t. ef. i · 2020. 9. 28. · with mag;nesium either low or marginal. it. was concluded that...
TRANSCRIPT
: . ).
• : .-:---+-I ·- -i~ ' .. •- • _,.~'.,. '. -;'; : • ··~_. •. .
THE UTILIZATION OF ESSENTIAL MINERALS .. BY PREADOLESCEN'l'
. GIRLS CONSl.JMING THREE LEVELS OF PROTEIN
by
Andal V ~ Raghavan . . I
.'thesis subt:tl.itted to the Graduate Faculty of the ·
Virginia Polytechnic Institute and State University .. Jr·.
in partial fulfillment of the :i;eqiliremehts for the degree of·
Master of Science
in
Human Nutrition and Foods
APPROVED:
t. ef. Ritchey/ Chairman
.... • 111 __ ,..,,
:R. P. ·· Abernathy I
March, 1974
Blacksburg, Virgi11:i,.a
-.----~~
TABLE OF CONTENTS Page
LIST OF TABLES . . • iii
LIST OF APPENDIXES • iv
ACKNOWLEDGEMENTS . . vii
INTRODUCTION ~ . • 1
LITERATURE REVIEW . . 2
EXPERIMENTAL PROCEDURES . • • • 11
RESULTS AND DISCUSSION . 17
SUMMARY AND CONCLUSIONS 35
LITERATURE CITED ·• . . . . . . . . ' . • 37
APPENDIXES • 41
VITA ••• • 76
ABSTRACT
ii
LIST OF TABLES
Table
1. Experimental design • , • • • . • . • . . . . • • . . • .
2. Intake and retention values of magnesium in preadolescent girls consuming three levels
Page
12
of protein , •. , . . . , • . . • • . • • • , • . . . . . • 18
3., Intake and retention values of calcium in preadolescent girls consuming three levels of protein . . . . . . . . . . . . . . . . e • • • • • • • 21
4. Intake and retention values .of manganese in preadolescent girls consuming three levels of protein . . . . • • . . . • . .
5. Intake and retention values of iron in preadolescent girls consuming three levels
23
of protein . . . . • . . . • • . . • . . • . 25
6.
7.
8.
9.
Intake and retention values of copper in preadolescent girls consuming three levels of protein • . • . • . •
Intake and retention values of zinc in preadolescent girls consuming three levels of protein • • . • . • . . • . • .
Intake and retention values of sodium in preadolescent girls consuming three levels of p~otein . . • . . • . .
Intake and retention values of potassium in preadolescent girls consuming three levels of protein • . . . • . • . . . • . . . . .
iii
27
30
32
•·"····~·· 34
LIST OF APPENDIXES
Appendix
1
2
3
4
5
6
7
8
9
Wet ashing procedure for food and fecal samples using nitric, perchloric acids and hydrogen peroxide
Procedure for wet ashing urine with nitric (HN03) acid and 30 percent hydrogen peroxide . . . . . . . .
Starting and ending heights and weights of subjects
. Daily mineral balance:. magnesium balance in preadolescent girls consuming three levels of protein (Period II) . • . . • . ; • . . . . •
Daily mineral balance: magnesium balance in preadolescent girls consuming three levels of protein (Period III) . . . . • . • , • . • .
Daily mineral balance: magnesium balance in preadolescent girls consuming three levels of protein (Perfod IV) . . • . • . . . . • •
Daily urinary loss of magnesium for Period .II
Daily mineral balance: calcium balance in preadolescent girls consuming three levels of protein (Period II) • . • . . • . • . • •
. . .
Daily mineral balance: cal.cium balance in preadolescent girls consuming three levels of protein (Period III) . ; ; . . . . . ~ · . . ti: .•- • • • • •
10
11
12
Daiiy mineral balance: calcium balance in preadolescent girls consuming three levels of protein (Period IV) . • . • . • . . . . • •
Daily urinary loss of calcium for Period II
Daily mineral balance: manganese balance in preadolescent girls consuming three levels of protein (Period II) . • . . • . • . , . • • •
13 Daily mineral balance: manganese balance in preadolescent girls consuming three levels of
Page
41
42
43
44
45
46
47
48
49
so 51
52
protein (Period III) • . . . . . • . . . • . • • . • . . • 53 .
14 Daily mineral balance: manganese balance in preadolescent girls·consuming three levels of protein (Period IV) . • • . • • • . • . . . • . . . • . . 54
iv
LIST OF APPENDIXES
Appendix . Page
15 Daily l.lrinary loss of manganese for Period II . . . . . . . 55
16 Daily·mineral balance: iron balance in. preadolescent girls consuming three levels of protein (Period II) . . . . . . . . . . . . . 56
17 Daily mineral balance: iron balance in preadolescent girls consuming three levels of protein (Period III) . . . . . . . . . . . 57
18 Daily mineral balance: iron balance in preadolescent girls consuming three levels of protein (Period IV) . . . . . 58
19 Daily urinary loss of iron for Period II 59
20 Daily mineral balance: copper balance in preadolescent girls consuming three levels of protein (Period II) . . . . . . . . . . . . 60
21 Daily mineral balance: copper balance in preadolescent·girls consuming three levels of protein (Period III) . . . . . ~ ·- . ·• . . 61
22 Daily mineral balance: copper balance in preadolescent girls consuming three levels of protein (Period IV) . . 62 . . .
23 Daily urinary loss of copper for Period II 63
24 Daily mineral balance: zinc balance in preadolescent girls consuming three levels of protein (Period II) . . . . . . . . . 64
25 Daily mineral balance: zinc balance in preadolescent girls consuming three levels of protein (Period III) . . . . . . . 65
26 Daily mineral balance: zinc balance in preadolescent girls consuming three levels of protein (Period IV) . . . . . . . ~
66
27 Daily urinary loss of zinc for Period II 67
28 Daily mineral balance: sodium balance in preadolescent girls.consuming three levels of protein (Period II) . • . . . . . . . . . . . 68
v
:.;
·k
LIST OF APPENDIXES
Appendix
29
30
31
32
Daily mineral balance: sodium balance in preadolescent girls consuming three levels of protein (Period III) . . . . . . . .
Daily mineral balance: sodium balance in preadolescent girls consuming three levels of protein (Period IV) • • • • • · • •
Daily urinary loss of sodium for Period II
Daily mineral balance: potassium balance in. preadolescent girls consuming three levels of protein (Period II) . . . . ·. . . . . . . . •
33 Daily mineral balance: potassium balance in preadolescent girls consuming three levels .of
Page
•· • • • • • ! 69
70
71
72
. protein (Period III) . • . • . . . • . . . . . • . . 73
34
35
Daily mineral balance: potassium balance in preadolescent girls consuming three levels of protein (Period IV) • • . • • • • • • • • •
Daily urinary loss of potassium for Period II • •
vi
74
75
./
ACKNOWLEDGEMENTS
The author wishes to express her indebtedness and gratitude to
for his valuable guidance and encouragement during
the course of this investigation.
Appreciation is expressed to and
for their helpful suggestions and counnents.
The author gratefully thanks
and other laboratory technicians for their help and cooperation.
Gratefulness is expressed to the Department of Human Nutrition and
Foods for providing financial assistance.
Last but not lea.st, the author expresses regards. to her husband,
1 who has indirectly contributed so much to this effort.
vii
INTRODUCTION
Minerals are essential nutrients for humans and animals, thus
their absorption and balance are very critical in nutrition. Mineral
absorption in man and animal depends on many interrelated factors, in-
cluding the rate of absorption of other nutrients, mineral availability,
source and quantity of protein and individual needs for the mineral.·
As protein malnutrition is one of the major world problems, the level
·Of protein seems to be a very important factor in mineral utilization,
One of the age groups affected by protein malnutdtion is gt"owing
children. There are several studies in which the effect of level of
protein on mineral absorption was tested, but there are very few meta-
bolic studies on preadolescent children. Thus it is important to
study the effect of level of protein on mineral balance as both are
critical for the growth of children.
Hence, a study was undertaken to find whether protein level affects
the absorption of minerals. The utilization of calcium, magnesium,
phosphorous, manganese, iron, zinc, copper, sodium and potassium were
studied in fi:l;teen preadolescent girls consuming low, medium and high
protein diets.
1
LITERATURE REVIEW
An interrelationship between dietary protein and several of the
minerals has been suggested by several studies. It was indicated that
the absorption of certain minerals could be affected by the quantity
of protein.
Most of the studies indicated that the level of protein could
affect the magnesium a~~orptian. But Schofield and Morrel (1) found
that preadolescent girls fed low p_rotein and low magnesium diets
utilized magnesium as well as subjects fed a high protein diet that
provided twice the intake of magnesium. The children received diets
with intake of protein ranging from 1 to 2.5 g of nitrogen per kg/day
with mag;nesium either low or marginal. It. was concluded that magnesium
utilization was independent of level of protein.
However, studies with adl,llts indicated significant relationship
between level of dietary protein and magnesium absorption. Maccane
et -~l (2) found in four .men that magnesium retention was slightly
better with· the high protein intake than with low protein intake. In
the study of Leverton and Linkswiler (3) with an intake of 11 g of
nitrogen and less than 250.mg magnesium, the subjects were in negative
nitrogen and magnesium balance. Hunt and Schofield (4) found that
adequate nitrogen was necessary for positive magnesium balance; in
their study with 20 g protein intake the subjects were in negative
nitrogen and negative magnesium balance. When the subjects were
brought to equilibrium with an increase in protein intake, they tended
2
3
·to be in positive magnesium balance. They suggested that at lower
protein intakes the utilization of dietary m11gnesium might have
improved as the requirements for magnesium for positive balance was
lowered. For proper absorption and positive magnesium balance there
must be an adequate level of magnesium in the diet in addition to
adequate dietary protein. Seeling (5) reported that both men and
women were in negative magnesium balance when their intakes were less
than 4 mg/kg/day. The effect of level of protein on magnesium require-
ment. has also been demonstrated in animals, Menaker and Kleiner (6)
showed that an adequate magnesium intake was necessary for new protein
synthesis and Menaker (7) showed that magnesium requirement increased
during protein synthesis as protein intake was increased. It was also
found that rats receiving low protein and adequate magnesium had a
greater weight gain than the rats on high protein and inadequate
magnesium diets. Vital et al (8) also found that raising the level of
dietary protein increased the severity of renal lesions which developed
after inadequate magnesium intake. Similar observation was made by
Bunce et al (9), when magnesium was limiting an increase in the dietary
protein level retarded growth and increased mortality in the chick and
restricted weight gain in rats. Colby and Fry (10) found that an in-
crease in dietary protein intakes of rat.s on low magnesium di.ets tended
to increase the severity of magnesium deficiency symptoms. High levels
of protein either. interfere with magnes.ium absorption or increase the
requirement for magnesium~
4
The Recommended Dietary Allowance (RDA) (11) for children between
the ages of 6 and 10 for magnesium is 250 mg per day. Schofield and
:Morrel (1) in their study maint.ained preadolescent girls in slightly
positive magnesium balance with an intake o.f 136 mg/day. But they
suggested the positive retention obtained was not enough to meet growth
requirements in this age group.
Combination of high calcium and high phosphorus levels were found
(12) to increase the severity of the magnesium deficiency. High calcium
intakes, by increasing the dietary ·requirements for magnesium, precipi-
tate magnesium deficiency in rats (12, 13, 14) and guinea pigs (15).
Metabolic studies in rats (16, 17) similarly have shown that increasing
the calcium intake increases magnesium loss. Seeling (5) suggests that
increasing the magnesium intake, particularly in the presence of an
adequate calcium intake, improves, rather than interferes with calcium
utilization. Briscoe and Ragan (18) found that adult men retained a ·
greater amount of calcium than magnesium. When the availability of
calcium or magnesium was increased, there was an increase in the
intestinal absorption of the other. , In a study with men, Johnston
et al (19) found that the dietary level of magnesium did not affect
the urinary and fecal calcium of the subjects. In addition to magnesium,
the quantity of protein was another variable. An increase in protein
intake from 48 to 141 g caused a highly significant increase in urinary
calcium. Calcium retention was observed to be better on q. low protein
diet. Hegsted {20) reported that protein quality and quantity affected
the calcium absorption. Increasing the amount of animal protein in a
,;_
5
low calcium diet. in man increased the urinary ca.lcium excretion. . . . '
Several other investigators also reported that increases in protein
intake.1we're accompanied by an increase in urinary calcium in adults . ' . . .
(21),. pre-school children: (22) and preadolescent girls (1). Maccane / .
. .. ' . . et al (2) attribut:ed the increase in urinary calcium to an increase
in absorption. Hal) and Lehmann (23) also. interpreted the increases .
in urinary calcium which followed protein feeding as indicat:i;ng an
increase in intestinal absorption. Pittman and. Kunerth (24, 25) in·
their studies found th.at the medium protein diet improved appreciably
the utilization of calcium .• The amino acid lysine and arginine have 45 ..
been shown to promote absorpt.ion of Ca in the rat (26). Hawks et al
(22) studied the effect of feeding two levels of protein, 3 and'4 g
protein per kg of body weight, on calcium absorption. They found that
.the higher protein diet had no effect on calcium absorption or retention.
