European Journal of
Eur J Clin Pharmacol (1982) 22:403-406
Placental Transfer of Clenbuterol Early in Human Pregnancy
O. Pelkonen, R. Tuimala, and A. Kauppila
Departments of Pharmacology and Obstetrics and Gynecology, University of Oulu, Oulu, Finland
Summary. After administration of clenbuterol 80 ~tg
p.o., a fl-adrenoceptor agonist, the concentrations of
(14C-labelled)clenbuterol in fetus, placenta and ma-
ternal plasma in 9 patients at 9-12 weeks gestation
were measured during therapeutic abortion. The time
interval between the administration and abortion
ranged from 120 to 280 min. The mean concentrations
of clenbuterol in maternal plasma, fetus and placenta
were 0.37 (range 0.22-0.56), 0.32 (0.14-0.48) and 0.91
(0.12-1.73) ng equivalents per ml or per gram of tissue
wet weight. The mean concentration ratio of clenbu-
terol between fetus and maternal plasma was 0.84
(6 cases); it did not vary with time. The concentration
of clenbuterol in three amniotic fluid samples ranged
from 0.06 to 0.18 ng/ml (mean 0.11). Maternal plasma
concentrations showed wide variability of the phar-
macokinetic phase at the time of abortion. The studies
indicate that clenbuterol crosses the placenta early in
human pregnancy and that it accumulates in the pla-
centa.
Key words: clenbuterol, pregnancy, placental trans-
fer; fetal concentration, placental concentration, am-
niotic fluid concentration, maternal plasma level, ma-
ternal-fetal concentration ratio
Clenbuterol I is an fl2-adrenergic receptor agonist,
which is useful in the treatment of asthma (Arzneim
Forsch, 26 (7 a): 1976). Asthma, being a common dis-
ease, often occurs concomitantly with pregnancy, so
the clinical evaluation of clenbuterol must include its
use in this circumstance. Another potential indication
i (clenbuter01 hydrochloride = 4-amino-a-[(tert.-butylamino)-
methyl]-3,5-dichlorobenzylatcohot-hydrochloride; trade name
Spiropent ®)
Clinical Pharmacology
© Springer-Verlag 1982
for clenbuterol during pregnancy would be as a toco-
lytic drug. The uterus contains fl2-receptors, stimula-
tion of which relaxes uterine muscle. Abortus immi-
nens is usually due to premature contraction of the
uterine muscle and at least in principle, clenbuterol
offers the possibility of preventing premature con-
tractions. On the other hand, because the fetus may be
endangered when the mother is exposed to drugs, it is
necessary to elucidate the fate of compounds which it
is intended to use in a pregnant subject.
As a preliminary investigation in elucidation of
the fate of clenbuterol in the pregnant mother, placen-
tal transfer of the compound during the first trimester
of human pregnancy was studied. From the results it
is clear that clenbuterol is able to cross the placenta
and to reach fetal tissues. The drug also accumulates
in the placenta, but the nature of this effect is not
known.
Materials and Methods
Patients
9 healthy women were studied, who came to abortion
for socio-medical considerations. Their mean age was
21 years (range 17-31 years) and mean weight 54.2 kg
(range 43.5-67.5 kg). The method of abortion was dil-
atation and curettage. Informed consent was ob-
tained from each subject. The protocol for the study
was reviewed and accepted after minor modifications
(e. g. time schedule of sampling) by the Ethical Com-
mittee of the Oulu University Central Hospital, and
by the Board of Medical Government. Clenbuterol
tablets 80 lxg (total radioactivity 14 aCi) were given to
patients before the operation. The mean dose was
1.5 ug/kg body weight (range 1.2-1.8 gg/kg). No ad-
verse effects were noted.
0031-6970/82/0022/0403/$01.00
404 O. Pelkonen et al.: Placental Transfer of Clenbuterol

Table 1. Placental transfer of clenbuterol early in human pregnancy

Subject Duration of Fetal A/D a Concentration of clenbuterol in

gestation weight interval
maternal Fetus Placenta Amniotic Ratio
[weeks] [grams] [mini
plasma fluid fetus/mother
[ng/mll [ng/g] [ng/g] [ng/ml]

