: . ).

• : .-:---+-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.