The Iron Content of Sweat in Normal Adults

MAJ. CHARLES A. COLTMAN, JR., Mc, USAF* AND NANCY J. ROWE, B.SC.t

J OHNSTON and Hagan’ were the first investi- sary. All-aluminum electrodes were used to avoid
contamination of the skin with iron, which was
gators to point out that in most studies of
present in small amounts in the brass electrodes used
iron metabolism up to 1949 no consideration
previously. Cellulose sponges for sweat collection
had been given to the loss of iron in perspira-
are unsatisfactory for study of iron content because
tion. Since that time periodic studies have
they adsorb some of the iron from standard solutions
been carried out in which the significance of the passed through them. Therefore, ve used plastic
iron composition of sweat was investigated. foam sponge$ which, after careful studs-, were found
Variations in the method and site of collection to have no effect on the iron content of the solutions.
and the technic of iron analysis (Table i) have All materials which contacted the skin during

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tended to create differences in the results ob- iontophoresis and the collection of sweat were
tamed. Because of the limited data available checked for iron contamination. The gauze sponges
used as electrode pads and for cleansing the skin
on the iron content of sweat in both healthy and
before and after iontophoresis were a particular
diseased subjects, ve attempted to apply a
source of difficulty. The- were macic iron-free by
simple technic used for stimulation and collec-
rinsing in demineralized distilled water until the
tion of sweat to the analysis of the iron com-
rinse water was iron-free. The usual precautions in
position of sweat. The technic and the results
cleaning glassware for iron analysis were observed.
of analysis of sweat from normal adults are de- The skin itself was inspected carefully and areas
scribed in this paper. with abrasions were aroided. Sites selected for
iontophoresis were rubbed vigorously with gauze
MATERIALS AND METHODS
sponges which had been moistened with deniineral-
Iron content of sweat, serum iron, hemoglobin ized distilled water to remove as much dirt and
and hemnatocrit were determined in twenty-eight desquaniated skin as possible. Before analysis the
normal adults (Table mm). Subjects whose serum sweat samples were centriftmged for 15 minutes at
iron levels were below 70 j.g. pe cent were not in- 3,100 r.p.m. to remove cellular material.
eluded in the studs’. Samples of bhood were obtained by venipuncture
Sweat was collected from the volar surface of both imiimediately after the collection of sweat. An au-
forearms, using the iontophoresis technic previously citmot of each sample was l)laCed in balanced oxalate
described by Coltman and Several modifi- anticoagulant for hemoglobin and hemnatocrit deter-
cations of this technic, which was devised originally mination, and the remainder was allowed to clot for
for electrolyte determinations in sweat were neces- serum iron analysis.
Iron analysis was performed using a microniodifi-
From the Ohio Tuberculosis Hospital and the Ohio
cation of the alpha-alpha’ dipvridvl mr,ethod, origin-
State University College of Medicine, Columbus, Ohio.
* Instructor, Department of Medicine, Ohio State ally described by y3 This technic was de-

University College of Medicine. Present address: Dc- ‘eloped by Scarlata and Moore4 for use with the
partment of Medicine, Wilford Hall, USAF Hospital, Beckman Spinco Ultramicro Analytical System. In
Aerospace Medical Division (AFSC), Lackland Air the present study, a Beckman Model DU spectro-
Force Base, Texas; t Clmemist, Ohio Department of photometer equipped with a microadapter assembly
Healthi amid Graduate Student, Department of Physi- and 0. 1 ml. absorption cells was used for colorimetry.
ology, Ohio State University. The iron content of sweat was determined ron-
This work was supported by Grant AM 04653-03 GM
tinely on 20!) lambda sample volumes, as used by
from the National Institutes of Health.
The contents of this paper reflect the views of the Scott Industrial Foam (Urethane), ‘/ inch thick
authors and are not to be construed as a statement of with 80 pores per lineal inch, niade to our specifications
official Air Force policy. by Merryweather Foam Latex Company, Akron, Ohio.

A merican Journal of Clinical Nutrition 270 Vol. 18, April 1966

 Iron Content of Sweat in Adults 271

TABLE I
Studies of Iron Concentration in Sweat

Iron

Study Source
Cell-Rich Sweat Cell-Free Sweat

Johnston, Hagan’ . . . 0.02-0.06 mg./L. (2-6) Whole body

Mitchell, Hamilton6 1-2 mg./L. (100-200) . . . Whole body
Johnston et al.8 . . . 0.27 mg./L. (27) Whole body
Adams et al.7 7.05 sg./cc. (705) 0.296 jig/cc. (29.6) Upper extremity
Foy, Kondi’4 0.3-6.0 mg./L. (30-600) 100-200 g./L. (10-20) ...