Schofield et al (1) reported the calcium metabolism in preadolescent
girls receiving varying levels of dietary protein with calcium intake
held constant. Calcium absorption, urinary excretion and mean retention
were the same in all the groups at different protein levels; thus the . ' '
changes in dietary protein had no significant. influence on ;calcium
retention of preadolescent children. Colby and Fry (10) in an experi-
ment with rats found that more calcium was retained when diets contained
high protein and high calcium levels than any other combination of pro-
tein and calcium levels. The. calcium b.alance was studied in the various.
experiments using large amounts of calcium. Since physiological factors
like worry, tension, temperature and humi4,ity have been shown to in-
·crease the calcium l()sses, one recommendation may not be sufficient for
6
all under different circumstances and environments (19, 27). The RDA
(11) for calcium is 800 mg for 7 to 10 year old children. Schofield
and Morrel (1) observed positive retentions of calcium in preadolescent
girls with an intake of about 1 gm per day.
Though the metabolism of manganese is somewhat similar to magnesium
the protein level does not seem to have a very significant effect on
·its absorption. Price et al (28, 29) in their study with preadolescent --girls 7 to 9 years old found that an increase in dietary nitrogen, either
as ammonium citrate or as the limiting amino acids, had only slight
effect on manganese balance (28). A similar effect was found by in-
creasing the protein occurring in natural food source (29). · Engel et al
(30) in their study in the preadolescent children receiving different
levels of protein did not find significant differences in absorption of
manganese. According to Lang.et al (31), the quantity of protein had
no effect on the absorption of manganese in adults. North et al (32)
reported that in collegE:l women there was no statistical differences in
the absorption of manganese in those receiving 37 gm of protein and
those receiving 76 g of protein. There is no recommended dietary
allowance for manganese at present. The occurrence of human deficiency.
has not been demonstrat!;ld. Results of studies indicate that absorption
is related to body size. Engel et al (30) estimated the manganese
requirement for equilibrium as 1.0 mg per day and they suggest 1.25 mg
per day as requirement of preadolescent girls. Better retentions were
found with intakes ranging from 2.13 to 2.43 mg per day (29). Comparable
retentions were achieved with 2, mg per day in another study ( 28). Bal-
ance studies of Emerson and Daniets (33) suggest that children should
receive 0.2 to 0.3 mg manganese per kilogram of body weight.
-'. ..
7
The level of manganese does seem to have an effect on the absorp-
tion of other minerals. Hartman il al (34), while working with lambs,
found that excessive levels of manganese depressed the absorption of
iron while increasing the copper stores in the liver. It was postu-
lated that excessive manganese antagonized the enzyme system that
oxidize or reduce iron at the site of absorption.
The effect of level of dietary protein on iron absorption is not
clear. In experiments with rats, Klav·in.s et al (35), demonstrated
that the level of protein in the diets affected iron absorption. Rats
fed 5 or 10 percent protein diets absorbed less iron than those fed 15
or more percent dietary protein. Lahey et al (36) concluded that low
serum concentrations in children suffering from kwashiorkor were not
due to deficient iron intakes but were secondary effects of inadequate
protein intake. However, the studies of Abernathy et al (37), with
preadolescent girls failed to demonstrate any consistent effect of
level of dietary protein on iron absorption when protein intake varied
from about 30 to 150 percent of the recommended allowance. But in this
study iron absorption was significantly increased when the daily pro-
tein intake was reduced by the removal of all milk from the diet. They
suggest that it was possible that the influence of dieta:i:-y protein on
iron was masked by other dietary factors or that a critical level of
protein intake beyond which an influence is exerted was not reached.
The Recommended Dietary Allowance (11) for iron for children be- .
tween the ages of 6 to 10 is 10 mg per day. Estimates of iron that
must be retained by children and preadolescent boys during growth
8
t . ·ranged from O. 2 to 10 mg per day. !$<;;,
Johnston (38) .estimated the daily "
. ~ .;
' i?
i.ron needs of 5 year old girls to be 0.3 nig for, growth, 0.3.mg for
maintenanc;:e and 0.1 mg for storage, and for iO year old girls to be
0.5 fug for growth, 0.4 mg for maint.enance and 0.4 mg for storage .
Abernathy et al (37) indicated ~hat when physiolOgical and dietary·
:f factors are favorable, daily iron intakes of 9 to 12 mg would be ~:;. :";; . . 1 sufficient to meet the needs of most 7 to 9 year old. girls, and intakes .·· ~
. ~ .. of _8 mg or less are not sufficient for most of the girls in this age
group. Since physiological and dietary factors necessary £6r good iron . . . . ' . . ·.. . . ,. .
·. :: absorption are not always favorable they advocate a.dietary allowance t ~-
of 12 mg per day for ch:l.ldren in this. age group.
'{ The level of .. p~otein does not seem to have an effect on copper . ~;
" balance. With preadolescent girls it was found that neither protein ) ; nor calcium level had any effect. on the copper absorption ( 28 ) ..
The .copper intake was marginal. Copper deficiency is extremely rare in
man and the copper provided by the ordinary d.iets seems to be adequate
for copper balance. In preadolescent girls 1.3 mg per day of copper
intake was adequate for copper balance (39, 30) Engel et al (30)
suggested a copper iuta,ke of .2·. 5 mg per day. for preadolescent girls.
They made a provision of 0.5 mg per day fOr sweat losses.· anc:l O. 7 mg . '
per day as a margin of safety. But positive retention of 0.48 to 0~77
mg were obtained in preac:lolescent gi~ls with copper intakes ranging
between· 1. 55 to 1. 70 mg per day ( 29 ·). · On a copper intake of l mg.
per day the girls were in slightly positive balance which was insignifi:-.
cant.
9
Copper retention is interrelated to the absorption and metabolism
of iron, magnesium and zinc. .Excessive levels of. orie mineral generally . ;. _· . . . .. ' . ·. . .
interferes with the absorption arid metabolism of other minerals. For.· -. . ·' ', . '
·~ · . instance;. (40) it was found that large intakes of zinc interfered with
;:-·i :..~ .
: :~. ·,-;,
·'-· ,'i
'.ii
the utilization. of copper by increasing its excretion.
The absorption of zinc was affected by the source and .level ·. o.f
dietary protein Price et al (29) studied: the effect of level of pro-
tein and calcitim on zinc· absorption in preadolescent girls. Three
<levels. of protein w~re fed. As the dietary protein was increased a
su.bstantial rise in zinc absorption and retention was noticed. The . .
variation of calcium employed in the study was without.effect on zinc
retention at either protein level. in another study (.28) ari increase
in dietary protein, either as ammonium c.itrate or synthetic limiting
'amino acids, appeared to itnpro~e zinc bala~ce primarily through a
reduction in fecal zinc; heowever zinc intake was marginal.
· The RDA (11) . for zinc is 10 mg per day. · .Price et al ( 28, 29) in
their experiments in.the preadolescent girls observed positive reten-
tion of 1.3 mg of zinc pe;- day with an intake of 4.5 mg per day. But
they thought that this may not meet the ne.eds of growing children, as. . .
t}ie sum of calculated allowanc.e for growth (0. 25 mg) and possible
derma.l and sweat losses (1: 75 mg) exceed the observe.d retention by o. 7 mg.
Scant attention is pa:ld to the effect~£ level of pro:t;~in on sodium
and potassium absorption. Though deficiencies of the.se minerals are. ·. .
.·.· .. rare in healthy persons, they are important as $()dium is concerned' with
the mainten·imce of osmot:l.c .equilibrium. and body fluid volume and potas-
sium with·cellular enzyme function. The differential concentration of
10
both these minerals across the cell wall that getermines the electrical
potential of the cell membrane thus cell excitability and nerve impulse.
It was observed that deficiencies of either cation retarded the growth
in animals (41).
Sodium occurs in many foods and sodium salts are often used during
food processing. The usual adult intake was calculated to be from 10
to 300 mcg per day or from 6 to 18 g of sodium chloride. The body con-
tent of sodium and its concentration in body fluids are under hemo-
static control; moderate loads are promptly excreted in the urine and
excretion quickly drops to very low levels when intake is reduced.
Deficiencies of healthy individuals are rarely encountered in the
absence of abnormal losses. Under normal circumstances, a sodium
chloride intake of 1 g per kilogram of water is recommended (11).
Potassium is also widely distributed in foods, the usual intake
being 50 to 150 mg per day (42). There is no recommended allowance.
Thus mineral absorption in humans and animals depend on man~
interrelated factors. The requirement of children for those minerals
has been based on positive mineral balance and retention for growth.
... ; :&
~ i?... .
. . EXPERIMENTAL PROCEDURE
The ~elationshi~ of protein level to .the uti,lizatfo11 of the
esseritial niinet~ls was studied by the human.balance technique. A ' . :·· . ·. •. . . . ·. . l ··metabolic study of. 35 days length was divided i~to four periods .. • .. The
·'j
first 3 days ·served as an initial adjustmeil;t: period during which all {. . ·, . '... .. . ·. .· ·.'.
the subjects were on a 30 gprotein diet. During the three experi-
riiental periods, each ten days in length,· following the adjustment: .
·period,· three diet~ry levels of .protein, (30 g, 60 g, and 90 :g) were. .·.
fed. The subjects were randomly allotted to three groups (A, B, C) of
five each and rotated through each of the three dietary.treatments
during the e:x:Perimentd periods (Table 1). ·
The intake of minetB.ls was riot brouglit to a given intake, but was
studied as it occurred with the normd dietary variations'. However,·
because the menus were similar, the m:i.ne:t;"al intakes did not va'ry wtd,ely
within a given protein leveL Calciilm. aµd iron were supplemEmted to
attain the recomnten~ed -daily intake.
Subjects ·
·Fifteen pr¢adolescertt girls ranging in age from 7 y~ars and 6 : .
.. months to 9 .Years a,nd 6 m<;mths wete the ~ntbjects·. Bef<:>re the. study,
each aubject Was given a ·physical examimltion by a physician to assure
. ' . . . . . .
to serve as a nutritional camp for the children.. The subjects lived in . .
the l!ome Management Resi.dence on the canipus of Virginia Polytechnic
... ·Institute and State Unive~sity and 4 counselors lived with the children.
11
12
,, TABLE 1 ~-
. EXPERIMENTAL DESIGN.
.. . Periods Diets Subjects
I 30 g protein All subjects on same diet
II 30 g protein Group A 60 g protein Croup B
.90 g protein Group c
III 30 g protein Group c ;: 60 protein ·Group A g ,·· 90 g protein Group B
IV 30 g protein Group B 60 g protein Group c 90 g protein Group A
. ,\
Ii>.,_. . . , . . . - . ... . . . . ~"'~_;
Recreational programs were provided throughout the · stuQ.y. A phydcian . . . ..
was on: call at all times. for emergencies during th~ study and a nurse ·. ·. .
checked the children daily for minor problems. Hef$ht and weight
records of th~ subjects were mairita]_ned. (Appendix 3).
Diet.
· .. Dfets contained foods that were generally acceptable .to preadoles-. . . ,'
cent chil<iren. With the. exception of protein the c:liets and the supple-
me11ts provide4 nutrient intakes sufficient to. meet or e~ceed RDA
stanQ.ards. A vitamin supplement containiiig18 mg of iron was given
throughout: the study. Wafers containing 232. mg of calcium were also
added to certain diets. Three wafers were ad.ded to ail three 30 g
protein level daily menus and one wafer was added to the X, z diets on
. the 60 g protein level. l'\lutrient content of foods .included in the· diet·.·
were calculated from Home and Garden Bulletin No .. 72. Calories were · · ' ; .. .. . . .
adjusted s.o each subject maintained or slightly gained weight, Three
levels of dietary :Protein, 30, 60, 90 g per day werefed.. Ion free
water was used for drinking and washing receptacles and in the labora~
; · tory procedures. The food was cooked in the diet' kite.hen in Wallace
. .. ·.,,
HalL I~ was weighed a:nd served :tn the diet kit~he.ll.. ·· A: cou.nselor ate
with the children t:o nµike certain that food and supple:rnents were con-·
sumed. When a child' spilled the food, replacements for that amourit
were provided.· . All £0.ods except p_erishables ~ere purchas~d in single ..
quantity lots before the study pegan •
14
Collection and Compositing of Samples . :'.;
~·. -
·.· :; ·Food. · ~
~ ...
Two portions of food were colle'cted at each meal and for 24 hour
food c0mposi:tes •. Margarine was not included in the general compos_ite' .
since it was ri.ot easily blended wi:th other foods. Capsules co:ntatning
At •. supplements were also not included in the general food composites.
the e!ld of each day the food collected for the whole day was blended
in a 5 quart ·waring blender using ion free water, and was brought Up
to 2200 g. !twas homogenized at speed 1 until the contents were
thoroughly mixed, then at speed 3 for one or .two minutes, then at top· .
. speed for on·e minute in order to homogenize the. fat th<::iroughly. A 220: g •.
aliquot of the.composite was ·weighed and was frozen. At the end of the
period, all the daily subsamples for that period were blended according
. ·} ·. to tlie procedure .described above. Samples for the determination· of
.
