Effects of feeding ultrahigh-temperature (UHT)-treated infant
formula with different protein concentrations or powdered formula,
as compared with breast-feeding, on plasma amino acids,
hematology, and trace element status1–3
Bo Lönnerdal and Olle Hernell
ABSTRACT The appropriate amount of protein to use in
infant formula is still under discussion. We found earlier that
protein digestibility is higher from ultrahigh-temperature
(UHT)-treated formula than from conventionally heat-treated
formula. In this study, we evaluated the nutritional, hematologic,
and biochemical effects of feeding infants whey-predominant
UHT-treated formula with 13 (UHT-13) or 15 (UHT-15) g pro-
tein/L as compared with a conventional, powdered, whey-pre-
dominant formula (PF) with 13 g protein/L from 6 wk to 6 mo of
age. Breast-fed infants served as control subjects. Growth was
assessed at monthly intervals and venous blood samples were
drawn at entry into the study and at 6 mo of age. At 6 mo, there
were no significant differences in weight gain or linear growth,
or hemoglobin, serum ferritin, zinc, or copper concentrations
among the groups. Blood urea nitrogen concentrations were low-
est for breast-fed infants; among the formula-fed groups the
UHT-13 group had the lowest values. All formula-fed groups had
higher plasma threonine concentrations than breast-fed infants.
Infants fed the UHT-13 formula had threonine values closest to
those of breast-fed infants. Concentrations of branched-chain
amino acids were similar in breast-fed infants and those fed
UHT-13 formula, whereas the other groups had higher values.
Plasma tryptophan concentrations were significantly higher in
the UHT-treated formula groups than in the other groups. Thus,
infants fed UHT-13 formula had metabolic measures similar to
those of breast-fed infants, possibly because of high protein
digestibility, or a difference in the protein source used. Iron,
zinc, and copper status was satisfactory in all groups. Selenium
status, as indicated by serum glutathione peroxidase activity,
varied with dietary selenium intake. Am J Clin Nutr
1998;68:350–6.
KEY WORDS Breast-feeding, infant formula, protein, trace
elements, amino acids, UHT, ultrahigh-temperature treatment,
powdered formula
INTRODUCTION
The protein concentration of mature breast milk is only
<8–9 g/L, whereas it is <10–13 g/L in early lactation (1–3). To
compensate for perceived lower digestibility of cow milk protein
and to achieve an adequate supply of all amino acids, most infant
formulas to date have had a protein concentration of 15–16 g/L
350 Am J Clin Nutr 1998;68:350–6. Printed in USA. © 1998 American Society for Clinical Nutrition
or higher. However, several studies have shown that plasma
amino acid patterns and blood urea nitrogen (BUN) concentra-
tions in infants fed whey-predominant milk formula with <13 g
protein/L are very similar to those of breast-fed infants (4–7).
Powdered formulas were used in all of these studies.
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During processing, infant formulas are heat treated to main-
tain the bacteriologic quality of the product. Traditionally, this
has been either retort sterilization for liquid products or high-
temperature, short-time (HTST) treatment before spray-drying
for powdered formulas. Ultrahigh-temperature (UHT)-treated
formula has become more commonly used in recent years. The
brief (2–3 s) treatment at 142 8C causes little protein denatura-
tion but ensures sterility of the product (as routinely tested by the
manufacturer). Compared with conventional retort sterilization
(118 8C for 10–15 min), this mild heat treatment has less harm-
ful effects on the digestibility of the protein. Our previous in
vitro studies (8) as well as in vivo studies in suckling rat pups (9)
showed that the protein digestibility of UHT-treated formula is
higher than that of powdered and retort-sterilized liquid formu-
las. We were therefore interested in evaluating the effect of UHT-
treated formula on plasma amino acids and BUN concentrations
in infants and comparing a lower protein concentration in such a
product with a powdered product with the same protein concen-
tration that had been spray-dried (< 60 8C). In Sweden and Fin-
land there are no liquid retort-sterilized milk formulas on the
market.
The amount of protein in infant formula has been shown to
affect the zinc status of infants; higher amounts of protein led to
lower serum zinc concentrations (10). It is likely that the low
serum zinc concentrations are caused by high plasma histidine
concentrations (10); a high protein intake results in high uptake
of histidine, which has been shown to result in zinc deficiency in
human adults through high urinary zinc losses (11). Free amino
acids, such as cysteine, have been shown to affect iron absorp-
tion, possibly by a chelation effect (12). Iron absorption has also
1
From the Department of Nutrition, University of California, Davis, and
the Department of Pediatrics, Umeå University, Umeå, Sweden.
2
Supported by a grant from Valio Ltd, Helsinki.
3
Address reprint requests to O Hernell, Department of Pediatrics,
Umeå University, S-901 85 Umeå, Sweden. E-mail: Olle.Hernell@pediatri.umu.se.
Received August 4, 1997.
Accepted for publication January 15, 1998.
 EFFECTS OF UHT-TREATED AND POWDERED INFANT FORMULAS 351

