UPTAKE, DISTRIBUTION, AND METABOLISM

OF INHALED ETHYLMERCURIC CHLORIDE

IN THE RAT 1
S. C. FANGand E. FALLIN
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COMMUNICATEDBY LOUISLYKKEN

The uptake and the tissue distribution of ethylmercuric chloride (EMC) by in-
halation and oral administration were studied in adult female rats. The pulmonary
uptake of EMC linearly increases with the time of exposure and is proportional to
the concentration of EMC vapor. With the exception of stomach, intestine, and hair,
the tissue distribution of 2 o 3 Hg from pulmonary uptake is quite similar to that from
oral administration. The biological halflife of EMC in the organs is longer from pul-
monary uptake than from oral administration. Kidney and liver convert EMC to in-
organic mercury irrespective of the routes of administration. A small amourit of
2°3Hg is found in the fetus, and the fetal liver accumulates more 2°3Hg than the
fetal kidney.
Alkyl mercurial pesticides were introduced during the 1940s for seed dressing and,
also, for control of fungus disease in plants. These mercury preparations usually contain
methyl mercury and, to a less extent, ethyl mercury. The short-chained alkyl mercury
salts vaporize easily at room temperature and poisoning occurs after exposure to these
vapors CSwensson t952, Trachtenberg 1969). Studies on the uptake and distribution
of mercury vapor through inhalation have been made in a number of animals (Gage
1961, Hayes and Rothstein 1962, Magos 1967, 1968, Berlin et a l . , 1969, Cassano
et al. 1969). However, detailed reports are not available regarding the respiratory uptake
of alkyl mercury compounds. The retention of dimethyl mercury after a single inhalation
exposure by mice under slight anesthesia has been reported (Ost]und 1969) but the
vapor concentrations used were not given.
This paper reports the uptake, distribution, and metabolism of ethylmercuric chloride
(EMC) in rats as the result of inhalation of EMC and, for comparison, ingestion of
the chemical.

Materials and methods

2°3Hg-labeled ethylmercuric chloride (EMC3°3Hg) was prepared by an exchange
reaction. One mmole of pure, recrystalized ethylmercuric chloride was dissolved in five

1Technical Paper No. 3496, Oregon Agricultural Experiment Station, Corvallis, Oregon.

347

Archives of Environmental Contamination and ToxicoLogy

Vol. 1, No. 4, 1973, © 1973 by
Snrin~er-Verlae New York Inc.
348 S . C . Fang and E. Fallin

milliliters of hot ethanol containing 0.5 ml of 6N HC1. Two mCi of 2°3Hg nitrate
was added and the mixture was allowed to stand at 50°C overnight. After cooling,
EMC-2O3Hg was crystalized out and was separated by filtration or by extraction with
benzene. The recovery of ethyhnercuric chloride was greater than 80%, and the initial
radioactivity of the final product was between 600 to 800 cpm per nmole depending
on the specific activity of 2°3Hg-nitrate used.

Adult female Wistar rats (between 2 and 6 months old, averaging 217+27 g for non-
pregnant, and 286-+9 g for pregnant animals) were used. Two to four rats were exposed
continuously to air containing EMC-203Hg vapor, of known concentration, in an
8-inch all-glass metabolism cage. Water was available but food was not given during
the time of exposure to avoid the consumption of EMC contaminated food. The flow
rate of air was maintained at 500 ml per minute during the experimental period.

Since EMC is quite volatile and its vapor pressure increases rapidly as the temperature
increases, air containing various concentrations of EMC can be produced by passing
the air, at a constant flow rate, over EMC crystals. The concentration of EMC in the
air is determined by the loss of EMC in the tube after a given volume of air has been
passed through the system. The tube containing the EMC crystals is immersed in a
thermoregulated water bath for precise control of temperature. Fig. 1 gives the relation-
ship of EMC vapor concentration in air that flows from the EMC vapor generator to
the temperature, under the experimental conditions used. When the log of EMC vapor
concentration is plotted against the temperature, a straight line is observed, indicating
that the temperature coefficient of the EMC vapor pressure can be expressed by the
Clapeyron equation. By maintaining the supply of EMC crystals in the generator between
100 to 150 mg and maintaining the temperature of water bath at an appropriate tempera-
ture (ranging from 25 ° to 50°C), it is possible to produce air containing various concentra-
tions of EMC, with reasonable accuracy. It is important that all connections between
the EMC generator and the metabolism cage be made of pyrex glass tubing with a
ball socket joint because the EMC vapor readily reacts with rubber tubing and, thus,
affects the vapor concentration. Also, the food pellets absorb uncertain amounts of EMC
vapor; so, the presence of food in the metabolism cage during the time of exposure
interferes with the measurement of pulmonary uptake of EMC. Therefore, the rats were
not provided with food during the inhalation test period.

