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Soy protein,

Richard Sandra ABSTRACT


tein human trinsic isolates subjects. radioiron on

phytate,

and iron absorption


B Reddy, Sean

in humans13
R Lynch,

F Hurrell, A Dassenko, The


Iron label

Marcel-A Juillerat, Manju and James D Cook effect of reducing


absorption was liquid-formula

the

phytate
was by

in soy-proexamined using in an

Materials
32 Preparation cxhy- Eleven

and

methods
isolates

nonheme-iron absorption in corn isolates

measured meals

ofsoybean-protein

different soybean-protein isolates were prepared from containing different batches of soy flour (Table ). Isolates 1 I-IV were egg white or one of three a isolates containing much of their native amount of phytate contents. Iron standard phytic acid (4.9-8.4 mg/g of isolate). Isolates V-VII were lowabsorption increased fourto fivefold when phytic acid was rephytate isolates (0.2-1 .0 mg phytic acid/g) in which the phytic duced from its native amount of 49-8.4 < to 0.01 mg/g of isoreduced by continuous acid-salt washing and ullate. Even relatively small quantities of residual phytate wereacid had been trafiltration. In isolate VIII the phytic acid was reduced < 0.01 to strongly inhibitory and phytic acid had to be reduced < 0.3 to mg/g by enzyme treatment. Isolate IX (< 0.01 mg phytic acid/ mg/g of isolate (corresponding <to 10 mg phytic acid/meal) enzyme treated and ultrafiltered. Finally, in isolates before a meaningful increase in iron absorption was observed. g) was both drolyzed corn starch, series ofsoy-protein oil, and either with different

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However, absorption
the egg inhibitory that from other these

even after removal ofvirtually from the soy-protein meal


white control. factor of factors products. It is concluded iron absorption to the

all the phytic acid, iron x and amount was still only half that of
that phytic in soy-protein poor acid is a major lates from isolates but

XI, by V and the

phytic adding VIII, batch

acid

was

restored

to approximately phytate Isolates and were isolates

its original

back sodium respectively. ofsoy flour

to the low phytate isoI and V were prepared fed in studyIsolates . 1

first

contribute

bioavailability

Am Soy-protein

J C/in

Nuir l992;56:573-8. isolate, phytate, iron

of ironII, VI, and X were made were fed in study 2. The third batch

from

the second

batch
were

ofsoy
made

flour
from 4.

and
the de-

remaining

KEY

WORDS

absorption

All
fatted water

of soy flour isolates ( 1-2 kg) soybean flour to water system that at native adjusted by

and were fed were prepared was 12000 first wt:wt)

in studies 3 and from commercial for 1 h in

soaked

deionized

(flour

ratio :7.5, 1

and

then

centrifuged
the

in
fibrous

Introduction
Soy protein in the United one-quarter is a major ingredient in infant formulas States where soy formulas now account of infant-formula sales1 ). The use of soy (

a continuous material. especially soybean for about which

X g at 37#{176}C remove to

For the milk was was recovered

phytate isolates (I-IV), the resulting to pH to precipitate 5.2 the protein, centrifugation, washed with deionized

is also
dairy-type and material

increasing
foods. for the excellent

in extended
Good functional development protein

meat
quality, of properties

products,
low make new cost,

baked
it an

protein goods,

water, neutralized and injection at 140


phytic acid raw contain 4.9-8.4 by in

with
#{176}C, and and mg

potassium
spray-dried. reason acid/g Those washing way is the

hydroxide,
The why our compared isolates and except

sterilized
washing native with in which ultrafiltration that after step phytate 9.0-1

by steam
removes isolates 7.0 mg/ phytate (V-VII)

plentiful attractive

supply, some foods

phytic

manufactured

One
an

potential
inhibitory

drawback
effect on

to the
iron

use of soy
in

protein
humans

is that
(3-7).