Hussain et al.’3 1.61 mg./L. (161) 0.44 mg./L. (44) Upper extremity
Moore’2 0.4-1.5 mg./L. (40-iSO) 0.346 mg./ml. (34.6)f Forearm
Hussain, Patwardhan” 1.15 mg./L. (115) 0.34 mg./L. (34) Upper extremity
Apte, \‘ankatachalam’5 330-622 pg./L. (33.0-62.2) 190-250 mg./L. ( 19-25) Upper extremity
Prasad et al.’6 120 ± 36 jzg.% 46 ± 11 g.% Upper extremity
Coltman, Rowe . . . 17.2 ± 7.8 g.% Forearm

I Figures in parentheses are those obtained by converting original values to micrograms per cent.

t Averaged from graph.

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Scarlata and Moore,4 but this volume could be re- sweat in the two samples did not differ by more
duced to 100 lambda if necessary by decreasing the than 5 zg. per cent with a mean difference of 2
combined volumes of the dipridl and sodium sul- g. per cent. Sweat volumes obtained from
fite reagents to 50 lambda. Handling smaller
the two arms were approximately equal and
volumes makes the procedure somewhat more
tedious, hut it is necessary to avoid dilution of
samples already low in iron content. Because larger
TABLE II
samples were available in our study, 0. 5 ml. of
Iron Content of Sweat
serum was used and the reagent volumes were in-
creased accordingly. A series of standard analyses
Iron (g.%)
Hemo.
was carried out with each determination, and a Subject. Total Hema-
globin
Age (yr.) Sweat tocrit
serum obtained commercially was used for quality and Sex (ml.) Sweat
(gm/mOO
(%)
ml.)
control. The serum iron value given by the manu- (av.) Serum

facturer was 107 .sg. per cent, and the value actually
obtained in a series of ten analyses was 105 zg. per I). P., 21, 51 0.75 12 154 15.1 43
J. p.. 23, 51 0.65 20 109 13.8 41
cent with a standard deviation of 2 tg. per cent. J. M., 24, 51* 0.35 17 100 14.0 42
J. H., 24, 51 0.55 17 139 16.0 49
J. 1)., 24, Si 0.65 23 109 15.8 48
RESULTS
E. S., 24, 51 0.45 29 109 14.0 39
P. SI., 25, 51 0.45 11 92 16.2 48
The results obtained in analysis of sweat J. B. . 25, 51 0.40 19 93 15.1 42.5
J. 1). , 27, 51 0.85 21 147 14.7 42
from twenty-eight normal adults are given in J. Mc., 45, M 0.45 27 132 14.0 42
Table II. In nineteen subjects the iron content J. w. , 63, M 0.45 2 100 15.5 48
G. 51. , 22, 51 0.50 37 127 13.8 40
of sweat was determined separately on samples C. G., 23, Si 0.50 14 100 14.0 40
S. B., 34, 51 0.45 14 153 14.9 42
from both arms. In seven subjects it was A. D., 23, Ft 0.35 20 101 13.7 40
necessary to poo1 equal amounts from each J. H., 23, F 0.45 18 136 13.8 40.5
S. S., 24, Ft 0.35 12 130 14.7 42
arm to obtain enough for analysis. In one F. H. 26, F
, 0.40 101 I :i .8 40.5
C. 1)., 27, Ft 0.30 11 72 12.8 37
woman the sweat obtained from one arm was 51. M., 34, F 0.40 25 137 13.2 42
H. 1). , 35, F 0.55 13 93 12.7 41
negligible but the sample from the other was B. I)., 39, Ft 0.25 7 13.6
94 41
adequate. Sweat was collected from only one M.G.,41,F 0.30 4 131 13.2 36
C. M. , 47, F 0.45 20 84 12.5 41
arm in one man becausc or a Iamlure in tne ion- L. P. , 58, Ft 0.35 25 152 14.7 40.5
S. I)., 21, Ft 0.35 18 101 13.8 41
tophoresis equipment. With these exceptions, 0. K., 31, F* 0.30 25 87 13.9 42
G. 51., 45. F 0.35 15 70 13.8 38
the values for the iron content of sweat listed
in Table II represent averages of the two sam-
1 Sweat collected from one arm only.
ples. In all subjects the iron composition of t Sweat pooled for analysis.
272 Coltman and Rowe

interest that Mitchell and Hamilton5 suggested

ISO that findings in their experiments rendered it
40
unnecessary to postulate ‘ ‘ physiological mecha-
I
nisms of unknown nature’ ‘ for limiting iron
130 I
absorption and excreting iron absorbed in ex-
bQ 20 cess of need.