In the first experiment, the rats were exposed to EMC vapor for 6, 12, 18, and
24 hours in order to determine the uptake and distribution of EMC in relation to the
time of exposure. In the second experiment, which was designed to study the uptake
and distribution of EMC by rats in relation to EMC vapor concentration, all the rats
were exposed to EMC vapor for 24 hours. Four pregnant rats were included in this
experiment to learn whether or not the 2°3Hg translocated to the fetus. In the third
experiment, which compared the route and rate of mercury elimination with the uptake
of EMC by inhalation and by oral administration, two groups of rats were either given
(by intubation) an oral dose of 3 /amole EMC-2OaHg in corn oil or were exposed
to EMC-2O3Hg vapor for 24 hours. Afterwards, the rats were housed individually in
metabolism cages allowing separate collection and measurement of Z°3Hg in the urine
and feces. At prescribed intervals, one or two rats from each group were killed and
 Inhalation Uptake of Ethylmercuric Chloride in Rats 349

the concentrations of ~'~'~Hg in various tissues and organs were determined. Some of
the blood, kidney, and liver samples were taken for determination of inorganic mercury,
according to the method of Clarkson and Greenwood (1970).

A "y-scintillation spectrometer (Technical Associates Model SM-10), equipped with

a two-inch NaI(T1) well detector, was employed for the measurement of radioactivity.
The mercury content in the sample was determined by simultaneously measuring the
radioactivity of the sample and of a sealed standard containing a known amount of
EMC-2O3Hg. The background was between 25 and 30 counts/minute and the counting
efficiency was approximately 40% for 2°3Hg. Each sample was counted for 10 minutes
and the count was corrected for background and the size of sample used. The probable
error, for most samples, was less than 1% and, for low-radioactivity samples, between 2
and 5%.

Results
The data obtained from the measurement of 2°3Hg contents of each organ and tissue
from rats, after exposing the rats to air containing 5 or 85 nmole EMC/1. for various
lengths of time are shown in Table I. It is apparent that the 2°3Hg concentration of

J
O
E
e-
o/o o
°/°
/o
ft
O

=o
O
o

o
> /o
III
O3
O
/
.J

0 I I i I I
20 30 40 50
o
Temperature, C

Fig. 1. Relationship of EMC concentration in air, after passage through the EMC vapor
generator, to the temperature of the generator.
 Table I. Concentration of 2°3Hg in Various Organs and Tissues of Female Rats Expose to Two
EMC-2°3Hg Vapor Concentrations (5 and 85 nmole/liter) for Various Lengths of Time

~°:~Hg content ~, nmole/g of fresh weight, after indicated exposure

5 nmole/liter 85 nmole/liter rao

Organ or tissue 6 hr 12hr 18 hr 24 hr 6 hr 12 hr 18 hr 24 "hr Regression Equation ¢3

Kidney 0.77 3.90 5.00 8.26 5.9 24.0 47.1 63.9 y= - - . 165+ .0387x
Liver 0.36 2,04 1,92 4.08 2.0 10,2 21.1 28. t y= - - . 104+ .0270x
Blood 0.42 2.37 1.87 3.92 2.7 12.1 32.9 45.0 y= --.116+.0311x
Lung 0.29 0.91 0.81 1.72 1.7 6,1 12,7 17,0 y= --.033+.0121x gZu