absorption

(2). in commercial g was removed it has Full-were prepared

isolates. acid-salt the same

the

precipitation

fat soy flour, textured soy flour, and isolated soy protein all protein the fraction was ultrafiltered in a two-step process at pH markedly reduce nonheme-iron absorption. The isolated protein 52 and pH 7 in the presence ofsodium chloride (9). has the greatest inhibitory effect (3). For isolates in which the phytate was removed by enzyme The nature ofthe substances in soybean products that inhibit treatment (VII and IX), the soybean milk was treated at pH 5.2
iron absorption to contain inhibitor is unclear. appreciable of iron However, quantities absorption absorption ofsoy-protein were meals in soy-protein ofphytate, in wheat products which bran (8). are is an From Nestec Ltd, Nestl#{233} Research Centre, Lausanne, Switzerland, Center, Kansas City, KS 66103. and AID Cooperative Agreement known important

The

present

study

was

designed

to define

the

role

the University of Kansas Medical of phytate and 2 Supported by NIH grant DK39246

in modifying nonheme-iron isolates in humans. A series fold variation in phytate absorption tein isolates radioisotopic Am J C/in

content

from soybean-protein DAN-S I S-A-00-7908-00. isolates with a 10003 Address reprint requests prepared. Nonheme-iron Centre, Vers-chez-les-Blanc, containing human these volunteers soy-pro- zerland. with a Received Accepted Society

to RF Hurrell, Nestec Ltd, Nestl#{233} Research PGBox 44, CH-l000 Lausanne 26, Swit-

from liquid-formula was then measured method. Nuir 1992:56:573-8. Printed

August 28, 1991. for publication March Nutrition

17, 1992. 573

in USA.

1992

American

for Clinical

574 TABLE
Analytical

HURRELL 1
data on soybean-protein isolates

ET

AL

Percent
Soy-isolate fractiont protein
%N
X

crude
Phytic mg/g acid Trypsin TIU/mg inhibitor N Iron mg/g Calcium mg/g

Inorganic
phosphorus mg/g

6.25

Native I IIi

phytate 96.4 87.7 8.4 7.2 4.9


0.41 0 1.1

5.3 5.3

0.145 0.175

0.278 0.563

0.1 1 0.21

IIIC
IVC

89.1
90.1 phytate

6.50.7

8.8
6.7

0.130
0.140

0.616
0.462

0.50
0.09

Acid-salt-reduced

V VP VIIC
Enzyme-reduced phytate

92.0 92.6
90.1

0.2 1.0
0.3 00l 0.01 99 3.7

0 0.1
0.01 0.01 0 02 0.2 without further

4.5 3.6
3.5 21.7 14.5 7.2 8.8 moisture

0.145 0.180
0.146 0.135 0.155 0.162 0.145 removal.

0.070 0.440
0.077 0.820 0.289 0.364 0.142

0.08 0.19
0.05 0.68 0.28 0.17 0.22

VIIIC
IXC Restored XL fl phytate

89.7
91.8 91.1 90.8 data the were same followed sodium sodium obtained superscript on the spray-dried were prepared

x1c11
S

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All analytical with

products

t Isolates

from

the same

batch

of soy flour.

t ; SE. Enzyme-reduced
II Produced #{182} Produced

by ultrafiltration. phylate phytate to isolate to isolate VI. VIII.

by adding by adding

with Finland)

phytase before

centrifuged, described
an additional

Helsinki, VII then was washed, neutralized, sterilized, and spray-dried above. For isolate IX the coagulum was subjected to
precipitation ofthe protein. Isolate ultrafiltration treatment to remove the

from Aspergillus

niger (Alko

Ltd.

lates,
and

and were

a control
meal corn

meal
were starch

containing egg white.