been shown to be affected by bovine milk proteins (13). It is pos- TABLE 1

sible that heat treatment may influence the release of free amino Composition of study formulas1
acids as well as the inhibitory effect of the milk proteins. It UHT-13 UHT-15 PF-4 PF-7
therefore appeared prudent to evaluate iron and zinc status of the
Protein (g/L)2 13 15 13 13
infants fed these products with different amounts of protein. In
True protein (g/L)3 13.1 15.3 11.7 11.7
addition, because the 2 UHT-treated formulas of interest were Fat (g/L) 35 35 35 35
manufactured in Finland, which is known as a country with a low Carbohydrate (g/L) 73 71 74 74
selenium intake (14), we were interested in assessing the sele- Calcium (mg/L) 350 350 460 460
nium status of these infants and comparing it with that of breast- Iron (mg/L) 7 7 4 7
fed infants. Zinc (mg/L) 3.5 3.5 4 4
Copper (mg/L) 0.1 0.1 0.4 0.4
Selenium (mg/L) 7 8 5 5
SUBJECTS AND METHODS 1
UHT-13 and UHT-15, ultrahigh-temperature-treated formula with
13 and 15 g protein/L, respectively; PF-4 and PF-7, powdered formula with
Subjects 13 g protein/L and 4 and 7 mg Fe/L, respectively.
2
Healthy, full-term infants were recruited from 3 well-baby Label claim.
clinics in Umeå, Sweden. The protocol for the study was
3
Total nitrogen2nonprotein nitrogen 3 6.25.
approved by the Ethics Committee on Research Involving
Human Subjects of the Medical Faculty, at Umeå University. groups were published earlier (15). The amino acid compositions
All infants were initially breast-fed and then allocated to of the formulas are given in Table 2.
breast-fed (BF) or formula-fed groups according to the mothers’
Blood urea nitrogen and plasma amino acids