Spleen 0.13 0.74 0.81 1.75 1.4 3.8 10.3 13,7 y= - - . 0 5 8 + .0126x
Heart 0,13 0,54 0.50 1.01 0.8 3,5 6.7 8.5 y= --,020+.0069x
Pancreas 0.14 0.62 0.53 1.00 1.6 3.5 5.5 8.0 y= - - . 0 0 8 + .0062x
Stomach 1.02 4.87 3.08 2.90 1.1 2.0 18,6 29.9 y= .067+.0213x
Intestine 0.29 1.04 0.81 1.28 0.7 4.4 6.7 6.8 y= - - . 0 0 5 + .0072x
Brain 0.02 0.07 0.06 0.17 0.2 0.6 1.2 1.7 y= - - . 0 0 5 + .0013x
Muscle 0.03 0.12 0.11 0.20 0.3 0.9 2.1 2,7 y= --.004+.0017x
Fat 0.03 0.08 0.08 0.22 0.3 1,0 1.2 1.7 y= .004+.0014x
Hair 60. 104. 107. 254. 983. 1845, 4045. 5805. y= --6.5+2.567x

aAverage result from two rats.

 Inhalation Uptake of Ethylmercuric Chloride in Rats 351

T a b l e II. Concentration of Z°aHg in Various Organs and Tissues of Female Rats

Exposed to Various EMC-2°aHg Vapor Concentrations for 24 hours

Organ 20a Hg content a, nmole/g fresh weight, after exposure

or to indicated vapor concentrations, nmole/liter
tissue 3 5 12 21 45 85
Kidney 7.4___.6 8.3 16.6±6.5 19.6±6.1 37.6±6.5 61.0±6.3
Liver 3.0± .2 4.1 7.9±3,0 8.0±2.5 17.6_+2.9 28.1±2.7
Blood 3.2± .3 3.9 8.9±3.8 10.6±2.5 25.6±4.1 44.9±2.0
Lung 1.3_+ .3 1.7 3.7_+2.0 3.9±1.2 8.1+_2.0 15.9±2.4
Spleen 1,3_+.2 1.8 4.0_+1.3 4.2±1.2 7.1_+1,6 13.3±1.7
Heart 0.9±.1 1.0 2.4_+0.8 2.9±1.1 5.2±1.5 8.5±1.0
Pancreas 0.8±.1 1.0 2.3±0.6 2.1±0.9 5.6±1.8 8.1±0.5
Stomach 1.5±.6 2.9 6.0±3.1 6.2_+2.0 15.3±4.7 25.7±4.1
Intestine 1 . 0 +- .7 1.3 3.9±3.0 2.9±1.2 5.2±1.4 7.5_+1.6
Brain 0.1-+.02 0.2 0.3±0.1 0.3±0.1 0.7_+0.3 1.4_+0.3
Muscle 0.2± .02 0.2 0.5±0.2 0.6_+0.1 1.4±0.3 2.5_+0.3
Fat 0.2±.1 0.2 0.3_+0.2 0.3_+0.1 0.9_+0.2 1.5±0.3
Hair 125 _+90 254. 507 _+96 1213± 165 2t 12± 183 5666_+387
Number o rats used
6 I 2 I 6. I 8 I 5

aMean _+_ standard deviation.

most organs and tissues increase with the time of exposure. The highest 2 o 3 Hg concentra-
tion is found in the kidney (excluding hair); it is about two-fold greater than those
found in the liver and blood. The lung, spleen, heart, pancreas, and intestine each
have a 2O3Hg concentration which is approximately one-fourth to one-half o f that
in the blood. The brain, muscle, and fat contain approximately one-twentieth o f the
z°aHg content in blood. The hair has extremely high radioactivity which increases linearly
with the time of exposure and the concentration o f EMC vapor. The binding of EMC
to hair probably is the result of chemical reaction because attempts to remove the radioac-
tivity from the hair by washing either with water or with organic solvent failed.

