Forensic Science International 237 (2014) e1–e5

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Forensic Science International

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Case Report

ICP OES and CV AAS in determination of mercury in an unusual fatal

case of long-term exposure to elemental mercury in a teenager
Teresa Lech *
Institute of Forensic Research, Westerplatte 9, 31-033 Krakow, Poland

A R T I C L E I N F O A B S T R A C T

Article history: In this work, a case of deliberate self-poisoning is presented. A 14-year-old girl suddenly died during one
Received 8 November 2013 of the several hospitalizations. Abdominal computer tomography showed a large number of metallic
Received in revised form 10 February 2014 particles in the large intestine. Analysis of blood and internal organs for mercury and other toxic metals
Accepted 15 February 2014
carried out by inductively coupled plasma optical emission spectrometry (ICP OES) revealed high
Available online 24 February 2014
concentrations of mercury in kidneys and liver (64,200 and 2470 ng/g, respectively), less in stomach
(90 ng/g), and none in blood. Using cold vapor-atomic absorption spectrometry (CV AAS), high levels of
Keywords:
mercury were confirmed in all examined materials, including blood (87 ng/g), and additionally in hair.
Elemental mercury
Münchausen syndrome
The results of analysis obtained by two techniques revealed that the exposure to mercury was
Teenager considerable (some time later, it was stated that the mercury originated from thermometers that had
Postmortem material been broken over the course of about 1 year, because of Münchausen syndrome). CV AAS is a more
ICP OES sensitive technique, particularly for blood samples (negative results using ICP OES), and tissue samples –
CV AAS with LOQ: 0.63 ng/g of Hg (CV AAS) vis-à-vis 70 ng/g of Hg (ICP OES). However, ICP OES may be used as a
screening technique for autopsy material in acute poisoning by a heavy metal, even one as volatile as
mercury.
ß 2014 Elsevier Ireland Ltd. All rights reserved.

1. Introduction dental amalgam exposure (mercury vapors outgassed from

Mercury (Hg) is a heavy metal of known toxicity. Intoxication
involving Hg has been linked to various sources of exposure, and
routes of administration of different chemical forms of the metal.
Elemental mercury (Hg0) is volatile at room temperature, slightly
soluble in water and easily oxidized to the mercuric ion (Hg2+)
when exposed to atmospheric oxidants, such as oxygen, ozone, and
chlorine [1,2]. Toxicity from elemental Hg varies with the dose and
the rate [3,4], and according to Ozuah, Risher et al., Lech et al., and
Caravati et al. [1,5–7], most commonly arises from inhalation of Hg
vapors in occupational settings (chemical, electronic and mining
industries, production of mercury switches, thermostats and other
instruments), but can also follow residential (e.g. from the
accidental breakage of mercury-containing devices such as
thermometers) and cultural exposure (elemental mercury is
believed to have enormous spiritual and magical powers) [1],
through ingestion of Hg bonded to organic moieties (methyl,
dimethyl, or ethyl mercury), primarily from seafood (entering the
brain, progressively demethylated to elemental Hg) [2–4,8,9] or
* Tel.: +48 12 6185749; fax: +48 12 422 3850.
E-mail addresses: tlech@ies.krakow.pl, tenialech@gmail.com
amalgams) [10]. Other rarer causes are: intravenous or subcuta-
neous self-injections with a large dose of metallic Hg (most often
for suicide attempts) [11–15], aspiration of Hg after ingestion [16]
or mercury intoxication due to topical application of an anti-
freckle cosmetic cream containing an enormous amount of Hg [17].
Acute elemental Hg poisoning is generally not common.
Children are particularly vulnerable to Hg intoxication [7,18–
24], which may lead to impairment of the developing central
nervous system (multiple hyperintense lesions in cerebral white
matter) [24], as well as pulmonary and nephritic damage
[18,19,21,23]. They are seldom exposed to high levels of vapor,
which occur mainly in occupational settings. A potential health
hazard is spilled elemental Hg in the house (at room temperature),
the consequences of which depend on ventilation conditions and
its difficult removal from the environment [7]. In the past, during
the first half of the 20th century, children were often treated with
teething powder that contained inorganic Hg in the form of
calomel, and acrodynia (painful limbs) was common [20].
A variety of techniques are available for determining total Hg,
including cold-vapor atomic absorption spectrometry (CV AAS)
[25,26], atomic absorption spectrometry with electrothermal
atomization in a graphite furnace (GF AAS) [26], inductively
coupled plasma optical emission spectrometry (ICP OES) [27] and

