0021-972X/84/5903-0551$02.
00/0
Journal of Clinical Endocrinology and Metabolism
Copyright © 1984 by The Endocrine Society
Age Changes and Sex Differences in Serum
Dehydroepiandrosterone Sulfate Concentrations
throughout Adulthood
NORMAN ORENTREICH, JOEL L. BRIND, RONALD L. RIZER, AND
JOSEPH H. VOGELMAN
Orentreich Foundation for the Advancement of Science, Inc., New York, New York 10021
ABSTRACT. In a cross-sectional study, serum dehydroepian-
drosterone sulfate (DS) concentrations were measured in 981
men and 481 women, aged 11-89, yr. The resulting data were
asymetrically distributed and were normalized by logarithmic
transformation and analyzed by 5-yr age grouping (e.g. 15-19 yr,
20-24 yr, etc.). The DS concentration peaked at age 20-24 yr in
men (logarithmic mean, 3470 ng/ml) and at age 15-19 yr in
women (log mean, 2470 ng/ml). Mean values then declined
steadily in both sexes (log mean at >70 yr of age, 670 ng/ml in
men and 450 ng/ml in women) and were significantly higher in
D EHYDROEPIANDROSTERONE sulfate (DS) is
the most abundant steroid in the circulation and
arises primarily by secretion from the adrenal cortex (1,
2). Cross-sectional and longitudinal studies showed that
the plasma DS level, while high in neonates, falls mark-
edly within months and remains low until age 7 yr. DS
increases during adrenarche, peaks between the ages of
15-25 yr, and then decreases steadily after the third
decade (3-9).
Previous reports have stressed the variability of the
concentration of DS in plasma, both within and between
age groups, and the difficulty in establishing normal
limits. For instance, Zumoff et al. (8) found significantly
higher values for men (n = 12) than women (n = 18)
over age 50 yr. This difference between the sexes was
not significant in adults below age 50 yr. In contrast,
DePeretti and Forest (3) found plasma levels of DS to
be significantly higher in adult men (aged 18-49 yr; n =
27) than in adult premenopausal women (aged 20-47 yr;
n = 25). Yamaji and Ibayashi (4) found a significant sex
difference only in the age group 21-30 yr, with higher
values for men (n = 25) than women (n = 20).
The above discrepancies might be due to a combination
Received January 18,1984.
Address requests for reprints to: Dr. Joel L. Brind, Biochemistry
Lab, Orentreich Foundation for the Advancement of Science, Inc., 910
Fifth Avenue, New York, New York 10021.
551
Vol. 59, No. 3
Printed in U.S.A.
men than women at ages from 20-69 yr. Analysis of 517 ran-
domly selected sera (from women) which had been stored frozen
for 10-15 yr gave results indistinguishable from values obtained
from fresh specimens. In a supplementary study, a longitudinal
analysis of weekly specimens from 4 normal men, aged 36-59
yr, revealed individual variability (mean coefficient of variation,
19%) and failed to demonstrate any monthly, seasonal, or annual
rhythmicity. Based on the above analyses, a table of normal
serum DS ranges for adult men and women is presented for use
as a clinical reference. (J Clin Endocrinol Metab 59: 551, 1984)
of small sample size and the variability of DS, producing
statistically inconclusive results. Using much larger sam-
ple populations we hoped to establish normal ranges and
sex differences for this hormone in adults. In addition,
to determine short term changes in DS, we studied the
longitudinal variation in the plasma DS concentration
by sampling a group of four men at weekly intervals for
a 2-yr period.
Materials and Methods
Subjects
Cross-sectional analyses. The following groups of patients from
a New York dermatological practice were studied: 1) 981 men
who were endocrinologically normal (based on normal serum
assays for T4, T 3 uptake, cortisol, testosterone, and sex hor-
mone-binding globulin) and were not taking medications
known to modify the level of DS (i.e. glucocorticoids or estro-
gens); and 2) 481 women who were endocrinologically normal,
were not taking glucocorticoids or estrogens, and were without
significant acne, alopecia, or hirsutism.
Data were analyzed by 5-yr age groups for men and women,
i.e. 15-19, 20-24, 25-29, 30-34, 35-39, 40-44, 45-49, 50-54,55-
59, 60-64, 65-69, and over 70 yr.
To evaluate the long term stability of DS in frozen serum, a
group of 517 randomly selected specimens from women was
assayed. These sera were taken from a collection that had been
552 COMMENTS JCE&M«1984
Vol59«No3

