Transfer of maternally administered magnesium sulfate

into the fetal compartment of the rat: Assessment of

amniotic fluid, blood, and brain concentrations
Mordechai Hallak, MD, and David B. Cotton, MD
Detroit, Michigan

OBJECTIVE: Our purpose was to determine whether parenteral magnesium sulfate crosses the rat
placenta and enters the fetal brain.
STUDY DESIGN: Twenty-eight pregnant female Long-Evans rats were divided into four groups, seven
animals in each group. These groups included the following: single saline solution injection group
evaluated after 20 minutes (control), single magnesium sulfate injection (270 mg/kg) evaluated after 20
minutes, and prolonged (2 and 4 hours) serum magnesium elevation (270 mg/kg loading and then 27
mg/kg every 20 minutes for maintenance). Each animal was killed, and maternal and fetal sera, amniotic
fluid, and specific brain areas were analyzed for levels of magnesium. Statistical analysis included
analysis of variance, multiple comparison procedure, and linear regression analysis.
RESULTS: All three regimens of maternal subcutaneous magnesium sulfate resulted in significantly
elevated maternal serum magnesium concentrations (p < 0.01). Fetal blood magnesium concentration
rose from 3.9 ± 0.3 to 4.9 ± 0.1 mg/dl after 2 hours of continuous injections (p < 0.05), and to 5.3 ± 0.0
mg/dl after 4 hours (p < 0.01). Amniotic fluid magnesium concentrations rose from 4.2 ± 0.2 to
5.1 ± 0.1 mg/dl after 4 hours (p < 0.05). Maternal blood magnesium concentrations were significantly
correlated with amniotic fluid concentrations (r = 0.59, P < 0.01). Four hours of maternal magnesium
sulfate treatment resulted in a 25% increase in magnesium concentrations in the fetal forebrain
(38.3 ± 2.3 to 47.9 ± 3.5 mg/dl/gm, p < 0.05). No significant changes in magnesium concentrations
were detected in hindbrain.
CONCLUSION: Magnesium sulfate injected subcutaneously into rats crosses the placenta within 2 hours
of sustained magnesium levels, enter the fetal blood-brain barrier, and concentrates in the forebrain. (AM
J OBSTET GYNECOL 1993;169:427-31.)

Key words: Magnesium sulfate, placental barrier, fetal rat blood-brain barrier

Magnesium sulfate is commonly used as a tocolytic
agent for preterm labor and for prevention and treat-
ment of eclamptic convulsions. The American College
of Obstetricians and Gynecologists has gone so far as to
recommend its use during labor and in the postpartum
period in every woman with the diagnosis of preeclamp-
sia-eciampsia. I
Magnesium has been shown to cross the placenta in
studies performed after prolonged maternal treatment
with magnesium sulfate!·6 Magnesium transfer from
maternal to fetal blood and amniotic fluid in a short-
From the Department of Obstetrics and Gynecology, Wayne State
University/Hutzel Hospital.
Received for publication july 27,1992; revised February 1,1993;
accepted March 8, 1993.
Reprint requests: Mordechai Hallak, MD, Department of Obstetrics
and Gynecology, Wayne State University, Hutzel Hospital, 4707 St.
Antoine Blvd., Detroit, MI 48201.
Copyright © 1993 by Mosby-Year Book, Inc.
0002-9378/93 $1.00 + .20 6/1/46958
term administration, however, has not been adequately
studied in in vivo model.
Several investigators have reported on the effects of
excess magnesium on the fetus and the newborn in-
fant. 7 . 13 At one extreme of the spectrum prolonged
(> 24 hours) maternal intravenous administration of
magnesium sulfate has been thought to cause hypoten-
sive infants with reduced responsiveness, depressed
reflexes, general weakness, and respiratory depression
requiring assisted ventilation. 12. 13 Magnesium sulfate
has also been found to affect the fetal biophysical
profile by decreasing fetal breathing movements and
fetal heart rate variability.g·ll Most of these fetal activi-
ties are controlled by the fetal brain. To affect these
activities, magnesium must cross the fetal blood-brain
barrier and concentrate in specific brain areas.
This study was initiated for the following reasons: (l)
to determine the concentrations of maternally admin-
istered magnesium sulfate in the fetal circulation, (2) to