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choice. To be recruited to one of the formula groups the infant
had to be completely weaned from the breast at 4 ± 2 wk of age.
Anthropometric measurements were taken monthly and a venous
blood sample (2–3 h postprandially, as observed by the research
nurse when collecting the sample in each infant’s home) at the
start of the study and at 6 mo of age when the study was discon-
tinued. Length was measured to the nearest 0.5 cm with use of a
Harpenden measuring device. Weight was measured to the near-
est 1 g by using an electronic balance. All recruited infants com-
pleted the study. Hematologic indexes were analyzed immedi-
ately. Plasma was separated within a few hours and stored frozen
at 220 8C until analyzed. Red blood cells were washed with
saline and frozen at 220 8C until analyzed.
Diets
Infants were either exclusively breast-fed or fed milk-based
(whey predominant, 60:40) powdered infant formula from 6 ± 2
wk of age until the end of the study at 6 mo of age. Three refer-
ence groups, 1 that was breast-fed (BF group) and 2 groups fed
powdered formula with 2 different amounts of iron (PF groups)
were part of a previous study performed just before this study
and in the same population (15). The powdered formula was
diluted with tap water according to the manufacturer’s instruc-
tions. No iron drops or solid foods were allowed. However, lim-
ited quantities of fruit purées (without iron) were allowed at
4–6 mo of age. These were provided by the investigators and
chosen to minimize interference with trace element status.
Infants were assigned to the 2 UHT-treated formula groups in a
randomized, double-blind design. Each group had ≥ 10 infants.
The 2 UHT-treated formulas, with 13 (UHT-13) and 15 (UHT-
15) g protein/L, respectively, were manufactured by Valio Ltd
(Helsinki), whey-adjusted with electrodialyzed whey, and pro-
vided in 200-mL, gas-impermeable packages. The composition
of the formulas is given in Table 1. The powdered formula con-
tained 13 g protein/L, whey-adjusted by ultrafiltered whey, and
was manufactured by Semper AB, Stockholm. Both of the UHT-
treated formulas contained 7 mg Fe as FeSO 4 per liter; the pow-
dered formula was manufactured with 2 different amounts of
iron and contained 4 (PF-4) or 7 (PF-7) mg Fe as FeSO 4 per liter.
The detailed hematologic and trace element data for the latter 2
Blood urea nitrogen (BUN) was determined by using a com-
mercial kit with urease (Sigma, St Louis). Proteins were sepa-
rated from plasma by sulfosalicylic acid precipitation (6%), and
free amino acids, including tryptophan, were analyzed in the
supernate with a Beckman 6000 amino acid analyzer (Beckman
Instruments Inc, Palo Alto, CA).
Hematologic and biochemical indexes
Hemoglobin was analyzed by the cyanomethemoglobin method,
mean corpuscular volume by Coulter counter (model STKR; Coul-
ter Electronics, Inc, Miami), serum iron and transferrin saturation
by commercial kits (Boehringer Mannheim, Indianapolis), serum
ferritin by radioimmunoassay (Diagnostic Products Corporation,
San Diego), and serum transferrin receptor by enzyme-linked
immunosorbent assay (R&D Systems, Minneapolis). Serum zinc
and copper concentrations were determined by atomic absorption
spectrophotometry (16). Serum glutathione peroxidase activity was
assayed by the method of Paglia and Valentine (17).
Statistical analyses
Statistical analyses were performed by using analysis of vari-
ance (ANOVA) with SAS (release 6.12; SAS Institute, Inc, Cary,
NC). Data were tested for residuals and no skewness was found.
When the ANOVA indicated significant group differences
(P < 0.05), multiple comparisons were performed by using
Tukey’s method to identify which groups differed (P < 0.05).
Values in tables and figures are given as means ± SDs. With the
number of subjects per group and the number of groups used in
this study we were able to detect weight differences > 7.4 g/d and
height differences > 0.20 mm/d with 80% power.
RESULTS
There were no significant differences in weight or length at
birth or at 6 wk (data not shown) or 6 mo of age among the
groups (Table 3), nor were there any significant differences in
weight or length gain during the study period. There were no
significant differences in any of the hematologic or biochemical
variables at the start of the study (6 ± 2 wk).
352 LÖNNERDAL AND HERNELL

TABLE 2 BF infants. Histidine concentrations were significantly lower in

Amino acid composition of study formulas1 infants fed either of the UHT-treated formulas than in infants in
Amino acid UHT-13 UHT-15 PF the PF group. There were no significant differences in leucine or
arginine concentrations among groups.
mmol/L
Plasma threonine was significantly higher in infants in the PF
Asparagine 9.69 11.19 8.88 group than in BF infants, whereas infants fed UHT-treated for-
Threonine 6.46 7.47 5.79 mula had intermediate concentrations (Figure 2). Glycine and
Serine 7.04 8.18 5.34 aspartic acid or asparagine concentrations were higher in infants
Glutamine or 20.05 23.04 15.99
fed UHT-13 than in BF infants, whereas glutamic acid or gluta-
glutamic acid
Proline 8.77 10.16 8.50
mine concentrations were higher in infants in either of the UHT-
Glycine 3.73 4.26 3.33 treated formula groups than in the PF group (Figure 3). There
Alanine 6.29 7.30 5.74 were no significant differences in concentrations of serine,
Cysteine 1.73 1.98 1.83 glycine, proline, lysine, taurine, or alanine.
Valine 7.08 8.19 6.85 There were no significant differences in hemoglobin concen-
Methionine 2.21 2.55 2.09 trations between the groups at 6 mo of age (Table 4). Serum fer-
Isoleucine 5.64 6.56 5.76 ritin concentrations varied considerably within groups, but the
Leucine 10.29 11.89 9.82 mean value was significantly higher for infants in the UHT-13
Tyrosine 2.81 3.26 2.53 than in the UHT-15 group. There was no infant in any group with
Phenylalanine 3.57 4.12 2.83
a serum ferritin value < 12 mg/L, which would indicate depleted
Histidine 2.26 2.58 1.70
Lysine 8.41 9.71 7.04
iron stores. Serum transferrin receptor concentrations in the BF
Arginine 2.30 2.64 1.89 group were significantly higher than those in the UHT-13 group;

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Tryptophan 1.08 1.22 1.01 the UHT-15 group had concentrations significantly higher than
Taurine 0.40 0.40 0.22 those in the PF-7 group. Serum zinc and copper concentrations
1
UHT-13 and UHT-15, ultrahigh-temperature-treated formula with 13 were not significantly different between groups (Table 4). Serum
and 15 g protein/L, respectively; PF, powdered formula. glutathione peroxidase activity was significantly higher in BF
infants than in all formula-fed groups.