http://dx.doi.org/10.1016/j.forsciint.2014.02.015
0379-0738/ß 2014 Elsevier Ireland Ltd. All rights reserved.
e2 T. Lech / Forensic Science International 237 (2014) e1–e5
inductively coupled plasma mass spectrometry (ICP-MS) [28,29] or
X-ray fluorescence, which enables nondestructive analysis of
mercury in solid samples (e.g. hair) [2].
In this paper, a case of deliberate self-poisoning by a teenager is
presented. Until now, such attempts mainly concerned adults
[30,31]. The aim of the study was to evaluate the effectiveness of
the ICP OES and CV AAS techniques in the determination of Hg in
body tissues and fluids in a case of long-term exposure to
elemental Hg at home and/or hospitals, most probably after
inhalation, partially by ingestion, and/or through the skin, but not,
however, after injection.
2. Case history
A 14-year-old girl suddenly died during one of many
hospitalizations, suffering from pains in the hypogastrium of
unknown origin, recurrent and persistent hematuria, as well as a
severe stress reaction of undetermined origin. Computer tomog-
raphy (CT/X-ray) examination of intestines (6 months before
death) revealed the presence of foreign bodies (possibly gunshot
pellets or mercury?). The toxicological effects mainly involved the
renal (renal colic, hematuria) and central nervous system
(psychological, adaptive) disorders. She had not passed stools
for several weeks, vomited, suffered from lumbalgia, kidney pains,
hematuria, and multi-organ failure. Toxicological analysis, per-
formed at hospital, for drugs, drugs of abuse and alcohols, was
negative. Some time after death it was stated (by the prosecutor
and a forensic psychologist) that she had inhaled unknown doses
of metallic Hg from broken thermometers for about 1 year (there
was no indication of the amounts of Hg involved), and, moreover,
she had probably also ingested some amounts of liquid, and/or she
had rubbed mercury into skin (permanent inflammatory state of
skin in the left hip), because she was suffering from Münchausen
syndrome.
Postmortem findings included cerebral edema to a great degree,
lung inflammation, congestion and edema of lungs, weak blood
supply of myocardium, erosive esophageal mucosa, hyperemia of a
part of the pancreas, intensive congestion of liver.
3. Materials and methods
3.1. Samples
Postmortem blood, hair and internal organs (stomach, liver,
kidneys) were taken from a young girl at a children’s hospital.
Samples of internal organs, except stomach, were homoge-
nized. Before determination of Hg by CV AAS, tissues (10 g, and in
the case of reference material – 0.5 g), blood and urine (5 mL), and
hair samples (0.25 g, washed with water and acetone, cut into
small pieces: 0.5–1 cm), were digested in duplicate with nitric(V)
and sulfuric(VI) acids (2 mL, and 10 mL, respectively) in closed
glass vessels.
Before determination of Hg by ICP OES, digestion of 2 mL of
blood, 5 mL of urine or 2 g of internal organ (in the case of reference
material – 0.5 g) with nitric acid (3 mL) and hydrogen peroxide
(1 mL) was performed by the use of a microwave oven (Ethos 1,
Milestone, Italy) with ten high pressure Teflon vessels, in
accordance with the following program: time 2 min, temperature
– 85 8C; time 4 min, temperature – 135 8C; time 5 min, tempera-
ture – 230 8C; time 15 min, temperature – 230 8C. The mineralized
samples were analyzed in glass vessels on the same day.
3.2. Instrumentation
The screening analysis of biological material for toxic metals
was carried out by inductively coupled plasma optical emission