.~.
nitrogen and minerals were frozen until used,·. Food (!Oi:nposites were wet
ashed using nitric., perchloric acids and 30% hydrogen pe.roxide
· (Appendix 1-).
Urine
1'wenty-four hou.:i; urine collections were ~de each day beginn£n:g · ..
with the second voiding after rising and ehding the riext day with tl:ie .
. first voiding. Collectio11s were made directly in opaque pyrex bottles . . . . ' ' . .
to which were added 10 mts of4 N HCL~· At the .end of ea~h 24.:,hour·
perfod the urine volumes were recorded. The total urine fc;>r the day. · ·
was diluted to 2000 mls wit;h fon free water. Two hundred mls were
15
measured and frozen until the end of each period. At the end of each
period; a period urine composite was made with 100 ml of urine from
each of the last fiv.e days samples. The urine samples were wet ashed
with nitric acid and 30% hydrogen peroxide (Appendix 2).
Feces
The feces was collected in Pyrex casserole, combined daily and
frozen• Fecal .markers (gelatin capsules containing 20 mg of brilliant
blue plus 175 mg methyl cellulose) .were given to each subject to ma.rk.
the feces for each experimenta,1 period. At the end of each period
partially thawed feces from the last five days was blended iri. a 5 quart ·
Waring blender. The weight was brought to 1000 g using ion free water.
Aliquot samples for analysis were frozen until used. A sample of feces
for each period was wet ashed using nitric and perchloric acids and
30% hydrogen peroxide (Appendix 1).
Nitrogen alanysis was done on daily urine samples, and on food
and feces composites for each period using a modified Kjeldahl-Grunning-
Arnold Method. Nitrogen and mineral balances were evaluated on the
basis of last 5 days in each experimental pericid. •
Analytical Procedure
The concentration of mine.rals in the food and fecal composites,
and in the urine can be measured in parts per million. In order to
avoid contamination from outside sources all equipment used in the col-.
lection and analysis of samples were acid washed prior to us.e. The
acid wash consisted of a 50-50 solution of concentrated nitric acid and
deionized water. All equipment was washed first .in the solution and
then rinsed twice with ion free water.
16
Mineral Determination
The concentration of each mineral was determined using a Perkin-
Elmer Model 305 Atomic Absorption Spectrophotometer according to the '
recormnended procedure. The solution containing the elements was
aspirated into the flame. This caused a population of ground st:at:e or
neutral atoms <t:o accumulate in t:he flame. A portion of the light: from
the appropriate cathode lamp is absorbed by the neut:ra.l atoms. Since
the concentration o:f: neutral atoms in the flame is proportional t:o
absorption, measurement: of absorption can be used t:o determine concen-
trat:ion of the metallic el.ement:s in a solution. Standard curves of
absorbance were plotted using prepared standards. The sample content:
was calculated according t:o the formula:
Element: (mcg/ml) = (sample absorbance) (std. concentration)
standard absorbance
The mineral samples for the determination of magnesium and calcium
were diluted with a 5% solution of lanthanum before they were read.
Food and fecal excretions were determined on the basis of each period.
Urinary excretion of minerals was determined on the basi;8 of each periad
except for Period II.. when daily samples were also analyzed separately
in order to determine daily variations. The balance figures were
obtained by subtracting urine and fec;.:tl values from the intake.
RESULTS AN]) DISCUSSION
Magnesium
Magnesium intakes of subjects on 30 g protein level in all the
three groups were •. below the Reconnnended Allowance of 250 mg per day
(Tabie 2). The mean intakes for all groups was about 176 mg, Twelve
out of fifteen subjects were in negative balance. Group A was in
negative balance at a.11 protein levels though magnesium intake was
higher on the medium and high protein levels than on the low protein.
The mean negative balance.obtained at the 60 g level was lower than
.that obtained at the 90 g and 30 g protein levels (Table 2). Group B
had better retention values at the 30 g protein level but had a mean
negative balance of -5.30 mg at the 60 g protein level, The subjects,
except one, were in positive balance, with a mean .retention value equal
to about 10% of intake. At the 90 g level the mean retention was about
5% of intake but two of the subjects were :in negative balance. At these
two levels magnesium intake was higher than the reconnnended level. The
subjects in· Group C had a mean negative balance of ~25 mg aP. the JO g
protein level. All the subjects in this group were in positive balance
on 60.g prote:in with a. mean retention value representing'about 27% of
th:is intake, 303.88 mg. Again three out of the five subjects were in
negative balance, and mean balance was also negative with an intake of
345 mg of i:nagnesil.lm. Individual day urinary value's indicated variation
in urinary losses. The overall mean intakes indicated .that an increase
in magnesium intake accompanied the increase in protein.
17
18 ·.,
TABLE 2
Intake and retention values (mg) of magnesium in preadolescent girls
consuming three levels of protein
Group and 30 g Protein 60 g Protein 90 g Protein Subject No. Intake Balanc.e Intake Balance Intake Balance
A 100 176 -134 226 -90 240 -43
101 176' -97 226 3 ~(
102 176 -47 226 -38 240 -239
103. 176 -61 226 ... 25 240 -46
. 104 176 -26 226 "".31 240 19
X A. 176" -73 226 -36 240 -77
B 105 . 163 -15 278 5 280 40
106 163 51 278 41 280 -13
107 163 .. 66 278 52 280 21
108 163 11 278 49 280 6
109 163 .. a 278 -6 280 20
X B. 163 .. 5 278 28 280 15
c llO 175 -2 304 90 345 -28
111 175 -6 304 59 345 -73
112 175 10 304 85 345 49
113 175 -33 304 85 345 .,.3
114 175 -93 304 95 345 6
x c. 175 -25 304 83 345 10
x .. 171 34 269 25 288 -17
· * Data missing for this period and• the subject.
19
1,,,,, With low protein intakes the utilization of magnesium w;is improved
and agrees with the findings of Schofield and Morrel (1 ) . Even on
such a low magnesium intake three of the subjects achieved positive
balance. The differences between the groups on 30 g protein level
with similar intakes of magnesium could be due to individual variation
·in their requirements for magnesium. The high negative mean balance
of Group A could also be due to the very high levels of calcium in the
diet as the calcium intake at that period, 2377 mg. Very high levels
of calcium, by increasing the dietary requirements for magnesium,
precipitated magnesium deficiency in rats (12) and guinea pigs (15).
At higher protein Levels an elevation in the requirement of magnesi'Um
was noticed as was suggested by different animal studies (13, 14) and
as suggested by Hunt and Schofield(l).
The 60 g protein level with an increased magnesium intake resulted
in improved retentions. The results indicated that 304 mg of magnesium
was required in order to bring all the subjects to positive balance at
this protein level since at an intake of 278 mg some of the subjects
were in negative balance though the mean retention for the group was
positive. This suggests that with medium protein intakes, the require-
ments ofpreadolescent girls may be above the recommended allowance of
250 mg. With protein intakes of 90 g, magnesium utilization was lowered,
suggesting that very high levels of protein were detrimental to magnesium
utilization.
20
Calcium
The calcium intakes were very high and most of the subjects were
in positive balance. Group A had mean positive balances corresponding
to 51% and 13% of respective intakes on 30 g and 60 g protein levels.
; At the 90 g protein level, two of the four subjects were in negative
, balance. Subject No. 102 was in negative balance which made the group
balance negative (Table 3). Group B was in positive balance at all
three levels. With comparable intakes of calcium on 30 g and 60 g
protein level they had mean retention values equal to almost 47% and
17% of their respective magnesium intakes. Unlike the other two groups
high retention values were observed at 90 g protein level also, about
29% of intake was absorbed. Group C had mean retention values correspond-
ing to 37%, 30% and 4% of their intakes on 30 g, 60 g and 90 g protein
levels, respectively.
These results suggest better retention with low protein i~takes
and decreasing retention of calcium with increases in protein in the
diet. These results cannot be compared with that of Pittmann and
Kunerth whe·re they indicated that ·better utilization of calcium with
medium protein than low protein, as the level of protein on their
low protein diet was much lower than present 30 g diet and their moderate
protein falls between the 30 g and 60 g protein levels in the present
study. This study differs from that of Hawks ~t al' s (22) where they
used only very high (3 g and 4 g/kg) protein diets, somewhat as the
90 g protein level in the present study. The high urinary excretion of
calcium with high protein intakes reported in several s.tudies (21, 22, 23)
is not applicable to this study as the calcium intakes were varied on
21
TABLE 3
Intake and retention values (mg) of calcium in preadolescent girls
consuming three levels of protein
Group and 30 g Protein 60 g Protein 90 g Protein Subject No. Intake Balance Intake Balance Intake Balance
A 100 2378 1256 1074 -51 1099 -35
101 2378 1190 1074 186 1099 "/(
102 2378 1120 1074 242 1099 -823
103 2378 nos 1074 131 1099 180
104 2378 1367 1074 191 1099 I 114
X A. 2378 1207 1074 140 1099 -141
B 105 n42 253 1279 196 1074 502
. 106 n42 302 1279 218 1074 233
107 n42 194 1279 138 1074 357
108 n42 60 1279 288 1074 208
109 n42 281 1279 160 1074 268
X B. n42 218 1279 200 1074 314
c no 1221 495 1404 611 1366 59
n1 1221 409 1404 340 13p6 -279
112 1221 499 1404 384 1366 321
113 1221 462 1404 318 1366 45
114 1221 391 1404 427 1366 156
--x c. 1221 451 1404 416 1366 . 60
x .. 1580 625 1252 252 nsCi 78
* Data missing for this period and the subject.
22
different protein levels. But with similar intakes the increased ex-
cretion of calcium on the 90 g protein diet than on lower levels was
evident; for example in Period II (Appendix 8). The calcium was better
utilized at the lower protein intakes and the absorption decreased with
increased protein.
Manganese
The manganese intakes were comparable to the standards set in
literature except for Group A at the 60 g and 90 g protein levels
(Table 4). Group A on the 30 g protein level were in positive balance
with a high intake, (3.51 mg) of manganese with much lower intakes on
60 g and 90 g protein diets the mean balances for Group A were negative.
On the 60 g protein level, negative mean balance equal were -.06 mg,
although only Subject No. 101 was in negative balance. The very high
fecal values of the mineral for this subject could result from problems
of the fecal composite. Except for one subject on 30 g protein level
Group B subjects were in positive balance. The intake of the manganese
was comparable on the three levels. In this group the retention values
increased as the protein intake increased from 30. g to 60 g, and then
decreased at the 90 g level. Mean retention values equal to 17%,
26% and 11% of intakes were found at the 30 g, 60 g and 90 g protein
levels, respectively. In Group C a similar pattern of retention was
found (Table 4).
The fact that the subjects obtained best retention at 60 g protein
level indicated that medium protein was beneficial to the utilization
<;'
"} . . ~· 23 '?: .. . '..· -;::
;':• t'l\I\!-. =:-:-:~ . :;•. TABLE 4 ..... ~
:,~
{ Intake and retention vaiues ·(mg) of ~:·
!\ manganese in preadolescent girls :if_
? consuming three levels of protein .::i"
r:; Group 'i:· '·i' ·and 30 gProtein 60 g Protein 90 g Protein .'£ ~~; .. ·Subject .... No. Intak~ Balance Intake · Balance Intake Balance \; '.(
A 100 3.51 0.43 2.00 -1.08 1.95 -1.14
~· ·'' 101 3.51 0.26 2.00 . 0.35 1.95 * °'' 'J ;,i:
:f.;
··~- 102. 3.51 -0.14 2 .• 00 0.19 1.95 ..;z.71
:j 103 3.51 -0.49 2.00 0.01 1,95 -:0.38 .. :;
~: ·104 3.51 o.76 2.00 0.25 L95 -0.40 ':J· ... ·1:
;; .. ).'"
X A. 3.51 -0.17 2.00 -0.06 1.95 -1.16 y
... B 105 2.74 0.39 2.47 0.73 2.51 1.13
106 2.74 0.99 2.47 0.26 2.51 . 0.34
107 2.74 0.70 2.47 0.50 2.51 0.81
108 2.74 .. -0.20 2.47 0.98 2.51 0.45
109 2.74 0.40 2.47 o. 71 2 .. 51 0.31
x B. i.74 0.46 2.47 .0.64 2.51 0.61
c 110 2.87 0.67 2.81 1.07 ' . 3.17 0.21 (
111 2.87 0.35 2.81 0.88 3.:17 0.07
112 2_.87 0.52 2.81 1.07 3.17 1.19
113 2.87 0.49 2.81 0.77 3.17 0.20
114 2.87 0.38 2.81 1.21 3.17 0.13 ·. '
x c. 2.87 . 0.48 2.81 1.00 3~17 0.36
' x . •. .• 3.04 .. 0.25 2.43 0.53 2.54 -0.16
· * Data missing for thi~ period and the subject.
24
~"" of manganese. These results cannot be compared with studies in the "
literature (28,29) as the levels of protein used in those studies were
lower than the medium protein (60 g) level in the present study.
