Legal Medicine 19 (2016) 67–71

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Legal Medicine
journal homepage: www.elsevier.com/locate/legalmed

Case Report

Detection of toluene in a body buried for years with a fatal cardiac

contusion
Toshiko Tanaka ⇑, Hiroaki Sato, Kentaro Kasai
Department of Forensic Medicine, Faculty of Medicine, University of Occupational and Environmental Health, Japan

a r t i c l e i n f o a b s t r a c t

Article history: This report aimed to present the postmortem finding of toluene in a homicide victim buried under the
Received 17 September 2015 ground for six years. The bones of the skull and limbs were exposed, and the remainder of the subcuta-
Received in revised form 23 February 2016 neous tissues, brain and heart had formed into adipocere. There were numerous fractures in the skull and
Accepted 23 February 2016
the anterior side of the ribs. A cardiac contusion extending into the cavity of the right ventricle was also
Available online 24 February 2016
observed. No other obvious injuries were identified on the body. The concentration of toluene in the bone
marrow within the head of the humerus was 58.4 lg/g. The cause of death was suspected as heart rup-
Keywords:
ture, possibly from a forceful impact or compression of the anterior chest under toluene intoxication. This
Toluene
Adipocere
report presents a rare case where toluene intake by a human was disclosed by autopsy even after several
Cardiac tamponade years of death.
Underground Ó 2016 Elsevier Ireland Ltd. All rights reserved.

1. Introduction The circumstance of the case was revealed subsequent to the

Toluene is used in a variety of applications in our daily lives and
has been frequently abused by teenagers and adults, who inhale
the vapors of paint and glue solvents for its intoxicating effects
[1]. The presence of toluene in postmortem autopsy cases has been
reported in several studies [1–15]; however, there is no report of
toluene detection in cases where autopsy was conducted several
years after death. In this report we present the autopsy case of a
toluene abuser who had been buried underground for six years.
Postmortem analysis revealed the presence of a high content of
toluene in the bone marrow. Toluene detection in a cadaver after
the passage of several years of death is rare.
autopsy. The victim had staggered due to thinner abuse on the
day of incident. The criminals had beaten and kicked the whole
body of the victim repeatedly with their fists and knees. The victim
had gone down and uttered a moan of pain pressing his hand to his
chest in the middle of the assault. His face and neck had turned to
be congested soon, and he had got to wheeze like asthmatically.
His respiration had become gradually weaker. His face, arms and
the tops of the foot had looked swollen when his breathing had
stopped forever. It had been less than one hour from the start of
the assault to his end.
3. Autopsy findings

2. Case history
A 20s-year-old man had been reported missing for six years. His
body was discovered at a depth of one meter below the damp sur-
face of the earth after the confession of the criminals. The place
located in the forest area facing the narrow animal trail that fol-
lowed the road of dead-end, where general public would not enter
easily. The body wore clothes. The autopsy was held one day after
the body was found.
⇑ Corresponding author at: Department of Forensic Medicine, School of Medicine,
University of Occupational and Environmental Health, Japan, Iseigaoka 1-1,
Yahata-Nishi, Kitakyushu 807-8555, Japan.
E-mail address: ttanaka@med.uoeh-u.ac.jp (T. Tanaka).
Almost all of the skin and muscles on his head and extremities
were lost, and the bones of the skull and limbs were exposed
(Fig. 1). The remainder of the subcutaneous tissues, brain and heart
had completely converted into adipocere. Sediment and mold
attached to all over the surface of the body. The remains of the
muscles also appeared to be formed in adipocere. Several fractures
were discovered on the skull surrounding the bilateral orbital rims
and the right side of the frontal bone of the skull. Articulate frac-
tures were also noted on both sides of the mandible. The brain
had turned into a mass of blackish-green clay with no sign of
bleeding. The skin and muscles in the upper part of the torso had
turned into adipocere resulting in the loss of several tissue details.
The costal cartilage was also lost under the soft tissues such as the
skin, fat and muscles. There were several fractures in the anterior

http://dx.doi.org/10.1016/j.legalmed.2016.02.005
1344-6223/Ó 2016 Elsevier Ireland Ltd. All rights reserved.
68 T. Tanaka et al. / Legal Medicine 19 (2016) 67–71

Fig. 1. The clinical appearance of the cadaver.

