TheAmerican Journal of Clinical Nutrition 3 3 : FEBRUARY 1980, pp. 227-231. Printed in U.S.A.

227
The effects of the dietary intakes of copper,
iron, manganese, and zinc on the trace element
content of human milk1-3
E. Vuori, M .D., M . Sc., S. M . M #{224}kinen, D. Sc., R. Kara, M . Sc.,
and P. Kuitunen, M .D.
ABSTRACT Fifteen breast-feeding mothers who volunteered for a study of trace elements in
human milk, kept two 7-day food records during the course of lactation. The 1st survey week
ranged from 6 to 8 weeks and the 2nd survey week from 17 to 22 weeks postpartum. The milk
samples submitted for analysis represented a sample of every feed during a period of 24 hr and
contained foremilk and hindmilk in equal proportions. The 7-day food records were handled by a
computing system to give an average daily food consumption and nutrient intake, but iron was the
only trace element given by the system. In order to estimate the trace element intakes of the
mothers, mixtures were prepared to represent the average daily food consumption. The trace
element concentrations of the mixtures and milk samples were analyzed using the atomic absorption
spectrophotometric method. The estimated average daily copper intake was 1.8 mg, the iron intake
16 mg, the manganese intake 5 mg, and the zinc intake 13 mg. The mean copper, iron, manganese,
and zinc concentrations of the milk samples were 0.36 0.07 mg/liter, 0.40 0.10 mg/liter, 4.5
1.8 tg/hiter, and 1.89 0.74 mg/liter for the 1st survey week and 0.21 0.07 mg/liter, 0.29
0.09 mg/liter, 4.0 1.5 jig/liter, and 0.72 0.44 mg/liter for the 2nd survey week. A significant
positive correlation (P < 0.01) was observed in the 2nd survey week between maternal manganese
intake and the manganese content of human milk. The intake of copper, iron, and zinc, at the
present level of intake, did not seem to affect the corresponding trace element levels in human
milk. Am. J. C/in. Nut,. 33: 227-231. 1980.
The composition of human milk is not
constant. Variations of trace element content
have been shown to occur from mother to
mother and also with the stage of lactation
(1-4). In our follow-up study of 27 mothers
a decrease in the iron, copper, and zinc con-
centration of human milk was associated with
increasing stage of lactation (5, 6). The man-
ganese concentration was at a minimum be-
tween the 4th to 5th month of lactation,
whereafter a rising tendency was observed
(E. Vuori, unpublished observations). In this
context a wide range of individual variation
in the trace element concentrations was con-
firmed.
One factor that may influence the trace
element content of human milk is the varia-
tion in the intake of these elements. The trace
element content of the diets of lactating moth-
ers has not been extensively investigated, and
correlations of trace elements other than iron
(7) between maternal diet and human milk
are lacking. Animal experiments (goats, cows,
ewes) have shown that if the total intake is
sufficient, nutritional factors play only a mi-
nor role; the increased intake of manganese
and zinc, however, increases the amount of
these elements in the milk of cows and ewes
(1-4, 8, 9).
In order to study the influence of the intake
of trace elements on the concentration of the
corresponding elements in human milk, the
copper, iron, manganese, and zinc concentra-
tions in the individual diets and milk samples
of 15 lactating mothers were analyzed twice
From the Department of Public Health Science.
University of Helsinki, Haartmaninkatu 3, SF-00290
Helsinki 29, The Department of Nutrition, University of
Helsinki, Viikki, B-tab, SF-00710 Helsinki 71, and The
Childrens Hospital, University of Helsinki, Stenb#{228}ck-
inkatu 11, SF-00290 Helsinki 29, Finland.
2 Supported by a grant from the Yrjo Jahnsson Foun-
dation, Helsinki, Finland.
Address reprint requests to: Dr. E. Vuori, Depart-
ment of Public Health Science, University of Helsinki,
Haartmaninkatu 3, SF-00290 Helsinki 29, Finland.