given (Fro-Dex, in studies

Three 2,

test 3, and Maize

meals 4. All test

a control

asmeals

fed as a semisynthetic IN), 36 g corn

liquid oil (Nugget

formula Brand,

containing
Products,

67

g hydrolyzed low-mo- Hammond,

American

by12 mL vanilla extract (McCormick V 200 mL deionized distilled water, and the enzyme-treated isolate VIII, respectively, before thex 6.25) derived from either a soy-protein isolate or egg white neutralization step. (Monarch Egg Corporation, Kansas City, MO). The calcium content of the soy-protein-isolate meals within each study was Analytical methods equilibrated by adding calcium chloride (CaCI2 2H2O) to raise the calcium content to 44 mg/meal in study 1, 19.2 mg in study Iron and calcium were determined by atomic-absorption 2, and 27.4 mg in studies 3 and 4. The amount of calcium (96 spectroscopy after dry ashing. Total nitrogen was determined mg/meal) in the egg white-control meal was not modified. by using an automatic nitrogen analyzer(Type NA Carlo1500; The test meals for studies 1-4 are described in Table 2. The Erba, Milan, Italy). Trypsin inhibitors were measured according test meals in study 1 included soy-protein isolate (I) containing to the method of Kakade et al (10) and trypsin inhibitor units its native phytic acid content and a low-phytate isolate produced (TIU) were expressed per milligram nitrogen.
adding sodium phytate back to the acid-salt-washed isolate
-

lecular-weight

compounds.

Isolates

and

XI

were

made

Stockton, CA), and Co. Baltimore,MD), and 30 g protein (nitrogen

Phytic

acid

was

measured

by

using

modification

of

by the

continuous

acid-salt

washing

and

ultrafiltration

(V).

Both

method of Makover (1 1) in which cerium replaced iron in theisolates were prepared from the same soy flour. The test meals precipitation step. Phytic acid was calculated from the phos- for the second study comprised a soy-protein isolate containing phorous content of the precipitate by using a factor 3.55. of its native amount of phytic acid (II), a low-phytate isolate proInorganic phosphorous was extracted from mg dried iso200 duced by continuous acid-salt washing and ultrafiltration (VI), late with 10 mL 1 mol H2SO4/L. Phosphorous was measured and the same isolate to which phytic acid had been added back immediately by using a microtitration plate assay based the on (X). Again, all isolates were prepared from the same soy flour. complex formation of malachite green with phosphomolybdate In the third study the test meals included a control soy-protein under acidic conditions (12). isolate containing its native amount ofphytic acid (III), an isolate from which the phytate had been removed by enzyme digestion Iron-absorption studies (VIII), and the same phytate-free isolate to which phytic acid back (XI). For study 4 the meals were a control Four iron-absorption studies were carried out in groups of 7- had been added isolate containing its native phytic acid (IV), a low-phytate isolate 9 human subjects. In study 1 the volunteers were fed two test produced by acid-salt washing and ultrafiltration (VII), and an meals, each containing one ofthe experimental soy-protein iso-

SOY

PROTEIN,

PHYTATE,

AND

IRON

ABSORPTION

575

TABLE 2 Effect of phytate

removal

on iron absorption

from

soy isolates
Absorpti on ratio vs meal D

Study, subjects, and mean age

Mean packed cell volume


%

Serum ferritin5 ig/L 59 (49-71) 38 (29-50) A B D A B C D A B C D A B C

Mealst

Iron

absorption5 % of dose

vs meal A

1, (6 M, 2 F), 24 y

44

2, (5 M, 4 F), 23 y

43

3, (7 M,

1 F), 23 y

45

68 (60-77)

4, (3 M, 4 F), 22 y

43

35 (28-45)

Isolate I (native phytate) Isolate V (A-S-reduced phytate) Egg white control Isolate II (native phytate) Isolate VI (A-S-reduced phytate) Isolate X (restored phytate) Egg white control Isolate III (native phytate) Isolate VIII (E-reduced phytate) Isolate XI (restored phytate) Egg white control Isolate IV (native phytate) Isolate VII (A-S-reduced phytate) Isolate IX (E-reduced phytate) D Egg white control