The accumulation o f 2°aHg in various organs and tissues, after exposing rats for
24 hours to air containing various EMC-2OaHg concentrations (ranging from 3 to 85
nmoles/1 .) is shown in Table II. The accumulation of z°aHg in various organs or tissues
increases with the increasing of EMC vapor concentration. The relative 2°3Hg accumula-
tion in each organ or tissue remains fairly constant throughout the range of concentration
used. However, when the results are analyzed in terms of EMC vapor available to
the rat, it is clear that, at tow EMC vapor concentrations, more EMC is taken up and
accumulated in the tissues per unit time per unit concentration of EMC than at high
vapor concentrations. This rate of uptake progressively decreases as the concentration
of EMC vapor increases (Table III). Since the initial event of pulmonary uptake of
any vapor is the striking of a gas molecule upon the surface of the lung, the frequency
352 S . C . Fang and E. Fallin

T a b l e III. Hourly Rate of 2°aHg Accumulation in Various Organs and Tissues of

Female Rats Exposed to Various Concenlrations of EMC-2°aHg Vapor

Organ Rate of 2°aHg accumulation, nmole/g fresh weight/nmole EMC/liter, after

or ex ~osure to indicated vapor concentration, nmole/liter
tissue 12 21 45 85

Kidney • 1027 .0692 .0576 .0389 •0348 ,0299

Liver .0417 .0342 .0274 .0159 .0163 .0138
Blood .0444 .0325 .0309 .0210 .0237 .0220
Lung .0181 .0142 .0128 .0077 .0075 .0080
Spleen .0181 .0150 .0139 .0083 .0066 .0065
Heart .0125 .0083 .0083 .0058 .0048 .0042
Pancreas .0111 .0083 .0080 .0042 .0052 .0040
Stomach .0208 .0242 •0208 .0123 .0141 .0126
Intestine .0139 .0108 .0135 .O058 •0048 .0037
Brain .0018 .0014 .0010 .0006 •0006 •0007
Muscle .0025 .0017 .0017 .0012 .0013 .0012
Fat .0024 .0018 .0010 .0006 •0008 .0007
Hair 1.74 2.11 1.76 2.40 1.96 2.15

of gas molecules to striking on a given surface is proportional to the pressure or the

concentration of the gas and this relation can be expressed by the Freundlich equation,
as follows: a
q=aC n

where q is the quantity of adsorbed substance, C is the concentration, a and n are

constants. By plotting the log of specific rate of accumulation against the log of concentra-
tion (Table III), a straight line is obtained, as shown in Fig. 2. At this range of EMC
concentration, the Freundlich equation fits the experimental data quite well and can
be utilized to estimate the pulmonary uptake of EMC and the deposition of mercury
in the tissues.

Table IV compares the 2°aHg accumulation in various organs and tissues of pregnant
and non-pregnant rats after exposure to air containing 12 nmole/1. EMC-203Hg vapor
for 24 hours, Although there are considerable differences in body weight between these
two groups o f rats, the average levet of 2°aHg accumulation in various tissues is quite
similar with the exception of the stomach. It is possible that this anomaly arises from
the supply of food given to the pregnant rats. A small amount o f 2°aHg is found in
the fetus and the level is comparable to those in the brain and muscle. Since the 2°aHg
concentration of the placenta is considerably higher than that found in the fetus, it
is reasonable to assume that the transport o f EMC through the placental barrier is slow.
However, partial conversion of EMC to inorganic mercury can also result in a difference
in the concentration of z°aHg in the placenta and in the fetus because the placental
transfer of mercuric mercury in mice is extremely slow (Berlin and Ullberg 1963, Suzuki
et al. 1967).
 Inhalation Uptake of Ethylmercuric Chloride in Rats 353

60 Kidney j O

-6
E
E
30
8 0///0//// Blood
r0
o
c~

0, l i | I I I I I I I
50 1O0

Vapor concentration(C), nmole/liter

Kidney O~ O
o~ .11A
E
8 O~ O...~ ~ . . -~/"
O
Blood
o1
3

| !
1 2

Log G
Fig. 2. Relationship of 2°aHg accumulation in the kidney and blood of rats to the EMC vapor
concentration of the air inhaled by the rats. Top: linear plot. Bottom: log-log plot.