side of the ribs shown with white arrows in Fig. 2. The spinous pro-
cess of the fifth thoracic vertebra was fractured, while the
adipocerous heart was located in the normal position. The left
and right panels in Fig. 3A show the anterior and posterior sides
of the heart. There was a contusion on the right ventricle that
extended from the anterior to the posterior side of the heart. The
ventricles of heart were horizontally sectioned into four blocks to
confirm the extent of the injury. The left panel in Fig. 3B illustrates
the topsides of the four sections of the heart, whereas the images
on the right panel demonstrate the bottom sides of the same sec-
tions. Arrows show the sites of dehiscence; the injury to the heart
was nearly linear in appearance and extended into the cavity of the
right ventricle. The total length of the split in the heart was slightly
greater than 15 cm. There was not any obvious contusion in the
right atrium or in the other area of the heart. No obvious splits
or contusions were observed either in liver or aorta. None of the
other primary organs could be determined in the thoracic cavity
or in the abdominal cavity.
4. Toxicological analysis
The remaining muscles of the body had been almost adipocere.
The inner content of the head of the left humerus was the only
sample of bone marrow available for toxicological analysis. The
surface of the humerus did not appear to be broken. The inner con-
tents had turned into a dry, oily and blackish material that adhered
to the inner walls of the humerus, and was scratched and collected
for analysis. The drug screening analysis was performed using the
NAGINATATM gas chromatography/mass spectrometry (GC/MS)

Fig. 2. The anterior side of the chest under the soft tissue. The soft tissue covering the breast was removed revealing rib fractures (white arrows).
 T. Tanaka et al. / Legal Medicine 19 (2016) 67–71 69

Fig. 3. The heart of the cadaver. (A) Anterior and posterior surfaces of the heart. The figures below depict a schema where the ruptured site is illustrated in black. (B) The
horizontal sections of the heart. The ventricles of heart were sliced horizontally into four blocks and arranged lining from top to bottom in the left panel; the anterior sides of
each block are placed upper and the posterior sides are placed lower; the topsides of the blocks are demonstrated. The right panel demonstrates the blocks putting upside
down of those seen in the left pane; the bottom planes of each block are demonstrated. L represents the left side of the heart, and R represents the right side of the heart.
White arrows indicate areas of dehiscence.