1.50 3.15 6.34 0.92 1.91 1.08 5.75 0.53 2.50 0.78 5.48 1.36 4.17 5.48 9.72

(1.10-2.06) (2.32-4.28) (4.72-8.5 )1 (0.65-1.32) (1.34-2.71) (0.75-1.54) (3.96-8.33) (0.41-0.68) (2.10-2.97) (0.52-1.15) (3.63-5.94) (0.94-1.98) (3.01-5.76) (4.16-7.21) (7.56- 12.5 )1

2.l0
-

0.24t 0.50
-

2.07 1.17
-

0.16* 0.33* 0.19*


-

4.75* 1.45
-

0.10* 0.46j 0.17*


-

3.06* 4.02
-

0.14* 0.4311 0.56

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Geometric

i (1

SE). phytate; E, enzyme-reduced phytate.

t A-S, acid-salt-reduced tP<0.0S P < 0.001.


IIP<

0.01.

isolate
treatment

from

which
followed

the
by

phytate
ultrafiltration

had
from differed

been
(IX).

removed
All

by enzyme second
isolates fed 3, and in

pair of meals was given on days 4. Only a single meal was fed on

16 and 1 7 in studies day 17 in study . The I

2,

studies 3 and 4 were produced As indicated above, isolate IX been The mean exhibited concentrations and denied the gastrointestinal was obtained tigation Human Center. Double absorption The meals sequential from were four radioiron separate water with administered and all Subjects subjected volunteer age of a wide 23 to an subjects y. range between a history from There ofiron 1 1 and additional

low-molecular-weight

compounds.
ranged were status 138

the same batch ofsoy flour. final blood sample was drawn 2 wk after the last test meal to from isolate VIII by having measure the increase in circulating red cell radioactivity. Raultrafiltration step to remove dioiron measurements were made on duplicate 10-mL samples of whole blood by a modification of the method of Eakins and in age from 20 to 3 1 y with Brown a ( I 5). Percentage absorption was calculated on the basis and All were are known 1 1 women. by serum in good to as reflected g/L. that They the blood of volume fen-itin and an assumed red health of8O% influence (18). estimated from cell incorporation height and for absorbed weight (16, radioactivity 17)

2 1 men

of disorders absorption ofiron. each volunteer

Percentage

absorption

values

were

converted

to

logarithms
to each antilostudy

Written, informed before beginning

consent before the inves- garithms

statistical analysis to recover the

and the results reconverted original units (19). Because

experimental Committee

procedures were approved at the University ofKansas labels meals between was the allowed extrinsic were consumed 0700 and used to measure by each 0900 after

by the contained several independent Medical paired t tests were used to test meals within the same iron mean subject. log absorption ratios

manipulations compare absorption


study differed by determining significantly

ofthe test meals, from selected


whether from zero. the

an

Results
3 (13) The results of the iron-absorption 2. In study of 1, subjects fed the the control soy isolate (I) with mg/g had a mean iron studies are shown in Table liquid-formula meal containing a native phytic acid content of of 1.5%, which increased

overnight fast and only h. All meals were labeled by adding 37 56FeCl3 in 0.01 to adjust 1, 6.4 On drawn (14), meals days the kBq mol total

for the subsequent label technique to a solution ofiron sufficient to 5.7 mg

59FeCl3 HC1/L iron

or 1 1 1 kBq 55FeCl3 containing a quantity ofeach meal

content

in study8.4

absorption

mg in study 2, and5.5 mg in studies 3 and 4. to 3. 1 5% (P = 0.01) when the phytic acid content of the soy the day preceding the first test meal,5 mL 1 blood was isolate was reduced to 0.2 mg/g by acid-salt washing and ultrafor the measurement ofpacked cell volume, serum femtin filtration (isolate V). In study there 2 was a similar twofold inand background radioactivity. The first and second test crease in iron absorption on feeding another low phytate isolate were 2 and labeled with either 59Fe or Fe 3 of the study. Blood (25 mL) incorporated red cell radioactivity. and was administered drawn on day on produced 16 turned acid was by to added acid-salt back. washing its In this and, original study, in addition, amount decreasing absorption when the the phytic rephytic acid approximately

to measure

A similarly

labeled

576

HURRELL
7.2 (isolate from II) 0.92% to to 1.0 mg/g
<

ET
abuJ

AL
0-a 0.7

from
sorption

(isolate 0.02).

