Verification of Fatal Gasoline Intoxication in
Confined Spaces Utilizing Gas-Liquid
Chromatography
ROBERT J. NELMS, JR., LCDR., MC,
USN,
RICHARD L. DAVIS, CDR., MC,
AND JAMES BOND, LT., MSC,
USN,
USN
Department of Pathology, Naval Hospital, Corpus Christi, Texas 7S419, Neuropathology
Branch,
Department of Pathology, Naval Hospital, San Diego, California, and Toxicology
Laboratory, Clinical Chemistry Division, Laboratory
Department,
Naval Medical School, Bethesda, Maryland
ABSTRACT
Nelms, Robert J., Jr., Davis, Richard L., and Bond, James: Verification of
fatal gasoline intoxication in confined spaces utilizing gas-liquid chromatography. Amer. J. Clin. Path. 53: 641-646, 1970. Two cases of fatal gasoline intoxication in confined spaces are presented, with a procedure for analysis of
hydrocarbons in tissue utilizing gas-liquid chromatography. Possible mechanisms causing death are discussed. It is suggested that with the advent of more
sensitive technics, which enable the investigator to detect and identify hydrocarbons qualitatively in the range of 0.1 mg. per Gm. of tissue, more useful
data may be accumulated to clarify further the dynamics of acute hydrocarbon intoxication in confined spaces.
to health caused by the fumes
of volatile hydrocarbons, particularly in
confined spaces,10 is well known; however,
there have been only a few reported cases
of fatal toxicity from gasoline fumes in
poorly-ventilated spaces that included autopsy findings or the results of toxicologic studies. 1 '"' 16 Furthermore, information about exact tissue concentrations of
gasoline that cause toxicity is lacking, principally because methods of analysis have
lacked the sensitivity to detect the minute
quantities usually involved.
A related problem with broader clinical
application is oral ingestion leading to
acute hydrocarbon toxicity, which occurs
most frequently in children. 11 ' 1S The morbidity under these circumstances results
from aspiration of the hydrocarbon, and
sometimes gastric contents, with resultant
T H E HAZARD
Received July 11, 1969; accepted for publication
September 5, 1969.
641
pneumonitis and hemorrhagic pulmonary
edema.3-6 Hydrocarbons with low boiling
points and high vapor pressures tend to
be more toxic when taken orally, apparently due to the inhalation of greater quantities of hydrocarbons in the vapor phase.
Hence the pathophysiologic mechanisms
simulate inhalation of toxic vapors in confined spaces.
Vapors in confined spaces constitute a
much more complex milieu than simple
ingestion, however, depending not only on
the relative toxicity and concentration of
the hydrocarbon in the atmosphere, but
also on the carbon dioxide and oxygen
tensions. Although effects on the central
nervous system and gastrointestinal effects
are transitory and rarely fatal in acute
toxicity, there is evidence that reversible
early central nervous system effects may
lead to behavioral aberrations which diminish the victim's ability to rescue him-
642
NELMS ET AL.
self.14 A further role is attributed to nervous
system changes by Gleason and associates,8
who state that the chief systemic reaction
to acute petroleum hydrocarbon poisoning
is central nervous system depression, with
death being due primarily to respiratory
arrest. Other features reported at autopsies
include subserous hemorrhages in the liver
and spleen, hemorrhages into the kidneys
and serous cavities, "cloudy swelling" and
fatty changes in the liver, renal edema
with damage to proximal convoluted tubules and glomeruli, and changes in the
brain, including hyperemia, edema, and
hemorrhages. 4
The purpose of this report is to present
two cases of fatal gasoline intoxication in
confined spaces, with autopsy and toxicologic findings, and the technical details of
the toxicologic procedure which utilizes
gas-liquid chromatography.
Report of Cases
Case 1. An 18-year-old Caucasian youth
was found overcome by fumes in an aviation pumping compartment aboard ship.
Upon removal from the space he was cyanotic, apneic, and without pulse or audible
heart beat. During attempted resuscitation,
the odor of gasoline was noted during expiration. The victim did not respond to
therapy. Areas of superficial skin reddening, blistering, and sloughing were noted.