software [16] along with the 7890 GC and 5975C mass systems. The bone marrow sample was also used for the analysis of vola-
b-Phenethylamine was the only material detected in the 100 mg tile hydrocarbon components. GC/MS was used for the analysis
bone marrow sample. using the modified method described by Kimura et al.[17]. The
70 T. Tanaka et al. / Legal Medicine 19 (2016) 67–71
bone marrow sample (100 mg) was placed in duplicates into a 10-
mL glass vial on an ice cooling mount. Then, 500 ll of cold distilled
water was added to each vial along with 1 lg of ethylbenzene-D10
that was mixed as an internal standard (IS) in 10 ll of methanol. A
standard solution was also prepared with 5% trironÒX-100 solu-
tion, and added to the blank bone marrow obtained from the wing
root of chicken which was commercially available. The vials were
sealed and heated at 60 °C for 20 min using a head space sampler
(Agilent, 7694), which was attached to the injection port of the
GC apparatus.
The reference materials were purchased from Wako Pure Chem-
ical Industries, Ltd. (Osaka, Japan), while other chemicals were pur-
chased from Nacalai Tesque, Inc. (Kyoto, Japan). The GC/MS
apparatus was a GCmate gas chromatography/mass spectrometry
(JEOL, Tokyo, Japan) equipped with a capillary column (DB-5,
0.53 mm i.d., 30 m length, 1.5 lm film thickness) (J&W Scientific,
Folsom, CA, USA) and attached with a deactivated capillary column
(0.25 mm i.d.  5 m length). The column temperature was pro-
grammed to hold at 45 °C for 3.5 min, elevate to 210 °C at 20 °C/
min, then immediately elevate to 260 °C at 30 °C/min, and remain
at the final temperature of 260 °C for 2 min. The injection port and
ion source were held at 260 °C. The scan mode (m/z 30–152) and
single ion-monitoring (SIM) mode were used for the qualitative
and quantitative analyses, respectively. In the qualitative analysis,
the prominent peak was identified by comparing the ratios of the
characteristic ions with those of the authentic standards: m/z 91,
92, 65, 51 for toluene and m/z 98, 116, 82, 70 for IS. The ions mon-
itored in the SIM mode were m/z 91, 92 for toluene, and m/z 98,116
for IS. The chromatograms in scan mode obtained from the bone
marrow of cadaver are shown in Fig. 4. The peaks of toluene and
IS were clearly visible in the chromatograms; no other peaks that
might be confused with objects in either the TIC (total ion
chromatogram) or the characteristic ions were noted in the
chromatograms. Both spectra of toluene and internal standard
were consistent with those of the authentic materials in the right
spectra (Fig. 4). The calibration curve of toluene in the bone
marrow obtained by using a spiking solution at a concentration
of 0–150 lg/g demonstrated the equation (Y = 0.0667 X
Fig. 4. Chromatograms and spectra of the bone marrow using GC/MS. The left panel illustrates the GC/MS chromatograms obtained from the bone marrow of the autopsy case
with the peaks (A and B). The two images on the right panel show the spectra obtained from the peaks A and B of the chromatogram, respectively. Each of the lower images of
the spectrum A and B shows the spectrum obtained from the standard material as noted in the image.
+ 0.00823) with a linearity of correlation coefficient (r = 0.996).
The detection limit of the system (LOD) was 0.06 lg/g and the
quantitation limit (LOQ) was 0.20 lg/g. The concentration of
toluene was 58.4 lg/g in the bone marrow obtained from the head
of humerus of the cadaver.
5. Discussion
The toxic levels of blood toluene have been reported in several
studies [1,10,18,19]. Reports of 136 persons who were either hos-
pitalized or arrested while intoxicated, showed blood toluene con-
centrations of 0.3–30 lg/mL and urine concentrations of 0–5.0 lg/
mL [1]. Three young men who died of acute toluene inhalation
were found to have postmortem blood concentrations of 50, 60,
and 79 lg/mL [9]. A 39 year old man who died while working in
an enclosed space had a postmortem blood toluene level of
270 lg/mL [20]. Eight male victims of ages 14–39 years who died
as a result of accidental or intentional acute exposure to toluene
fumes exhibited the following toluene levels in postmortem spec-
imens; 10–48 lg/mL in blood, 10–182 lg/g in brain, and 1–5 lg/
mL in urine [10–15]. The interpretation of blood data and those
of the bone marrow have proven to be difficult. Tominaga [21]
reported that the distribution of inhaled toluene largely varied
by case; however, bone marrow aspirate levels were about twice
as high as blood levels in 7 abusers. In the present study, half of
the bone marrow concentration of toluene was within toxic and
fatal levels. The postmortem levels of toluene would be higher than
the antemortem levels if it remained for a longer period in the bone
marrow when compared to the other components such as water,
lipids, and proteins. On the contrary, the postmortem levels of
toluene would be lower than the antemortem levels if it remained
in the cadaver for a shorter period of time. The postmortem
dynamic changes in toluene levels, over a period of several years,
have not been documented in a cadaver, so far. Incidentally,
toluene was determined by head space sampling, in the present
study. However, as the bone marrow was very oily the values
obtained may be inaccurate. Thus, we could not confirm the effect
of toluene on the cause of death in the present case despite its
 T. Tanaka et al. / Legal Medicine 19 (2016) 67–71
nearly toxic or fatal levels combined with a history of antemortem
toluene usage.
There were a lot of fractures in the skull and ribs. The presence
of bleeding or contusion could not be determined in the brain,
although cardiac contusion was apparent. The heart lay at the nor-
mal position in the thoracic cavity. The contusion was blunt and
ran nearly straight into the right ventricle. There were no sharp
injuries in the soft tissues of the intercostal ribs or on the heart.
There were no obvious contusions in the other region of the heart.
Right atrium is generally easier to be damaged than right ventricle
in the metamorphic process or some artificial postmortem strength
applied on the body, because the anterior wall of right atrium is
thinner and locates forward in the chest. It’s almost inconceivable
that some metamorphic effect or a certain postmortem external
strength caused the contusion only in the right ventricle. It was
plausible that the contusion in the right ventricle was diagnosed
as occlusive. The causes of occlusive heart contusion include;
steering wheel injury when the driver’s chest is violently pressed
against the car handle in a traffic accident, the pressure of a large
machine, the kick of horse, and the hitting of a ball to the chest wall
[22]. The heart is sandwiched between a strong sternum and the
spine; a powerful force can cause injury to the pericardium, coro-
nal artery, cardiac valves, aorta, and cardiac muscle [23,24]. It is
generally reported that a huge external pressure on the diastole
can induce a heart rupture, whereas external pressure on the sys-
tole can induce an atrioventricular valve rupture. The fracture of
the anterior ribs in the present case indicates that the rupture of
the right ventricle was caused by a forcible impact on the chest
wall during a late diastole under a certain level of toluene intoxica-
tion. The most common feature of blunt traumatic cardiac rupture
is cardiac tamponade [22], leading to uncontrolled blood leakage
and fullness to the pericardial cavity, and finally, death. The frac-
ture of the spinous process of the fifth thoracic spine can also
remind the scene where the back is pressed against a solid plane
such as the floor or wall. This presumable process accorded well
with a sequence of symptoms described in the section of case his-
tory such as a moan of pain pressing hand to chest, congestion of
face and neck, wheeze, and edema of arms and the top of the foot.
The process was the most convincing mechanism about the death.
In summary, we were unable to disclose the direct effects of
toluene on the death of the man in the present study, although
considerable toluene intake was suspected before death. However,
it was clear that the cardiac injury was fatal, as it could not have
been formed after his death. Therefore, death may have been
caused due to cardiac tamponade with rupture of the right ventri-
cle caused by assault and battery on the anterior part of the chest.
The time elapsed from his death until autopsy was estimated to be
five to six years because of the disappearance of the costal cartilage
and the complete formation of systemic adipocere.
71
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