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THE DIETARY PROCEDURES LEMF'LOYED
IN CGNDUCTlNG A NON- CUMCAL
METABOUC BALANCE STUDY

Thesis for the: Begs-'99 a? M. S.

.MlCHIGAN STATE UNIVERSITY

:Caron-n Mae Friedemann
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THESIS

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ABSTRACT

THE DIETARY PROCEDURES EMPLOYED IN CONDUCTING A
NON-CLINICAL METABOLIC BALANCE STUDY

by Carolyn Mae Friedemann

Non-clinical metabolic balance studies have long been employed to
evaluate the nutritional requirements of human beings. The dietary
procedures to be used in such studies have not been well defined. A
non-clinical metabolic balance study in which the dietary procedures
were evaluated was undertaken to investigate bread as a primary source
of protein for man. This paper concerns only the dietary procedures
involved in such a study. A later presentation by Miss S. Bolourchi
will discuss the clinical aspects of the study.

The subjects chosen for the experiment were twelve male college
students. The control period and the experimental period were twenty-
one and fifty-one days in length, respectively. The amount of protein
in both diets was limited to approximately 65 grams per day. During the
control diet small amounts of animal protein were served, whereas, in
the experimental regimen no animal protein was allowed. In the experimental
diet 90 to 95 per ceflzof the protein was derived from wheat products.

A seven day menu cycle was devised for each period. Tables of food
composition were used to calculate the nutrients in the diets. A diet
which was a duplicate of those served to the subjects was collected each

day for analysis. Nitrogen, fat, calories, and moisture were determined

in the dietary samples.

Ihe diets
pie. These in
during the ex:
foods such as
intake of thes
The amount of

The food
St‘de- All m:
for the study.
perishable 1:6
perishable ite

T0 be Ce:
0f the diet I}
Z‘éiority of t}

UPOD C03;
Yal‘JQS, ta.

Die:

itcurate for I

Protein Vaer:

200d gI’Oup . -

ESE analvzed ‘

‘-

The diets contained mixed dishes and desserts such as cake and
pie. These increased palatability and acceptability of the diet consumed
during the experimental period. To provide an extra source of calories,
foods such as hard candies were provided on an ad libitum basis. The
intake of these was permitted only as a supplement to the regular diet.
The amount of these foods consumed was measured.

The food purchased was of the same variety and brand throughout the
study. All non-perishable items were purchased in sufficient quantity
for the study. The perishable items were purchased biweekly. The non-
perishable items were stored in the dry storage areas, whereas, the
perishable items were stored in a refrigerated walk-in or a freezer.

To be certain that the diet collected for analysis was representative
of the diet the subjects consumed, the samples were collected when the
majority of the subjects were served.

Upon comparing the calculated values for the diets to the analyzed
values, tables of food composition were not reliable for fat. They are
accurate for protein. The nitrogen was calculated from the computed
protein values with the aid of the specific conversion factors for each
food group. The calculated values for nitrogen were within 5 per cent of
the analyzed values in 71 per cent and 86 per cent of the control and
experimental diets, respectively. The diet analyzed in Egtg was in better
agreement with the calculated values for nitrogen than when the
individual items of the same diet were analyzed and totaled.

The differences between the analyzed and calculated calories in

the experimental and control diets were statistically significant.

THE DIETARY PROCEDURES EMPLOYED
IN CONDUCTING A NON-CLINICAL
METABOLIC BALANCE STUDY

BY

Carolyn Mae Friedemann

A THESIS

SubmittedAto

Michigan State University

in partial fulfillment of the requirements
for the degree of

MASTER OF SCIENCE

Department of Foods and Nutrition

1965

The author
assistance and
extended to the
and Nutrition 1'

Appreciati
in the organizs
extended to Mr.

A special
his patience, h

A thank yo

AéfiCulture

Y V
£11.17

ser’ices for E5

ACKNOWLEDGEMENTS

The author gratefully acknowledges Dr. Olaf Mickelsen for his
assistance and guidance throughout the study. Gratitude is also
extended to the staff and graduate students in the Department of Foods
and Nutrition for their help, concern, and encouragement.