VI) Adding

increased back

iron phytic

L9 1 % (P

acid to an amount of99 which was not significantly


>

mg/geduced r different produced


isolate

iron absorption to 1.08%, from its original value (P by enzyme


had a phytic

0.6
0.5

0.2).

A low
vestigated

phytate
in study

isolate

treatmentwas inacid content of

3. This

0.4

OOl mg/g, compared with 0.2-1.0 mg/g in the low-phytate 0.3 isolates produced by acid-salt washing and ultrafiltration (Table 0.2 1). In this study (Table 2), reducing the native phytic acid content 0.1 ofthe control isolate (III) from 6.5 to 0.01 mg/g (isolate VIII) increased iron absorption almost fivefold from 053% to 2.50% 0.0 0 2.0 4.0 6.0 8.0 10.0 (P < 0.00 1). Again, adding back phytic acid to 3.7 mg/g decreased PHYTIC ACID Cmg I g soy isolate I iron absorption to 078%, whichwas not significantly different from its original value (P > 0.2). FIG 2. Relationship between phytate content (mg/g) of soy-protein Study 4 compared directly a low-phytate isolate produced byisolate and mean iron absorption 1 SE ofeach soy-protein-isolate meal relative to its respective egg white control meal. (Data compiled from acid-salt washing (isolate VII) with a similar isolate produced studies 1-4 as listed in Table 2). Approximate phytic acid content (mg) by enzyme treatment (isolate IX). Both isolates were ultrafiltered by multiplying the phytic acid content (mgi to remove the low-molecular-weight compounds. Mean iron per meal can be calculated g) of the soy-protein isolates by 33. absorption from the control isolate (VI) containing 49 mg phytic
acid/g washing was 1.36%. increased Reducing absorption phytic acid to 4. 17% (P
<

to 0.3 0.001)

mg/g by acid-salt whereas re-

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to the egg white control. Thus, in Figure 2, iron absorpacid to 0.01 mg/g by enzyme treatment in- relative tion from meals containing the different soy-protein isolates relcreased absorption to 5.48% (P < 0.05). In this study there was the egg white-control meal fed in the same no significant difference in iron absorption between these twoative to that from subject (relative absorption 1.0) is compared with the phytic low-phytate isolates (P > 005). However, when all absorption the acid content ofthe isolates. At phytic acid contents between 9.9 ratios ofthe low-phytate isolates relative to their respective conand 3.7 mg/g, iron absorption relative to the egg white control trols were combined (Fig 1), it is seen that acid-salt washing to varied randomly from 0. 10 to 024. Only after produce isolates with 0.2-1.0 mg phytic acid/g increased iron was low and acid was reduced to 0.3 mg/g was there a substantial absorption 2.3-fold, whereas enzyme treatment to give isolates phytic in iron absorption to 0.43-0.56 ofthe egg white control. with 0.01 mg/g phytic acid produced a significantly greater increase Relative iron absorption can also be compared with ap-the 4.4-fold increase (P < 0.01). phytic acid content of the meal. Because the hydroThe mean iron absorption from the egg white-control meal proximate contained no measurable phytic acid, the soywas 6.34%, 5.75%, 548%, and 9.78%. respectively, in studies 1- lyzed corn starch isolates were the only phytic acid-containing components 4. As absorption from the egg white-control meal was measured protein of Each meal contained 30 g crude protein from the in all subjects, it is possible to compare the absorption from the meal.

ducing

phytic

different

meals

between

studies

by

comparing

their

absorption

test

isolate,

and,

because

the

protein

contents

of

the

isolates

50

differed slightly (Table 1), is equivalent to 33 g isolate per meal. The phytic acid content per meal can be obtained by multiplying the phytic acid content of the isolate (mg/g) by 33. It can be seen that a substantial increase in iron absorption occurred only after phytic acid was reduced <to 10 mg/meal.