4.00-

1
1
3.75- 1
3
1 2 1 1 1
1 5 4 1 1
2 3 3 3 1
3 7 4 2 3 2 1
3.50- 4 5 10 1 2 2
1 6 6 7 2 1 2
1 1 9 2 3 6 1
2 2 4 3 3 2 1 2 1
3 2 2 3 6 5 4 3
3.25- 2 6 4 6 4 2 1 1
log DS 4 4 2 2 3 1 1 1
in 4 2 4 4 1 4 1
serum 2 1 1 1 2 3 3 2 3 1 1
(ng/ml) 2 1 3 3 5 2 6 3 3 3 1
3 > 0 0-
3 1 3 2 3 2 2 3 2
FIG. 1. Scattergram of log serum DS us. 3 3 5 l 3 2 2 4 4 2
age in normal women. Data points show 4 4 2 2 2
the number of subjects at each measured 3 1 1 7 6 4 3 1 1
2 2 2 4
value. 2.75- 1 1 5 1 2 1 1
1 1 1 1 3 3 2 1
5 1 2 3
1 1 1 1 1 1 1
1 1 1 2 2 1 2
2.50- 2 3 1
2 2
1 1 1 1 1
1 4 2 1
1 1 1
2.25-
1 1
1
1 1 1
2.00

t i .... i i
i
20-24 30-34 40-44 50-54 60-64 >70
T5-19 25-29 35-39 45-49 55-59 65-69

AGE GROUP (years)

gathered 10-15 yr ago as part of a multiphasic health screening
program by The (Kaiser) Permanente Medical Group (Oak-
land, CA). The sera had been frozen immediately and were
maintained without thawing at —20 C until assay.
Longitudinal analysis. DS concentrations were measured in the
sera of four men at frequent intervals for 2 yr to determine
short term variability and possible cyclical variation. Two of
the subjects were in their late 30s, and two were in their late
50s. Blood was drawn weekly after an overnight fast between
0730 and 0830 h, and the serum was stored at -20 C and
subsequently analyzed in a single assay.
RIA of serum DS: Serum DS levels were measured by the
direct RIA method of Buster and Abraham (10), except that
the radiolabeled ligand was [1,2-3H]- instead of [7-3H]dehy-
droepiandrosterone ([7-3H]DHEA). Duplicate aliquots of se-
rum were diluted 1:2000 to a final volume of 0.5 ml and
incubated overnight with 6000 cpm [1,2-3H]DHEA (in 0.1 ml)
and 0.1 ml antiserum (no. 1490-S-1502#7, Radioassay Systems
Laboratories, Carson, CA). The unbound steroid was removed
with dextran-coated charcoal, and an aliquot of the supernatant
was counted. The cross-reactivity of the antiserum with uncon-
jugated (free) DHEA was 100%; the results, therefore, represent
the sum of serum DHEA and DS. Although there was a minor
degree of cross-reaction with other steroids, they did not con-
tribute more than 0.2 ng/ml to the assay values because of the
relative abundance of DS. The sensitivity of the method was
0.04 ng/assay tube, equivalent to 160 ng/ml serum (11). Sera
which contained less than 400 ng/ml were reassayed at a 1:500
dilution (detection limit, 40 ng/ml). The intraassay coefficient
of variation never exceeded 7%. The interassay coefficients of
variation (n = 18) were 20% at 600 ng/ml, 9% at 2100 ng/ml,
and 12% at 4800 ng/ml.
Results
In all age groups, DS was not distributed normally,
but was skewed to higher values. Others found this
pattern and stated that their data assumed a more nor-
 COMMENTS 553