Appreciation is extended to Miss Simin Bolourchi for her help
in the organization and direction of this study. Thanks is also
extended to Mr. David Anderson for his technical assistance.

A special thank you to Mr. Larry Garber, my husband-to-be, for
his patience, helpful ideas, and continued reassurance.

A thank you is extended to the United States Department of
Agriculture Human Nutrition Research Division of Agricultural Research

Services for making this study possible.

ii

r

SIZE OF LI

[=54

General

Cl

Ft

TABLE OF CONTENTS

REVIEW OF LITERATURE . . . . . . . . . . . . . . . .
General Considerations . . . . . . . . . . . .
Clinical versus Non-Clinical Balance Study
Objectives of the Study . . . . . . . . .
Experimental Design . . . . . . . . . . .
Control and Experimental Periods . .
Transitional Period . . . . . . . . .

Length of the Study . . . . . . . . .
Collection Period . . . . . . . . .

Number of Subjects . . . . . . . . .

Selection of Subjects . . . . . . .

Menus and Diets . . . . . . . . . . .

The Personnel . . . . . . . . . . .
Evaluation of Results . . . . . . . . . .
Dietary Management . . . . . . . . . . . . .
Purchase and Storage . . . . . . . . . . .
Number of Menus . . . . . . . . . . . . .
Composition of the Diet . . . . . . . . .
Selection of the Menu . . . . . . . .

Caloric Requirement . . . . . . . .

Food Preparation . . . . . . . . . . . . .
Calculation of Mixed Dishes . . . . . . .
Weighing and Measuring the Food . . . . .

Formula Diets . . . . . . . . . . . . .

iii

Page

to

454‘

10

10

12

12

13

14

14

15

15

17

18

18

TABLE OF CONTENTS (Cont.)

Dietary Analysis . . . . . . . . .
Collection of the Sample . .
Components of the Sample . .
Containers for the Collection
Weight of the Diet Sample
Preparation for Analysis
Storage of the Sample . .

Drying of the Sample . .

Two Methods For Determining Food Composition

Food Composition Tables . .

Conversion Factors . .
Calories . . . . . . . .
Protein . . . . . . . .

Calculation versus Analysis
Protein and Nitrogen . .
Fat . . . . . . . .
Fat and Calories . . .

Protein Fat and Calories

Individual Items versus Total Diet

Individual versus Average Diet Analyses

Variation in Analyses of Identical Diets

Differences in Analyses of Duplicate Samples.

iv

Page
19
20
20
22
22
22
23
24
25
26
26
26
28
29
29
31
31
32
34
35
35

36

TABLE OF CONTENTS (Cont.)

Page

EXPERIMENTAL PROCEDURE . . . . . . . . . . . . . . . . . . 38
Metabolic Balance Study . . . . . . . . . . . . . . . 38
Selection of Subjects . . . . . . . . . . . . . 38
Control and Experimental Regimens . . . . . . . 39
Personnel . . . . . . . . . . . . . . . . . . 39
Calculation of Composition of diets and recipes. 40
Purchase and Storage of Food . . . . . . . . . . 41
Scales for Weighing . . . . . . . . . . . . . . 42
Preparation of the Food . . . . . . . . . . . . 42
Nutritive Value of Diets . . . . . . . . . . . . 43

Food Energy Computation . . . . . . . . . . . . . . 43
Protein Conversion Factors . . .p. . . . . . . . .'. 44
Weight Control of Subjects . . . . . . . . . . . . . 44
Items Allowed ad libitum . . . . . . . . . . . . . 45
Trial Period . . . . . . . . . . . . . . . . . . 45
Analysis of Diets . . . . . . . . . . . . . . . . . . 45
Collection Time . . . . . . . . . . . . . . . . 45
Individual Food Item . . . . . . . . . . . . . . 46
Collection of Sample for Analyses . . . . . . . 46
Preparation of the Sample for Analyses . . . . 46