427
428 Hallak and Cotton
ascertain the time needed for accumulation of magne-
sium in amniotic fluid, and (3) to determine whether
elevated fetal serum magnesium concentrations were
associated with increased magnesium levels in brain
parenchyma and, if so, whether there was a predilection
for specific brain areas.
Material and methods
Sampling procedure. Timed pregnant female Long-
Evans rats were obtained from Harlan Sprague Dawley
(Indianapolis). Rats were individually housed in poly-
carbonate boxes in an environmentally controlled vivar-
ium under 12-hour-light and 12-hour-dark cycles. An-
imals were fed ad lib throughout the experiments. The
animal care and experiments were approved by the
Wayne State University Animal Investigation Com-
mittee.
On 19 to 20 days' gestation, animals were anesthe-
tized with 100 mg/kg of sodium pentobarbital adminis-
tered subcutaneously. After adequate anesthesia oc-
curred, the abdominal wall was opened and the uterine
horns exposed bilaterally. Each gestational sac with its
placenta was separated individually, the sac opened,
and the amniotic fluid collected into a tube. Maternal
blood was also aspirated at this stage and analyzed for
magnesium concentrations. All samples were kept on
ice until the end of the experiment, at which time
magnesium concentrations in each sample were deter-
mined.
All fetuses from one horn were used for blood collec-
tion. The blood was aspirated from the fetal body trunk
after decapitation. Three to four fetuses were used to
collect one sample. Fetuses from the contralateral horn
were used to determine brain magnesium concentra-
tions. To clean the brain vasculature from blood, each
fetal chest was opened and the animal perfused trans-
cardially with 2 to 3 ml of normal saline solution, after
which the brain was dissected and analyzed for magne-
sium. The forebrain (including cortex, hippocampus,
and caudate nucleus), and the hindbrain (including
cerebellum, pons, and medulla oblongata) were sepa-
rated, weighed, and homogenized in 0.25 ml of saline
solution. Magnesium concentrations in maternal serum,
fetal sera, and amniotic fluid (aU in milligrams per
deciliter), forebrain, and hindbrain (in milligrams per
deciliter/gram of wet brain weight) on each animal were
determined by a blinded investigator. Magnesium con-
centrations were analyzed colorometrically (Eastman
Kodak, Rochester, N .Y.). The amount of magnesium
present in a sample was measured by reflectance spec-
trophotometry at 630 nm.
Study design. Twenty-eight rats were divided into
four groups. Control animals (n = 7) had a subcutane-
ous injection of normal saline solution, followed 20
minutes later by the sampling procedure. In the second
August 1993
Am J Obstet Gyneco1
group the sampling procedures were performed at 20
minutes (n = 7) after a single 270 mg/kg subcutaneous
irUection of magnesium sulfate. In the third and fourth
groups the effects of prolonged elevation of serum
magnesium concentrations were evaluated. In these
groups each animal received a 270 mg/kg loading dose
of subcutaneous magnesium sulfate followed by 27
mg/kg every 20 minutes for 2 hours (group 3, n = 7)
and for 4 hours (group 4, n = 7) before the sampling
procedure.
Magnesium sulfate doses were chosen on the basis of
previous studies. 14• 15 The magnesium sulfate was recon-
stituted in sterile saline solution to a concentration of
90 mg/m!. The order of magnesium sulfate versus saline
solution administration was randomized across animals.
All animals were killed at the conclusion of the study.
Statistical analysis. Magnesium concentrations in
maternal blood, fetal blood, amniotic fluid, and fetal
forebrain and hindbrain were compared among the
various groups. One-way analysis of variance was used
to test for differences among groups, and then a mul-
tiple comparison procedure (Scheffe and Tukey tests)
was applied. Linear regression analysis was used to test
the correlation between magnesium concentrations in
maternal or fetal blood and amniotic fluid. All values
are reported as the mean ± SEM. A P value of < 0.05
was considered statistically significant.
Results
A total of 28 rats and 364 fetuses were used in this
study. Maternal subcutaneous magnesium sulfate injec-
tions resulted in significant elevations of magnesium in
fetal blood, amniotic fluid, and brain.
Fetal blood and amniotic fluid magnesium concen·
trations (Table I). All three regimens of maternal sub-
cutaneous magnesium sulfate administration resulted
in significant elevations of maternal serum magnesium
concentrations (p < 0.01). A single maternal injection
of magnesium sulfate (group 2) resulted in an 18%
increase in magnesium concentration in fetal blood.
Continuous injections for 2 hours (group 3) resulted in
a 25% increase (p < 0.05) and for 4 hours (group 4) in
a 36% increase in magnesium concentration in fetal
blood (p < 0.01). In the control group the fetal serum
magnesium concentrations were 36% higher than the
concentrations in the mother, and amniotic fluid con-
centrations were 47% higher. This relative proportion
was changed remarkably in the various treatment
groups. There was a significant correlation between
maternal and fetal serum magnesium concentrations
(p < 0.05); however, the correlation coefficient was low
(r = 0.43).
Amniotic fluid magnesium concentrations rose by
21% after 4 hours of maternal injections (p < 0.05).
The correlation between maternal serum and amniotic
Volume 169, Number 2, Part 1 Hallak and Cotton 429
Am J Obstet Gynecol