At 6 mo of age, mean BUN concentrations in infants in the

UHT-15 and PF groups (13 g protein/L) were significantly
higher than those in breast-fed infants (Figure 1). Among the
formula-fed groups, infants fed UHT-13 had the lowest mean
BUN concentration, which was significantly lower than that in
the UHT-15 and PF groups, but not significantly different from
that in the BF group.
Concentrations of the branched-chain amino acids valine and
isoleucine were significantly higher in infants fed UHT-15 than
in BF infants (Figure 2). Infants fed UHT-13 had protein con-
centrations closest to those of BF infants. Tryptophan concentra-
tions were significantly higher in both of the UHT-treated for-
mula groups than in the BF and PF groups. Plasma tyrosine
concentrations were significantly higher in infants fed UHT-15
than in infants fed UHT-13. Infants in the UHT-15 and PF groups
had plasma phenylalanine concentrations significantly higher
than those in BF infants. Methionine concentrations were signi-
ficantly higher in both groups fed UHT-treated formula than in
DISCUSSION
The capacity of an infant formula to meet the protein require-
ment of infants can be assessed in several ways (18). A basic cri-
terion, of course, is that the infants consuming the formula grow
normally. For formula-fed infants, this is usually interpreted as
having a growth pattern commensurate with the National Center
for Health Statistics standards (NCHS; 19). It has been argued
that these standards were established largely on the Fels data,
which were obtained for infants fed formulas with higher energy
and protein contents than those currently used. One consequence
of this reasoning is that attempts are being made to establish sep-
arate growth standards for breast-fed infants (20). A revision of
the NCHS standards for formula-fed infants has not yet been
made, although Guo et al (21) published reference data on weight
and length of infants ≤ 2 y of age. In our study, all 3 groups of for-
mula-fed infants showed growth patterns that were not signifi-
cantly different from those of breast-fed infants and in agreement

TABLE 3
Weight and length of breast-fed (BF) infants and infants fed the various formulas at birth and 6 mo of age1
UHT-13 UHT-15 PF-4 PF-7 BF
(n = 6 M, 5 F) (n = 5 M, 6 F) (n = 4 M, 6 F) (n = 6 M, 4 F) (n = 5 M, 5 F)
Weight (g)
Birth 3737 ± 359 3403 ± 466 3263 ± 393 3634 ± 438 3623 ± 463
6 mo 8298 ± 1164 8001 ± 745 7527 ± 870 7790 ± 1178 7344 ± 934
Length (cm)
Birth 51.4 ± 1.9 51.3 ± 3.7 49.8 ± 1.6 50.8 ± 2.1 50.5 ± 2.0
6 mo 69.2 ± 3.3 67.3 ± 2.4 66.2 ± 1.6 66.9 ± 3.5 65.9 ± 1.9
Weight gain (g/d) 25.3 ± 5.2 25.5 ± 2.6 23.7 ± 4.1 23.1 ± 5.0 20.7 ± 4.4
Length gain (mm/d) 0.99 ± 0.12 0.87 ± 0.20 0.91 ± 0.11 0.89 ± 0.16 0.86 ± 0.11
1–
x ± SD. UHT-13 and UHT-15, ultrahigh-temperature-treated formula with 13 and 15 g protein/L, respectively; PF-4 and PF-7, powdered formula with
13 g protein/L and 4 and 7 mg Fe/L, respectively.
 EFFECTS OF UHT-TREATED AND POWDERED INFANT FORMULAS 353