: The iron intakes were very high on all protein levels because of
the supplement. Group A had a mean positive balance representing 3.45%
and 2.17% of their intakes on 30 g and 60 g protein diets, respectively.
At the 90 g protein level, the group had a mean negative balance; two
of the 4 subjects were in negative balance.. Group B. achieved a mean
positive retention at all three levels of protein intake. Retentions
represented 8.7%, 6.43% and 21.11% of intakes on the low, medium and
high protein levels, respectively. Group C had mean retention values
equal to 1/3 of their intake on 30 g level and the retention reduced
with an increase .in protein. · On 60 g and 90 g protein levels they had
mean retention values representing about 24.18% and 5.04% of intakes
respectively.
This data indicates that individual retentions vary according to
individual needs. The prior intakes of subjects ~ight also have an
effect on their balances. In two groups increases in protein intakes
were accompanied by a decrease in iron utilization. But the third group
showed exceptionally good retention on high protein level. Generally,
the iron utilization was better on low protein than with moderate protein
diets.
:)
~- . . :~
-'.~ .
Group and Subject No.
A 10.0
lOl
102
103
104
X A~
B 105
106
107
108
109
X B•
c 11.0
111
112
113
114
x c. x . ~
25:.
TABLE 5
Intake and retention values (mg) of iron in preadolE!scent girls
consuming three levels of protein
30 g protein
Intake
31.55
31•55
31.55
31.55
31.55
31.55
29.85
29.-85
29.85
29.85
29.85
29.85
29. 61 ..
29.61
29.61
29.61
29.61
29. 61
30.34
Balance
2.13
2.13
0.30.
-3.48
4.35
1.09
0.89
5.88
' 3.10
-0.20
3.33
2.60
5.26
12.37
10.04
12.16
10.31 ·'
10.03.
4.57
60g Protein
Intake
29.41
29.41
2~.41
29 .. 41
29.41
.. 29 .41
32.33
32.33
32.33
32.33
32.33
32 •. 33
33.33
33.33
. 33.33
33.33
33.33
33.33
3L69
Balance
-0.61
-1.87
-0.60
2.10
0.37
o.63
3.45
-1.08
3 •. 71
6. 71
-2.40
2.08
10.95
5.58
11.10
6.84
5.84
~.0.6
3.32
* Data missing for this· period and the subject.
90. g Protein
Intake Balance
31·.24 1.79.
* 31.24 .-35,07
31.24 7.58
3L 24 -1. 59··.
31.24 -6.82
28.98 9.21
28.98. 0.86
28.98 12.10
28.98 3.89
28.98 4.52
38.98 6.12
35.50 0.97
~s.50·· -0.45
35.50
35.50
35.50
35.50
31.91
3.29
1.26
3~88
l_.79
o.36
26
Copper
Copper intakes ranged from 730 mcg to 1444 mcg per day. On the
30 g protein diet and an intake of 1234 mcg, all the subjects in Group A
were in positive balance; the mean retention equaled 19% of intake.
With a lower copper intake on the 60 g diet and still lower on the 90 g,
they were in negative balance (Table 6). Group B with comparable
copper intakes on the three levels absorbed about 18% and 26% of their
copper intakes on 30 g and 60 g protein diets, respectively;, but on the
· 90 g protein level they were in negative balance. Group C on 30 g pro-
tein ievel had mean negative balance of -73 mcg with an intake of 957 mcg
of copper. Two out of the five subjects were in positive balance on 60 g
and 90 g protein levels. They had mean retention values representing
19% and 27% of their intakes, respectively.
The results obtained with Group A were not conclusive as the copper
intake was lower on the higher protein diets. It is not clear whether
this difference was due to protein level or due to the low levels of
copper in the diet. The results in another group on comparable intakes
on all the three levels indicated better utilization of copper with
medium protein diet than with the low protein diet and utilization
decreasing at the higher protein level. The data from the third group
though not conclusive indicated that with high copper intakes, good
copper retentions can be obtained at high protein level also.
On the whole it can be suggested that medium protein enhances
copper utiHzation;less than 1000 mcg of copper intake was not sufficient
for copper balance. Very high protein intake might affect copper balance
27
TABLE 6
Intake and retention values (mcg) of copper in preadolescent girls
consuming three levels of protein
Group and 30 g Protein 60 g Protein 90 g Protein Subject No. Intake Balance Intake Balance Intake Balance
A 100 1234 63 930 -227 730 ..,399
101 1234 378 930 -232 730 *
102 1234 248 930 -156 730 -977
103 1234 209 930 -184 730 -362
104 1234 290 930 -109 730 -350
X A. 1234 238 930 -182 730 -522
B 105 1122 181 1070 218 1002 78
106 1122 335 1070 224 1002 -85
107 1122 202 1070 282 1002 73
108 1122 155 1070 503 1002 -141
109 1122 161 1070 182 1002 -160
X B. 1122 207 1070 282 1002 -47
c 110 957 -216 1081 294 1444 455
111 957 -93 1081 191 1444 348
112 957 -149 1081 203 1444 459
113 957. 26 1081 122 1444 168
114 957 68 1081 228 1444 507
x c. 957 -73 1081 208 1444 387
x .. 1104 124 1027 103 1059 -61
* Data missing for this period and the subject.
28
adversely when copper is present in moderate levels in the diet, but
enhances the absorptiOn of copper when the intakes of copper are as
high as 1444 mcg. For instance with such an intake of copper all the
subjects were in positive balance and had very good percentage of reten-
tions,· These data indicate that when copper is present in moderate
(1000 mcg) levels in the diet, protein enhances its absorption till a
certain level and higher protein intake after that level is detrimental
. to copper absorption by increasing the copper requirements. But in
copper deficient diets even medium.protein appeared to cause higher
negative balances (for instance if we compare Group A on 60 g and
Group C on 30 g protein level diets with comparable copper intakes,
Group A had higher negative values). So it. indicates that with proper
copper intakes protein might have an effect on copper absorption.
These results cannot be compared with that of Price et al's studies
(28,29) where the levels of protein used were lower than the levels
used in the present study. The highest level (48 mg) in their study
is somewhere between the lower and medium protein levels in this study.
Even the copper intakes were higher.than the highest intake used in the
present study. The copper intakes ranged from 1550 mcg to 1700 mcg per
day. In another study the copper intakes were 999 mcg but the protein
level used were lower than the present 30 g protein diet.
So it can be concluded that there is a strong possibility that
moderate protein might improve the .utilization of copper when the
mineral is present in moderate levels in 1100 m:cg in the diet. And it
can also be concluded that high protein intakes are detrimental to
29
copper absorption when copper is present in low levels in the diet
but enhances the copper retention when the diet contains more than
1450 mcg of copper.
Zinc
The zinc intakes for all the groups on )0 g protein level diets
were lower than the intakes on 60 g and 90 g levels. All the subjects
in Group A, except for one, were in negative balance at all three
levels. With similar intakes on 60 g and 90 g protein levels, the mean ..
negative balance was lower on 60 g protein diet than on 90 g (Table 7).
Group B had mean negative balances on both 30 g and 60 g, less on 30 g
than on 60 g protein diets. But on the 90 g diet the mean negative
balance of the group was -0.02 mg with an intake of 9.53 mg. Three
out of five subjects were in positive balance. Group C on 60 g protein
level absorbed about 33% of their intake (10 mg) o.f zinc. But with.
higher intakes subjects were in negative balance.
These results are in agreement with RDA, as the zinc requirements
of preadolescent girls seemed to be about 10 mg. It was not possible
to compare the retention on 30 g with that obtained on 60 g and 90 g
protein diets as the zinc intake was low at this level. The data
obtained from Groups A and C with comparable intakes on 60 g and 90 g
levels suggested that very high levels of protein.might interfere with
zinc absorption. The utilization ,of zinc seemed to be improved with·
moderate protein (60 g) compared with a high protein (90 g) intake a,s
indicated by higher retention values. The differences between Groups . .
(B and C) on 90 g protein diet could be due to individ\lal variations in
requirements for zinc.
.. , ... ,.
. -
30 ' < ... ~·
~-,.
::,;'
TABLE 7
Intake and retention valu.es - (mg) of .•z-inc in preadolescent girls,
.·· consumfo.g three levels of protein
Group. and 30 ~ Protein 60 g Protein 90 g J;>rotein Subject No. Intake Balance l;ntake ]3alance Intake Balance
A 100 4.77 -4.50 6.87 ... 2 .63 6.35. -2.55
101 4.77 -5_.44 6.87 .. 0.90 6.35 * 102 4.77 -4.98 6.87 -1~04 6.35 -14. 21
103 4. 77 -3.45 6.87 -1. 79. 6.35 -3.20
104 4,77 -3.64 6.8} 0.44 6.35 -4.58
X A. .4.77 -4.40 6.87 -1.19 6.35 -6.14
B 105 4.23 ---0.13 7.75 -2 ._55. 9.53 1.06
106 4.23 0.33 7.75 -2.06 9.53 -1.14
' 107 4.23 -0.56 7.75 -0.66 9.53 0.87 ']'
108 4.23 -2.09 7.75 -0.91 9.53 0.59 -
109 4.23 - -0.38 7.75 -1.82 9.53 -l.49
x B. 4.-23 -0.69 7.75 -1.60 9.53 -0.02
c llO- 4.13 -2.73 10.03 3.97 ll •. 61 .. 3.52
1ll -4.13 0.24 10.03 2. 73 11;61 -2.75 8 .,,
112 4.13 .. 1.36 10.03 3.11 11.61 0.29 :·:t ll3 4.13 -2.33 10.03 3.3i 11 .. 61 -2.51 •.;.
114 4.13 -2.10 10.03 3.55 H.6J .-2.77 ~·.
-1.66 10.03 3.34 n.61 ... 2 .25 x c. 4.13
x .. 4.38 -2.25 8.22 0.18 9.16 ·2.80
* Data missing for this. period and the subject.
c: .;,
'
31
These results cannot be compared with that of Price et al' s . -·. -.-
. . studies (28, 2'9) since as the protein level used in these studies were
from 25.g to 46 g El,nd were lower than the int;ermediate protein level . . . ,' ..
(60 g) in the present study.
Sodium·
Sodium 'intakes were above .2 g and were quite varied. On the 30 g .
protein level the intakes of so<;lium were lower than the intakes at: the
other two levels. At this level with cotiiparable i~takes of sodium for · · ·
•the three groups, the mean balances differed from group to group. In···
Group A two subjects on 30 g, two on 60 g and one on 90 g protein levels
were in positive balance. All other subjects were in negative balance.
Group A.with closer intakes on 60 g and 90 g protein dieti:; had lower
f mean negative balance on 60 g diet (Table 8) ~ . In Group B th.e mean . 'r ;\~
/ balances on all three levels were positive •. !n this group, the best ·'
. . .
' . retentions were obtained with the 60 g protein diet. The retention . .
~ . values were lo~er on; 30 g and lowest on · 90 g level diets. . The mean .·· · ·
retention of the .gro~p was about 7%,, 9% and 3% of t.heil;' respe~tive
intakes on 30 g, 60 g and 90 g protein levels. Group C had mean reten-
tions corresponµing to 3%, 18% at1d 15% on.the three levels ip. inci:;easing·
order of protein.
The results obtained from Groups B and C indicate that a better
per7entage of retention was obtained at 60 .. ~ than ,at 30 g protein level. . .
On the 60 g levels, subj~ct;s .with equal and also even on lower sodium
intakes than that on 90 g level had better retention values than on
90 g protein level. These data suggest that sodium is well .absorbed
TABLE 8
Intake and retention values (mg) of sodium in preaciolescent girls
consuming three levels of. protein
Group.·.•. and Subject No.
30.g Protein 60 g. 1,'rot.ein 90 g Protein
A 100
lOL
102
103
104
In.take
2216
2216
22l.6
2216
2216
XA. 2216
B . 105 2003
· 106 2003
107 2003
108 2003
109 2003
X B. 2003
c 110 2252
lll 2252
112 2252.
113 2252
114 2252
x c~ 2252
x .. 2157
··Balance
-106'
.. 1253
-16
232
137
-548
608
46
.690
-134
132
389
382
510
170
..;1123.
66
-1
Intake .· Balance Intake Balarrce •.··
2815
2815
2815
2815
2815·
2815
2956
2956
2956
2956
2956
2956
3505
3505
3505
3505
3505
3505
3092
-195
.;746
94
..;126
-147
2725
2725
2725
2725
2725
2725
.281 · 3476
398 .· 3476
604 3476
·· 121
-56
270
569
223
780··
725
.. 1883
636
253
3476
3476
3476
4265
4265
4265
4265·
. 4265
4265
348.9
-254
* -878 ..
-1149
730
·-388·
123
.. 186
···· 119
1
465'
104
I 113.0
672
8.37
469
117
645
120 ..
· * Data. missing for t]i{s period and the subject.
33
at medium protein level. When sodium intakes are high, comparable
retention can be achieved, but when sodium intakes are low, very high
protein in the diet seemed to be detrimental to sodium utilization. i
.Sodium seems to be better absorbed at the 30 g .protein level than at
the very high protein level.