20#{149} Discussion 10
S.

The

observation led may be effect an

that Widdowson

consumption and inhibitor

of wheat McCance of

bran (20) to nonheme

reduces suspect food

iron that iron

I
5
S

absorption phytate absorption. its inhibitory

important

Subsequent
but

investigations
other studies

with
have

bran
yielded

have

confirmed
contradictory

I..

#{149}1#{149}
1
.5
I

results as to the inhibitory nature of phytate study (21) the reduction of phytate in wheat to have no effect on nonheme-iron absorption
.
S

specifically. In one bran was reported and monoferric


bran (22), (8) was

phytate,

which

represents

half

the

iron

in wheat

reported ACID-SALT ENZYME


FIG 1. Iron-absorption ratios between reduced phytate isolates and their corresponding control isolate containingits native acid content. Acid-salt-reduced isolates contained 0.2-1.0 acid/g and enzyme-reduced isolates contained 0.01 mg phytic phytic adding

to be well

absorbed.

In other
did improve rolls inhibited

studies,

the

reduction

of
and doseas

soy-protein dependently phytic absorption mg phytic iron

acid in wheat bran phytic acid to wheat

iron absorption iron absorption

(23).

The
from

role
soy

of phytate
products was

in modifying
even more

nonhemeunclear

acid/g.

Short

horizontal

lines represent

mean

values.

neither (6) nor

the removal of phytate from soy a twofold variation in the phytate

flour by acid washing content of soybeans

SOY

PROTEIN,

PHYTATE,
(24) suggest isolates. influenced

AND
(28). made

IRON Our
from

ABSORPTION indicate
soy

577

produced nonheme-iron

under

different absorption. findings, inhibitory

growing however, factor

conditions

results formula.
still

that

iron

absorption
would be similar

from isolate,
absorption.

formulas
to that from

phytate-free moderately

isolate inhibitory

The present acid is a major

now strongly in soy-protein

that phytic milk-based are Removal whey

Phytate-free

soy-protein
to iron

casein,

and
The in-

hibitory nature of whey would appear to be due to its high calofphytic acid to 0.01 mg/g ofisolate increased iron absorption fourto fivefold whereas adding back the phytic acid reduced cium and phosphorous contents and not to its protein compoiron absorption to its original low value. Our results also dem- nent (29). The inhibitory nature of casein and phytate-free soy onstrate that relatively small amounts of phytic acid can still isolate on the other hand is due probably to the binding of iron strongly centration mally absorption. in low failed in soy a meal amounts to products to
<

inhibit in 0.3

iron isolates mg/lOO latter of


(8,

absorption must be figure acid a beneficial reduced g to ensure 5 mg Fe. could

and

that

the 1 .0 mg/g

phytic and increase for earlier

acid opti-

conto tides

insoluble from iron-binding

peptides casein are

in peptides

the

duodenum. that from contain soy could

The serine be

iron-binding phosphate those containing

pep(29).

to
<

those

a meaningful The effect necessity why

in iron The

The

corresponds

to10 mg phytic

acid
these phytic studies

a high
very

proportion

of carboxylic

acid

groups.

13

containing phytic
24).

explain

demonstrate

of reducing

acidReferences
1.

Beard

et al (24)

reduced

By modifying the phytic

the growing acid content


cooked phytic

conditions, of soybeans

from 7.04 to 3.76 mg/g. or pur#{233}e meals in providing phytate They not

They fed the -220 mg

beans acid in

as a soup 2. the high-

meal
showed increase

and
that iron

I 10 mg phytic
reducing absorption phytic

acid
relative

in the
acid to by

low-phytate
these amounts reference

meal.
did meal.