spectrometry (ICP OES), with the use of an iCAP 6300 emission
spectrometer (Thermo Electron Corp., Waltham, MA, US), allowing
the recording of the full emission spectrum of the sample:
166.250–847.000 nm with the help of a charge-injection device
(CID). The fundamental features of the spectrometer and measur-
ing conditions applied in the analysis were the following: Echelle
type monochromator, radio frequency – 27.12 MHz, radial/axial
plasma observation. The instrument was calibrated against a
multi-element standard (23 elements: Ag, Al, B, Ba, Bi, Ca, Cd, Co,
Cr, Cu, Fe, Ga, In, K, Li, Mg, Mn, Na, Ni, Pb, Sr, Tl, Zn), in the range
0.01–2 mg/mL, and a mercury standard. Linear regression analysis
for Hg gave a regression coefficient (R2) of >0.999 for emission
lines of l = 184.950 nm, 194.227 nm, and 253.652 nm – in the
concentration range up to 10.0 mg/mL of Hg.
Total mercury was determined by means of cold vapor atomic
absorption spectrometry (CV AAS) using an atomic absorption
spectrometer Solar MQZe equipped with vapor system VP100
Continuous Flow Vapor Accessory and a mercury absorption cell
(Thermo Electron Corp., Waltham, MA, US). Hg2+ ions were reduced
by means of 0.5% (m/v) sodium tetrahydroborate(III) in 0.5% (m/v)
sodium hydroxide in the presence of 5% (v/v) hydrochloric acid
(from 30% HCl) to elemental mercury Hg0. The linear range of
calibration was up to 200 mg/L of Hg.
3.3. Reagents
All reagents were of analytical grade (concentrated acids were
Suprapur quality) from Merck (Darmstadt, Germany).
The calibration solutions for Hg (and other toxic metals) and
spiked samples were prepared with 1000 mg/L of Hg (Hg(NO3)2) in
2 M (w/v) HNO3 (Merck), and 1000 mg/L of ICP multi-element (Ag,
Al, B, Ba, Bi, Ca, Cd, Co, Cr, Cu, Fe, Ga, In, K, Li, Mg, Mn, Na, Pb, Sr, Tl
and Zn) standard solution IV (Merck).
Certified reference materials: NIST Bovine Liver 1577b
(Gaithersburg, Maryland), CRM Seronorm: Whole Blood L-2, and
Trace Elements Urine (SERO AS, Billingstad, Norway) for mercury
determination by ICP OES; and NIST Bovine Liver 1577b
(Gaithersburg, MD), CRM 185 Bovine Liver (BCR, Geel, Belgium)
and Seronorm Trace Elements Urine (SERO AS, Billingstad, Norway)
– for analysis by CV AAS, were used throughout.
All aqueous solutions were prepared using deionized water
(18.2 MV, NANOpure Diamond Barnstead, Dubuque, Iowa, US).
4. Results and discussion
The overall procedures for analysis of biological material for
metals, particularly mercury, were assessed by using reference
materials and spiked samples (CV AAS).
4.1. ICP OES
The limits of detection (LOD) and quantification (LOQ)
measured (n = 20) for the replicates of the blank (which had been
taken through the digestion procedure before analysis), accepted
to be three SD (LOD) and ten SD (LOQ) at the emission lines of Hg,
were as shown in Table 1 about 20 ng/L and 70 ng/L (blood), 7 ng/L
and 22 ng/L (urine), respectively, and LOD and LOQ in tissues was
about 23 ng/g and 70 ng/g of Hg at the most sensitive line for Hg
(l = 194.227 nm).
The intra-day precision (RSD), determined on the basis of
results obtained for 3 different samples of digested liver – each
tested 10 times for Hg – was below 6% at the level of 2400 ng/g of
Hg. The inter-day precision was not evaluated because samples
should be analyzed for mercury on the same day that they were
prepared. The accuracy of the whole procedure (digestion in a
microwave system and determination by ICP OES) was assessed
 T. Lech / Forensic Science International 237 (2014) e1–e5 e3