FIGURE 1. Mean (± SD) blood urea nitrogen concentrations of infants
fed different formulas and breast-fed (BF) infants at 6 mo of age. Number
of infants per group: BF = 10; powdered formula with 7 mg Fe/L and 13
g protein/L (PF-7) = 10; ultrahigh-temperature-treated formula with 13 g
protein/L and 7 mg Fe/L (UHT-13) = 11; ultrahigh-temperature-treated
formula with 15 g protein/L and 7 mg Fe/L (UHT-15) = 11. Groups with
infants fed the UHT-13 formula had plasma amino acid concen-
trations that were similar to those of BF infants, whereas several
amino acids in infants fed the UHT-15 formula were higher than
in BF infants. This was particularly notable for the branched-
chain amino acids (valine and isoleucine), but also for tyrosine
and phenylalanine. It has been suggested that high concentra-
tions of branched-chain amino acids may be responsible for the
elevated concentrations of C-peptide found in the urine of infants
fed formula with high protein concentrations (24). This, in turn,
may be indicative of elevated serum insulin concentrations, the
long-term effects of which are not yet known. In any case, it
appears that a protein concentration of 15 g/L is unnecessarily
high, at least for infants > 6 wk of age. In one of our previous
studies we showed that formula with a protein concentration of
15 g/L resulted in significantly higher BUN and several plasma
amino acid concentrations in infants than did a formula with 13
g/L or breast milk at 2, 4, 8, and 12 wk of age (6). This finding
agrees with those of several other studies (4–7).
A concern with reducing the protein concentration of whey-
predominant formula has been that the amino acid tryptophan
may become limiting. Some studies have shown significantly

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different superscript letters are significantly different, P < 0.05.
with NCHS standards and the reference data by Guo et al (21).
Fomon et al (22) suggested that a safe protein-energy ratio for
milk formula is 1.44 g/100 kcal (418 kJ) from 56 to 83 d of age
and 1.25 g/100 kcal from 84 to 111 d of age. The lowest protein
concentration used in our study was 4.64 mg/kJ (1.94 g/100 kcal),
which is considerably above these minimum amounts.
Another way to assess the adequacy of a formula to supply
amino acids to growing infants is to analyze plasma amino acid
concentrations in a fasting state, usually 2–4 h postfeeding (23).
It is generally assumed that plasma amino acid concentrations of
formula-fed infants should be similar to those of breast-fed
infants and that significantly lower or higher amounts may be
considered as possibly inadequate or excessive. In our study,
lower plasma tryptophan concentrations in infants fed formulas
with 13 g protein/L than in breast-fed infants (4, 5, 7). It should
be cautioned, however, that several of these formulas had a crude
protein concentration of 13 g/L, but the true protein concentra-
tion was (or was likely to have been) considerably lower because
of a high nonprotein nitrogen content in the whey protein
sources used in the formulas (25). The UHT-treated formulas
used in this study had a low nonprotein nitrogen content (10%)
and, consequently, the true protein concentration was closer to
the crude protein concentration of 13 g/L. This amount of protein
resulted in plasma tryptophan concentrations that were not lower
than those of the BF infants. In fact, plasma concentrations of all
amino acids in the infants fed the formula with 13 g/L were sim-
ilar to or higher than those of the BF infants. We therefore do not
believe there is any reason to fortify this type of formula with
tryptophan, as was done in some studies (7, 26, 27).

FIGURE 2. Mean (± SD) plasma concentrations of essential amino acids in infants fed different formulas and breast-fed (BF) infants at 6 mo of
age. Number of infants per group: BF = 10; powdered formula with 7 mg Fe/L and 13 g protein/L (PF-7) = 10; ultrahigh-temperature-treated formula
with 13 g protein/L and 7 mg Fe/L (UHT-13) = 11; ultrahigh-temperature-treated formula with 15 g protein/L and 7 mg Fe/L (UHT-15) = 11. a Signi-
ficantly higher than the BF group, b significantly different from the PF group, and c significantly higher than the BF and PF groups, P < 0.05.
354 LÖNNERDAL AND HERNELL

FIGURE 3. Mean (± SD) plasma concentrations of nonessential amino acids in infants fed different formulas and breast-fed (BF) infants at 6 mo
of age. Number of infants per group: BF = 10; powdered formula with 7 mg Fe/L and 13 g protein/L (PF-7) = 10; ultrahigh-temperature-treated for-
mula with 13 g protein/L and 7 mg Fe/L (UHT-13) = 11; ultrahigh-temperature-treated formula with 15 g protein/L and 7 mg Fe/L (UHT-15) = 11. a

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Significantly higher than the BF group, b significantly different from the PF group, c significantly higher than the BF and PF groups, d significantly
higher than the UHT-13 group, P <0.05.