Potassium
The potassium intakes were high at all protein levels and almost
all subjects were in positive balance. Group A had mean retention
values equal to 27, 67 and 37 percent of their potassium intakes on
the 30, 60,90 g protein levels, respectively. The best percentage of
retention was at the 60 g protein level, with a very high potassium
intake. Group B with comparable intakes on all the 3 levels had
about 40, 35 and 76 percent of their intakes as mean retention on
30, 60, 90 g levels. Group C had mean retention values representing
77, 35,·32 percent of their intakes with the mean retention decreasing
with an increase in protein.
There ~as.no similar pattern of absorption in the three groups.
Each group obtained good retention values at different protein levels
and the mineral was absorbed well at all protein levels •. At 90 g
protein level best mean retention values were obtained with an intake
of 2844 mg of potassium, and a further increase in the mineral decreased
the percentage of retention •. Protein level does not appear to have any
effect on the potassium utilization.
34 ·'
.':. ;;"""' ~1 TABLE 9
Intake and retention vall,les (mg) of potassium in preadolescent girls
consuming three levels o~ protein
..'."
Group j arid 30 g Protein 60 g Protein 90 g Protein
Subject No. Intake Balance Intake Balance Intake Balance·.
A 100 2457 1062 3333 2250 2831 12so·
101 2457 -542 3333 1701 2831 * 102 2457· 1033 3333 2335 2831 560
103 2457 850 3333 2417 2831 575
104 2457 945 3333 2481 2831 1_769
. ·:;:~ x A~ 2457 670 3333 2237 2831 1046
B 1.05 2252 605 2998 780 2844 2179
106 2252 1299 2998 1250 2844 2063
107 2252 690 2998 1607 284-4 2159
108 2.252 1269 2998 851 ·2844 2266 ~~
109 2252 727 2998 742 2844 2179
X B• 2252 918 2998 1046 2844 2169
c llO 2452 1922 2808' 664 3352 1377
lll 2452 1967 2808 1302 3352 915
ll2 2452 2010 2808 1087 3352· L278
ll3 2452 1943 2808 926 3352·. . 968
114 2452 1563 2808 969 3352 683 ., ~ x c. 2452 1881 2808 990 3352 1044
x •• 2387 ll56 3046 1424 3009 1420
* Data missing for th~s period and subject
.·\'
. . J
., ., ·:r
SUMMARY AND CONCLUSIONS
The utilization of magnesium, calcium, manganese, iron, copper,
zinc; sodium and potl:lssium in preadolescent girls consuming diets with
30 g, 60 g, and 90 g protein was studied. Fifteen preadolescent girls
participated in the 35 day balance study. Samples of food, urine and
feces were wet ashed and the mineral concentrations were determined by
atomic absorption spectrophotometery.
The data on magnesium indicated better utilization of the mineral
on low protein intakes when the mineral was present in low levels in
the diet. Increases in protein level increased the requirement for the
mineral and the high protein intake was not beneficial to the utiliza-
tion of magnesium.
Calcium was better utilized at low protein intakes; there was
decreasing retention with an increase in protein except when both pro-
tein and calcium are present in high levels in the diet. Manganese
utilization was best with low protein intake and high protein diet
caused an ~ncrease in manganese requirement.
The re.sults obtained for iron differed from group to group. Iron
utilization was better with low protein than with medium protein intakes.
The data on copper indicate a strong possibility of improved utili"-
zation of copper with med.ium protein intake when copper was present in
moderate levels. Intakes less than 1 mg of copper we.re not enough for
positive balance. High protein was detr.imental to copper retention with
moderate copper intakes, but enhances copper utilization when copper was
present in the range of 1450 mcg of copper.
35
' ..
36
~~ Zinc utilization was improved in the medium protein diets compared
to the high protein. Sodium was also better utilized with .medium pro-
. tein than. with high protein intake. Comparable r~tention was ·obtained ~ ~~ .
with high protein intake when sodium intake was also high but. high
protein was detrimental if sodium intake was low. Protein level did .. .
not have any effect on the potassium absorption.
. LITERATURE CITED
1, Schofield, F.A. and E. Morrell, Calcium, phosphorus and magnesium. Fed. Proc. 19: 1014, 1960.
2~ Maccane, R.A.; E.M. Widdowson and H. Lehnman. The effect of protein intake on the absorption of calcium and magnesium. Biochem. J. 36: 686, 1942.
3. Leverton, R.M. and H. Linkswiler. Magnesium requirement of young women receiving controlled intakes. J. 'Nutrition. 74: 33, 1961.
4, Hµnt, S.M. and F.A. Schofield. Magnesium balance and protein intake level in adult human females. Am. J. Clin. Nutr. 22: 367, 1969.
5. Seeling; M.S. The requirement of magnesium by the normal adult: Sunnnary and analysis of published data. Am. J. Cli~..!...-Nutr.
14: 342, 1964.
6. Menaker, W. and I. S. Kleiner. Effect of deficiency of magnesium and other minerals on protein synthesis. ProS!_ So~; Exper. Biol. and Med. 85: 377, 1952.
7. Menaker, W. Influence of protein intake on magnesium requirements during protein synthesis. Pro. Soc, _Exper. Biol. and Med. 85: 149, 1954.
8. Vitale, J.J., P.L. White, M. Nakamura, D.M. Hegested, N. Zamcheck and E.E. Hellerstein. Interrelationship between experimental hypercholestremia, magnesium requirement a.nd experimental athero-sclerosis. J. Exper. Med. 106: 757, 1957.
9. Bunce, G.E., P.G. Reeves, T.S. Oba and H.E. Sauberlich. Influence of the dietary protein level on the magnesium requirement. J. Nutr. 79: 220, 1963.
10. Colby, R.W., and C.M. Fry. Effect of feeding high levels of protein and calcium in rat rations on magnesium deficiency syndrome. Am. J. Phi sol. 166: 408, 1951.
U. Recommended Dietary Allowances 9th ed. rev. Natl. Acad. Sci. Res. Council Puhl. Washington, D.C. ,1974.
12. Colby, R.W. and C.M. Fry. Effect of feeding various levels of calcium, phosphorus and magnesium to rats. Am. J. Phisol. 166: 209, 1951.
37
38
13. Tufts, E.V. and D.M. Greenberg. The biochemistry of magnesium deficiency. The minimum requirements for growth, gestation and lactation and the effect of dietary calcium level thereon. J. Biol. Chem. 122: 715, 1937.
14. Watchois, E. and R.A. and R.A. Mccance. Subacute magnesium deficiency in rats. Biochem. J, 31: 1379, 1937.
15. O'Dell, B.L., E.R. Morris and W.O. Ragan. Magnesium requirements of guinea pigs and rats: Effect of calcium and phosphorus and symptoms of magnesium deficiency. J. Nutr. 70: 103, 1966.
16. Forbes, R.M. Effect of dietary calcium: magnesium: phosphorus ratio on mineral utilization in rat. Fed, Proc. 20: 293, 1961.
17. Alcock, N. and L Macintyre. Interrelationship of calcium and magnesium absorption. Clin. Sc. 22: 185, 1962.
18. Briscoe, A.M. and c. Ragan. Effect of magnesium on copper metabolism in man. Amer. J. Clin. Nutr. 19: 296, 1966.
19. Johnson, N.E., E.N. Alcantara and H. Linkswiler. Effect of level of protein intake on urinary and fecal calcium and calcium and calcium retention of young adult males. J. Nutr .• 100: 1423, 1970.
20. Hegested, D.M., I. Moscoso and C. Collazes. calcium requirement of adult men. J. Nutr.
A study of the minimum 46: 181, 1925.
21. Knapp, G.L. Factors influencing the urinary excretion of calcium in normal persons. J. Clin. Invest. 26: 182, 1947.
22. Hawks, ,:1.E., M.M. Bray, M.O. Wilde and M. Dye. The interrelation-ship of calcium, phosphorus and nitrogen in the metabolism of pre-school children. J. Nutr. 24: 283, 1942.·
23. Hall, T.C. and H. Lehman. Experiments on the practicability of increasing calcium absorption with the protein derivatives. Bibchem. J. 3.8: 117,' 1944.
24. Kunerth, B.L. and M.S. Pittman. A long time study of nitrogen, calcium and phosphorus metabolism on a low protein diet. J. Nutr. 17: 161, 1939.
25. Pittman, M.S. and B.L. Kunerth. A long time study of nitrogen, calcium and phosphorus metabolism on a low protein diet. J. Nutr. 17: 175, 1939.
39
26. Wasserman, R.H., C.L. Comar.· and M.M. Nold. The influence of amino acids and otherorganic compounds on the gastrointestinal absorption of calcium45 and strontium89; J. Nu tr. 59: 371, 1956.
27. llegsted, D.M. Present knowledge of calcium, phosphorus and magnesium. In: Present.Knowledge in Nutrition. New York: The Nutrition Foundation, Inc .• , 1967 Chapter XXX:I, p. 147.
28, Price, N.O. and G.E. Bunce, Effect of nitrogen and calcium on balance of copper, manganese, and zinc in preadolescent girls. Nutritional Reports International. 5: 275, 1972.
29. Price, N.O., G.E. Bunce and R.W. Engel. Copper, manganese and zinc balance in preadolescent girls. Am •. J. Clin. Nutr. 23: 258, 1970.
30, Engel, R.W., N.O. Price and R.F, Miller. Copper; manganese, cobalt and molybdenum balance in preadolescent girls. J. Nutr; 92: 197, 1967.
31. Lang, V .M., B.B. North and L.M. Morse. Manganese metabolism in college women consuming vegetarian d:i,ets. J. Nutr. 85: 132, 1965. .
32. North, B.B,,. J.M. Leichseuring and L.M. Norris. Manganese metabolism in college women. J. Nutr. 72: 217, 1960. ·
33. Emerson, G.J. and A.L. Daniels .. A study of manganese retentions in children. J. Nutr. 8: 497, 1934.
34. Hartman, R.H., G. Matrone and G.H. Wise. Effect of high dietary manganese on hemoglobin formation. J. Nutr. 57: 429, 1955.
35. Klavins; J.V., T.D. Kinney and N. Kaufman. The influence of dietary· protein on iron absorption. Br. J. Exper. Path. 43: 172, 1962.
36. Lahey, M.E., M. Behar, F. Viteri and N.S. Scrimshaw. Values for copper, iron and iron"'binding capacity in the serum inkwashiorkor. Pediatrics. 22: 72, 1958.
37. Abernathy, R.P., J. Miller, J. Wentworth and M. ·speirs. Metabol:i.c patterns in preadolescent children. XII. Effect of amount and source of dietary protein on absorption of iron. J. Nutr. 83: 265' 1965.
38. Johnston, F .A. Iron requirements of children. J. Am. Dietet. A. 29: 758, 1953.
40
39. Cartwright, G.E. and M.M •. Wintrobe. The question of copper defiCiency in man. Amer. J. Clin. Nutr. 15: 94, 1964.
40. Magee, A.C. and G. Matrone. Studies on growth, copper metabolism and iron metabolism of rats fed high levels of zinc. J. Nutr. 72: 233; 1960.
41. Forbes, G. B. Sodium. In Mineral Metabolism. C. L. Comar and F. Bronner (eds.), Academic Press, New York (1962), Vol. 2, Part B, Chap. 25.
42. Widle, W.S. Potassium. In Mineral Metabolism, .C.L. Comar and F. Bronner (eds.), Academic Press, New York (1962), Vol. 2, Part B, .Chap. 25.
APPENDIX 1:
Wet ashing procedure for food and fecal samples using
nitric., perchloric acids and hydrogen peroxide
1. Using .3 grams of fecal samples and 10 grams of food sample, add . . . I
15 mls of nitric (HN03) acid in a covered beaker, begin reduction
on a hot plate at 300 degrees centigrade. Reduce to dryness.
2. Ad.ding 15 mls of HN03 at a time,. reduce .until a white ash is formed.
The sample will go from a black ash to a brown crust to a yellow
crust to a white ash. · This indicates removal of fat.
3. Add 10 mls of nitric and 5 mls of 30% hydrogen peroxide till a
pure white ash is obtained.
4. Add 5 mls of HN03 and keep in the hot plate for 2 or 3 minutes.
Remove from the hot plate and add slowly 3 mh of perchloric acid
(HCL04). Replace on the hot plate and reduce to dryness. Repeat
until a white crystal is formed on the bottom of the beaker. ,
5. Dissolve crystals with boiling deionized water and 3 mls of
hydrochloric acid (HCL). Transfer contents to a 25 nil volumetric
flask and d:i.lute to volume into deionized water.
5. Dilute to acceptable concentrations.
41
APPENDIX 2: _
Procedure for wet ashing urine with nitric (HN03) .acic}. and 30 percent hydrogen peroxide
1. Evaporate 50 mls of c}.ilute urine with 20 mls of HN03 to dryness ..
Place on a hot plate until a black ash is obtained.
2. Add 20 mls of HN03 and swirl until the black ash dissolves. Cover
with a water glass and put back on the hot plate. A brownish
crust will form.