Several fetuses were taken from two litters and dissected, .and the mercury depositions
in the liver, kidney, lung, and heart were measured. The results are shown in Table
V. The accumulation of mercury in the fetal liver is highest among the four organs
measured, all of which are considerably less than the corresponding organs of the dams.
Approximately 88% to t00% of the radioactivities, respectively, in the liver of the
dams and fetuses are extracted with HCl-benzene, and this extractable radioactivity
consists entirely of EMC-2OaHg (based on thin layer chromatography).
 354 S.C. F a n g a n d E. FMlin

Table IV. Accumulation of "°aHg in Various Organs and Tissues of Pregnant and
Non-Pregnant Rats Exposed to EMC-"°aHg Vapor Concentration of
12 nmole/titer for 24 Hours

Organ 2°aHg content ~, nmole/g fresh weight

or

tissue Pregnant h Non-pregnant b

Kidney 25.2 ± 4.0 16.6 ± 6.5

Liver 6 . 7 ± t.1 7.9 ± 3.0
Blood 8.8 ± 1.4 8.9 ± 3.8
Lung 5.1 ± 0 . 9 3.7 ± 2.0
Spleen 4.2 ± 0.4 4.0 ~ 1.3
Heart 2.5 -± 0.2 2.4 ± 0.8
Pancreas 2 . 9 ± 0.2 2.3 ± 0.6
Stomach 2.2 ± 0.4 6.0±3.1
Intestine 1.7 ± 0.3 3.9 ± 3.0
Brain 0.4±0.1 0.3+0.1
Muscle 0.6_+0.1 0.5 ± 0.2
Fat 0.9 ± 0.4 0.3 ± 0.2
Placenta 2.7 ± 0.3
Fetus ~ 0.39 ± 0.04
Hair 519 507 ~c 96

~'Mean + standard deviation.

~Number of rats used: pregnant = 4; n o n - p r e g n a n t = 6.
CAverage weight = 3.55 g; N = 18.

T a b l e V . Comparative 2°aHg Accumulation in Carcass and Several Organs of Two

Pregnant Rats and their Fetuses Exposed to EMC-2°aHg Vapor Concentration of
12 nmole/liter for 24 Hours

Carcass 2°aHg contenC, nmole/g fresh weight

or Rat 1 Rat 2
organ Dam Fetus b Dam Fetus b

Carcass 0.35 ± .03 E 0,34 ± .03

Liver 5,32 3.42 ± . 15 5.93 2.38 _+ .40
Kidney 33.2 1.05 ± .89 20.2 0,89 ± .12
Lung 6.30 1,46 ± .35 5.75 1.17 ± .61
Heart 2.63 t.02 ± .21 2.51 1.56 ± .48

aMean ± standard deviation (fetus only).

bNumber of fetus used: rat 1 = 9; rat 2 = 6.
 Inhalation Uptake of Ethylmercuric Chloride in Rats 355

In order to determine the rate of loss of EMC in organs and tissues, a group of
16 female rats were exposed to EMC vapor (12 mmole/1.) for 24 hours and then sacrificed
at various times after exposure. The result is shown in Table VI. The 2°3Hg content
in the kidney increases slowly from 10.4 nmole/g (fresh weight) to 76.8 nmole/g in
14 days, whereas that in the liver, blood, heart, stomach, and intestine shows a decline.
The changes of 2°3Hg level of the brain, muscle, and fat are insignificant.
For comparison of tissue distribution of EMC as a result of pulmonary uptake and
oral administration, a group of female rats was given a single oral dose of 3 /xmole
of EMC in corn oil, and the animals were sacrificed at various elapsed times after
dosing. The results are shown in Table VII. All organs, with the exception of the
stomach and intestine, show an increase to a maximum between 1 to 3 days after dosing
and then declines steadily. The 2°3Hg level in the kidney shows a continuous increase
from 21.1 nmole/g (fresh weight) at 6 hr after dosing to 189.0 nmole/g at 10 days.
This result is quite similar to that found with pulmonary uptake. The level of 2°3Hg
in the stomach and intestine decreases very rapidly during the first 4-day period. This
is mainly the result of redistribution (Table VII) and, to a lesser extent, of fecal elimination.
Initially, the level of 2°3Hg in the hair is tow and this level increases continuously
with time to 14 days. On the contrary, the level of Z°3Hg in the hair of rats exposed
to EMC vapor for 1 day is very high and this radioactivity appears to decline during
the 14-day post-exposure period. When the log of 2O3Hg concentration of the organs
and tissues are plotted against the elapsed time after dosing, it is evident that the rates
of change of the 2°aHg content in certain groups of organs or tissues are quite different
(Fig. 3). The 2°aHg remains in the tissues or organs much longer from pulmonary
exposure than from oral administration.