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2 2 8 V UORI ET A L .
duri ng the course of l actati on. A l though
many excel l ent revi ew arti cl es have been pub-
l i shed on the nutri ti on of l actati ng mothers
(10-12) no speci al i nterest has been pai d to
the extra l oss of trace el ements, other than
i ron and zi nc, whi ch the l actati ng mother
provi des f or the i nf ant i n her mi l k. I t i s
unknown whether the i ntake of trace el e-
ments f rom an ordi nary W estern di et i s suf -
f i ci ent or i f a need f or speci f i c suppl ementa-
ti on exi sts.
M ateri al s and methods
A l l the study mothers were pri mi para and bel onged
to a group of mothers col l ected f rom a materni ty ward
(I Department of Obstetri cs and Gynaecol ogy, Uni ver-
si ty Central Hospi tal , Hel si nki ) f or trace el ement studi es
of human mi l k (6). The ori gi nal group consi sted of 30
mothers of whi ch 27 were abl e to suppl y mi l k sampl es.
Di etary data were obtai ned f rom two 7-day f ood records
kept by the mothers accordi ng to i nstructi ons (13). The
f i rst record (1st survey week) was kept f rom 6 to 8 weeks
and the second record (2nd survey week) f rom 17 to 22
weeks postpartum. Duri ng the 2nd survey week 15 moth-
e r s we r e s t i l l b r e a s t - f e e d i n g a n d wi l l i ng to keep record.
Th e s e mo t h e r s f o r m t h e p r e s e n t s t u d y g r o u p . Al l mo t h e r s
bel onged to the two hi ghest soci al groups i n the three
grade soci al groupi ng accordi ng to Rauhal a (14). The
mean age was 28 years (range 24 to 35 years), the mean
hei ght was 164 cm (range 154 to 170 cm), and the mean
wei ght bef ore pregnancy 54.4 kg (range 47 to 62 kg) and
af ter del i very 59.1 kg (range 54 to 71 kg). A t the 1st
survey week the mean wei ght was 57.6 kg (range 52 to
67 kg) and i n the 2nd survey week 55.5 kg (range 48 to
65 kg).
M i l k al i quots of 8 ml were obtai ned at the begi nni ng
and at the end of each f eed duri ng a peri od of 24 hr and
pool ed to one sampl e (6). M ost mothers del i vered a
sampl e duri ng the survey week. I f an actual mi l k sampl e
was not obtai ned, the trace el ement concentrati ons i n the
mi l k were esti mated wi th the hel p of data obtai ned f rom
sampl es col l ected i n the precedi ng or f ol l owi ng week.
The mi l k sampl es were di vi ded i nto f our groups accord-
i ng to the quarti l e to whi ch the mi l k bel onged at the
survey week f or the cal cul ati on or correl ati ons. The data
on the trace el ement concentrati ons of human mi l k were
establ i shed i n the f ol l ow-up study of 27 mothers (5, 6,
manganese; E. V uori , unpubl i shed observati ons). The
actual val ues coul d not be used because the stage of
l actati on af f ects the trace el ement content of mi l k and
al l mothers were not i n the same stage of l actati on duri ng
the survey week.
The 7-day f ood records were handl ed accordi ng to
the computi ng system programmed f or a Burroughs 6700
computer at the Computi ng Centre of the Uni versi ty of
Hel si nki to gi ve the average dai l y f ood consumpti on and
nutri ent i ntake of the mothers (15, 16). The average i ron
content of the di ets was the onl y trace el ement gi ven by
thi s system. Theref ore, a mi xture representi ng the cal -
cul ated average dai l y f ood consumpti on f or both survey
weeks separatel y was prepared f rom the f oodstuf f s
bought f rom supermarkets i n the Hel si nki area. The
mi xtures were homogeni zed wi th a K enwood chef . A
701/A homogeni zer used i n the study of the mi neral
el ement composi ti on of Fi nni sh f oods and known not to
cause contami nati on (17). The trace el ement anal yses
were made f rom ash sol uti ons by the f l ame atomi c
absorpti on spectrophotometri c method (dry ashi ng 450
C) (5, 6). The accuracy of the method was tested usi ng
as a control Bovi ne L i ver (Standard Ref erence M ateri al
no. 1577, suppl i ed by the Nati onal Bureau of Standards,
W ashi ngton, D.C.). The preci si on was establ i shed by
repeated anal ysi s of pool ed human mi l k (5, 6).