Table 1 Table 3
LOD and LOQ of Hg in blood and urine samples by ICP OES. Accuracy study for Hg using SRM seronorm trace elements urine, bovine liver
1577b, and CRM 185 bovine liver (n = 5) by CV AAS.
Wavelength (nm) Blood (n = 20) Urine (n = 20)
Sample Certified value Found Recovery
LOD (mg/L) LOQ (mg/L) LOD (mg/L) LOQ (mg/L)
(mg/L) (mg/L) (%)
184.950 0.013 0.043 0.0048 0.0160
Seronorm Trace 0.0403  0.0026 0.0410  0.008 101.7
194.227 0.021 0.070 0.0066 0.0220
Elements Urine
253.652 0.321 1.07 0.0402 0.134
Bovine Liver 1577b (0.003)a 0.003  0.001a 100.0
CRM 185 Bovine Liver 0.044  0.003a 0.046  0.004a 104.5
a
Table 2 mg/g, in parentheses – noncertified value.
Accuracy study for Hg using SRM seronorm whole blood L-2 and seronorm trace
elements urine, bovine liver 1577b, and CRM 185 bovine liver (n = 5) by ICP OES.
Table 4
Sample Certified value (mg/L) Found (mg/L) Recovery (%) Postmortem tissue distribution of mercury (n = 3) in a fatal case of poisoning in a
teenager.
Seronorm whole 0.0087  0.0013 Not detected 0
blood L-2 Sample Concentrations of Hg (ng/g)
Seronorm trace 0.0403  0.0026 0.032  0.008 80
ICP OES CV AAS
elements urine
Bovine liver 1577b (0.003)a Not detected 0 Blood Not detected 87  8
a Stomach 90  20 150  12
mg/g, in parenthesis – noncertified value.
Liver 2470  137 2680  45
Kidney 64,199  1200 56,070  180
Hair: 0–3 cma Not tested 1037  28
using reference standards – CRM Seronorm: Whole Blood L-2,
20–25 cma Not tested 964  23
Trace Elements Urine, and SRM Bovine Liver 1577b (n = 5). The
a
results obtained are summarized in Table 2. Distance from scalp.