Autopsy revealed a normal-looking young
man, 5 feet, 10 inches tall, weighing 150
pounds, who had multiple circular "burns"
of the left arm, right scapula, and buttocks. The larynx, trachea, and smaller
bronchi were edematous and filled with
blood-tinged sputum. The lungs were
slightly congested, with focal hemorrhages.
Microscopic examination of the lungs
revealed marked alveolar capillary congestion, intra-alveolar edema, and focal acute
hemorrhage. There was acute inflammation
within many bronchial lumina, and the
mucosa of larger bronchi showed marked
capillary dilatation with abundant sub-
Vol. 53
mucosal chronic inflammation. Mild centrilobular congestion of hepatic sinusoids was
noted. Sections of cerebrum, cerebellum,
basal ganglia, and pons showed generalized
vascular congestion and edema. There were
early ischemic changes in occasional neurons, more prominent in the basal ganglia
and pons. A section taken from the
"burned" area of skin showed absence of
surface epithelium with no capillary congestion or inflammation.
Toxicologic studies of various tissues revealed aviation gasoline, with a spectrum
identical by gas chromatography to that
sampled in the chamber. The gasoline concentration in the brain was estimated to
be 0.3 to 0.4 mg. per Gm. of tissue.
Blood-gas studies of arterial blood, performed on a 30-hr. postmortem specimen,
were: 0 2 saturation 63.5%; P 0 2 33 mm.
Hg; P c 0 2 100 mm. Hg; pH 6.8; hematocrit
47%. In the inferior vena cava the 0 2 saturation was 7%; P 0 2 6 mm. Hg; P 0 0 2 100
mm. Hg; pH 6.8; hematocrit 40%.
Case 2. An 18-year-old Caucasian youth,
repairing a leak in an aviation gasoline
filter room on board ship while at sea, was
overcome by gasoline fumes in company
with another man, who was able to summon help before losing consciousness. The
youth could not be revived, although his
partner recovered.
Autopsy revealed a young man, 5 feet, 9
inches tall, weighing 160 pounds, with variably-sized red to red-brown excoriated lesions of the skin on all extremities and scattered small lacerations and contusions. The
pericardial sac contained sanguineous fluid,
and an epicardial hematoma was present,
both probably associated with resuscitative
attempts. The epiglottis, larynx, and tracheobronchial tree showed marked erythema and roughening of the mucosa. T h e
lungs were heavy, with diffuse congestion
and hemorrhage. The liver and kidneys
appeared congested grossly.
Microscopic examination of sections of
the lungs showed marked alveolar capillary
May 1970
VERIFICATION OF FATAL GASOLINE INTOXICATION
congestion with extensive intra-alveolar
hemorrhage and many pigment-laden septal cells. The pigment was Prussian-bluenegative. Capillary congestion was present
in all organs. No abnormalities in the central nervous system were noted.
Qualitative analysis, by gas chromatography, of blood, urine, gastric contents,
lung, liver, and kidney, yielded a spectrum
identical to that obtained from aviation
gasoline. Quantitation revealed, in the
liver, 0.7 mg. per Gm. of tissue, and in the
lung, 0.4 mg. per Gm. of tissue.
Toxicologic Procedure
The method employed in the Toxicology
Laboratory, Naval Medical School, National Naval Medical Center, Bethesda, to
measure volatile compounds relies upon
the rapid partition which takes place between a volatile substance in solution and
the air above it. Harger and co-workers9
have measured alcohol concentrations in
the liquid phase from the analysis of the
air above the liquid. Goldbaum and associates 7 have modified the technic, using
gas-liquid chromatography to identify and
measure complex mixtures of volatile hydrocarbons.