Duri ng l actati on 60 ? of the mothers used i ron sup-
pl ementati on regul arl y and an addi ti onal 20 ?i occasi on-
al l y.
Resul ts
The mean energy and cal cul ated i ron i n-
takes were si gni f i cantl y (P < 0.05) l ower i n
the 2nd survey week compared to the 1st one.
The l owest i ndi vi dual i ntakes of copper, i ron,
and zi nc were 1.1, 9.7, and 9.3 mg, respec-
ti vel y. The manganese i ntake showed the
wi dest range f rom 2.3 to 9.4 mg (Tabl e 1).
The correl ati on between cal cul ated and
chemi cal l y anal yzed average dai l y i ntakes of
i ron was si gni f i cant (n = 30, r = 0.636, P <
0.001). The zi nc and i ron contents correl ate
si gni f i cantl y wi th the energy content of the
di ets (Tabl e 2). The concentrati ons of zi nc,
copper, and i ron i n human mi l k were si gni f i -
cantl y hi gher i n the 1st survey week than i n
the 2nd survey week (Tabl e 3).
The correl ati on between the manganese
content i n the maternal di et and the man-
ganese l evel i n human mi l k was si gni f i cant i n
the second survey week (n = 15, r = 0.650, P
<0.01). A posi ti ve correl ati on was f ound al so
i n the 1st survey week, al though i t remai ned
j ust bel ow the si gni f i cant l evel . There were
no si gni f i cant correl ati ons between the cop-
per, i ron, and zi nc contents of the di et and
the correspondi ng trace el ement l evel s i n
mi l k. Those mothers who used i ron suppl e-
mentati on regul arl y di d not di f f er f rom the
rest of the mothers wi th regard to the i ron
content of thei r mi l k.
Di scussi on
I ron was the onl y trace el ement gi ven by
the computeri zed program that was used to
cal cul ate the average dai l y f ood consumpti on
and nutri ent i ntake f rom the 7-day f ood rec-
ords. Si nce a more comprehensi ve program
was not avai l abl e, mi xtures representi ng the
cal cul ated average f ood consumpti on were

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TA BL E 2
The l i near regressi on anal ysi s of the trace el ement
content of the di ets wi th respect to energy i ntakes
a x, ener y i ntake (kcal ); y, trace el ement i ntake (mg);
n = 30. P < 0.001. P < 0.01.
TA BL E 3
The concentrati ons of trace el ements i n human mi l k;
the 1st survey week ranged f rom 6 to 8 and the 2nd
survey week f rom 17 to 22 weeks postpartum.
a The
weeks.
<0.001.
P<0.01.