Per Cent Meisture . . . . . . . . . . . . . . . 47
Nitrogen Determination . . . . . . . . . . .. 47

Fat Determination . . . . . . . . . . . . . . . 48

Food Energy Determination . . . . . . . . . . . 48
Ihnalysis of Data . . . . . . . . . . . . . . . 48

    

RESCUS #20 DZ"
Requiremcz
length 0f
Rap?“t b"
ImportanCt
SuperviSiC‘

Purchase an

 

maintenaace
ReliabiliEF
iaiability '
Calculation

Calculation

Individual ‘
When Must a
Various C011

Moisture L05

SMSRY AND CONCI

1%I‘LRE CITED

ID“! ,
(UL;
DIX .

Recipes Used

TABLE OF CONTENTS (Cont.)

RESULTS AND DISCUSSION . . . . . . . . . . . . . . . .
Requirement of Transitional Period . . . . . . .
Length of Study . . . . . . . . . . . . . . . .
Rapport between Personnel and Subjects . . . . .
Importance of Trial Period . . . . . . . . . . .
Supervision of Subjects . . . . . . . . . . .
Purchase and Preparation of Food . . . . . . . .
Maintenance of Subjects' Weight . . . . . . . .
Reliability of Scales . . . . . . . . . . . . . .
Riiability of Food Tables . . . . . . . . . . .
Calculation of Caloric and Nitrogen Content . .

Calculation versus Analysis of Nitrogen, Fat and
Calories

Individual Components versus Total Analysis . .
When Must a Diet be Analyzed . . . . . . . . . .
Various Collection Times During the Meal . . . .
Moisture Losses Due to Storage Containers . . . .
SUMMARY AND CONCLUSIONS; . . . . . . . . . . . . . . .
LITERATURE CITED .. . . . . . . . . . . . . . . . . .
APPENDIX . . . . . . . . . . . . . . . . . . . . . .

Recipes Used in Balance Study . . . . . . . . . .

vi

Page
67
67
67
67
68
69
69
71
72
74

75

79
92
95
98
104
108
113
121

123

RULE

10

ll

12

13

   

Compositi
The compo:

The compo:

Menus for :
Menus for t
The validit
C021Parison

0f the dict
COUDariSOn

factors for
0f f00d co:
compariSOn

fat and c
SErVice

‘i
‘14

COEIpariSOn
fat) and CE
COIIEcted C

ays

individual
diets

CompariSon
lets C0116

COntrol

l

HZ~

~(

1
t

E
fit

T: Q.‘

TABLE

10

11

12

13

14

LIST OF TABLES

Composition of the diets . . . . . . . . . . . . . . . .
The composition of the foods comprising the control diets .

The composition of the foods comprising the experimental
diets

Menus for the control period . . . . . . . . . . . . . .
Menus for the experimental period . . . . . . . . . . .

The validity of the scales employed in the metabolic
kitchen . . .

Comparison of two methods to determine the energy value
of the diets . . . . . . . . . . . . . . . .

Comparison of dietary nitrogen calculated using various
factors for converting the protein values listed in tables
of food composition with the analyzed nitrogen values. .

Comparison of calculated and analyzed values for nitrogen,
fat and calories collected at two times during the meal
Semice O O O O O O I O O O O O O O O 0

Comparison of calculated and analyzed values for nitrogen,
fat, and calories in the seven experimental diets
collected during four successive weeks . . . . . . . . .

Per cent fat in three separate diets on three different

days 0 O O O O O O O O O O O O O O O O O O O 0

Comparison of Calculated versus analyzed values of
individual foods that were prepared in three experimental

diets O O O O O O O O O O O O O O O O O O O O 0

Comparison of duplicate analyzed values of the experimental
diets collected during four successive weeks . . . . . .