Apparatus and Operating Conditions
The instrument used in this laboratory
is a Perkin Elmer Model 154 gas chromatograph, equipped with a hydrogen-flame
ionization detector. The inert carrier gas
used is helium. A full-scale Leeds and
Northrup Speedomax G, 0 to 5 mv. per
sec. response, recorder with a chart speed
set at 30 in. per hr., is used with the
model 154. The chromatographic column
used to measure volatile hydrocarbons is
a stainless steel tube of 4.5 mm. internal
diameter, six feet long, packed with 60-80
mesh Chromasorb W, diatomaceous silica,
impregnated with 28.5 Gm. of Ucon Oil
550X * per 100 Gm. of Chromasorb W.
* The Ucon Oil 550X, liquid phase was obtained
from Analabs Inc., Hamden, Connecticut 06518.
643
T h e column was preconditioned for 4
hr. at 125 C. prior to use.
The operating conditions of the gas chromatograph for the determination of volatile compounds were: column temperature
100 ± 2 C ; both injection port and detector, approximately 50 C. higher than
column temperature; inlet pressure of
hydrogen and air, 15.5 p.s.i. and 30 p.s.i.,
respectively, at the pressure gauge; carriergas flow rate approximately 60 ml. per min.
Analytic Procedure
After removal from the freezer, the tissue was allowed to thaw partially at room
temperature, in order to permit easier handling of the specimens. After thawing, but
while still cold, 20.0 Gm. of liver tissue
and 20.0 Gm. of lung tissue were placed
in separate Waring blenders. An equal
quantity (20.0 ml.) of distilled water was
added to each and the tissues were thoroughly homogenized. Aliquots of the homogenates weighing 2.0 Gm. were placed
in clean 10 ml. red-top vacutainer tubes
and the rubber stoppers replaced. The
tubes were placed in a 50-C. water bath
until the temperature of the specimens
reached the temperature of the bath; this
normally took about 20 min. After equilibration was complete, the needle of an airtight syringe outfitted with a two-way stopcock was inserted through the rubber stopper of the sample container. The stopcock
was opened, 4.0 ml. of the vapor over the
sample was drawn into the syringe, and the
stopcock was closed. The needle was disengaged from the syringe and the syringe
attached to a two-way stopcock on the
injection port of the gas chromatograph.
Then the stopcock was opened and the
sample injected into the injection port of
the column, following which the stopcock
was closed. As the sample was being injected, a stopwatch was started, to time the
various peaks displayed on the recorder as
the volatile components passed through the
column to the detector. Thus, the time re-
644
NELMS ET
quired for each component to pass through
the column to the detector served as a
means of qualitative identification. Because
peak height is proportional to concentration of the particular component, the peak
height also can be used for quantitation.
T o provide positive identification and
quantitation of the suspected gasoline, a
control specimen was prepared using normal liver tissue. A 20.0-/xl. aliquot of the
suspected gasoline t was added to 20.0 Gm.
of normal liver, using a 5O7J. syringe. This
tissue with the known gasoline concentration was used as a standard, being prepared
and handled exactly like the unknown
tissue.
Quantitation and Results
The gas chromatographic patterns of the
unknown liver and lung tissues had the
same component peaks and retention times
as the control tissue with aviation gasoline
added. As depicted in Figure 1, the relative
retention times of the five components of
the unknown specimens, compared with
the components of the control specimen,
were sufficient proof of the identity of aviation gasoline present in the suspected tissues. The component peak with the retention time of 1 min. and 30 sec. was selected
for quantitation. The following formula
was used to estimate the amount of gasoline present in the tissues:
Peak height of unknown _ .
. .„ . , . _
2
X (cone, sttl.) X 0.7 =
Peak height of standard
milligrams of gasoline per Gram of tissue.
Cone. std. = concentration of standard =
1.0 /J. perGra.
The density of the aviation gasoline, 0.7,
was used as a multiplication factor in order
to convert microliters per Gram to milligrams per Gram. Using this technic, the
f Aviation gasoline 115/145 (mil. F5572D) was
obtained from the Naval Air Station, Patuxent
River, Maryland.
Vol. 53
AL.
aviation gasoline found in the second victim (Case 2) amounted to 0.7 mg. per Gm.
of liver and 0.4 mg. per Gm. of lung.