MO 9 ND b 220 0.22 ND 0.71 - -

HP 10 ND 265 0.30 ND 0.84 - -
KI 11 ND 180 0.42 BS~ 0.91 0.t0 -
EN 11 0.66 120 0.50 0.48 1.73 - 0.96
MM 9 0.89 180 0.27 0.14 0.12 0.06 0.52
MM 12 2,53 220 0.39 0.30 0.90 - 0.76
JP I1 3.02 280 0.27 0.45 0.60 - 1.67
AK 12 4.15 180 0.56 0.31 0.91 - 0.56
JM 11 5.05 180 0.38 0.21 1.49 0.18 0.54

a interval between administration ofclenbuterol and operation

b ND means that the fetat weight could not be determined because of disruption of fetal structures, so the fetal concentration of clenbuterol
could not be measured in two cases (MO and HP)
BS means that experimental counts from the recovered fetal parts did not differ from background, i. e. counts were below the limit of detec-
tion (less than 0.05 ng/g)

diately centrifuged. The plasma was separated and

0-6
stored at - 20 °C until analysed, which was about one
month after sampling. The fetuses and placentas were
also frozen and thawed immediately before the analy-
0"5
sis. In three cases it was possible to sample amniotic
E
tO fluid, and it was preserved like the plasma samples.
r~ /
n 0-4
Determination of Ctenbuterol
O1
c 0"3
The plasma concentration of clenbuterol was mea-
sured by mixing plasma 1 ml with scintillation fluid
ocr 10 ml (Pico-Fluor, Packard Instrument) and counting
uJ 0"2
the radioactivity in a Packard TriCarb liquid scintilla-
tion spectrometer; the counting efficiency was 61% to
Z 87%. Variation in counting efficiency was mainly be-
L:J
_J
0.1
tween-sample variation and within-sample variability
was small. For unknown reasons the efficiency was
much lower with amniotic fluid samples, being only
30% to 35%.
1 2 3 4 5 The placental concentration was determined by
TIME AFTER ADMINISTRATION homogenising a piece of placenta in 4 volumes of 0.3
(hours) M TCA and centrifuging the mixture. A 2-ml sample
Fig. 1. Maternal plasma concentrations of clenbuterol before abor- of the supernatant was mixed with 18 ml scintillation
tion. The last sample was taken at the time of abortion. Values are fluid and counted; the counting efficiency was be-
expressed as n g / E q of clenbuterol, because unchanged clenbuterol
and its metabotites could not be measured separately in individual tween 66% and 91%. The pellet was digested with sol-
samples ubilizer (Soluene 100, Packard Instrument) and
mixed with scintillation fluid. The pellet did not con-
tain any detectable radioactivity, and so if any radi-
The 14C-labelled clenbuterol was a gift from the oactivity were protein-bound, it was transferred to the
manufacturer Boehringer, Ingelheim. supernatant during the TCA treatment.
The fetal content of clenbuterol was measured in
essentially similar way as that of placenta. Fetuses
Samples were homogenized in TCA, the homogenate centri-
Blood samples (10 ml) were taken from a maternal pe- fuged, a portion of the supernatant mixed with scintil-
ripheral vein before and at abortion. The samples lation fluid and counted. Counting efficiency varied
were collected in heparinised plastic tubes and imme- from 62% to 89%.
O. Pelkonenet al.: PlacentalTransferof Clenbuterol
Direct digestion of the fetal sample with solubiliz-
er and counting the solution decreased the counting
efficiency to about 45% to 60%, but otherwise gave
very similar results.
Results
Maternal plasma concentrations of clenbuterol be-
fore and at abortion are presented in Fig. 1. Although
doses calculated on the basis of body weight, did not
vary very much, plasma concentrations i h after ad-
ministration varied from less than 0.1 to 0.6 ng Eq/ml
plasma. This large variation is a consequence of dif-
ferences in the pharmacokinetic phases between dif-
ferent patients, as can readily be seen in Fig. 1. In
some patients (e.g. AK, HP and KI) absorption
seemed to be essentially complete 1 h after adminis-
tration and the plasma concentrations were already
declining, whereas in other patients (e.g. JM and
MMi) clenbuterol concentrations at one hour were
lower than at later time points, indicating continuing
absorption.
The concentrations in the various samples are list-
ed in Table 1. The mean concentrations of clenbuterol
in maternal plasma, fetus and placenta were 0.37,
0.32, and 0.91 ng Eq/mt or per gram tissue wet weight.
The concentration of clenbuterol in three amniotic
fluid samples ranged from 0.06 to 0.18 ng/ml. The
mean concentration ratio of clenbuterol between fe-
tus and maternal plasma was 0.84 (six cases), and it
showed no sign of dependence on time, at least not
between 120 and 280 rain after administration.
Discussion
The passage of clenbuteroi through the placenta is a
relatively rapid phenomenon, because the concentra-
tion ratio between fetus and maternal plasma was
close to unity between 2 and 4,5 h after administration
of the compound, and during this phase the ratio did
not exhibit any time-dependence. The reason why the
ratio was lower than unity (0.84) is not known. With
other compounds, when plasma concentrations are
compared, the decreased binding ability of fetal plas-
ma proteins seems able to account for the difference
(Mirkin 1975). The concentration of clenbuterol in
the three amniotic fluid samples available for study
was about 30% of that in fetus or maternal plasma.
With several other substances it has been demonstrat-
ed that amniotic fluid concentrations near birth are
similar to those in cord and maternal blood (Waddell
and Marlowe 1976). The probable reason for the low-
er amniotic fluid concentration of clenbuterol is that
405
at such an early period of gestation the fetal excretory
functions responsible for the transfer of drugs to am-
niotic fluid are not fully developed.
Clenbuterol is completely absorbed from the gas-
trointestinal tract, and the maximum plasma level is
achieved two to three hours after dosing (Zimmer and
Bucheler 1976). Thus, it is interesting to find that ab-
sorption by pregnant women was very variable
(Fig. t). In some women absorption was already ap-
parently complete 1 hour after administration, be-
cause at that time the maximum plasma level had
been achieved. In other women, even 3 to 3.5 h after
administration of clenbuterol the plasma concentra-
tion was still increasing. Again, the exact reason is not
known, but explanations can be sought from two di-
rections. First, absorption by pregnant women may be
different, and there is evidence from other substances
in favour of this interpretation (Krauer et al. 1980).
Second, there is also a possibility that factors associat-
ed with the surgical operation, such as premedication
or fasting, might have affected the absorption of den-
buterol.
The mean placental concentration of clenbuterol
was about three times higher than that of maternal
plasma or an homogenate of fetal tissue. Apparent ac-
cumulation of digoxin in the placenta has previously
been demonstrated (Allonen et al. 1977). The nature
of the accumulation is not known, but it is reversible,
because denbuterol can be extracted from a placental
homogenate by organic solvents. As to any possible
clinical effect of the placental retention, it has been
demonstrated that t-stimulation with other adrener-
gic agonists has not resulted in any change in intervil-
lous placental blood flow in animals (Greiss 1972), or
in humans (Kauppila et al. 1978). No study ofclenbu-
terol appears to have been published.
The effects of denbuterol in the fetus must de-
pend on the presence oft-receptors in appropriate fe-
tal organs. In late pregnancy fl-adrenoceptors are
present in the fetal heart, and an increase in fetal heart
rate, as well as fetal cardiac arrhythmias, have been
reported after maternal infusion of ritodfine (Miller et
al. 1976). In vitro experiments with isolated human fe-
tal tissues have demonstrated that B-adrenoceptors
are well developed early in gestation, and conse-
quently dosedependent responses to maternally-ad-
ministered clenbuterol can be expected. Human fetal
studies in vitro have suggested quantitatively similar
response to those of the adult, but it is not known
whether in vitro findings correctly mimic the in vivo
situation (McMurphy and Boreus 1971). Van Petten
(1975) claimed that the ovine fetus in vivo was more
sensitive than the mother to certain fl-adrenoceptor
agonists. In view of these considerations, evaluation
of intravenous clenbuterol as an tocolytic drug re-
406 O. Pelkonen et al.: Placental Transfer of Clenbuterol