• Control D MgS04-20min • MgS04-2hr MgS04-4hr

*
mg/dL/g

Forebrain Hindbrain
Fig. 1. Magnesium concentrations in various areas of fetal brain after maternal subcutaneous
administration, Results are presented as mean ± SEM, Asterisk, magnesium sulfate (MgS04) at 4 hours
> control, p < 0,05, Four conditions include control, saline solution; magnesium sulfate-20
minutes, single injection of magnesium sulfate (in the last two conditions the sampling procedure was
performed 20 minutes after injection); magnesium sulfate-2 hours, injections every 20 minutes for
2 hours; magnesium sulfate - 4 hours, injections every 20 minutes for 4 hours,

Table I. Maternal-fetal transfer of magnesium into the fetal blood and amniotic fluid (mean ± SEM)
Prolonged treatment

Control Acute treatment 2 hr I 4 hr

Maternal blood (mg/dl) 2,9 ± 0,2* 12,9 ± 1.0 1004 ± O,S 9.1 ± 1.0
Fetal blood (mg/dl) 3,9 ± 0,3 4,6 ± 0,1 4.9 ± O.lt 5.3 ± O.Ot
Amniotic fluid (mg/dl) 4,2 ± 0,2 4,S ± 0,2 4.S ± 0.2 5.1 ± O.It

*Control < acute, 2 and 4 hours; p < 0.01.

tControl <2 hours I.p < 0.05),4 hours I.p < 0.01).
tControl < 4 hours I.p < 0.05).