The analyzed values for nitrogen, fat, and calories for the

control diets collected at two time intervals during the
serVin-g periOd O O O O O O O O O O O C O O O O O O O O 0

vii

Page
50

51

55
59

63

73

76

78

80

82

91

93

96

98

LIST OF TABLES (Cont.)
TABLE Page
15 The analyzed values for nitrogen, fat, and calories for
the experimental diets collected at two time intervals

during the serving period . . . . . . . . . . . . . . . 100

16 Analysis of individual meals collected at different
serVing times 0 O O O O O O 0 C O O O O O O 103

17 Moisture determinations on diet samples . . . . . . . 106

viii

5*]

PISTE

1 Effects or

 

 

 

LIST OF FIGURES
FIGURE

1 Effects on drying with time . . .

ix

REVIEW OF LITERATURE

A metabolic balance study will be defined as an investigation of the
intake and output of a nutrient or nutrients by human beings. This can
occur by keeping constant all possible factors other than the one that
is being explored(Comfort, 1957). Such a study requires the precise
control and measurement of both intake and output. Because this is such
a vast subject, this review of literature will consider only the first
phase; that of intake. This presentation is divided into three sections.
The first section will concern the general principles involved in
conducting a metabolic balance study; the second will present the factors
involved in the dietary management of such a study; the third will
compare the two methods employed to determine the nutrient value of the
diets.

I. General Considerations

Clinical versus Non-Clinical Balance Study

 

A metabolic balance study may be carried out in a clinical or a
non-clinical situation. In the former there is constant supervision of
the subjects, whereas, in the non-clinical study the only supervision
the subjects receive is generally during meals and the various tests to
which they are subjected. The site of the clinical study is the hospital,
whereas, the university or a comparable institution composes the non-
clinical location. Bauer and Aub (1927), are the forerunners of today's
modern research metabolic unit in a hospital. They stressed that the
uniqueness of such study is the control of all phases involved. Since
there is more supervision in a hOSpital, there is inevitably increased

control. Coons (1930) conducted a metabolic study, using as her

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subjects pregnant women who were consuming a self-chosen diet in their
homes. The only supervision entailed in this study was visits made to
the individual's home during the serving of the meals. Less control of
the study occurs, and thus more variables enter into the non-clinical
study.

There are a number of factors which further distinguish a
clinical study from a non-clinical study. One of these is cost. The
clinical situation is more expensive than the non-clinical. The
difference in cost may be a factor ranging from four to five fold
depending on the location of the study. Furthermore, in most cases,
the subject in a clinical situation cannot perform physical activity
at a normal level. Since physical exercise influences the deposition
of fat and the development of muscle, a lack of it could play an
influential role in the results of the study. The application of these
findings to normal adults might thus be questioned. The third comparison
of the two sites for a metabolic balance study is the psychological
aspects of the environment. In the hospital the emphasis is primarily
upon illness. The environment created in a non-clinical situation is
more typical of home life. McKay g£.§l.,(l942) report that it is
important that the participants maintain their usual curricular and
extracurricular activities and continue their ordinary manner of life
during a balance study.

Schottstaedt and co-workers (1958) conducted a study to determine
the effects of naturally occurring stressful situations on a group of
patients in a social setting of a metabolic ward. It was discovered

that there were metabolic changes in the urinary excretion of water,
sodium, potassium, calcium, nitrogen and creatinine during situations

of stress. Interpersonal difficulties were the most common source of

3

tension which produced the metabolic changes. They concluded that a meta-
bolic ward is not necessarily a natural environment, but adequate
interpretation of metabolic deviations must include an appraisal of the
participant's life situation and emotional state at the time of the
experiment. Thus, for normal adults the clinical study is limited and,

consequently the results may be of limited value.