Some concern has also been raised that elevated plasma threo-
nine concentrations in infants fed whey-predominant formula may
reach excessive concentrations that may affect transport of other
amino acids through the blood-brain barrier (28). In our study, the
threonine concentrations of the infants fed the UHT-13 formula
were not significantly different from those of BF infants whereas
infants fed UHT-15 had higher concentrations of threonine. It does
not seem likely that this plasma threonine concentration would
have any negative effects on these infants; however, the lower
amount of protein would make this even less likely.
BUN concentrations have been used as a crude indicator of
protein excess (29). When infants receive protein intakes that are
higher than their metabolic needs, serum urea concentrations
rise. It has been shown in several studies that infants fed formu-
las with high concentrations of protein have BUN concentrations
that are considerably higher than those of breast-fed infants (10,
30, 31). In our study, infants in the PF and UHT-15 groups had
significantly higher BUN concentrations than BF infants. Infants
fed UHT-13 formula had values closest to those of the BF
infants. Note, however, that the differences in BUN were small
and there were no significant differences between the 3 formula-
fed groups. In fact, all formula-fed infants in our study had BUN
values considerably lower than those reported in infants fed for-
mula with high amounts of protein. Infants in the PF group had
somewhat higher concentrations of BUN and some amino acids
than those in the UHT-13 group, even if the 2 formulas had sim-
ilar amounts of protein. It is possible that the different heat treat-
ments that these 2 formulas received affected protein digestibil-
ity and amino acid absorption differently; alternatively, the 2
different whey protein sources used may explain the difference.
The powdered formula was made of ultrafiltered whey, which
has considerably higher nonprotein nitrogen and urea concentra-
tions than the electrodialyzed whey, which was used in the UHT-
treated formulas. This higher urea concentration may explain the
higher BUN concentrations of infants in the PF group. It is not
likely that the formula-fed groups differed in relevant character-
istics (although this was not a strictly randomized design)
because the characteristics of the 3 groups were not different at
the start of the study.
Overall, the major difference in indexes of protein metabolism
was between the UHT-15 and BF groups, whereas the differ-
ences between the UHT-13 and PF groups and the BF group
must be considered minor. There were no significant differences
in hemoglobin concentrations among groups. There were, how-
ever, several infants in each group that had hemoglobin values
< 107 g/L, a suggested cutoff for anemia in this age group (32),
but none of them had low serum ferritin concentrations or were
iron deficient as judged by multiple criteria. Except for infection,
anemia due to causes other than iron deficiency is highly
unlikely in this population. Limited data from infants in the third
National Health and Examination Survey suggest that hemoglo-
bin values < 105–107 g/L may be physiologic at < 12 mo of age
(P Dallman, personal communication, 1996) and not necessarily
indicative of iron deficiency. A recent study from the United
Kingdom also found a relatively large percentage (23%) of 8-
mo-old infants to be considered anemic (33). On the basis of data
from a large number of infants (n = 1175), these authors sug-
gested that a more appropriate cutoff at this age would be 97 g/L.
It is likely that new cutoff values for hemoglobin will be estab-
lished for infants < 1 y of age. Serum ferritin concentrations were
satisfactory in all groups and no infant had a value < 12 mg/L,
which would indicate depleted iron stores.
We also used serum transferrin receptor concentration as an
index of iron status. It has been shown that circulating concen-
trations of transferrin receptor are correlated with cellular iron
needs in adults and that concentrations are elevated in situations
of iron depletion (34, 35). This index was used previously to
diagnose iron deficiency in adult humans (36) and has several
advantages, such as not being affected by infection. In our study,
breast-fed infants had the highest concentration of serum trans-
ferrin receptor at 6 mo of age. It is known that exclusively breast-
fed infants usually have satisfactory iron status at 6 mo of age
 EFFECTS OF UHT-TREATED AND POWDERED INFANT FORMULAS 355