3. Add 20 mls of HN03 , dissolve crust evaporate on hot plate. Repeat
this step until a white ash with a slightly yellowish tinge is
obtained.
4. To the white ash add 15 ml of HN03 and 5 ml of 30 percent hydrogen
peroxide slowly, Add as much hydrogen peroxide as needed to obtain
a white crystal. A clear colorless liquid precedes the formation
·of the white crystal.
5. To the white crystal add l:>oiling deionized water and 3 ml of HCL.
Wash the sides of the beaker with deionized water and dissolve the
crystals.
6. Pour into a 25 rnl volumetric flask and dilute to volume. Dilute
to desired concentrations.
42
Subjects
100
101
102
103
104
105
106
107
108
109
llO
111
112
113
114
43
APPENDIX III
Starting and ending weights and heights of the subjects
, : 'Height· '.in cm Period I Period IV Subjects Day I Day II
132.0 135 100
143.5 144.5 101
133.5 134.5 '102
128.5 128.5 103
128 127.5 104
133 133.5 105
139 140 106
125.5 126 107
123.5 124.5 108
137.5 138 109
133 133 110
139 140 111
127 128 112
137 138 .5 113
p6.5 128 114
Weight in kg Period I Period IV Day I Day II
.31.4 32
39.5 38.4
25.8 26.6
25.6 26;3
24.1 24.8
31.0 '30.6
26.6 27.6
23.0 ' 23. 6
23.4 23.2
32.1 32.3
32;8 33.8
31.1 31.8
24.2 24.4
38.2 36.9
24.4, 27.9
Protein Level
30 g
60 g
90 g
44
APPENDIX 4
Daily mineral balance:* magnesium balance in preadolescent girls
consuming three levels of protein (Period II) (mg)
Subject No. Urinary Fecal Total Intake
100 118. 72 191.38 310 .10 176.10
101 137.47 135.56 273.03 176.10
102 115. 60 107.70 223.30 176.10
103 109.35 127.70 237.05 176.10
104 106.22 95.43 201.65 176.10
105 124.97 148.57 273,54 278.14
106 93. 72 143.14 236.86 278.14
107 112; 47 113. 98 226.45 278.14
108 93. 72 135.52 229.24 278.14
109 131. 22 153.31 284.53 278.14
llO 124.97 247.66 372. 63 345.06
111 156.22 261. 93 418.15 345.06
112 131. 22 165.09 296,31 345.06
113 137.47 210.83 348.30 345.06
114 124.97 213.68 338.65 345.06
* calculated on period basis
Balance
-134.00
-96.93
-47.20
-60.95
-25.55
4.60
41.28
51.69
48.90
-6.39
-27.57
-73.09
48. 75
-3.24
6.41
Protein Level
60 g
90 g
30 g
45
APPENDIX 5
Daily mineral balance:* .magnesium balance in preadolescent girls
consuming three levels of protein (Period III) (mg)
Subject No. Urinary Fecal Total ~ Intake
100 124.96 191.38 316. 34 226.29
101 70.25 153.39 223.64 226.29
102 132.78 131.51 264.29 226.29
103 117 .16 134.53 251. 69 226.29
104 119.34 138 .16 257.50 226.29
105 105.44 134.79 240.23 280.16
106 128.87 164.22 293 .09 280.16
107 121.84 137.65 259.49 280.16
108 97.62 176.66 274.28 280.16
109 85.91 174.64 260.55 280.16
110 78.09 98.59 176.68 17 5. 06
lll 93.72 87.31 181.03 175.06
112 93. 72 71.31. 165.03 175.06
113 123.41 85.03 208,44 175.06
114 171.84 96.61 268.45 175.06
* calculated on period basis
Balance
-90.05
2.65
-38.00
-25 .40
-31. 21
39.93
-12.93
20.67
5.88
19.61
- l. 62
:..s. 97
10.03
M33,38
-93.39
Protein Level
90 g
30 g
60 g .
46
APPENDIX 6
Daily mineral balance:* magnesium balance in preadolescent girls
consuming three levels of protein (Period IV) (mg)
Subject No. Urinary Fecal Total Intake
100 91.22 191.38 282.60 239.59
102 74.97 403.37 478.34 239.59
103 122.47 163.15 285.62 239.59
104 50.00 . 171.07 221.07 239.59
105 84.35 93.54 177 .89 162.97
106 50.00 61.60 111.60 162.97
107 74.97 154.35 229.32 162.97
108" 37.50 114.34 151.84 162.97
109 78.10 92.61 170 .. 71 162.97
·110 94.97 118. 88 213.85 303.88
111 124.97 119.61 244.58 303.88
112 109 .35 109.64 218.99 303~88
113 91.22 127.43 218.65 303.88
114 87 .47 121. 35 208,82 303.88
* calculated on period basis
Bala.nee
-43.01
-2,38.75
-46.03
18.52
-14. 92
51.37
-66.35
11.13
- 7. 74
90.03
59.30·
84.89
85.23
95.06
Subject Day Number 6
100 19.50
101 32.50
102 43.00
103 57 .oo 104 22.00
105 118. 72
106 103. 72
107 74.97
108 74.97
109 93. 72
110 97.47
111 118. 72
112 153.40
113 143. 72
114 93. 72
47
APPENDIX 7
Daily urinary loss of magnesium for period II
(mg)
Day Day 7 8
106.22 124.97
75.60 53.11
112.47 50.61
81.22 78.11
87.47 62.47
193.72 112 .47
187.48 122.47
124.97 68.74
87.47 87.47
97 .47 156.22
62.50 87.47
.187.47 143.72
150.00 68.74
137.50 137.47
99.97 143.72
Day Day 9 10
148. 72 42.50
78.74 36.00
149.97 52.00
134.35 88.00
112 .47 49.50
109.97 99.80
84.99 174.97
137 .47 78.73
140.60 87 .48
109.35 91.22
93. 72 103.10
134.35 162.47
153.10 137.47
124.97 137.47
118. 72 125.60
Protein Level
30 g
60 g
90 g
48
APPENDIX 8
Daily mineral balance:*. , calcium balance in preadolescent girls
consuming three levels of protein (Period II) (mg)
Subject No. Urinary Fecal Total Intake
100 32.13 1089.65 1121. 78 2377. 77
101 74.98 1112. 61 1187 .59. 2377~77
. 102 40.69 1216.87 1257. 56 2377. 77
103 57.12 1215.52 1272. 64 2377. 77
104 84.27 926.07 1010.34 2377. 77
105 223.20 859.37 1082 •. 57 1278 .88
106 39.98 1020.49 1060.47 1278.88
107 196.42 944.02 1140.44 1278.88
108 48.55 942.48 991.03 1278.88
109 142.84 975.96 1118 .80 1278.88
110 160. 71 1146.11 1306.82 1365.98
111 '160. 71 1484.23 1644.94 1365.98
112 142.85 902.07 .1044. 92 1365.98
113 339.27 982.14 1321.41 1365.98
.. 114 171.42 1038.99 1210.41 1365.98
* calculated on period basis
Balance
1255.99.
1190 ,18
1120. 21 .
1105 .13
1367.43
196.31
218.41
138.44
287.85
160.08
59.16
-278.96
321.06
44,57
155~57
Protein Level
60 g
90 g
30 g
49
APPENDIX 9
Dailymineral balance:* ·calcium balance in preadolescent. girlS
consuming three levels of protein (Period HI) (mg)
Subject No. Urinary Fecal Total Intake
100 35.70 1089. 65 1125.35 1074~35
101 35. 70 852.65 888.35 1074.35
102 39.27 792.60 831.87 1074.35
103 64.26 878.76 943.02 1074.35
104 51.05 832.57 883.62 1074.35
105 128.52 443.28 571.80 1073.85
106 60.47 779. 96 840.43 1073 .85
107 99. 95 . 617.37 717. 32 1073.85
108 64.26 801.33 865.59 1073.85
109 46.40 759.75 806.15 1073.85
110 46.41 679. 37 725. 78 1220.89
111 28.56 783.67 812.23 1220,89
112 42.84 678.92 721. 76 1220.89
113 137.44 621. 93 759.37 1220.89
114 67.82 762.18 830.00 1220.89
* calculated on period basis
Balance
-51.00
186.00
242.48
131.33
190.73
502.05
233.42
356. 53
208.26
267.70
495.11
408.66
499 .13
461.52
390.89
Protein Level
90 g
30 g
60 g
50
APPENDIX 10
Daily mineral balance~'r calcium balance in preadolescent girls
consuming three levels of protein (Period IV) (mg)
Subject No. Urinary Fecal Total Intake
100 44.64 1089.32 1133. 96 1098.60
102 35. 71 1885.99 1921. 70 1098.60
103 104.64 813.10 917.74 1098.60
104 53.57 930.16 983.73 1098.60
105 104.64 784.36 889.00 1141. 86
106 35. 71 804.05 839.76 1141.86
107 68.93 879.34 948.27 1141.86
108 17.86 1064.04 1081.90 1141.86
109 68.93 792.07 861.00 1141. 86
:110 68.93 724.23 .793.16 1403. 72
111 65.35 998.51 1063.86 1403. 72
112 65.35 954.16 1019~5.l 1403.72
113 149.28 936.08 1085.36 1403.72
114 68.93 907.89 976.82 1403. 72
* calcblated on p~riod basis
Balance
-35.36
-823.10
180.86
114.87
252.86
302.10
193.59
59.96
280.86
610.56
339.86
384.21
318.36
426.90
.. 51
./
@"-·
APPENDIX 11
Daily urinary loss of calcium for period. II
(mg)
J .:!
~-5 Subject Day Day Day Day Day ~ Number 6 7 8 9 10 ( '-t
i i·. .,_, 100 14.26 196.38 392.80 62.46 16.07 ~-
101 35.68 71.42 32.14 124.95 89.28
102 20. 68 . 178.53 223.17 67.82 24.28 :.i ~ 103 48.54 374.96 53.57 124.96 58.57
104 57.83 133.88 71.42 178.53 56.78
105 223.20 446.37 133.89 205.31 157.10
106 267.84 71.42 40.16 276.74 80.32
107 . 133 .92 142.81 121.39 214.24 116.03
108 53.57 93.71 98.17 . 103 .53 53.53 .
. 109 196.42 151.74 107.10 187.45 160.67
110 214.27 80.34 133.87 151.74 160.67 •.'
~: 111 178.56 166.03 196.38 178.53 223.16
~; 112 178.56 142.85 89.26 182:99 178.52
'· 113 481.76 607.11 357.08 303,52 441.90
114 205.31 214.26 f 160.67\ 214.24 258.89
-""··
.}
Protein Level
30 g
60 g
90 g
52
APPENDIX 12
Daily mineral balance:* manganese balance in preadolescent girls
·consuming three levels of protein (Period II) (mg) . .
Subject No. Urinary. Fecal Total -Intake
100 0.003 3.08 3.08 3.51
10.1 o.03 3.22 3.25 3.51
102 0.03 3.62 3.65 3.51
103 0.01 3.99 . 4.00 3.51
104 0.03 2. 72 ..
2.75 3.51
105 0.05 1.69 1. 74 2.47
106 0.01 2.20 2.21 2.47
107 0.07 1.90 1. 97 2.47
108 0.01 1.49 1.30 2.47
109 O.Ql 1. 75 1.76 2.47
110 0.03 2.93 2.96 3.17
111 o.o4 3.06 3.10 3,17
112 0.04 1.94 1.98 3.17
113 .. 0.04 2.93 2,97 3.17
114 0.04 3.00 3.04 3 •. q
* cdculated on period bas.is '
Balance
0,43
0.26
-0.14
-0.49
0.76
0.73
0.26
0,50
Q.98
o. 71
' o. 21
Q.07
1.19
0.20·
0.13
Protein Level
60 g
90 g
30 g
53
APPENDIX 13
Daily mineral balance:* manganese balance in preadolescent girls
consuming three levels of protein (Period III) (mg)
Subject No. Urinary Fecal Total Intake
100 o.oo 3.08 3.08 2.00
101 o.oo 1. 65· L65 2.00
102 o.oo 1.81 1.81 2.00
103 o.oo 1. 99 . 1. 99 2.00
104 0.00 1. 75 1. 75 2.00
105 o.oo 1.38 1.38 2.51
106 0.00 2.17 2.17 2.51
107 o.oo 1. 7 1.7 2.51
108 o.oo 2.06 2.06 2.51
109 o.oo 2.20 . 2. 20 2.51
110 0.00 2.20 2.20 2.87
111 0.00 2.52 2.52 2.87
112 0.00 2.35 2.35 2.87
113 0.00 2.38 2.38 2.87
114 o.oo 2.49 2.49 2.87
* calculated on period basis
Balance
-1.08
0.35
0.19
0.01
0.25
1.13
0.34
0.81
0.45
0.31
0.67
o. 35 .