Objectives of the Study

The objectives of a metabolic balance study are defined as the
questions which the study plans to answer. The objectives of the study
determine its length which includes the control and experimental
periods, the requirement of a transitional period between the control and
experimental regimens, the number of personnel needed to carry out the
study (Horwitt 25.31., 1949), the selection of the subjects
(Leichsenring e£_al? 1958), the choice of foods, the method of food
cookery, the type of storage required for the food, the composition of
the menu or menus (Sampson ggflgl., 1952), the precautions to be employed
in collecting the food for analysis, and the chemical determinations to
be performed.

The researcher in designing the objectives should take into con-
sideration money and personnel available to conduct the study
(Leichsenring g£,§l,, 1958). These two factors influence the number of
subjects, the number and/or type of analytical methods to be employed,

the menus and the preparation of the food.

 

A netabol

These two peri
the phase duri
cedures involv
intake of the .
period. The 0
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Experimental Design
Control and Experimental Periods ‘

A metabolic balance study should be composed of two periods.
These two periods are control and experimental. The control period is
the phase during which the subjects are becoming adapted to the pro-
cedures involved in a balance study, the dietary regimen and to the
intake of the nutrient that is going to be studied in the experimental
period. The control period should serve as a period in which the
subject and the investigator develop rapport.

In addition to the preceding, Leichsenring 35.31., (1958) suggest
that a trial period be carried out prior to the study in which every-
thing but the analytical work is performed. They claim that such a
period helps to clarify procedures and also assists in developing a
rapport between the researcher and the subject. However, since the
control period serves as the phase during which the subjects are
attaining equilibrium of the nutrient to be investigated, it could
also serve as the trial period. The results of the control period
serve as the norurwith.which the values from the experimental period
are compared. The two cycles may either occur simultaneously pr
sequentially.

Reifenstein 2;.gl., (1945) and Comfort (1957) report that a
metabolic balance study may follow one of two plans. The first plan
compares the finding of one individual who is on a control diet with
the results of another subject who is on an experimental diet. The
second arrangement consists of a comparison of the results of a subject
when he is on a control diet with the results of the same subject when

he is on an experimental diet. Another experimental design is reported

by Bricker a

  
    

on the cont:
way through
By this near
during the e:
to compare tlf
the experimen

a control for

 

and which nig‘r

There arc
HOT Citéd in C
involves the u
control diet t

control diet I:

5
by Bricker EE.El°’ (1949). In this plan half the subjects are started
on the control diet and the other half on the experimental diet. Half
way through the experiment, the regimens for the two groups are switched.
By this means, it is possible to compare the results for the same subject
during the experimental and control periods. It is furthermore possible
to compare throughout the entire duration of the study, the subjects on
the experimental regimen with those on the control. This also, provides
a control foreny environmental factors that might influence the study
and which might change during its course.

There are other experimental designs for a balance study which are
not cited in the literature but which require discussion. The first
involves the use of two separate groups; the first is maintained on the
control diet throughout the experiment; the second group is on the
control diet to establish equilibrium and then proceeds to the
experimental diet. The results of the experimental diet are compared
with the control group. The advantage of this design can be explained
by an example. In the bread study conducted at Michigan State
University, Spring, 1964, there was a weight loss in most of the subjects
during the experimental period. In this experiment the results of a
subject on the control diet were compared with the results of the same
subject on the experimental regimen. If a control group had been
employed simultaneously with the experimental group, it would have been
possible to determine with considerable certainty whether the weight
losses exPerienced by the subjects were due to the increased physical
activity which presumably occurred with the advent of warm weather.

(See Section Results and Discussion, p.71 ). However, for such a design
additional personnel and facilities for food storage, preparation and

Serving are required. The two groups should be served in separate areas

 

 

in Of!

diet i

Ira

contro
betvee
contro
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or diet

three 1

6

in order to maintain the emotional stability and integrity of each group.

Another experimental design involves the use of an experimental
diet which is preceded and followed by a control regimen.