To ascertain that the difference in rate of change of 2°3Hg content might be derived
from a difference in the rate of biotransformation of EMC in the tissues, samples of
kidney, liver, and blood were analyzed for inorganic mercury. The results (Fig. 4)
show that the percent of inorganic mercury in these samples increases with the elapsed
time after dosing, indicating that such transformation is a continuous biological process.
Higher conversion is consistantly observed in the kidney samples than that in the samples
of liver and blood, regardless of whether the EMC uptake is from inhalation or from
oral administration. However, the slight differences observed in the liver and blood
and no difference in the kidney of EMC conversion would suggest that the difference
in the rate of change of the 2°3Hg content is not linked to EMC conversion.

Discussion
The results found in this study indicate that 2°3Hg accumulation in organs or tissues
from'~pulmonary uptake of EMC-2O3Hg vapor increases linearly with the time of
exposure and, also, proportionately with the vapor concentration. With the exception
of stomach, intestine, and hair, the relative amounts of 2°3Hg deposition in various
organs and tissues are quite similar either from inhalation of the vapor or from oral
administration. For example, the ratios of 2°3Hg in the blood to that in the brain are
26.0-+4.I (pulmonary, N=50), 26.8+2.8 (oral, N =16), and 26.9-+8.2 (subcutaneous
(sc) injection, N= 24 for both EMC and EM cyst.; Takeda et al. 1968). The kidney:brain
 Table V I . Change of Concentration with Time of 2°aHg in Various Organs and Tissues of Female Rats After Exposure
to EMC-2°aHg Vapor Concentration of 12 nmole/liter for 24 Hours

Organ 2°3Hg content a, nmole/g fresh weight, at indicated elapsed time after ex insure, days
OF
tissue 0 1 2 3 5 7 10 14

Kidney 10,4 20.5 12.8 25.2 27.5 34.8 66.4 76.8

Liver 5.3 3.5 2,4 5.1 3,6 2.8 3,1 3.1
Blood 4,6 5,5 2.5 4.8 4.3 4.2 4.3 3.4
Lung 1.7 3.6 1.4 2.7 2.7 2.1 2.8 1.7 0"Q
Spleen 2.6 2.4 1,3 2.4 2.0 -9.? 2,3 1.6
Heart 1.6 2.1 0.9 1.4 1.4 1.2 IA 0.9
Pancreas 1,7 1,8 0.7 1.8 1.4 1.1 1.6 1.2
Stomach 2.7 0.3 0.2 1.1 0,3 0.4 0.2 0.3
Intestine 1,7 0.8 0.4 0.9 0,5 0.4 0.5 0.5
Brain 0,2 0.3 0.2 0,3 0.3 0,3 0,4 0.4
Muscle 0.3 0.5 0.4 0.7 0.4 0.4 0.6 0.4
Fat 0.2 0.4 0.4 0.5 0,4 0.4 0.4 0.3
Hair 298 462 446 519 314 216 232 230
Number of rats used
2 I 2 I 2 I 2 I ' 1 j 2

aAverage value, if 2 rats were used.

