American Journal of Obstetrics and Gynecology (2006) 194, e35e38

www.ajog.org

Effects of menstrual cycle and oral contraceptive use on

serum levels of lipid-soluble antioxidants
Prabhudas R. Palan, PhD,a,* Amy T. Magneson, MD,a Monique Castillo, RN,a
James Dunne, MS,b Magdy S. Mikhail, MDa

Departments of Obstetrics and Gynecologya and Pathology,b Bronx-Lebanon Hospital CenterdAlbert Einstein
College of Medicine, Bronx, NY

Received for publication August 9, 2005; revised October 13, 2005; accepted November 15, 2005

KEY WORDS Objective: The purpose of this study was to examine the influence of menstrual cycle and oral
Menstrual cycle contraceptive use on serum levels of lipid-soluble antioxidants.
Oral contraceptive use Study design: In this cross-section study, nonfasting blood samples were collected twice from 10
Coenzyme Q10 healthy premenopausal women during the follicular phase (between days 8 and 11) and the luteal
Antioxidant phase (between days 18 and 22) of their same menstrual cycle. In addition, blood samples from 15
premenopausal women who used oral contraceptive for at least 6 months and 40 women who did
not use oral contraceptive were collected randomly at any day of the menstrual cycle. Serum levels
of coenzyme Q10, a-tocopherol, g-tocopherol, b-carotene, a-carotene, and lycopene were deter-
mined using high pressure liquid chromatography.
Results: Serum coenzyme Q10 and a-tocopherol levels were significantly lower during the follic-
ular phase compared with the luteal phase of the same menstrual cycle (P ! .05). Oral contra-
ceptive use also significantly decreased coenzyme Q10 and a-tocopherol (P ! .001). Other
antioxidant levels were comparable.
Conclusion: Alterations in coenzyme Q10 and a-tocopherol levels during the menstrual cycle and
in oral contraceptive users should be taken into consideration, concerning the future antioxidant
research in premenopausal women. Further studies are needed to investigate the potential role of
endogenous and exogenous ovarian hormones on oxidative stress in women.
2006 Mosby, Inc. All rights reserved.

Oxidative damage caused by oxygen free radicals has
been implicated in the pathogenesis of a large number of
chronic diseases.1 Lipid-soluble antioxidants coenzyme
Presented in part at the 53rd Annual Clinical Meeting of the Amer-
ican College of Obstetrics and Gynecologists, San Francisco, CA, May
7-11, 2005.
* Reprint requests: Prabhudas Palan, PhD, Department of Obstet-
rics and Gynecology, Bronx-Lebanon Hospital Center, 1650 Grand
Concourse, Floor 5th, Bronx, NY 10457.
E-mail: daspalan@aol.com
Q10 and a-tocopherol can neutralize free radical and
are thus postulated potentially to decrease the risk of
major diseases, such as cancer and cardiovascular dis-
ease (CVD).2,3 Ovarian hormones, primarily estrogens,
possess antioxidant properties and have been postulated
to protect against CVD.4 Although the uctuations in
ovarian hormone levels during the menstrual cycle in-
creasingly are believed to play a signicant role in the
cause of many disorders and diseases in women, several
studies were conducted to determine whether plasma

0002-9378/$ - see front matter 2006 Mosby, Inc. All rights reserved.
doi:10.1016/j.ajog.2005.11.032
e36 Palan et al

Table Serum levels of antioxidants during the follicular and the luteal phases in the same premenopausal women and in women who
used OCs and control subjects
Menstrual phase OC
Follicular: Group 1 Luteal: Group 2 Users: Group 3 Nonusers: Group 4
Antioxidant (n = 10) (n = 10) (n = 15) (n = 40)
Coenzyme Q10 (mg/dL) 0.50 G 0.3* 0.60 G 0.2 0.38 G 0.1y 0.60 G 0.2
a-Tocopherol (mg/L) 4.60 G 1.0* 5.90 G 1.8 4.70 G 0.9y 6.13 G 1.3
g-Tocopherol (mg/L) 1.09 G 0.4 1.22 G 0.7 1.22 G 0.3 1.11 G 0.5
b-Carotene (mg/dL) 27.9 G 6.2 28.7 G 3.8 31.0 G 3.8 28.6 G 4.7
a-Carotene (mg/dL) 3.9 G 2.3 3.8 G 2.1 3.6 G 1.4 3.9 G 2.6
Lycopene (mg/dL) 35.5 G 13.2 38.0 G 11.1 32.2 G 9.2 29.6 G 14.0
Values are given as mean G SD.
* P ! .05, group 1 vs group 2, by Wilcoxon paired-sample test.
y
P ! .001, group 3 vs group 4, by the Student t test.