number of sampl es i s 15 f or both survey
Si gni f i cantl y hi gher i n the 1st survey week, P
C Si gni f i cantl y hi gher i n the 1st survey week,
TRA CE EL EM ENTS A ND L A CTA TI ON 229
TA BL E 1
The average dai l y energy and trace el ement i ntakes of l actati ng mothers and recommended dai l y di etary
al l owances (or esti mated requi rements)#{ 176}
Intake per day
R ecom m ended 1st survey w k 2nd survey w k
Energy (kcal )
M ean 2500-2600 2429 2125
SD 375 287
Range 1 9 7 6 - 3 2 2 6 1 5 6 9 - 2 5 8 8
Copper, anal yzed (mg)
M ean ( 1 . 8 ) 1 . 8 8 1 . 7 3
S D 0 . 6 2 0 . 5 5
Range 1.15-2.87 1.08-3.17
I ron, cal cul ated (mg)
M ean 16 15W 13.4
SD 2.4 2.6
Range 12.2-19.9 8.6-18.0
I ron, anal yzed (mg)
M ean 16 16.5 14.7
SD 3.3 4.6
Range 1 1 . 7 - 2 2 . 8 9 . 7 - 2 7 . 0
M anganese, anal yzed (mg)
M ean (2-3) 4.45 5.49
S D 1 . 1 3 1 . 7 5
Range 2.48-6.68 2.34-9.42
Zi nc, anal yzed (mg)
M ean 2 5 1 3 . 7 1 2 . 8
S D 2 . 7 2 . 8
range 1 0 . 0 - 1 9 . 4 9 . 3 - 1 8 . 6
a The 1st survey week ranged f rom 6 to 8 and the 2nd survey week f rom 17 to 22 weeks postpartum. The number
of mothers i s 15. Ref erence 23. Ref erence 25. d Si gni f i cantl y hi gher i n the 1st survey week (P <
0.05). Ref erence 26.
Trace element
R egression line
y - a + ba
r
Copper
I ron, cal cul ated
y 0.84 + 0.0004x
y = 4 . 2 2 + 0.0047x
0 . 2 6 2
0.596
I ron, anal yzed y = 3.39 + 0.0054x 0 . 4 7 8
M anganese y = 4.38 + 0.0003x 0.061
Zi nc y = 3 . 5 9 + 0.0042x 0 . 5 5 4
prepared and anal yzed. W hi te (18) has
poi nted out that anal yzed val ues do not nec-
essari l y agree wi th cal cul ated val ues, whi ch
represent averages of i ntakes on a country-
wi de basi s and are f ounded on a year-round
consumpti on. I n the present study i t was
possi bl e to compare cal cul ated and chemi -
cal l y anal yzed esti mates of the dai l y i ntake of
i ron. The correl ati on between the two meth-
ods was si gni f i cant (P < 0.00 1); the anal yzed
val ues bei ng about 7% hi gher on the average
than the cal cul ated ones.
A ccordi ng to W asl i en (19) the average cop-
per i ntake f rom a sel f -sel ected W estern di et
has typi cal l y ranged f rom about 1.7 to 2.4
mg, the i ron i ntake f rom about 11 to 15 mg,
the manganese i ntake f rom about 0.9 to 7.0
mg, and the zi nc i ntake f rom about 11 to 15
mg. Si mi l ar val ues f rom New Zeal and have
been recentl y reported by Guthri e and
C opper Iron M anganese Z inc
m g /l i t er t g /t i i er m g /l i t er
1st survey week
M ean 0.36 0.40 4 . 5 1.89
SD 0.07 0.10 1.8 0.74
2nd survey week
M ean 0 . 2 1 0 . 2 9 4 . 0 0 . 7 2
S D 0 . 0 7 0.09 1.5 0.44

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2 3 0 V UORI ET A L .
Robi nson (20) f or copper, manganese and
zi nc i ntakes. Hasunen et al . (21) i n Fi nl and
cal cul ated the average dai l y i ntakes of cop-
per, i ron, manganese, and zi nc of women
over 15 years of age to be 2.8, 14, 4, and 14
mg, respecti vel y. The present mean i ntakes
were about 16 mg f or i ron, 5 mg f or man-
ganese, and 13 mg f or zi nc are comparabl e
wi th earl i er i nternati onal and Fi nni sh data.