Inhalation Uptake of Ethylmercuric Chloride in Rats 357

b
qb

~S
¢3
358 S . C . Fang and E. Fallin

0 0-~'~ 0,, k:~-o.11o

0"%... 0
1-
~'%~. k= -0.109 ~ 0
z.~.!
E
E
8
-i-
e4 k= 0.0267 ~1~
o
,._l

A- _
I U ~

•" ' ' " . O k=-00310

k=-0,107 "°° ,
.°
I
I" -- m "'D

k= -0.0281
! ! !
5 10 15
Days

Fig. 3. 'Rate of change of 2°3Hg content in certain groups of organs and tissues of female rats
after a single oral dose of 3 ~moles EMC-2O3Hg or after 24 hours of inhalation of air containing
12 nmole/liter of EMC-~3Hg. Solid symbols pertain to pulmonary uptake; open symbols pertain
to oral administration. © ©: average values of liver and blood; A A : average values
of lung, spleen, heart and pancrease; [] ... L]: average values of muscle and fat.

ratios are 53.0+11.1 (pulmonary, N = 5 0 ) , 56.1-+8.6 (oral, N = 8 ) , 4 6 . 8 + 1 7 . 2 (sub-

cutaneous injection, N = 24; Takeda et a l . , 1968). The blood:brain ratio decreases gradu-
ally as the post administration time increases, because of a greater rate of clearance
in the blood than in the brain. On the other hand, the kidney:brain ratio increases with
the increase of post administration time. Differential rates of Hg clearance in the brain
and the blood after intravenous (iv) injection of methylmercury are reported by Swensson
and Ulfvarson (1968) and Norseth and Clarkson (1970). In comparison to other alkylmer-
cury compounds, the blood:brain ratios for methyl mercury hydroxide are 11.7+3.5
(Ulfvarson 1%9) and 53.3_+9.2 for n=butyl mercury chloride (Takeda et al. 1968);
the kidney:brain ratios are 17.3_+ 8.0 for methyl mercury hydroxide (iv; Ulfvarson 1969)
 Inhalation Uptake of Ethylmercuric Chloride in Rats 359

100

1-00
O~
Kidney

• O
•/
E . . . . [] . . . .
o 50
O/ / [] Liver
g /
/

/~1' B

~/" ~Z~/'k~ Blood

/A
/
~ I I I I 1 I I
2 4 6 8 10 12 14
Days
Fig. 4. Rate of change of percent of 2°aHg present as inorganic mercury in the kidney, liver, and
blood of female rats after single dose of 3 /xmoles EMC-2O3Hg or after 24 hours of in-
halation of air containing 12 nmole/liter of EMC-Z°aHg. Solid symbols pertain to oral
administration; open symbols pertain to pulmonary uptake. O O: kidney, U 1 - - D : liver,
/x ZX: blood.

and 4 7 . 9 _+ 15.6 for n--butyl mercury chloride (sc; Takeda et al. 1968). This suggests
that the deposition of mercury in the brain possibly is inversely related to the length
of alkyl chain.

An extremely large amount of EMC is bound to the hair and the binding increases
either with the increasing time of exposure or with increasing EMC vapor concentration.
The level of EMC-2O3Hg in the hair declines at a rate quite similar to those of internal
organs. The mechanism of elimination of EMC from the hair is not known. Since elimina-
tion of methyl mercury through the hair is one of the major routes of elimination
(Berglund 1969, Ostlund 1969) and since an extremely high concentration of EMC
is bound to the hair when the rats are exposed to EMC vapor, the elimination of EMC
through this route possibly is hindered. As a result, different rates of elimination in
the organs are obtained between those receiving a single oral dose and those exposed
to EMC vapor.
360 S . C . Fang and E. Fallin

Analysis of liver, blood, and kidney for inorganic mercury reveals that kidney converts
a greater percentage of EMC to inorganic mercury; no significant differences are found
between the organs obtained from pulmonary uptake and oral administration. The values
on percent conversion of EMC to inorganic mercury in liver, kidney, and blood agree
very well with those reported by Miller et al. (1961), using the Sprague-Dawley strain
rats, and they are higher than those reported by Takeda and Ukita (1970), using male
Donryu rats. A difference in metabolism of mercurial compounds also has been reported
in different strains of mice (Miller and Csonka 1968) and chicks (Miller et al. 1959,
1967).

Acknowledgment

This study was supported in part by grant ES-00210 from the U.S. Public Health
Service, Institute of Environmental Health Science.