0.52
0.49
0.38
:Protein Levei
90 g
30 g
60 g
54
.APPENDIX 14
Daily mineral balance:* manganese balance in preadolescerit girls
~onsuming three levels of protein (Period IV) (mg)
Subject No. Urinary Fecal Total Intake
100 0.01 3.08 3.09 1.95
102 0.02 4.14 4.66 1.95
103 0.01 2.32 2.33 1. 95
104 0.00 2.35 . 2. 35 1.95
105 0.00 2.35 2.35 2.74
106 o.oo 1.75 1. 753 2.74
107 0.003 2.03 2 •. 033 2.74
108 0.00 2.94 2.94 2. 74
109 0.004 2.34 2.34 2.74
llO 0.005 1. 73 1.735 2,81
111 0.003 1.93 1.933 2.81
112 0.00 1. 74 1.74 2.81
113 0.00 2.04 2.04 2.81
114 0.00 1.60 1.60 2.81.
* calculated on period basis
Balance
-1.14
-2.71
... o .. 38
.,.0.40
0.39
0.99
0,70
-0.20
0.40
1.07
0.88
1.07
-0. 77.
1.21
Subject . Number
100
101
102
103
104
105
106
107
J.08
109
110
111
112
113
114
Day 6
0.012
0.025
0.010
0.025
0.053
0.050
0.057
0.017
0.017
0.028
0.014
0.00
0.014
0.10
0.042
55
APPENDIX 15
Daily urinary lo.ss of manganese for period II
(mg)
Day Day 7 8
0.043 0.043
0.036 0.036
0.107 0.086
0.086 0.021
0.043 0.021
0.057 0.021
0.029 0.036
0.029 0.050
0.071 0.078
0.036 0.014
0.047 0.036
0.064 0.071
0.099 0.043
0.086 0.014
0.043 0.043
·----------------·-·
Day Day 9 10
0.032 0.021
0.014 0.029
0.039 0.029
0.061 0.021
0.047 0.053
0.057 0.021
0.086 0.032
0.025 0.018
0.007 0.029
0.057 0.071
0.043 0.050
0.039 0.086
0.043 .0.043
0.043 0.086
0.064 0.029
• J
. ·3 :-: t,~
'.~ ';.'."
~ 2; .... ·, ;'.;:
;~ ·~
~t
~( ., -1:.
Protein Level
30 g
60 g
90 g.
..:.·.-.:· · .. ·'· -.
56
I
APPENDIX 16
. .·. :Qaily mineral ba!anc~:* , iron balance in preadolescent girls
c()nsuming .three level.s of protein (Perio.d II) (mg) . . .
Subject No. Urinary Fecal Total .. Intaite
·.
100 0.28 29.14 29.42 3L55
101 0.19 29.23 29.42 31.55
102 o.o 31.25 31..25 31.55
103 o.13 34.90 35.03 31.55
104 0.19 27.01 27.20 31 .• 55
105 0,38 28.50 28~88 32·.33
106 0.07 33.34 33.41 32,33
107 0.38 28.24 28.62 32.33
108 0.09 25.53 25.62 32.33
l09 0.3.8 34.5 34.73 32.33
110 0'.12 34.41 34.53 35.50
111 o.2s 35.70 35.95 35.50
112 0 .13 32.08 32.21 35.50
113 0.44 33.80 34~24 3s.so 114 0.19 31.62 31 ~81 35.50
* ~alculated on l'eriod basis
"· / _/
Balari.ce
2.t3
2.13
Q.;30 ' .• .:.).48
4~35
3.45 ..
-1.08
3.71
6 .• '71
.. 2.40.
0.97
..Q.45
3~29.
1.26
3.88
: ' .. ·,.
:-'•:.
·. 57·
'·
APPENDIX l.7
. Daily mineral balance:* iron bS,lance in pteadoiescent girls .
consuming three levels of protein (Period III) . (mg) .
Protein Subject .. Level No.; Urinary ·Fecal Total ·:rntake Balance ·
60 g 100 0~88 29 .. 14 30.02 29.41 -0.61
101 0.31 27.23 . 27 .54 29.41· 1,:,87
102 0.28 29.73 30.01 29.41 -0.60.
103 0.47 26.84 27.31. 29.41 2.10.
104 0.38 28.66 29.04 29.41 o.37
90 g 105 0.47 19.30 19.77 28.98 9.21
106 0.25· 27.87 28.1;2 .28.98 o.86
107 0.16 16. 72 16.88 28.98 12.10
108 0.25 24.84 25.09 28.98 3.89
109 . 0~20 24.26 24,46 28.98 4~52 ..
30 g 110· 0.18 24.17 24.35 29.61 . 5.26
111 0.19 17.05 · 17 .24 29 ~61 .. 12.37
112 0 .03. 19.54 19.57 29.61 10.04
113 0.38 17.07 17.45 29.61 12.16
114 0 .5 0 18.80 19.30 29.61 10.31 ·
. ; * calculated on period basis
58
APPENDIX 18
Daily mineral balance:* iron balance in preadolescent girls
)] - consuming three levels of protein (Period IV) (mg)
.'~ '
Protein Subject Level No. Urinary. Fecal Total Intake Balance
90 g ~~
100. 0.31 29.14 29.45 31.24 1. 79 .
102 0.21 66.10 66.31 31.24 ~35 .07 ..
103 0.31 23.35 23 .• 66 . 31.24·· 7.58
104 0,38 32.45 32.83 31.2·4 -1.59
30 g 105 0.25 ' 28. 71 28.96 29.85 0.89
106 0.46 23.51 . 23. 97 29.85 5.88 .·
107 0.41. 26.34 26.75 29.65 3.10
108 o.o 0 30.05 30.05 29.85 -0.2
109 0.38 26.14 26.52 29.85 3.33 .3: ;,
60 g ,110 o.59 21. 79 22.38 33 ,33 . 10. 95.
111 0.53 27.22 27.75 33.33 5.58
112 0.50 21. 73 22.23 33.33 11.10 .;
113 0.38 26.11 26.49 33~33 6.84
114 0.34 27.15 27 .49 . 33.33 5.84.
:.:,
.* caleulated on period basis
Subject Day No. 6
100 0.188
101. o.ooo
102 0.250
103 0.125
104 . o. 281
105 0.125
106 0.175
107. 1.000
108 0.113
. 109 0.000
110 0.188
111 0.016
112 0.219
113 0.063
. 114 0,031
59
APPENDIX 19
Daily urinary 16ss of iron for Period II
(mg)
Day Day 7 8
1.37 0.281
0.125 0.25
o .. 113 0.281
0.094 0.063
0.063 0.109
0.313 0.031
0.313 0.250
0.156 0.375
0.313 0.375
0.125 0.875
0.031 0.438
0.438 0.438
0.563 0.125
0.563 0.250
0.188 0.438
Day Day 9 10
0.812 1. 375.
0.875 0.438
0.625 0.500
0.438 0.406
0.813 0.750
0.25
0.50 0.500
0.375 0.375
0.250 0.313
0.563 0. 750.
0.563 o.469
0.348 o.781
0.375 o.344
1.875 o.438
0.875 o.5oo
60
APPENDIX 20
. Paily mineral balance:* . . . . . . ' . . copper balance in pread(>lescent; girls
consuming three levels of protein· (Period, II)· (mcg)
Protein Level
60 g
90 g
30 g
61
APPENDIX 21
Paily mineral balance:* copper balance in preadolescent girls
consuming three levels of protein (Period III) (mcg)
Subject No. Urinary Fecal Total Intake
100 126.54 1070.74 1197.28 930.49
101 93.20 1069.59 1162. 79 930.49
102 93.24 992.88 1086 .12 930.49
103 74.93 1039.69 1114 .62 930.49
104 83.25 956.50 1039.26 930.49
105 80.87 843.30 924.17 1001. 95
106 74.93 1011. 63 1086.56 1001;95
107 59.94 869.38 929.33 1001.95
108 63.27 1079.69 1142. 96 1001.95
109 53.28 1108. 97 1162.25 1001. 95
110 63.27 1109 .60 1172.87 957.20
111 53.28 996.82 1050.10 957~20
112 59.94 1046.25 il06,19 957.20
113 74.93 856.39 931.22 957.20
114 66.6 822.19 888.79 957.20
*calculated on period basis
Balance
-226.79
-232.30 '
-155.63
-184.13
-109. 77
77. 78
-84.61
72.62
-141.01
-160.30
-215.67
-92.90
-148.99
25.88
68.41
. \
~··
.· ~1 '~ . . ~~ ..
'.''.'
1 90 g
.,.;. .
~ 30 g
62
APPENDIX 22
Daily mineral balance:* copper balance in preadolescent girls
consuming three levels of protein (Period IV) (mcg)
· Subject No.
100 .·
102
103
104.
105
106
107
108
109
Utina:ry
58.28
53.28
59.94
33.30
83.21
76.59
66.60
53.28
.83. 25
Fecal Total Intake
1070. 74 1129.02 729. 72
1653.39 1706,67 729. 72
1032.10 1092.04 729. 72
1046.62 1079.92 729. 72
858.32 . 941. 53 1122.19
711.06 787.65 1122.19
854.09 920. 6'9 1122.19
913.62 966 •. 90 . 1122,19
877. 53 960.78 1122.19
60 g 110 56.61 730.81 '787.42 1081.38
111 66.37 824.03 890.40 1081.38 ..
112 66.60 811.30 877.90 1081.38 .;.~ 113 .83.25. 876.56 ·959.81 1081.38
114 58,63 794,39 853,02' 1081.38
; * calcuiated Qn period basis
.(
Balance
;_399. 30
-976 •. 95'
... 362.32
-350;20
180.66
334.54
201.50
155.29
161.41
293.96
190.98
203.48
121.57
228.36
Subject Day Number 6
100 133.20
101 59.20
102 141.53
103 126.53
104 74.93
105 158.18
106 199.80
107 74.93
108 266.40
109 66.60
110 59.94
111 66.60
112 66.60
113 82.25
114 59.94
63
APPENDIX 23
Daily urinary loss of copper for period II
(mcg)
Day Day 7 8
133.20 91.58
78.56 104.89
74.93 56.61
63.27 83.25
66.60 91.58
126.54 66.60
109.89 63.27
116 .55 59.94
. 59.94 74.93
91.58 99.90
74.93 149.85
66.60 133.20
74.93 133.20
83.25 116 .55
66.60 141.53
Day Day 9 10
166.54 76.59
133.20 96.52
141.53 149.85
166.50 . 78.06
106.56 166.50
91. 575 108.23
133.20 133.2
86.58 73.26
109.89 149.85
91. 575 99.90
106.56 186.48
73.26 115. 62
109.89 99.90
158.18 89.91
149.85 74.93
:r.
... 64
APPENDIX 24 · ... ·•
. Daily mineral balance:* . zinc . balance in preadolescent girls
consuming three.· levels of protein (Period II) (mg) .
. cy,;.,,:l, .•··
i ....... ~ ............... ,.........~ ....... -----~..--....,..._,.......,.......,.......,.......,.......,.......,...;.__,..~....,...~~~....,...__,..__,.. __ ....,...~ Protein
;:~ ,, Level
30 g
60 g·
90 g
'·~ ~' .
Subject No.
100.
101
102
103
104
105
106
107
108
109
110
111
.112
113
114
Urinary
o. 71
0.78
0.88
0.62
0.64
0.60
0.40
1.18
0.64
1.07
1.10
0.66
1.22
2.02
'3.54
: * calcµlated on period basis
Fecal Total Intake Balance
8.56 . 9'.27 4,77 -4.50
9.43 10.21 4. 77 .-5.44
8•87 9.75 4. 77 .;.4.98
7.60 8,22 4.77 -3.45 .
7. 77 8.41 4. 77 -3. 64 .