Discussion
The following information, if known,
would more fully document these cases:
(1) the atmospheric concentration of the
particular hydrocarbon that would be toxic
if inhaled, (2) the actual concentrations of
hydrocarbon in the confined spaces, (3) the
concentrations of oxygen, carbon dioxide,
and carbon monoxide in the confined
spaces at the times of exposure, (4) the tissue concentrations of the particular hydrocarbons known to cause toxicity or death,
and (5) the levels of blood gases in the
victim immediately after his demise.
Petrol, or gasoline, is a mixture of paraffins, cycloparaffins, and aromatic hydrocarbons. In 1941 Machle 12 stated that the oral
fatal dose was 7 Gm. per kg., and that inhalation of atmospheres with greater than
10,000 p.p.m. would lead to rapid death
in animals. Goodman and Gilman 8 state
that concentrations above 2,000 p.p.m. are
rapidly toxic, but in instances of prolonged
exposure not more than 300 p.p.m. should
be allowed, according to Sterner.15
Not enough information about tissue
concentrations is available to permit any
conclusions about toxic levels due to the
difficulty of performing sufficiently sensitive
analyses. In the inhalation death of a 3year-old child reported by Ainsworth, 2 one
ml. of petrol was distilled from 195 Gm.
of lung (equivalent to 5 mg. per Gm.), but
none could be isolated from stomach (15
ml.), liver (200 Gm.), or brain (200 Gm.).
Wang and Irons 1 6 reported a case of death
following a 5-min. exposure in an airplane
wing tank, and an unidentified volatile
material was extracted from the brain by
steam distillation. In contrast, in the present cases, petrol was determined qualita-
May
1970
VERIFICATION OF FATAL GASOLINE INTOXICATION
tively and quantitatively in the range of
0.3 to 0.7 mg. per Gm. of tissue, levels onetenth those detected by Ainsworth (5 mg.
per Gm.). It is also of interest that the two
cases of Ainsworth and Aiden 1 - 2 were complicated by bullous excoriations of the skin
similar to those found in both of the above
cases.
Blood-gas studies of the first victim were
clone (Case 1), but not until 30 hr. postmortem. Although the results at first glance
suggest anoxia with carbon dioxide retention and acidosis, the equal C 0 2 tensions
in arterial and in venous blood demonstrate postmortem diffusion of C 0 2
throughout the body fluids and suggest
that postmortem oxidative processes may
have contributed to the low oxygen tension and pH.
Some generalizations about the above
cases can be hazarded. At least some, if
not all, of the toxic morphologic changes
were the result of the inhaled hydrocarbons. The hemorrhagic pulmonary edema
testifies to this, being more severe than the
congestive changes expected in the presence of anoxia alone. However, the combination of a hypoxic atmosphere with
hemorrhagic pulmonary edema may have
been overwhelming, when separately they
might not have been fatal. Alcohol or sedatives, if present, might have contributed to
the hypoxic state or potentiated hydrocarbon toxicity. Although toxicologic studies
were not performed, there was no historic
evidence of ingestion.
645
T i m * (minutei)
STD. LIVER
1.0 /i LITER / G M OF LIVER
AVIATION
GASOLINE ,
The central nervous system effects, even
if reversible, might have rendered the victims unable to escape the hostile atmosphere. If respiratory arrest was a factor,
it cannot be established without knowing
experimentally or from clinical experience
what concentrations in the brain are likely
to produce such arrest.
FIG. 1. Gas chromatographic comparison of aviation gasoline in tissue from patient's lung and liver
and a control tissue containing 1.0 pi. per Gm. of
aviation gasoline (Case 2).
J^,
Time (pimutai)
-9-
646
NELMS ET
The extent to which each of the above
mechanisms contributed to death, and in
what order, remains speculative. With further improvements in technics, data more
useful for identifying tissue levels that produce symptoms may be gathered. Hopefully, these findings will clarify further the
dynamics of hydrocarbon intoxication in
confined spaces when they are combined
with the results of more refined studies in
the future.
AL.
7.
8.
9.
10.
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