Transitional Period

Reifenstein gg‘al., (1945) report that if the same subjects on the
control diet are placed on the experimental diet a transitional periOd
between the two phases may be required. This is only the case if the
control diet is high in the substance which is extremely low or absent
in the experimental diet. The analytical results obtained when the
subject first starts on the experimental diet should not be considered
as conclusions for the experimental phase. A lapse of time must occur
before a new equilibrium is established (Reifenstein_g£_gL, 1945).
Coons (1930) reports that during this period no collections of excreta
or diets should occur. This length of time has been reported to be
three to five days (Coons, 1930). However, the exact length of time
depends on the difference in the nutrient content of the two diets. A
fifteen day period preceded the ten day collection period in studies
conducted by McKay gg'a1., (1942). This study was concerned with the
effect of different levels of milk upon the calcium, phosphorus and
nitrogen metabolism in college women. They found this length of time
necessary in order for the women to establish equilibrium on the
experimental regimen.

Length of the Study

The length of a balance study is very important because of the
extreme daily variations manifested in the magnitude of retention or
loss of the various nutrients (Donelson 25.31., 1931). It is further

reported by Donelson 35.31., (1931) that a balance study which includes

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only a few days registers fluctuations in metabolic processes which a
longer period tends to conceal. Both periods should be sufficient in
length for the subjects to attain equilibrium for the nutrient under
consideration. The length of the control period is influenced by the
amount of change between the control regimen and the diets of the
subjects prior to the study.

Collection Period

 

Within both phases, control and experimental, of a balance study
there are collection periods. The length of this period usually varies
according to the number of menus designed for each period. Young $5.21.,
(1953) in studying reducing and post-reducing maintenance on a moderate
fat diet employed a seven day diet, thus, a seven day composite period.
A composite sample represents the total collection for the balance
period. This would be made up separately for food, urine, and feces. A
six day collection period was used in studying the metabolic patterns in
preadolescent children (So. C00p. Series Bull. No. 94, 1964).

Schofield 2E.2l°’ (1956) employed a four day collection period. Bauer
and.Aub (1927) used a three day collection period in their studies of
inorganic salt metabolism. In each of the studies previously mentioned,
the number of menus was equal to the number of days in the collection
period.

Number of Subjects

No report was found in the literature that describes the ideal
number of subjects to be employed in metabolic balance studies. The
number of subjects range from.one (Sherman, 1920) to one hundred twenty-
four (MeKaypgg‘gl., 1942). The extent to which they represent a sample

Of the normal population has not been discussed by the investigators.

econo
vould
elimi
the i
(Laid
a thor
nutri:

free c

1933).

Selection of the Subjects

The emphasis of the study will determine the age, sex, health, and
socio-economic status of the subjects to be selected (Leichsenring g£.§1.,
1958). At a university today, varied cultural backgrounds Which mean
assorted dietary habits may play a more important role than the socio-
economic status. The subjecnswith food dislikes or allergies which
would interfere with their complete consumption of the diets should be
eliminated. The researcher must also consider such characteristics as
the integrity, reliability and the native ability of the individual
(Leichsenring 25.21., 1958). The subject prior to selection should have
a thorough physical examination to assess his general health and
nutritional status (Leichsenring 35.21., 1958). The subject should be
free of disease. These are broad generalizations which have to be
supplemented by specific instructions to the examining physician.
Individuals with physical handicaps should be omitted from any balance
study which attempts to evaluate normal individuals. The subjects
selected should be as uniform as possible insofar as their body weight
is concerned. For normal individuals an adherence of 5 per cent of
their ideal weight should be followed. In addition to height-weight
tables for evaluating a subject's weight, a researcher may wish to
employ anthropometric measurement such as those of skin folds.

Upon selection, the subject should be presented a clearly written
statement which includes his obligations, the directions for the
collection of excreta, and if applicable, the directions for the
collection of food (Leichsenring 35.31., 1958). Preceding the study
the research director should obtain each subject's permission to enable
him.to use any data as a result of the investigation (Leichsenring 25.21.,

1958).