4.2. CV AAS
LOD and LOQ estimated for Hg (l = 253.652 nm), measured as
replicates of the blank (n = 20), were 0.3 and 1 mg/L of Hg (blood,
urine), and 0.19 and 0.63 ng/g of Hg (tissues), respectively.
Recovery of Hg, assessed using spiked urine samples (with
addition of 1, 2, and 5 mg/L of Hg), was 89.9%, 95.7%, and 99.2%,
respectively. The results for the accuracy study of Hg are presented
in Table 3. Intra-day precision (RSD) was below 3.70% at the level of
1.53 mg/L of Hg (urine).
Analysis of blood and internal organs in the described case for
mercury and other toxic metals carried out by ICP OES revealed
high concentrations of Hg in kidneys and liver (64,200 and
2470 ng/g, respectively), less in stomach (90 ng/g), and none in
blood (small amounts close to the LOQ are not presented) (Table 4).
Other heavy metals (e.g. Pb, Cd, Zn, Cu, Mn, Cr) in internal organs
and blood were at concentrations which are usually found in
normal cases; Tl, Ba, and As were not detected. Using CV AAS, high
levels of Hg were confirmed in all examined materials, including
blood (87 ng/g), as well as in hair (Table 4). The results of analysis
obtained by the two techniques revealed that in this case, the
exposure to mercury was considerable. The concentrations of Hg in
the investigated material exceeded the normal levels by many
times and were comparable to the toxic mercury concentrations
reported by other authors. One additional and noteworthy finding
was that CV AAS is a more sensitive technique, particularly for
blood samples (negative results using ICP OES), with LOQ: 0.63 ng/
g of Hg (CV AAS) and 70 ng/g of Hg (ICP OES) for tissues. However,
ICP OES may be used as a screening technique for autopsy samples
in an acute poisoning by a heavy metal, even one as volatile as
mercury. In the present case, lack of brain and lung tissues for
analysis made it impossible to indicate the exact way (or ways) of
poisoning – to the author’s best knowledge (the records of the
proceedings in the case), the most probable route for intoxication
with mercury in the young girl was the respiratory tract, but
ingestion cannot be excluded, as well as introduction through the
skin.
The mercury content in hair samples was also relatively high –
about 1.0 mg/g near the scalp – in comparison with the mean
mercury levels in hair of non-occupationally exposed inhabitants
of Warsaw or Kraków (Poland) were 0.15–0.17 mg/g [15]. The
mercury content in hair (mean and median) for a non-exposed
group of children (n = 40) in Spain was 0.76 mg/g, lower than the
value of 1 mg/g that the WHO considers as normal for populations
that do not consume fish with high methylmercury content [32].
Pesch et al. [33] reported that the geometric mean and median