3.

their would the

Our
content on
<

results
of iron

would
a meal iron absorption

also
from but

suggest
220 that to

that
by

decreasing

the
have

phytic
little

acid 4.
effect acid S. to
6.

1 10 mg decreasing would

phytic

10 mg/meal,

absorption

be increased

substantially.

Enzyme
than isolates mg/g. the was

treatment
acid-salt

was more
washing

effective

at removing
with ultrafiltration, compared in addition

phytic

acid
giving

combined

7.

with 0.01 Acid washing acid,

mg phytic acid/g with ultrafiltration, a variety

with 0.2-1.0 to removing com8.

phytic

removes

oflow-molecular-weight

pounds,

which

could

also

influence

iron

absorption.

To

inves-

Witherly SA. Soy formulas are not hypoallergenic. Am I Clin Nutr 1990:1:70S-6. Erdman 1W, Fordyce El. Soy products and the human diet. Am I Clin Nutr 1989:49:725-37. Cook ID, Morck TA, Lynch SR. The inhibitory effect ofsoy products on nonheme iron absorption in man. Am I Clin Nutr 1981:34: 2622-9. Morck TA, Lynch SR, Skikne BS, Cook ID. Iron availability from infant food supplements. Am I Clin Nutr 1981:34:2630-4. Morck TA, Lynch SR. Cook ID. Reduction ofthe soy-induced inhibition of nonheme iron absorption. Am Clin Nutr 1982:36:219-28. I Hallberg L, Rossander L. Effect of soy protein on nonheme iron absorption in man. Am I Clin Nutr l982;36:5l4-20. Derman DP, Ballot D, Bothwell TH, et al. Factors influencing the absorption of iron from soya-bean protein products. Br I Nutr 1987;57:345-53. Hallberg L, Rossander L, Skanberg A-B. Phytates and the inhibitory effect ofbran on iron absorption in man. Am I Clin Nutr 1987;45:
988-96.

Downloaded from www.ajcn.org by guest on July 18, 2011

Dc Rham 0, Jost T. Phytate-protein interaction in soybean extracts and low phytate soy protein products. Food Sci 1979;44:596-600. I 10. Kakade ML, Rackis II, McGhee IE, Puski G. Determination of absorption. Absorption from the meal containing the enzymetrypsin inhibitor activity in soy products: A collaborative analysis reduced phytate isolate VIII (Table 2) was 2.50% compared with of an improved procedure. Cereal Chem 1974;5 1:376-82. 5.48% for the egg white-control meal. Absorption from soy iso- 1 1. Makover RU. Extraction and determination ofphytic acid in beans. late VIII relative to the egg white-control meal was thus 0.46. Cereal Chem 1970;47:288-9S. Absorption from the enzyme-reduced phytate isolate (Table IX 12. Van Veldhoven PP, Mannaerts GP. Inorganic and organic phosphate 2) subjected to an additional ultrafiltration was 5.48% compared measurements in the nanamolar range. Anal Biochem 1987;161:45-8. ID, Layrisse M, Martinez-Ton-es C, Monsen E, Finch CA. with 9.72% from the egg white-control meal in the same subjects. 13. Cook Food iron absorption measured by an extrinsic tag. I Clin Invest Absorption of soy isolate IX relative to the control meal was

tigate this possibility we a further acid-salt washing indicate that the

subjected an enzyme-treated with ultrafiltration. Our step did not further

isolate to results would 9. improve iron

ultrafiltration

0.56.

The

phytic studies

acid

content

of both different

isolates protein

was OOl sources

mg/g. incorpo14.

I 972:1:80-1S.