 

 

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9

Horwitt 35‘21., (1949)selected their subjects according to the
following procedure. They first selected 50 per cent more subjects
than were required by the study. The subjects were observed for two
months on a dietary regimen similar to that which was to be employed in
the study. They evaluated the subjects'attitudes, cooPerativeness, and
eating habits. The individuals deficient in any of these respects were
eliminated. This study was conducted in a mental hospital which makes
this arrangement possible.

To eliminate subjects who may have emotional problems to the
extent that they would be unreliable, irresponsible, and dishonest,
Young, (personal communication, 1964), suggests the use of the Bell
Adjustment Inventory Examinationl. The total score of the Stanford
examination is divided into four parts--home, health, social, and
emotional. Young indicates that before subjects are selected for her
metabolic balance studies they must score an excellent or good
emotional score. The selection of the participants on the basis of the
emotional score eliminates those who are not able to withstand the
restrictions that are involved in a controlled feeding experiment
(Young, personal communication, 1964).

Menus and Diets

The control and experimental diets should contain the same come
ponents with the exception of the nutrient that is to be investigated
during the experimental phase. Furthermore, since the control period

Serves as the time when the subject is becoming adapted to the dietary

‘

This is a publication of the Stanford University Press and is
distributed by the Psychological Corporation, 522 Fifth Avenue,
New York 18, New York. The examination is available in two forms,
an adult form and a student form.

 

10

regimen, the two diets should contain similar types of food insofar as
possible. For example, if the balance study is to evaluate a low
thiamine intake on college men, the control diet should be the same as
the experimental regimen. The experimental diet would require supplemen-
tation in order for the diet to fulfill the Recommended Dietary Allowances
(National Research Council, 1964). Ideally, the only difference between
the two diets would be the change in the amount of thiamine supplementa-
tion during the experimental diet.

The diets and menus are further discussed in the following section
(II. Dietary Management).

The Personnel

In reviewing the literature regarding the personnel required to
conduct a balance study, there is only slight reference to this topic.
The people that are employed for any phase of the study must be
intelligent, willing to follow detailed instructions precisely and be
accurate in their work perfonmance (Leichsenring st 21., 1958). The
number of personnel required to supervise thirty subjects in a study to
determine the riboflavin requirements of adult subjects, included a
dietitian, two cooks, and a nursing attendant or equivalent (Horwitt 25.21.,
1949). This paper did not report the number of employees used to perform
the analytical techniques as demanded by the balance study.

Evaluation of Results

This discussion will be limited to the results of the nitrogen
balance studies. In any balance study the researcher is faced with the
intake and output of a Specified nutrient which is relatively large when
compared to the amount retained. Very few investigators have carefully

evaluated the over all significance of positive nitrogen equilibrium.

11
Leverton st 31., (1956) report that because of the presence of inherent
variations, both human and mechanical, in human metabolism studies an
evaluation of the results require careful consideration.

A nitrogen equilibrium is defined as a zone in which the excretion
and intake closely approximate each other, rather than a single point at
which they are numerically identical (Leverton E£.El°’ 1956). Therefore,
in their studies of amino acid requirements, the subjects were in
nitrogen equilibrium.when the difference between the intake and
excretion did not exceed 5 per cent.

Rose (1957) comments that irregardless of the accuracy in controlling
the amount of nitrogen that a subject consumes there is a fluctuation of
nitrogen from day to day. Perhaps the explanation is a result of
slight alterations in the daily rate of metabolism or excretion or in
the degree of muscle tone (Rose, 1957). In establishing the amino acid
requirements of human aduhts, Rose (1957) induced a negative nitrogen
balance. The intake of the amino acid under investigation was then
increased until a slight but distinct positive nitrogen balance, as
measured by the average for a period of several days, was achieved. The
greatest amount required to achieve the positive balance in an individual
was defined as the amino acid requirement for normal man.

