures, due to low solubility and possible co-precipitation
of barium sulfate. Currently, preference is given to wet digestion
by nitric acid or a nitric acid –hydrogen peroxide mixture in a
microwave system, which was used in this study. In the proced-
ure for the ICP-OES analysis of serum, Rahil-Khazen et al. (27)
used similar sample digestion (serum: HNO3 –H2O2 in a ratio of
3:2:1). Mauras and Allain (24) merely diluted samples of blood
Table IV.
Concentrations of Ba in Blood and Urine in a Man Exposed to BaCO*3
Sample Concentrations of Ba (mg/L)
On admission to hospital
Blood 160
Serum 460
Urine 1,458
*A dash indicates that the sample was not analyzed.
for healthy adults. Mauras and Allain (24) obtained a mean value
of 4.3 + 1.4 mg/mL of Ba for urine (n ¼ 13). Urinary levels of Ba,
determined by ICP-MS in 1,437 samples collected from US
patients by Komaromy-Hiller et al. (28), were 3.5 + 2.2 mg/mL
of Ba within a range of 1.0 –7.0 mg/mL pf Ba, and they were
similar to the reference values from a control population, as
reported by Minoia et al. (26). The relevant values obtained in
the study using the proposed procedure ranged up to 24 mg/mL
for urine (mean: 4.43 + 4.60 mg/mL of Ba; median: 2.20 mg/mL
of Ba).
Probably due to glass contamination, Rahil-Khazen et al. (27)
for serum (n ¼ 141) obtained by using ICP-OES a median value
of 60.4 mg/mL of Ba (i.e., 0.44 mmol/mL of Ba in the range of
0.22– 0.71 mmol/mL of Ba); this is similar to the results
obtained by Goullé et al. (29) by ICP-MS: median, 111; range,
30– 154 mg/mL of Ba.
Limited data have been published on blood barium concen-
tration. Minoia et al. (26) reported levels below 10 mg/mL
of Ba; Heitland and Koster (31) obtained by using ICP-MS a
median of 0.8 mg/mL of Ba (range: 0.17 –1.9). The barium con-
centration in the blood of living persons estimated in the
present study is within these ranges. Moreover, as established
in a previous study (unpublished data), the results obtained
for postmortem blood and urine samples of non-poisoned
After 11 months
people (n ¼ 63) sent for analysis were usually higher. This may
6.1 be because of possible contamination during sampling (the
—
4.9
samples were collected using different kinds of devices, includ-
ing those made of glass), and/or putrefaction effects in blood:
41.8 + 32.0 (mean + SD), 36.0 (median), 0.2–168 (range) mg/mL

Table V.
Concentrations of Ba in Blood and Urine in People Not Exposed to Ba

Material Concentrations of Ba (mg/L)

Current study Maurias and Allain Schramel et al. Minoia et al. (26) Rahil-Khazen et al. Komaromy-Hiller et al. Goullé et al. Heitland and Köster
(ICP-OES) (24) (ICP-OES) (25) (ICP-OES) (ET-AAS/ICP-OES) (27) (ICP-OES) (28) (ICP-MS) (29) (ICP-MS) (30–31) (ICP-MS)
Blood 3.34 + 2.20* ,1 — 1.2 + 0.6* — — 0.8*
(0.2 –8.9)† (0.47 –2.9)† (0.17 –1.9)†
1.88‡
Serum — — — — 60.4‡ — 111‡ —
(30.2–97.5)† (30 –154)†
Urine 4.43 + 4.60* 4.3 + 1.4* 4.5 +4.2* 2.7 + 1.5* — 3.5 + 2.2* 0.89‡ 1.96*
(0.2 –24)† (1.8 –7)† (0.2 –12.7)† (0.25 –10.1)† (1.0 –7.0)† (0.17 –3.85)† (0.1– 14)†
2.20‡ 3.0‡

*Arithmetic mean + SD.

†
Range.
‡
Median.

224 Lech
of Ba; and urine: 70.6 + 95.1 (mean + SD), 21.3 (median),
0.7 –213 (range) mg/mL of Ba.
The concentrations in blood, serum and urine in cases of ex-
posure or acute poisoning with barium compounds are usually
considerably higher. In the described case of acute poisoning,
probably by inhalation of barium carbonate, levels reached 160
(blood), 460 (serum) and approximately 1,500 (urine) mg/mL
of Ba. Inhalative exposure to soluble Ba compounds in a large
group of welders resulted in a median urine level up to
101.7 mg/mL of Ba (13). Mauras and Allain (24), in an acciden-
tal poisoning, found concentrations of 260 and 280 mg/mL of
Ba in blood and urine, respectively. According to Zschiesche
et al. (13), only plasma concentrations exceeding 10 mg/L and
urine levels higher than 20 mg/L may be assumed to reflect ele-
vated Ba exposure. In cases of the ingestion of Ba compounds,
the levels are usually higher. Hung and Chung (14), in a case of
poisoning by a mixture of cadmium and barium, reported
values in serum of 341 mg/mL of Ba (normal: 30 –200 mg/mL
of Ba); Rhyee and Heard (4), in a case of the ingestion of
“snake” fireworks, reported 20 –100 mg/mL of Ba in serum and
5,600 mg/mL of Ba in urine. In a fatal case after oral administra-
tion of barium sulfate for contrast radiography (due to intrava-
sation), the concentrations in blood reached the following
values: 370 mg/mL (39 days after incident) and 150 mg/mL (41
days after incident), and 440 mg/mL in cerebrospinal fluid (39
days after incident) (22). In a suicidal poisoning with barium
chloride, autopsy barium levels were: 9,900 mg/mL (blood),
6,300 mg/mL (urine) and 8,800 mg/mL (bile) (17).
Conclusions
On the basis of this study, it can be concluded that ICP-OES is
both reliable and sensitive; thus, it is a current technique of
choice for the determination of Ba in body fluids in clinical and
forensic toxicology. This study showed LOD and LOQ of
0.11 mg/mL of Ba and 0.4 mg/mL of Ba in solution (l ¼
455.403 nm), respectively. The accuracy of the whole proced-
ure (as a relative error) was 4% (blood) and 7% (urine), and
can be affected by matrix dilution. The recovery of the method
for samples digested in a microwave system was 76 –104%
(blood) and 85–101% (urine).
The concentrations of Ba in biological specimens in non-
poisoned people are low: 3.34 + 2.20 mg/mL of Ba for blood
and 4.43 + 4.60 mg/mL of Ba for urine; in poisoning, the levels
of Ba are considerably higher.
This method allows the analysis of blood, serum and urine
for Ba in people suspected of having been poisoned with Ba
compounds. It was applied in a case of poisoning of a man at
the workplace (contact with powdered BaCO3), probably by in-
halation. The concentrations of Ba, in mg/L, were 160 (blood),
460 (serum) and 1,458 (urine) on admission to hospital, and
6.1 (blood) and 4.9 (urine) after 11 months.
Acknowledgments
This research was supported by research project No. N N404
189136 funded by the Ministry of Science and Higher Education
in Poland. The author would like to thank Dr. Bozena Wrzosek
from the Provincial Hospital, Cardiology Department, in Radom
Application of ICP-OES to the Determination of Barium in Blood and Urine in Clinical and Forensic Analysis 225
and Dr. Patrycja Krawczyk from the Nofer Institute of
Occupational Medicine in Lodz, Poland, for providing antemor-
tem blood and urine samples for barium determinations.
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