Table 5
Reference concentrations of mercury in blood and internal organs by different authors (arithmetic mean, median, range).

Material Reference concentrations of Hg (ng/g)

Drasch (1994) [26] Hać (2000) [34] You (2000) [27] Bárány (2002) [35] Lech (2004) [36] Goullé (2005) [37] Goullé (2010) [29]
Germany Poland Korea Sweden Poland France France
(CV AAS) (CV AAS) (ICP OES) (ICP-MS) (CV AAS) (ICP-MS) (ICP-MS)

Blood 150  120 1.1a 1.6  1.2 3.0a

1.2  0.6 1.4a (0.94–77.6)
(<0.7–6.1) (<6.5)
Stomach 2.0  1.0
1.8a (<4.5)
Liver 51  59 29.9  22.0 220  220 15.5  9.5 45a
12.9a (10–170)a
(2.6–65)
Kidney 170  235 68.6  92.3 330  310 35.9  31.9 77a
28.2a (20–440)a
(3.2–170)
Hair 1200  810 149  65 660a
155a (310–1660)
a
Median.
e4 T. Lech / Forensic Science International 237 (2014) e1–e5
were 0.18 mg/g, and the concentrations ranging between: lower
than 0.06 up to 1.7 mg/g, respectively, for mercury content in the
hair of 245 German children (8–10 years old).
There is, generally, not much data on current reference values
for mercury in the human body, particularly in internal organs.
Some levels of Hg in human organs, blood and hair in non-
intoxicated and non-exposed persons, according to the literature
and own data, are quoted in Table 5. The whole blood and urine
concentrations of mercury are low [38–40]. The reference level of
Hg (geometric mean) in U.S. children (aged 1–5 years) is 0.32 mg/L
in blood, and the 95th percentile is 1.2 mg/L. In urine the reference
values are 0.254 mg/L (in young children, aged 1–5), and 0.358 mg/
L (in young people 12–19 years), and are similar to background
mercury levels in blood and urine in German children [21,40]. The
reference values for Hg in the blood of children (6–12 years old)
in the German population given by Wilhelm et al. [40] are
1.0 mg/L (previously 1.5 mg/L), and 0.7 mg/L in urine (lowered
from 1.4 mg/L).
Although children are not typically exposed to mercury in
active workplaces, some domestic instruments (e.g. thermo-
meters) may be a source of toxic mercury [7,9,24,41].
Acute poisoning with mercury always has dramatic conse-
quences due to the cellular effects in different vital organs [42];
however, chronic poisoning may also be dangerous. In this case,
death was probably a result of multi-organ failure in the course of
chronic intoxication by mercury. The fact that the young girl
sometimes heated elemental mercury in order to develop
symptoms of toxic effects was significant (she had been searching
for such information on numerous websites). Caravati et al. [7]
described a few cases of acute elemental mercury poisoning in
children and adults with symptoms of chemical pneumonitis and
several deaths after exposure from vapors from heating liquid
mercury in an open pan on a kitchen stove; e.g. a 7-month-old girl
died from respiratory failure after exposure to elemental mercury
vapor from heated elemental mercury at her home. Asano et al. [9]
stated that exposure to more 1–2 mg/m3 of elemental exposure
vapor for a few hours causes acute chemical bronchiolitis and
pneumonitis, 2 h after exposure, lung injury appears (hyaline
membrane formation), and finally, extensive pulmonary fibrosis
occurs. Clinical findings correlate always with the duration of
exposure, the concentration of mercury in blood and urine, and the
survival time after exposure, but there is no correlation between
pathological findings and the levels of mercury in tissues.
Liquid metallic mercury is poorly absorbed from the gastroin-
testinal tract [4,18]. Cases of mercury poisoning where mercury
was deposited in diverticulas, fistulas or abscesses in the
alimentary tract, however, have been described. Although
systemic absorption of metallic Hg was probably minimal in
uncomplicated cases, such situation might be harmful if stasis
occurs long enough for the elemental mercury to be converted to
toxic mercuric compounds [4,18]. In the available literature, there
is no data on concentration of Hg in tissues after inhalation or/and
additional ingestion of metallic mercury. For comparison and to
indicate a significant difference in organ distribution, however, the
following postmortem tissue concentrations of Hg in a fatal
intentional poisoning with mercury chloride by ingestion (simul-
taneously with alcohol, tricyclic antidepressants, and potassium
cyanide) in a 36-year-old woman can be cited: mucosa of stomach
– 6624 mg/g, kidney – 284.3 mg/g, liver – 65.1 mg/g, spleen –
56.3 mg/g, lung – 38.5 mg/g, myocardium – 16.9 mg/g, brain –
1.3 mg/g [42].
Elemental mercury is still a current potential toxic substance in
the modern world, because, as Mercer et al. [41] have already
mentioned, it is difficult to eliminate mercury from personal care
products, household items, medications, and vaccinations. This
leads, in consequence, to toxic exposure to elemental mercury in
the home environment, an example of which is a 3-year-old girl
with acrodynia [41] and the described case of acute poisoning of a
teenager with mercury.
5. Conclusions
Assessment of tissue mercury content was a valuable contri-
bution to ascertainment of the cause of ‘‘sudden’’ death of the
young girl.
The results of analysis obtained by two methods (ICP OES, CV
AAS) proved that in this case, exposure to mercury was
considerable, and it could have been the cause of death of the
14-year-old girl. A screening analysis of biological material for Hg
carried out by ICP OES revealed high concentrations of Hg in kidney
and liver (64,200 and 2470 ng/g, respectively), less in stomach
(90 ng/g), and in blood <LOQ; the concentrations of Hg exceeded
the normal levels many times. Other heavy metals (Pb, Cd, Zn, Cu,
Mn, Cr) in internal organs and blood were at concentrations which
are usually found in normal cases; Tl, Ba, and As were not detected
(ICP OES). Using CV AAS, high levels of Hg were confirmed in all
examined materials, including blood (87 ng/g), and additionally in
hair; this is a more sensitive method, particularly for blood
samples (negative results obtained by ICP OES); LOQ: 0.63 ng/g of
Hg (CV AAS), and 70 ng/g of Hg (ICP OES). The main conclusion is
that CV AAS has been a method of choice in determination of
mercury in a wide range of biological material, in comparison with
ICP OES, which may be used as a screening technique for autopsy
material in a case of acute poisoning by a heavy metal, even one as
volatile as mercury, but not for hair or blood.
Conflict of interest
There are no financial or other relations that could lead to a
conflict of interest.
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