Miles LEM, Lipschitz DA, Bieber CP, Cook ID. Measurement of serum femtin by a 2-site immunoradiometric assay. Anal Biochem rated into the same liquid-formula meals as administered in the 1974;6 I:209-24. present investigation demonstrated that soy-protein isolate was 15. Bothwell TH, Charlton RW, Cook ID, Finch CA. Iron metabolism the most inhibitory ofthe protein sources tested. Iron absorption in man. Oxford: Blackwell Scientific, 1979. from a soy-protein-isolate mealwith its native phytic acid content 16. Wennesland R, Brown E, Hopper I, et al. Red cell, plasma and was 020 ofthe egg white-control meal (3) compared with 0.31 blood volume in healthy men measured by radiochromium (Cr) for wheat gluten, 0.40 for whey protein (25), 0.55 or 1 08 for cell tagging and hematocrit: influence ofage, somatotype and habits ofphysical activity on variance after regression ofvolumes to height casein (3, 25), 1 .90 for bovine serum albumin (26), 3.00 for beef and weight combined. I Clin Invest 19S9;38:106S-77. muscle (27), and 3.53 for the protein-free meal (26). In the pres17. Brown E, Hopper I, Hodges IL, et al. Red cells, plasma and blood ent study, removal ofvirtually all the phytic acid from soy-protein volume in healthy women measured by radiochromium cell-labelling isolates increased iron absorption from 0. 10-0.24 to 0.55 of and hematocrit. I Clin Invest 1962;4l:2182-90. the absorption from the egg white-control meal, indicating that 18. Hosein F, Marsaglia G, Finch CA. Blood ferrokinetics in normal even after the removal of phytate, soy protein itself is still relman. I Clin Invest 1967:46:1-9. atively inhibitory to iron absorption. 19. Cook ID, Layrisse M, Finch CA. The measurement ofiron absorpEarlier studies have shown that, at the same amount of ascorbic tion. Blood 1969:33:421-9. acid in the formula, iron absorption from milk-based infant for20. Widdowson EM, McCance RA. Iron exchange of adults on white mulas is two to three times higher than that from soy formulas and brown bread diets. Lancet 1942;l:588-9l. Earlier comparing

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RF, Lynch SR. Trinidad TP, Dassenko SA, Cook ID. Iron Simpson KM, Morris ER, Cook ID. The inhibitory effect of bran 26. Hun-elI on iron absorption in man. Am I Clin Nutr 198 l;34:l469-78. absorption in humans: bovine serum albumin compared with beef 22. Morris ER, Ellis R. Isolation ofmonoferric phytate from wheat bran muscle and egg white. Am I CIin Nutr l988;47:l02-7. and its biological value as an iron source in rats. J Nutr l976;l06: 27. Cook JD, Monsen ER. Food iron absorption in human subjects. III 753-60. Comparison of the effect of animal proteins on nonheme iron absorption. Am I Clin Nutr 1976:29:859-67. 23. Hallberg L, Brune M, Rossander L. Iron absorption in man: ascorbic acid and dose dependent inhibition by phytate. Am I Gin Nutr 28. Gillooly M, Torrance ID, Bothwell TH, et al. The relation effect of ascorbic acid on iron absorption from soy-based and milk-based 1989;49: 140-4. infant formulas. Am I Clin Nutr l984;40:522-7. 24. Beard IL, Weaver CM, Lynch S, Johnson CD, Dassenko S, Cook RF, Berrocal R, Lynch SR. Dassenko SA, Cook ID. The ID. The effect of soybean phosphate and phytate content on iron 29. Hun-elI bioavailability. Nutr Res l988;8:345-52. influence ofbovine milk proteins on iron absorption in man. In: 25. Hun-eli RF, Lynch SR, Trinidad TP, Dassenko SA, Cook JD. Iron Hercberg 5, Galan P, Dupin H, eds. Recent knowledge of iron absorption in humans as influenced by bovine milk proteins. Am J and folate deficiencies in the world. Paris: INSERM, 1990: 265-74. Clin Nutr l989;49:546-52.

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