fluid magnesium concentrations was found to be highly
significant (r = 0.S9, P < 0.01). No significant correla-
tion was detected between fetal blood and amniotic
fluid magnesium concentrations.
Fetal brain magnesium levels (Fig. 1). Magnesium
concentrations were increased in the fetal forebrain,
including the cortex and hippocampus. The concentra-
tions of magnesium in the forebrain increased only
slightly in groups 2 and 3. However, only prolonged
magnesium sulfate treatment for 4 hours (group 4)
reached statistical significance with a 2S% increase in
magnesium concentrations above the control group
(from 38.2 ± 2.3 to 47.9 ± 3.S mg/dVgm, p < O.OS).
Magnesium concentration changes in the hindbrain
were not statistically different among all groups (the
largest change was from 41.0 ± 2.1 in control to
46.7 ± 2.9 mg/dVgm in group 4).
Comment
This study demonstrates that peripheral magnesium
sulfate injections to the pregnant rat result in signifi-
cant elevations of magnesium in fetal blood, amniotic
fluid, and fetal brain. Fetal blood magnesium concen-
trations rose by 2S% and 36% after 2 and 4 hours,
430 Hallak and Cotton
respectively, of continuous maternal peripheral magne-
sium administration. Magnesium concentrations in the
fetal forebrain were increased by 25% and reached
statistical significance after 4 hours of sustained mater-
nal levels. No significant changes in magnesium con-
centrations in the hindbrain were found.
Baseline magnesium concentrations were found to be
higher in the fetal than in the maternal serum!' 16- 1.
Active maternal-fetal transfer of magnesium has previ-
ously been demonstrated in the in situ perfused rat
placenta!' 18 Our study in the pregnant rat, and human
data, also indicate higher baseline concentrations of
magnesium in fetal as opposed to maternal serum. '9
These data suggest a putative placental pump that may
be responsible for the gradient of magnesium across
the placenta.
In the face of increasing maternal magnesium con-
centrations the fetus will equilibrate, and eventually
exceed, maternal levels.· ' s However, this process de-
pends on the length of time of sustained maternal
magnesium levels. Others have demonstrated that after
prolonged (hours) maternal treatment with magnesium
sulfate for preeclampsia umbilical cord magnesium con-
centrations ranged from 70% to 100% of maternal
concentrations."s In another study, when magnesium
sulfate was given for 4 to 28 days, both amniotic fluid
and umbilical cord blood magnesium concentrations
exceeded the maternal levels .s Our study shows that
after 20 minutes of maternal magnesium administra-
tion, although maternal concentrations were increased
fourfold, the fetal concentrations were not significantly
changed. It took 2 hours of sustained maternal magne-
sium elevation to reach a significant change and 4
hours for a further increase in the fetal serum magne-
sium concentrations. This may be attributed to a pro-
tective effect of the placenta on the fetus.
Amniotic fluid magnesium concentrations were found
to be significantly increased only after 4 hours of ma-
ternal treatment. The correlation between fetal blood
and amniotic fluid magnesium concentrations was not
significant as opposed to the correlation between ma-
ternal serum and amniotic fluid, which was found to be
highly significant. This suggests the possibility that
magnesium transfer into amniotic fluid is more through
the membranes than across the placenta, at least during
the first 4 hours of maternal administration.
The accumulation of magnesium in the fetal com-
partment can potentially affect its behavior and even
jeopardize fetal well-being. Severe neonatal conse-
quences from perinatal hypermagnesemia have been
described by Lipsitz and English . ' 2 . 13 The clinical man-
ifestations of neonatal hypermagnesemia reported in-
cluded severe depression requiring endotracheal intu-
bation and ventilation, weak cry, and absent reflexes
that necessitated intravenous alimentation because of
August 1993
Am J Obstet Gyneco1
lack of the sucking reflex. However, most of the litera-
ture today agrees that the administration of magnesium
sulfate to the mother for anticonvulsant or tocolytic
effects does not usually pose a risk to the fetus or
newborn. 3• 7 Transient effect on fetal breathing move-
ments and fetal heart rate variability have been
shown.'· 11 However, the newborn situation as reflected
by Apgar scores is usually not affected by maternal
magnesium sulfate therapy in the standard doses rec-
ommended for preeclampsia. s
Many of the fetal activities reported to be affected by
the excess of magnesium are controlled by the fetal
brain. In a previous study we disproved the argument
that magnesium does not cross the maternal blood-
brain barrier. 15. 20. 21 We showed a significant deposition
of magnesium in the cortex and hippocampus of adult
rats after 2 hours of sustained magnesium blood eleva-
tion. ' 5 In the current study we demonstrated a similar
effect on the fetus . As in the adult, increased magne-
sium concentrations were demonstrated in the fetal
forebrain but not the hindbrain . We believe that mag-
nesium exerts its anticonvulsant activity at least partially
by blockade of the excitatory amino acid N-methyl-D-
aspartate receptors. The highest density of these recep-
tors is found in the hippocampus (stratum radiatum),
followed by the cerebral cortex, striatum, and thalamus,
all of which are various components of the forebrain. 22
In conclusion, magnesium sulfate injected subcutane-
ously into rats crosses the placenta within 2 hours of
sustained magnesium levels, crosses the fetal blood-
brain barrier, and concentrates in the fetal forebrain.
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