x x Stewart et al.6 were the first investigators to

110 I

use radioiron in the study of iron excretion;

a’
it
they concluded that insignificant amounts of
.
radioiron were present in the washings of dog
x
a I
skin after the administration of radioactive
#{149} I
I
a xx XI x iron. Mitchell and Hamilton3 found that as
1J x
much as 6.5 mg. of iron is lost in sweat per day.
. x
Impressed by their report, Adams et al.7 studied
the iron content of both “cell-rich sweat” and
50
‘ ‘cell-poor sweat” obtained by centrifugation
x -MALES
and found a significant difference between
#{149}- FEMAL.CS

them. In all their determinations, “cell-poor

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10 3
SWEAT IN.MlORN4S % 40 sweat” contained less than 0.64 pg. per cc.;
since this was below the level of accuracy of
FIG. 1. Relationship of serum iron to iron content of
sweat. their technic, they concluded that the iron
content of “cell-poor sweat’ ‘ was negligible.
the values given . represent the total amount of The “cell-rich sweat” contained high concen-
sweat obtained from each subject. trations of iron, related primarily to desquama-
Although precautions were taken to avoid tion. Johnston et al.8 studied the calcium and
contamination, the iron content of sweat varied iron loss in the perspiration of four college
considerably, between 2 and 37 jsg. per cent. women and found a mean iron loss of v.27 mg.
The lowest values must be interpreted in view per L. of sweat. These investigators found no
of the sensitivity limitations of the analytical evidence of an increase in the amount of iron
method. It is possible that the values 2 and 4 excreted in perspiration following the intake of
tg. per cent represent actual iron levels of zero. additional iron. The results of their study
The mean iron content of sweat for the group showed that the amount of iron lost in perspira-
was 17.2 tg. per cent, with a standard deviation tion was small but of sufficient magnitude that
of 7.8 jig. per cent. The mean value for male the average woman who engages in average
subjects was 18.6 j.g. per cent, that for females activity under average conditions of tempera-
15.7 zg. per cent ; the difference between them ture and humidity must increase her iron in-
is not statistically significant. No correlation take to compensate for these losses. They
was evident between serum iron levels and iron proposed several possibilities but came to no
content of sweat (Fig. 1). conclusion about the difference between their
results and those of Mitchell and Hamilton.1
COMMENTS
Dubach et al.,9 who studied the problem of
The initial conclusions of Johnston and iron excretion by administering isotopic iron
Hagan’ were that the iron concentration of intravenously, detected only minute traces of
perspiration was negligible. Shortly there- isotopic iron in the large volumes of sweat col-
after, Mitchell and Hamilton5 . became inter- lected. The radioiron content of sweat from
ested in the dermal excretion of various ele- one patient with hypoplastic anemia was some-
ments including iron and demonstrated that the what higher than that in sweat from three
sweat gland is an important means of iron ex- normal people.
cretion. They concluded that 37 per cent of Foy and Kondi’#{176}were the first to investigate
total iron loss is through the skin. It is of some the etiologic importance of dermal iron loss in
 Iron Content of Sweat in Adults 273

the microcytic anemia o certain areas of thermal sweat varied from 0.63 to 1 .8 mg. pe
India. They found that the sweat might con- L. of sweat, vith a mean value of 1 15 per L.
.

tam from 0.3 to 6 jig. per ml. iron, and that of sweat. These values are somevhat lower
under conditions of maximum sweating (up to than those reported by Mitchell and Hamilton,5
10 L. per day) this might be an important Adams et al.7 and Foy and Kondi,’4 but if the
source of iron loss. Because the quantity of higher values are assumed to be representative
iron present is related to the cell content of for Indians, as much as 13 mug. of iron can be
sweat, desquamation and regeneration rates in- lost per day under conditions of maximal sweat-
fluence iron loss. Foy and Kondi’#{176} empha- ing. Hussain et al.’3 considered cutaneous
sized the confusion generated because some iron loss one of the possible contributory factors
workers have estimated the iron content of in the genesis of iron deficiency anemia in the
cell-free sweat after filtration or centrifugation tropics.
and others have used sweat containing epithe- Recently, Apte and Vankatachalamn’5 studied
hal cells. Hussain and Patwardhan” ana- healthy male volunteer subjects between the
lyzed the cell-free sweat of seventeen female ages of twenty-one and forty and found that
subjects suffering from niicrocytic hypochromic the iron content of their ‘ ‘ cell-rich’ ‘ sweat was
anemia and found no iron ; furthermore, the greater thami that of their “cell-free” sweat, but