mal distribution when expressed in logarithmic form (12, indistinguishable from those obtained from fresh sam-
13). [While this procedure effectively normalizes the ples.
data, considerable scatter is still evident (Fig. 1).] Table
1 shows the antilogs of mean log DS values and the 2-yr longitudinal study of serum DS concentrations
statistical analyses. The calculated distribution of the To eliminate interassay variation, frozen specimens
logarithm of the DS values is shown in Fig. 2. The limits were thawed and assayed in the same assay for each
shown in this figure are condensed in Table 2 to give subject. The results (Table 3) showed slightly greater
proposed normal ranges for serum DS levels in men and variability in the younger subjects, but sample size was
women throughout adulthood. too small to determine the significance of this finding.
The relationship of age and sex to the serum DS The correlation coefficients for the time distribution of
concentration is clear. For men, there was a peak at ages the DS values of these four men revealed no significant
20-24 yr, while for women, it occurred between 15-19 yr. monthly, seasonal, or annual rhythms. Computation of
Mean values then declined until the seventh decade, the autocorrelation function for these men showed no
falling to about one seventh and one fifth of the maxi- significant rhythms. These functions were obtained by
mum for men and women, respectively. Mean values for calculating the correlation coefficient of each man's se-
men were significantly higher than those for women at ries of values for various time offset intervals (14).
all ages from 20-69 yr of age (P < 0.001, by Student's t
test; Table 1). The decline in serum DS was significant Discussion
when taken in 10-yr intervals, except for ages 20-29 yr
The normal ranges that we propose include approxi-
in men and ages 15-19 and over 50 yr in women. After
mately 88% of our normal population. Although 95%
menopause, there were no significant differences between
confidence limits are more desirable, in the current study
age groups in women.
there were islands or subpopulations of low values de-
DS values obtained from specimens (from women) that tached from the main body of data, and we do not view
had been stored frozen for 10-15 yr were statistically these as normal. This feature was most pronounced in
TABLE 1. Significance of differences in serum DS between age groups women. While we eliminated subjects with conditions or
taking medication that might affect DS concentrations,
Significance of it is possible that there is an effect from conditions that
difference be-
Age Age DS logarithmic tween means 10 we did not identify or that these subnormal DS levels
group range n mean (ng/ml) yr apart represent subclinical endocrine abnormalities. Several
investigators found, for example, that DS tends to be
Groups P
significantly reduced in breast cancer patients (8). Fur-
Men thermore, Bulbrook et al. (12), in a prospective study,
A 15-19 182 2570 A-C <0.001
B 20-24 216 3470 B-D <0.2
have demonstrated subnormal urinary excretion of the
C 25-29 151 3320 C-E <0.3 DS metabolites androsterone and etiocholanolone up to
D 30-34 82 3020 D-F <0.001 9 yr before the diagnosis of breast cancer. In this regard,
E 35-39 70 2660 E-G <0.001 the establishment of reliable normal limits for serum DS
F 40-44 63 2280 F-H <0.001 is essential.
G 45-49 59 1910 G-H <0.1
This study confirms earlier reports of the age depend-
H 50-54 53 1580 H-J <0.05
I 55-59 40 1280 I-K <0.01 ence of DS in serum. Caution must be observed, however,
J 60-64 27 1030 J-L <0.01 in drawing any inferences about long term individual
K 65-69 20 830 changes in DS level from cross-sectional studies. For
L >70 18 670 example, if high DS levels were a survival risk factor,
Women
then the cross-sectional decrease in DS concentration
A 15-19 29 2470 A-C <0.2
B 20-24 54 2310 B-C <0.01 with age could be accounted for by the continuous selec-
C 25-29 63 2040 C-E <0.001 tion of individuals with lower levels rather than by the
D 30-34 52 1730 D-F <0.1 summation of longitudinal decreases (15). To differen-
E 35-39 54 1420 E-G <0.001 tiate between these hypotheses, a long term, longitudinal
F 40-44 45 1140 F-H <0.001 study has been started in this laboratory to establish
G 45-49 51 910 G-I <0.05 individual patterns of serum DS in relation to age, dis-
H 50-54 39 740 H-J <0.2
I 55-59 28 610 I-K <0.1 ease, and survival. The long term stability of DS in frozen
J 60-64 36 520 J-L <0.6 serum, which we have shown, is a necessary prerequisite
K 65-69 19 470 for such a study.
L >70 11 450 In spite of the variability of serum DS concentrations
554 COMMENTS JCE&M«1984
Vol59«No3

FIG. 2. Serum DS concentrations us.

age in men (O) and women (•). Data
points are antilogs of mean log DS val-
ues. Shaded areas delineate normal
ranges for men (am) and women (ESS). SERUM
Normal limits for each age and sex group DS
were computed by the equation: mini- (pg/ml)
mum or maximum = antilog [mean of
log DS values ± (1 + log 2) SD] corre-
sponding to the log equivalent of the
mean ± 2 SD and including about 90%
of measured values. To obtain smooth
curves for means and limits, curves were
computer fitted to the general equation:
y = K Exp [ax3 + bx2 + ex + d], where
y is the DS value in micrograms per
milliliter and x is the mean age of the
group.

i
20-24 30-54 40-44 50-54 60-64 >70
15-19 25-29 35-39 45-49 55-59 65-69
AGE GROUP (years)

TABLE 2. Proposed normal ranges for serum DS (calculated using log- TABLE 3. Means and variations of serum DS during a 2-yr period in
normal distribution) four men sampled weekly

Age range Range Age Mean SD CV

(yr) (ng/ml) (yr) (ng/ml) (ng/ml) (%)
Men 1 36 3442 750 22
15-39 1500-5500 2 39 3324 713 21
40-49 1000-4000 3 57 1820 263 14
50-59 600-3000 4 59 1202 220 18
>60 300-2000
CV, Coefficient of variation.
Women
15-29 1000-5000 of this sexual dimorphism remain to be determined.
30-39 600-3500
40-49 400-2500
The lack of monthly, seasonal, or annual rhythms in
>50 200-1500 serum DS in normal men is noteworthy and, to our
knowledge, has not been reported previously. A lack of
significant circadian rhythmicity in normal men has,
within and among individuals, we were able to demon- however, been described previously (18). In a recent
strate a significant difference between the sexes at all paper, Read et al. (19) demonstrated a clear circadian
ages from 20-69 yr (P < 0.001). It is our belief that rhythm in salivary DS. While the latter researchers
previous investigators failed to demonstrate this differ- acknowledge a lack of correlation between salivary and
ence consistently because of sample sizes that were too plasma DS levels, they also cite Zumoff et al. (20) for
small relative to the large variation in normal values. "recognizing... the marked circadian fluctuation of
Previous studies suggest that this sex difference in serum plasma DS." However, this citation is erroneous, the
DS levels results from a difference in the DS clearance cited reference referring not to DS, but, rather, to circa-
rate (16, 17), but the biological and clinical significance dian fluctuation of unconjugated DHEA.
 COMMENTS
Acknowledgment
The authors wish to thank Dr. William Rosner of the Department
of Medicine, Columbia University College of Physicians and Surgeons
and St. Luke's-Roosevelt Hospital Center, New York, NY, whose
consultation on this paper was uniquely valuable.
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