a-tocopherol levels uctuate by phase of the menstrual
cycle.5-7 However, relatively little research has been
focused on menstrual cyclerelated changes in lipid
soluble antioxidant coenzyme Q10. We investigated
the inuence of menstrual cycle phase and oral contra-
ceptive (OC) use on serum levels of coenzyme Q10, a-
tocopherol, g-tocopherol, b-carotene, a-carotene, and
lycopene in healthy premenopausal women.
Material and methods
The Institutional Review Board approved the study
protocol. In this cross-section study, 65 nonpregnant
women with regular menstrual cycles, who attended the
gynecology clinics at the Bronx-Lebanon Hospital Cen-
ter, Bronx, New York, between January 2002 and
November 2004 were recruited, with informed consent.
All subjects came from the same catchment area and
had a similar socioeconomic composition and were not
using any medications or hormonal contraceptives.
Women who consumed coenzyme Q10 or multivitamin
supplementation or women who had irregular men-
strual cycles were excluded from the study. No dietary
restrictions were imposed on any of the subjects. Ten
healthy women (median age, 33 years; range, 28-44
years) with regular menstrual cycles (27-29 days) were
studied at 2 time points during the same menstrual cy-
cle. Nonfasting venous blood samples were obtained
in the follicular phase (between days 8 and 11) and in
the luteal phase (between days 18 and 22) of their men-
strual cycle. Subjects in this group were not taking any
form of OC. In addition, 55 healthy premenopausal
women (median age, 36 years; range, 22-42 years)
were recruited. Of these 55 subjects, 15 women were us-
ing Ortho Tri-Cyclen-Lo (norgestimate/ethinylestradiol,
0.18 mg; 0.025 mg [7 days], 0.215 mg/0.025 mg [7 days],
0.25 mg/0.025 mg [7 days]) for at least a 6-month dura-
tion, and 40 women did not use OCs. A nonfasting
peripheral venous blood sample (10 mL) was collected
randomly at any time of the menstrual cycle from
non-OC users and also from OC users during the ste-
roid medication and placebo phase. Serum was sepa-
rated by centrifugation within 1 to 2 hours and stored
at 70(C for no O7 days. Serum levels of coenzyme
Q10, a-tocopherol, g-tocopherol, b-carotene, a-carotene,
and lycopene were measured by the high pressure liquid
chromatography methods, as described previously.8,9
The coecients of variation were !8% for all nutrients.
Serum 17b-estradiol and progesterone levels were mea-
sured by the radio immunoassay method in the clinical
laboratory at our institution. Statistical analyses were
performed with the Student t test, and by Wilcoxon
paired-sample test between the luteal phase and follicu-
lar phase. Signicance was dened by a probability
value of !.05.
Results
A total of 65 non-smoking premenopausal women were
enrolled in this study. Most of the study subjects were
Hispanic (60%) and black (38%) women, and most of
them were representatives of an inner-city population.
Of these 65 women, 10 women who were not taking any
OC participated in the menstrual cycleantioxidant
study. In these 10 women, serum levels of estradiol
increased from 84 G 20 pg/mL in the follicular phase to
205 G 60 pg/mL in the luteal phase (P ! .01), and
progesterone levels changed from 0.7 G 0.3 ng/mL to
6.0 G 2 ng/mL (P ! .01). Progesterone levels in all
subjects during the luteal phase were O3 ng/mL.
Among the remaining 55 women, 15 women were OC
users for a minimum period of 6 months, and 40 women
did not use OCs. Signicantly lower serum levels of co-
enzyme Q10 and a-tocopherol were detected during the
follicular phase of the menstrual cycle, when compared
with the luteal phase, in healthy premenopausal women
(P ! .05, Wilcoxon paired-sample test).
g-Tocopherol, b-carotene, a-carotene, and lycopene
levels were comparable between the follicular and luteal
phases of the menstrual cycle. OC use signicantly
Palan et al
decreased serum levels of coenzyme Q10 and a-tocoph-
erol (P ! .001, by the Student t test) in premenopausal
women. Serum levels of g-tocopherol, b-carotene,
a-carotene, and lycopene were comparable between OC
users and nonusers (Table).
Comment
We examined the inuence of ovarian hormones during
the follicular and luteal phases of the menstrual cycle on
serum levels of lipid-soluble antioxidants in healthy
premenopausal women. Our data demonstrate signi-
cantly lower coenzyme Q10 and a-tocopherol levels during
the follicular phase, when compared with the luteal phase,
in women with regular menstrual cycles, whereas other
antioxidants (g-tocopherol, b-carotene, a-carotene, and