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iron content of cell-rich sweat from these sub- significantly lower than levels reported by
jects was significantly lower than that of six other workers. The iron content of “cell-free”
healthy female subjects. These workers re- sweat was nearly the sanie as that reported by
ported that after forty-two days of iron therapy other workers, and variations in the location of
the average iron content of ‘ ‘ cell-rich’ ‘ and collection of sweat produced variations in iron
“cell-free” sweat increased to volumes within content. It appears that this variable makes
normal range. it difficult to extrapolate the iron loss from one
Moore,tm2 in a report on the extension of area to total body loss. Continuous thermal
radioiron studies, reaffirmed that in 24 hours stress did not result in a diminution in the con-
05 to 1 mg. of iron is lost by sweating at a centration of iron in sweat, and Apt and Van-
maximum rate. As Moore pointed out, one katachalam concluded that a significant
should recognize the error in determining the amount of iron was lost during artificially in-
24 hour dermal loss of iron in sweat by multi- duced thermal sweating and that factors such
plying the value obtained from a collection as environmental temperature and regional
over a brief period of time from a small portion body sweating largely determined the magni-
of the body by an estimated surface area. tude of the iron loss. Prasad et have fur-
Hussain et al.,’3 in considering the discrepancy ther confirmed the importance of sweat as a
between iron excretion values obtained by source of iron loss.
chemical analysis and those obtained by radio- In Table I it is shown that there is a signifi-
isotope methods, suggested that although radio- cant difference between “cell-rich” and “cell-
iron administered to subjects appears readily in free” sweat in all studies reported although the
the secretion of the sweat glands, it may take values vary markedly. However, most of the
much longer to arrive in the cellular layers of values obtained for iron in “cell-free” sweat are
the epidermis. These workers studied the iron of the same order of magnitude. The results of
content of insensible perspiration from a small studies reported by Foy and Kondi’4 and Apte
area of the upper limb in sixteen healthy men and Vankatachalam’4 are strikingly similar to
and found that the average iron loss for an 8 those found in our study. The problems of
hour period was 2.1 g. with a range from 0.92 extrapolating local sweating and exfoliation to
to 3.48 Mg. If it is assumed that this loss was the whole body dermal iron loss have not been
identical from area to area, total iron loss for solved by this technic.
the whole body surface through insensible The sweat obtained in this study was essen-
perspiration would be 0.13 mg. per 24 hours. tially cell-free because the skin preparation re-
The iron content of the “cell-rich” portion of moved the cells; some cells adhered to the
274 Coitman and Rowe

sweat collection sponge and centrifugation re- subjects, with particular

reference to calcium and
moved the remainder. iron. J. Biol. Chem., 178: 345, 1949.
6. STEWART, W. B. , SNOWMAN, R. T. , YUILE, C. L.
The technic described herein constitutes a
and WHIPPLE, G. H. Radioiron excretion by the
simple, rapid method for the collection and skin and kidney of dogs. Proc. Soc. Exper. Biol.
analysis of the iron content of “cell-free” sweat & Med., 73: 473, 1950.
and promises to be valuable in the study of 7. ADAMS, W. S., LESLIE, A. and LEVIN, M. H. The

this parameter during disturbances of iron dermal loss of iron. Proc. Soc. Exper. Bio. &
Med., 74:46, 1950.
metabolism.
8. JOHNSTON, F. A. , MCMILLAN, T. J. and EVANS,
E. R. Perspiration as a factor influencing the re-
SUMMARY
quirement for calcium and iron. J. Nutrition,
A simple, rapid method for the collection and 42:285, 1950.
analysis of the iron content of sweat is de- 9. DUBACH, R. , MOORE, C. V. and CALLENDER, S.
Studies in iron transportation and metabolism.
scribed. A review of the literature reveals that
Ix. The excretion of iron as measured by the iso-
there is considerable variation in the iron con-
tope technique. J. Lab. & C/in. Med., 45: 599,
tent of sweat when iron lost in both exfoliation 1955.
of epidermis and sweating is considered. There 10. Foy, H. and K0NDI, A. Nutritional and intestinal
is reasonable agreement on the iron content of factors and iron
losses in the genesis of tropical
anemias. Lancet, 1 : 423, 1956.

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cell-free sweat, and in this study it amounts
11. HUSSAIN, R. and PATWARDHAN, V. N. Iron content
to an average of 17.2 g. per cent in the normal of thermal sweat in iron-deficiency anemia.
male or female adult. Lancet, 1: 1073, 1959.
12. MOORE, C. Iron metabolism and nutrition. In:
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Fed. Proc., 8: 387, 1949. MACMAR!, S. Dermal loss of iron in healthy

2. COLTMAN, C. A. and ATWELL, R. J. Sweat collec- Indian men. mndwn J. M. Res., 48: 235, 1960.
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1957. Trop. Med., 60: 105, 1957.
4. SCARLATA, R. W. and MOORE, E. W. A micro- 15. APTE, S. V. and \‘ANKATACHALAM, P. S. Factors

method for the determination of serum iron and influencing dermal loss of iron in human volun-
serum iron-binding capacity. C/in. Chem., 8: teers. Indian J. M. Res., 50: 817, 1962.
360, 1962. 16. PRASAD, A. S., SCHULERT, A. R., SANSTEAD, H. H.,
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ma! excretion under controlled environmental gen content of sweat in normal and deficient
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