References

Berglund, F.: Experiments with rats relating to the toxicity of methyl mercury com-
pounds. Nord. Hyg, T. 50, 118 (1969).
Berlin, M., and S. Ullberg: Accumulation and retention of mercury in the mouse, III.
An autoradiographic comparison of methylmercuric dicyandiamide with inorganic
mercury. Arch. Environ. Health 6, 610 (1963).
Berlin, M., J. Fazackerly, and G. Nordberg: The uptake of mercury in the brains of
mammals exposed to mercury vapor and to mercuric salts. Arch. Environ. Health
18, 719 (1969).
Cassano, G. B., P. L. Viola, B. Ghetti, and L. Amaducci: The distribution of inhaled
mercury (Z°3Hg) vapors in the brain of rats and mice. J. Neuropath. Exp, Neurol.
28, 308 (1969).
Clarkson, T. W., and M. R. Greenwood: Selective determination of inorganic mercury
in the presence of organomercurial compounds in biological material. Anal,
Biochem. 37, 236 (1970).
Gage, J. C.: The distribution and excretion of inhaled mercury vapor. Brit. J. Ind.
Med. 18, 287 (1961).
Hayes, A. D., and A. Rothstein: The metabolism of inhaled mercury vapor in the rat
studied by isotope techniques. J. Pharmacol. 138, 1 (1962).
Magos, L.: Mercury-blood interaction and mercury uptake by the brain after vapor
exposure. Environ. Res. 1, 323 (1967).
Magos, L.: Uptake of mercury by the brain. Brit. J. Ind. Med. 25, 315 (1968).
Miller, V. L., G. E. Bearse, Y. Kimura, and E. Csonka: Strain differences in mercury
retention by chicks. Proc. Soc. Exptl. Biol. Med, 100, 301 (1959).
 Inhalation Uptake of Ethylmercuric Chloride in Rats 361

Miller, V. L., P. A. Klavano. A. C. Jerstad, and E. Csonka: Absorption.. distribution,

and excretion of ethylmercuric chloride. Toxicol. Appl. Pharmacol. 3, 459 (1961).
Miller, V. L., D. V. Larkin, G. E. Bearse, and C. M. Hamilton: The effects of dosage
and administration of two mercurials on mercury retention in two strains of chickens.
Poultry Sci. 46, 142 (1967).
Miller, V. L., and E. Csonka: Mercury retention in two strains of mice. Toxicol. Appl.
PharmacoI. 13, 207 (1968).
Norseth, T. and T. W. Clarkson: Studies on the biotransformation of 203Hg-labeled
methyl mercury chloride in rats. Arch. Environ. Health 21, 717 (1970).
Ostlund, L.: Studies on the metabolism of methyl mercury and dimethyl mercury in
mice. Acta Pharmacol. 27, Suppl. 1, 1 (1969).
Swensson, A.: Investigation on the toxicity of some organic mercury compounds which
are used as seed disinfections. Acta. Med. Seand. 143, 365 (1952).
Swensson, A., and U. Ulfvarson: Distribution and excretion of various mercury com-
pounds after single injections in poultry. Acta Pharmacol. 26, 259 (1968).
Suzuki, T., N. Matsumoto, T. Miyama, and H. Katsumma: Placental transfer of mercuric
chloride, phenylmercury acetate and methyl mercury acetate in mice. Industr. Health
g, 149 (1967).
Takeda, Y., T. Kunugi, O. Hoshino, and T. Ukita: Distribution of inorganic, aryl,
and alkyl mercury compounds in rats. Toxicol. Appl. Pharmacol. 13, 156 (1968).
Takeda, Y., and T. Ukita: Metabolism of ethylmercuric chloride-Z°3Hg in rats. Toxicol.
Appl. Pharmacol. 17, 181 (1970).
Trachtenberg, I. M.: The chronic action of mercury on the organisms. Current aspects
of the problem of micromercurialism and its prophylaxis. Zoorov'Ja, Kiev 1969,
292 pp. (In Russian, German translation.)
Ulfvarson, U.: The effect of the size of the dose on the distribution and excretion of
mercury in rats after single intravenous injection of various mercury compounds.
Toxicol. Appl. Pharmacol. 15, 1 (1969).

Manuscript received November 8, 1972; accepted January 5, 1973