The average dai l y i ntake of copper (1.8 mg)
f rom maternal di ets i s, however, 36% l ess
than the esti mate of Hasunen et al . (21). Thei r
observati ons are based partl y on the f ood
tabl e val ues and partl y on chemi cal anal yses
(22). By cal cul ati ng the copper i ntake accord-
i ng to the method of Hasunen et al . (21),
c o p p e r wo u l d b e a s h i g h a s 3 mg / d a y f o r o u r
maternal di et. On the basi s of the present
resul ts we concl ude that Hasunen s esti mate
f or copper seems to be too hi gh.
Usi ng the Recommended Dai l y A l l ow-
ances revi sed by the Food and Nutri ti on
Board i n 1973 (23), the average i ntakes of
energy and i ron are sl i ghtl y bel ow the rec-
ommended val ues f or l actati ng mothers. Zi nc
i ntake was on the average about hal f of the
r e c o mme n d e d v a l u e ( 1 3 v e r s u s 2 5 mg ) , a n d
no mother reached the recommended l evel ,
the greatest i ntake bei ng 19.4 mg. Cal cul ati ng
f rom the l i near regressi on equati on (Tabl e 2)
the zi nc i ntake of 25 mg coul d be reached
onl y i f the average energy i ntake were about
5000 kcal /day, whi ch i s f ar too much f or the
la c ta tin g mother. I t has been shown th a t th e
z i n c b a l a n c e o f a d u l t s i s n e g a t i v e i f t h e i n t a k e
of zi nc i s about 6.5 mg/day (24). A l l our
mothers were wel l above thi s l i mi t, the l owest
i ntake bei ng 9.3 mg. Onl y approxi matel y 50%
of the mothers (8 /1 5 ) had a di etary i ron
i ntake above the recommended val ue of 16
mg i n t h e 1 s t s u r v e y we e k a n d o n e - t h i r d ( 5 /
15) i n the 2nd survey week. Neverthel ess,
60% of the mothers used i ron suppl ementa-
ti on regul arl y, whi ch gave 100 to 200 mg of
i r o n a d a y .
Of f i c i a l r e c o mme n d a t i o n s f o r t h e i n t a k e s
of copper and manganese have not been
gi ven. The W orl d Heal th Organi zati on Ex-
pert Commi ttee (2 5 , 26) h a s suggested 30 tg/
kg per day f or an adul t mal e f or copper and
2 to 3 mg of manganese f or adul ts, but no
d a t a a r e a v a i l a b l e f o r t h e c o p p e r a n d ma n -
ganese need of l actati ng women. Usi ng the
above cri teri a, the manganese i ntake of al l
our mothers was above the gi ven l evel , but i t
i s obvi ous that nearl y hal f of the mothers (7/
15) had copper i ntakes bel ow the i ndi cated
l i mi t i n the 1st survey week and two-thi rds
(10/15) i n the 2nd survey week.
W i th the excepti on of manganese the trace
el ement contents i n the human mi l k of the
present study are i n accordance wi th those
f ound earl i er (1-6, 27). The concentrati on of
manganese i n the present study appears to be
somewhat l ower than presented earl i er (27).
The coef f i ci ents of correl ati on showed that
the i ntake of manganese af f ects the man-
ganese content of human mi l k. I t i s notewor-
thy that the correl ati on between manganese
content i n the maternal di et and i n the mi l k
was greatest when i ntake f rom the di et
showed the wi dest range, namel y i n the 2nd
survey week.
W e concl ude f rom the present study that
the i ntake of copper, i ron, and zi nc at the
present l evel of i ntake has no ef f ect on thei r
concentrati ons i n human mi l k. Si mi l arl y, the
i ron suppl ementati on does not af f ect the i ron
content of mi l k. K armarkar and Ramakri sh-
nan (7) al so have f ai l ed to demonstrate a
correl ati on between the i ron content of the
di et and human mi l k. Previ ousl y i t has been
reported that the i ron content of mi l k f rom
vari ous speci es i s resi stant to changes i n the
l evel of i ron i ntake (1).