lycopene) were not altered signicantly. We also studied
the eects of exogenous ovarian hormones on levels of
the same antioxidants in a separate subgroup of premen-
opausal women who were OC users for at least a 6-month
duration. The results show that the use of OC signi-
cantly lowered the serum levels of coenzyme Q10 and
a-tocopherol, compared with the levels in non-OC users.
g-Tocopherol, b-carotene, a-carotene, and lycopene
levels were comparable between both groups. Similarly,
we observed a decrease in serum levels of coenzyme Q10
with the use of hormone replacement therapy (unpub-
lished data). The limitations of our present study include
small sample size and the fact that all blood samples were
nonfasting. Blood samples from OC users were collected
irrespective to placebo or the steroid medication phase,
and blood samples from non-OC users were taken ran-
domly at any day of the menstrual cycle. This may explain
the dierences between serum levels of a-tocopherol in
non-OC users and the levels that were obtained in the fol-
licular phase. It is noteworthy that, unlike water soluble
antioxidants where plasma levels are related directly to
daily intake, lipid-soluble antioxidants levels are related
directly to lipid stores and not inuenced appreciably by
uctuations by daily intake.10
Several reports have demonstrated a nonsignicant
uctuation in plasma levels of a-tocopherol by phase of
the menstrual cycle. Lanza et al6 reported signicantly
lower (12%) plasma a-tocopherol levels during menses
(days 1 and 2) than during the luteal phase in subjects
with controlled-diet conditions, but not in a free-living
group. Recently, Reimer et al7 reported 12% lower se-
rum a-tocopherol levels during the follicular compared
with the luteal phase. We observed 28% lower serum
levels of a-tocopherol in the follicular phase compared
with the luteal phase of the menstrual cycle (Table).
The uctuation in plasma levels of vitamin E has
been reported in premenopausal OC users.11-14 We
found a 23% lower serum level of a-tocopherol in OC
users, which is comparable with a 20% decrease that
e37
was reported by Aftergood et al11 in plasma a-tocoph-
erol level in young women who used OCs. They also
reported signicantly lower plasma a-tocopherol levels
in OC users who received vitamin E supplements.
Reimer et al7 reported an association of high-energy
intake in the luteal phase, compared with the follicular
phase. The lower energy intake in the follicular phase
likely explains lower intake of several key nutrients com-
pared with the luteal phase. This may have a signicant
impact on serum levels of lipid-soluble antioxidants. Die-
tary interventions may be important for womens health,
especially among young OC users, because of the impact
of diet, nutrition, and weight patterns on many conditions,
which include CVD, the leading cause of death in women.
Although the relation between estrogen and CVD
events remains unclear, there is a substantial body of
observational studies that points toward a cardioprotec-
tive role for estrogen in healthy women.4 Estrogen, which
acts through 1 of its receptors, stimulates COX-2 produc-
tion, which boots prostacyclin, prostaglandin I2, which in
turn protects the heart from atherosclerosis in female mice
by restraining both oxidant stress and platelet activa-
tion.15,16 Oxygen free radicals can cause oxidative damage
to lipids and proteins and contribute to the development
of CVD.17 Lipid-soluble antioxidants are important
natural cellular defense agents that protect lipids and
protein molecules from oxidation.1 Coenzyme Q10 and
a-tocopherol are lipid-soluble free radical scavengers
that are located together in cell membranes capable of
neutralizing oxygen free radicals and are thus, postulated
potentially to decrease the risk of CVD.2,3
The precise molecular mechanisms of action of anti-
oxidants remain poorly understood. Dietary antioxi-
dants are protective agents that can counteract oxidative
stress and potentially re-establish a healthy cellular
redox balance. Our ndings demonstrate the varying
eects of endogenous and exogenous ovarian hormones
on lipid-soluble antioxidant levels and should be taken
into consideration in future antioxidant research. If our
ndings are conrmed by larger studies, women who
receive OCs may be considered for coenzyme Q10 and/or
a-tocopherol supplementation. The potential value, if
any, for coenzyme Q10 and a-tocopherol supplementa-
tion in OC users and the eect of menstrual cycle phase
on oxidative stress deserve further investigation.
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