TABLE 4
Hematologic and biochemical indexes of study groups at 6 mo of age1
UHT-13 UHT-15 PF-4 PF-7 BF
(n = 6 M, 5 F) (n = 5 M, 6 F) (n = 4 M, 6 F) (n = 6 M, 4 F) (n = 5 M, 5 F)
Hemoglobin (g/L) 110 ± 5 111 ± 7 111 ± 8 108 ± 6 109 ± 6
Serum iron (mmol/L) 9.0 ± 3.9 10.3 ± 3.4 14.0 ± 3.8 12.8 ± 7.1 13.0 ± 9.4
Serum ferritin (mg/L) 74 ± 39a 44 ± 16b 46 ± 18a,b 60 ± 34a,b 69 ± 47 a,b
Serum transferrin receptor (mg/L) 2.0 ± 0.5a 2.9 ± 0.5b 2.6 ± 0.4a,b 2.1 ± 0.4a 3.0 ± 0.9b
Serum zinc (mg/L) 0.64 ± 0.14 0.65 ± 0.13 0.88 ± 0.15 1.01 ± 0.06 0.94 ± 0.13
Serum copper (mg/L) 0.75 ± 0.20 0.86 ± 0.10 1.01 ± 0.15 0.96 ± 0.05 0.88 ± 0.19
Glutathione peroxidase activity 16 ± 4a 18 ± 5a 22 ± 3a 21 ± 8a 32 ± 6b
(nmol NADPH · min21 · mL21)
1 –
x ± SD. Means with different superscript letters are significantly different, P <0.05. UHT-13 and UHT-15, ultrahigh-temperature-treated formula with
13 and 15 g protein/L, respectively; PF-4 and PF-7, powdered formula with 13 g protein/L and 4 and 7 mg Fe/L, respectively; BF, breast-fed.

(37, 38) but that they are at risk of iron deficiency at 9 mo of age
(37). Thus, it is possible that the elevated concentrations of
serum transferrin receptor are an early indication of increased
cellular needs for iron at this age, although there are still no ref-
erence values for infants. The lowest concentrations of serum
trations in the group fed the UHT-treated formula with 13 g pro-
tein/L suggest that iron was well utilized from this formula.
We are most grateful to Margareta Henriksson for her enthusiastic man-
agement of the infants and the blood sampling, and to the participating par-
ents and infants. We also appreciate the statistical expertise of Janet M Peer-

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transferrin receptor were found in the group with the highest iron
fortification (7 mg/L) and in infants fed the UHT-treated formula
with 13 g protein/L and 7 mg Fe/L. This indicates that iron was
well utilized from the UHT-treated formula with the lower
amount of protein. Hurrell et al (13) showed that milk proteins
can have a negative effect on iron absorption in adults and it is
possible that phosphopeptides or other components are responsi-
ble for the inhibitory effect.
There were no significant differences in serum zinc or copper
concentrations among groups. In an earlier study, we found lower
serum zinc concentrations in infants fed follow-up formula with a
high protein concentration (10). We speculated that low zinc status
was induced by increased urinary losses of zinc caused by high
concentrations of serum histidine, a known chelator of zinc. In the
present study, however, protein concentrations were much lower
than in the previous study and we did not find any increases in
serum histidine concentrations. In our study, all groups had similar
serum copper concentrations. We found differences in the activity
of serum glutathione peroxidase between the BF and the formula-
fed groups, most likely reflecting their different selenium intakes.
It is known that dietary intake of selenium is well correlated with
serum glutathione peroxidase activity (39). The selenium concen-
tration of breast milk from Swedish women is <15 mg/L (40),
which is higher than that of unfortified powdered formula (5 mg/L).
This is reflected by a significantly higher serum glutathione perox-
idase activity (15). When the formula was fortified with selenium
to a total amount similar to that of breast milk, glutathione peroxi-
dase activity also became similar to that of breast-fed infants. In
this study, the UHT-13 and UHT-15 formulas were not fortified
with selenium and contained 7 and 8 mg/L, respectively. The activ-
ity of glutathione peroxidase in these 2 groups was similar to that
of infants fed the unfortified powdered formula.
In conclusion, it appears that feeding UHT-treated formula or
powdered formula with a protein concentration of 13 g/L results
in satisfactory growth, hematology, trace element status, and
plasma amino acid patterns as compared with breast-fed infants.
Concentrations of several amino acids and BUN in infants fed
this amount of protein are closer to those of breast-fed infants
than are corresponding values from infants fed formula with 15
g protein/L. In addition, low serum transferrin receptor concen-
son and the technical assistance of Shannon Kelleher and Lalith
Jayawickrama.
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