I n ani mal experi ments a def i ci ent copper
i ntake i s ref l ected by subnormal l evel s of
copper i n the mi l k. However, the addi ti on of
copper to a di et al ready adequate i n thi s
el ement has l i ttl e i nf l uence (2, 8). I n ani mal s
the manganese and zi nc content of mi l k can
be rai sed by i ncreasi ng the manganese and
zi nc i ntake (3, 4, 9). I f a si mi l ar mechani sm
exi sts i n humans, i t can be concl uded that the
i ntake of copper i n the present study was
suf f i ci ent to prevent di mi ni shed l evel s i n
mi l k, and that the i ntake of zi nc was not hi gh
enough to i ncrease the zi nc l evel s i n mi l k.
The manganese i ntake, however, was suf f i -
ci entl y hi gh to af f ect the manganese content
of mi l k.
W i ddowson (12) has cal cul ated th e extra
burden of i ron and zi nc that l actati on bri ngs
to th e mother. A ccordi ng to her cal cul ati ons
(3rd month, 950 ml of mi l k a day) mothers
l ose 1.5 mg of i ron and 5 mg of zi nc a day i n

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TRA CE EL EM ENTS A ND L A CTA TI ON 2 3 1
the mi l k. A ppl yi ng si mi l ar cal cul ati ons to our
data, copper, i ron, manganese, and zi nc l osses
are def mi tel y l ower, bei ng about 0.31, 0.34,
0.004, and 1.0 mg, respecti vel y (5, 6, man-
ganese; E. V uori , unpubl i shed observati ons).
The concentrati on of copper, i ron, and zi nc
i n human mi l k i n our study are i n agreement
wi th earl i er i nternati onal data, but the man-
ganese concentrati on i s apparentl y l ower (27).
Thus there i s probabl y no need to suppl ement
maternal di et wi th trace el ements, even
though the i ntake of zi nc and copper i s some-
what l ower than the recommended val ues.! ]
Ref erences
1. UNDERW OOD, E. J. Trace El ements i n Human and
A ni mal Nutri ti on (3rd ed). New Y ork: A cademi c
Press, 1971, pp. 25-26.
2 . UNDERW OOD, E. J. Trace El ements i n Human and
A ni mal Nutri ti on (3rd ed). New Y ork: A cademi c
Press, 1971, pp. 73-74.
3. UNDERW OOD, E. J. Trace El ements i n Human and
A ni mal Nutri ti on (3rd ed). New Y ork: A cademi c
Press, 1971, pp. 1 8 2 - 1 8 3 .
4. UNDERW OOD, E. J. Trace El ements i n Human and
A ni mal Nutri ti on (3rd ed). New Y ork: A cademi c
Press, 1971, p. 217.
5. SuM Es, M . A ., E. V u0RI AND P. KUI T UNEN. Breast
mi l k i ron-a decl i ni ng concentrati on duri ng the
course of l actati on. A cta Paedi at. Scand. 68: 29,
1979.
6. V uoRl , E., AND P. K UI TUNEN. The concentrati ons of
copper and zi nc i n human mi l k-a l ongi tudi nal
study. A cta Paedi at. Scand. 68: 33, 1979.
7 . K ARM ARK AR, M . G., AND C. V . RA M A K RI SHNA N.
Studies on human lactation. Relation between the
di etary i ntake of l actati ng women and the chemi cal
composi ti on of mi l k wi th regard to pri nci pal and
certai n i norgani c consti tuents. A cta Paedi at. Scand.
49: 599, 1960.
8. ARCH I BAL D, J. G. Trace el ements i n mi l k: a revi ew-
part I . Dai ry Sci . A bstr. 20: 712, 1 9 5 8 .
9. ARCH I BAL D, J. G. Trace el ements i n mi l k: a revi ew-
part I I . Dai ry Sci . A bstr. 20: 799, 1958.
10. FI L ER, L . J., JR. M ater nal nutr ition in lactation. Cl i n.
Peri natol . 2 : 3 5 3 , 1 9 7 5 .
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