9.70 10.30 7.75 ,;.2.55
9.35 9.81 7.75 .;2.06
7.23 8.41 7.75 -0.66
8.02 8.66 7,75 -0.91
8.50 9,57 7.75 -1.82
14.03 15.13 11.61 ~3.52
13.70 14.36 11.61 .. 2. 75
.10.10 11.32 11. 6.1 0.29
12.10 14.12 11.61 -2.51
10.84 14.38 ll.61 -2.77
· .... '
Protein
65
APPENDIX 25
Daily mineral balance:* zinc balance in preadolescent girls
consuming three levels of protein (Period III) (mg)
Subject Level No. Urinary Fecal Total Intake
60 g 100 0.84 8.56 9.40 6.87
101 o. 74 7.03 ·7. 77 6.87
102 0.84 7.07 7.91 6.87
103 0.74 7.92 8.66 6.87
104 0.60 5.83 6.43 6.87
90 g 105 0.68 7.79 8.47 9.53
106 0.79 9.88 10.67 9.53
107 0.47 8.19 8.66 9.53
108 0.43 8.51 8. 9.4 9.53
109 0.40 10.62 11.02 9.53
30 g 110 0.65 6.21 6.86 4.13
111 0.56 3.33 3.89 4.13
112 0.54 4.95 5.49 4.13
113 0.95 5.51 6.46 4.13
114 0.94 5.29 6.23 4.13
* calculated on period basis
Balance
-2.53
-0. 90
-1.04
-1. 79
.44
1.06
-1.14
0.87
0.59
. -1.49
-2.73
0.24
-1.36
-2.33
-2.10
Protein
66
APPENDIX 26
Daily mineral balance:~• . zinc balance in preadolescent girls
· consuming three levels of protein (Period IV) (mg)
Subject Level No. Urinary Fecal Total Intake
90 g 100 0.34 8.56 8.9 6.35
102 0.41 20.15 20.56 6.35
103. 0.28 9.27 9.55 6.35
104 0.38 10.55 10.93 6.35
30 g 105 0.50 4.46 4.96 4.23
106 0.32 3.58 3.90 4.23
107 0.38 4.41 4. 79 4.23
108 0.20 6.12 6.32 4.23
109 0.20. 4.41 4.61 4.23
60 g .110 0.40 5.66 6.06 10.03
'111 0.41 6.89 7.30 10.03
112 0.94 5.98 6.92 10.03
113 0.70 6.02 6. 72 10.03
114 0.42 6.06 6.48 10.03
* calculated on period basis
Balance
-2.55
-14.21
-3.2
-4.58
-0.73
0.33
-0.56
-2.09
-0.38
3.97
2.73
3.11
3.31
3.55
Subject Day Number 6
100 0.34
101 0.08
102 0.45
103 0.19
~ 104 0.59
105 0 .. 44
106 0.49
107 0.23
108 0.47
109 0.55
110 0.52
111 0.44
112 0.68
113 0.68
114 0.49
67
APPENDIX 27
Daily urinary loss of zinc for Period II
(mg)
Day Day 7 8
0.52 0.56
0.58 0.56
0.65 0.46
0.38 0.28
0.43 0.58
0.88 0.58
1.12 0.56
0.60 0.28
0.80 0.44
0.68 0.52
0.54 0.78
0.84 0.60
1.10 1.36
0.80 0.68
0.82 o. 72
Day Day 9 10
0.64 0.37
0.76 0.24
0.64 0.48
0.68 0.46
0.52 0.84
0.60 0.76
0.64 1.28
0.60 1.16
0.42 0.52
0.64 0.42
0.64 o. 72
0.59 0.88
0.84 0.78
1.40 0.73
0.62 0.90
Protein Level
. 30 g
60 g
90 g
68
APPENDIX 28
Daily mineral balance: ·k sodium balance in preadolescent girls
consuming three levels of protein (Period II) (mg).
Subject No. Urinary Fecal Total !ntake
100 2310.55 12.11 2322.66 2216 .46 .
101 3374.05 95.23 3469.28 2216.46
102 2217.00 15.20 2232.20 2216.46
103 1966.80 17 .67 1984,47 2216.46
104 2060.55 19.16 2079.71 2216.46
105 2623.05 51. 78 2674.83 2955 .71
106 2529.30 28.22 2557.52 2955. 71 •
107 2310.55 40.86 2351 •. 41 2955.71
108 2810.55 24.60 2835 .15 2955. 71
109 2998.05 13.15 3011. 20 2955. 71
HO . 3123.05 12.22 3135. 27 2955.71
lll 3554.85 38. 33 . 3593.18 . 2955. 71
. l12 3404.30 23.65 ·3427,95 4265.33
113 3779. 30 17.33 3796.63 2955. 71
l14 4123.05 25.70 4148. 7 5 2955. 71
~'(. calculated on period basis
Balance
.-106. 20
-1252.82
;.;.15; 74
231.99.
.136.75
280.88
398.19 .
604.30
120.56
-55.49
l130 .06
6 72 .15
837.38
468.70
l16. 58
Protein Level
60 g
90 g
30 g
.-.,l.''
. 69
APPENDIX 29 .
Daily mineral balance:* solfiuni. balance in preadolescent girls ..
consuming three levels of protein (Peribd III) (mg)
subject No. Urinary Fecal ·Total lnt:ake
100 2998 .. 05 12.11 3010.16 2815.21
101 2498.05 77.09 2575.14 2815.21
102 3529.30 31.98 3561. 28 : ·2815 .21
103·. 2685.62 35.83. 272L45 2ais~.21.
104 2873.05 68.41 2941.46 2815.21
105 3328.05 25.41 3353.46 3476.48
106 3544.93 f17;91 3662.84 3476.49
107 3341 ~80. 16.19 ~357. 9'9. 3476.48
108 3435.55 40.41 3475.96. 3476.48 :
..
109 2996.80 15 .11 .3011.91 3476.48.·
110 1841.80 21. 2.2 1S63,03 2251.78
111. 1841~80 27 .94 1869;74 . 2251.78
112 1701.18 40.17 1741.35 ·2251.78
113 2060.55 "21. 65 2082.20 2251 .. 78
.114· 3310.55 64;59 3375.14 2251. 78
* ~alculat~d bn perio4 basi~
.Bil lance. ·.
..
.-194.95 :
·-240.07
•746.07
.···93.16 \.''
.. · ..... 126. 25
. 12.3.02 ....
-186.36 :
, . ...
118.49 '
: ~0.52 .·
464~57
38S.76 .
382.03 : . •·,'
510A3
1~9.5'8 ..
.,.1123 ~ 36
·~ .. '. .
. ' .. ,·.:'•'
Protein Level
90 g
30 g
60 g
70
APPENDIX 30 ',.
Daily mineral balance:* sodium balance in preadolescent girls
consuming three levels of protein (Period IV) (mg)
Subject No. Urinary Fecal Total Intake
100 2966.80 12.11 2978.91 2724.58
102 3498.05 104.70 3602.75 2724.58
103 3841. 80 31.81 3873.61 2724.58
104 1966.80 28.10 1994.90 2724.58
105 2560.04 17.88 2577.92 2003.13
106 1310.55 84. 75 1395.30 2003 .13
107 1905.30 52.26 1957.56 2003.13
108 1248.05 64.65 1312.70 2003.13
109 2123.05 13.58 2136.63 2003.13
110 2904.30· 31. 71 2936.01 3505.40
111 3237.01 45.57 3282.58 3505.40
112 2685 .55 39.79 2725. 34 3505.40
113 2748. 05 32.07 2780.12 3505.40
114 2504.34 118.58 2622.92 3505.40
* calculated on period basis
Balance
-254.33
-878.17
-1149.03
729.67
-547.79
607.83
45.57
690.43
-133.50
569.39
222.83
780.06
725. 29
882.49
Subject Day No. 6
100 . 622.99
101 1154.25.
102 1185. 50
103 1966. 75
104 810.50
105 2841. 75
106 1560.50
107 2404.25
108 2622.99
109 2685.50
1io 3466.75
111 2872.99
112 . 3404.25
113 . 3530. 25
114 . 3372.99
71.
APPENDIX 31
Daily urinary loss of sodium for Period II
(mg)
Day Day 7 8
2138.62 1435.50
1591. 75 1497.99
2685.50 599.56
1060.50 1248.01
2435.50 1529.25·
2498.00 2747.99
3810.50 2904.25
2591.75 1935.50
2341. 7 5 2405.25
1966. 75 2810.49
2435. 50 2341. 75
3966.75 2185. 50
2872.99 1622.99
1623.00 3279.25
3154.25 3122.99
Day Day 9 10
2966.75 1310.50
3060.50 1247.99
2748.00 1091.75
3091.75 1091. 75
3435 .50 .. 1716.75
3435.50 2544.87
3122. 99 3435.50·
3091. 7 5 2123.00
3060.50 2466.75 -
2872. 99
3310.50 2185,50
3997.99 3123.00
4310.50 3435.50
4060.50 3279.25
4060.50 3185.49
Protein Level
30 g
60 g
90 g
72
APPENDIX 32
Daily mineral balance:* potassium balance in preadolescent girls
consuming three levels of protein (Period II) (mg)
Subject No. Urina~y Fec::al Total Intake
100 1197.19 197.68 1394.87 2457.30
101 2454.81 544.68 2999.49 2457.30
102 1249.25 175. 04 2999.65 2457.30
103 1301.31 306.27 1607.58 2457.30
104 1301. 31 210.75 1512.06 2457.30
105 1874.00 344.48 2218.48 2998.45
106 1353.38 394.59 1747.97 2998.45
107 1093.06 ·298.56 1391 •. 62 2998.45
108 1821. 94 325.34 2147.28 2998.45
109 1874.00 382.17 2256.17 2998 .45
110 1561.63 412.81 1974.44 3351. 70
111 1974.96 462 .11 2437 .07 3351. 70
112 1665.75 407.95 2073.70 ·3351,70
113 2082. 25 . 301. 21 2383.46 3351. 70
114 2186. 38 482.28 2668.66 3351.70
* calculated on period basis
Balartce
1062.43.
- .542 .19
1033.01
849.72
945.24
779.97
1250.48
1606.83
851.17
742.28
1377 .26
9t4.63
1278.00 .
968.24
683.04
Protein Level
60 g
90 g
30 g
73
APPENDIX 33
Daily mineral balance: ·k potassium balan,ce in preadolescent girls
consuming three lev.els of protein (Period III) (mg)
Subject No. Urinary Fecal Total Intake
100 884.81 197.68 1082.49 3332.67
101 1145 .13 486.55 1631. 68 3332.67
102 728.63 268.59 997.22 3332. 67
103 676.56 238.82 915.38 3332.67
104 728. 63 122.70 851.33 3332.67
· 105 567.43 98.12 665.55 2844.34 '
106 624.50 156.76 781. 26 2844.34
107 624.50 60.62 685.12 ·2844. 34
. 108 416.25 162.. 21. 578.46 2844.34
109 520.38 144.93 665.31 2844.34
110 416.25 114 .45 530.70 2452 .. 33
111 416.25 68. 71 484.96 2452.33
112 312 .13 130.29 442~42 2452.33
113 468.31 40.91 509.22 2452.33
114 780.69 ·108.94 889.63 2452.33·
* ~alculated on period basis
Balance
2250.18
1700. 99.
2335.45
2417.29
248I.34
2178.79
2063.08
2l59.22 ..
2265.88
2179.03
1921.63
1967;37
2009.91
1943.11
1562.70'
Protein Level
90 g
30 g
60 g
74
APPENDIX 34
Daily mineral balance:* potassium balance in preadolescent girls
consuming three levels of protein (Period IV) (mg)
Subject No. Urinary Fecal Total. Intake
100 1353 .38 197.68 1551.06 2831.00
102 1665.75 605.49 2271. 24 2831.00
103 1821.94 433. 79 2255.73 2831.00
104 780.69 281. 31 1062.00. 2831.00
105 1405.20 241.31 1646.51 2252.00
106 728.63 224.69 953. 32 2252.00
107 1249.25 312.82 1562.07 2252.00
108 520,38 462.94. 983.32 ·. 2252.00
.109 1197.19 328.13 1525.32 2252.00
110 1874.00 270.04 2144.04 2808.38
111 985,58 521.19 1506. 77 2808.38
112 '1249. 30 . 472 .46 1721. 76 2808.38
113 1561. 63 320.54 1882 .17 2808.38
114 1374.80 464.88 1839.68 2808.38
* calculated on period basis
Balance
1279. 94
559.76
575.27
1769.00
605.49
1298.68··
689.93
1268.68
726. 68
664.34
. 1301 .. 61
.1086. 62
926 . .21
968.70
SubJect Nurn er
100
101
102
103
104
105
106
107
108
109
110
111.
112
113
114
75
APPENDIX 35
Daily urinary loss of potassium for period II
(mg)
Day Day Day 6 7 8
260.17 1510.17 1145.58
833.08 1614.34 1041.26
1718. 50 1770.58 833.08
1'666.41 551.83 1041.42
728.92 1301.84 1249.74
2499.74 1666.42 1666.42
2135.15 2083.08 1666.42
1041.42 1614.34 1666.40
1978.92 1666.42 1197.66
1093.50 1353.92 1666.41
1770. 58 1041.42 1145. 59
1978. 91 2395.58 1353.92
2187.24 1562.25 728. 92
2499.75 1249.76 1614.34
1562.26 1458.08· . 1}14. 66
Day 9
1666.42
1562.24
1510.16.
1614.33
1666.42
1614.33.
1666.42
1815. 62.
1458.08
1978.91
2030.99
2343.49
2030. 99
2395. 58
2187.24
' '
Day 10
1041.41
572 .66
1041.40
1093.49
987. 90
1510 .16
1874.74
1041.41
1249.74
1249.74
1770. 57
1458.08
1458.08
1770.57
1822.66
The vita has been removed from the scanned document
THE UTILIZATION OF ESSENTIAL MINERALS BY PREADOLESCENT.
GIRLS CONSUMING THREE LEVELS OF PROTEIN
by
Andal Ra.ghavan
(ABSTRACT)>
The effect of three levels of protein on the utilization of
magnesium, calcium, manganese, iron, copper, zinc, sodium and potassium
was studied in preadolescent girls. Minerals were determined in excreta
and food following a wet ash by absorption spectrophotometric procedures.
The minerals magnesium, calcium and manganese were better utilized with
low protein intakes. The utilization of copper, zinc, and sodium were
.enhanced on moderate protein intakes. A high protein diet was detri•
mental to the utilization of these minerals when present in low levels
but enhanced the utilization of copper, and did not affect sodium utili-
zation adversely, The effect of protein on iron al::isorption was not clear.
Protein level did not affect the potassium utilization.