quires a combined pharmacodynamic and pharma- Miller FC, Nochimson DJ, Paul RH, Hon EH (1976) Effects ofri-
cokinetic study. todrine hydrochloride on uterine activity and the cardiovascular
system in toxemic patients. Obstet Gyneco147:50-55
Mirkin BL (•975) Perinatal pharmacology: Placental transfer, fetal
Acknowledgement. The labelled clenbuterol was a gift localization, and neonatal disposition of drugs. Anesthesiology
from Boehringer Ingelheim. 43:156-170
Van Petten GR (1975) Pharmacology and the fetus. Br Med Bull 31 :
75-79
Waddell WJ, Marlowe GC (1976) Disposition of drugs in the fetus.
References In: Mirkin B (ed) Perinatal pharmacology and therapeutics.
Academic Press, New York, pp 119-268
Allonen H, Kanto J, Iisalo E (1977) The foeto-maternal distribution Zimmer A, Bucheler A (1976) Einmalapptikation, Mehrfachappli-
of digoxin in early human pregnancy. Acta Pharmacol Toxicol kation und Metabolitenmuster yon Clenbuterol beim Men-
39: 477-480 schen. Arzneim Forsch 26:1446-1450
Arzneimittel-Forschung(Drug Res.) (•976) Themanummer: Clen-
buterol. 26: Nr. 7 a
Greiss FC Jr (1972) Differential reactivity of the myoendometrial Received: June 2, 1981
and placental vasculatures. Am J Obstet Gyneco1112: 20-30 in revised form: September 28,1981
Kauppila A, Kuikka J, Tuimala R (1978) Effect of fenoterol and accepted: October 2,1981
isoxsuprine on myometrial and intervillous blood flow during
late pregnancy. Obstet Gyneco152: 558-562
Krauer B, Krauer F, Hytten FE (•980) Drug disposition and phar- Olavi Pelkonen, M. D.
macokinetics in the maternal-placental-fetal unit. Pharmacol Department of Pharmacology
Ther 10:301-328 and Obstetrics and Gynecology
McMurphy DM, Boreus LO (1971) Studies on the pharmacology University of Oulu
of the perfused human fetal ductus arteriosus, Am J Obstet SF-90 220 Outu 22
Gyneco1109:937-942 Finland