Walker (1962) states that the results of nitrogen balance studies
'may reveal an excessive accumulation of nitrogen which over a prolonged
period of time would be an impossibility. This may be due to a loss
of the nitrogen other than through the urine and the stools, especially
losses through the skin and in certain circumstances significant
changes in body composition. This fallacy was illustrated by Walker

(1962) from a long-term balance study on malnourished Bantu adults

 

cond

prot
cont
accm

if d

bala:
intal
in lo
error

SOUtc

12
conducted by Holmes (1954). The observations from this study disclosed
a consistently high retention of nitrogen which was, in some cases, as
much as ten grams per subject per day. The diet contained 100 grams of
protein and 3000 to 4000 calories per day. Occasionally the protein
content was increased to 200 grams per day. Such retentions were
accompanied by much lower gains in body weight than is to be expected
if the subject was storing ten grams of nitrogen daily.

There are three sources of error that may influence the nitrogen
balance of an individual (walker, 1962). They are 11) inaccuracy in
intake and output collections and methodological error31—particular1y
in long-term balance studies which are liable to lead to cumulative
errors, (2) lack of knowledge regarding losses thru the skin and other
sources, (3) lack of methods for precisely determining body composition.

II. Dietary Management

The choice of foods for a metabolic balance study is determined by
the facilities available for proper storage, season of year, method of
preparation, and purpose of study.

IPurchase and Storage

Sampson g; 21., (1952) suggest that the entire lot of food with the
exception of fresh food be purchased from a single source in quantity
sufficient for the duration of the study. This requires adequate
storage space. The freezer space per subject is a minimum.of five cubic
feet.

As often as is possible and facilities permitting, all food items
Sfuiuld be prepared at one time and frozen until needed
(30. Coop. Series Bull. No. 94, 1964). The diets described in this

blllletin contained such items as bread, fruit-flavored ice, cupcakes,

13
and other desserts. Prior to the experiment, these items were purchased
and/or prepared for the length of the study. The designated portions of
the items were weighed and stored in a freezer.

The food selected for a balance study should undergo minimal change
in composition during storage (Sampson gE.§1., 1952). The canned or
frozen foods are not altered significantly in composition if stored
properly, however, upon storage perishable items are changed. If the
determination of vitamins is the objective of the study, the perishable
items should be stored for short periods of time (Sampson 25.31., 1952).
Hawks gt 21., (1937) state that perishable foods should be purchased
for one balance collection period. This eliminates the variability in
composition which occurs in food when purchased on a day to day basis.

Marble (1939) reports that all food used should be in season
throughout the experiment. This statement is most important in selecting
fresh foods. However, this factor would also apply to canned or frozen
food if there is inadequate storage space. This problem can be
alleviated by ordering the same lot and brand of food in adequate amount
at the beginning of the study and arranging with the food supplier or
the proprietor of storage facilities for storage of the items until
needed.

Number of Menus

 

The number of menus employed in a balance study usually is the
same as the number of days in the collection period. Presson (1955)
suggests that a trial period prior to the actual study should be employed

to determine the most palatable diet for the subject. The useci one

diet

error

ideal

defi:

to ti

. be we

leads

accep

HERE

‘l5—;£§A;n=llu’

i'z-i .._ 9;; ’17:“?

L; \ - four

 

accep

is no

 

l4
diet according to Presson (1955) eliminates excessive calculations and
errors which may occur when using many menus. The use of one menu is
ideal, but is not always possible. In order to achieve a better
definition of intake Ahrens g£.a1., (1954) advises the use of only one
to three menus. Bassett and Van Alstine (1935) report that one menu may
be well tolerated by children but, frequentlx,when presented to adults
leads to refusal and smuggling of food. To achieve palatability and
acceptability of the diet by the subjects in their studies, a six day
menu was required. A factor to be considered is that their study was
four months in length. Any study of this duration increases the non-
acceptance of one menu, whereas, for short term studies a one day menu
is more likely to be tolerated.

Composition of the Diet

 

The composition of the menus should not only consider the purpose
of the study but also, the subject's eating habits and caloric requirements
(Sampson SE 31., 1952). The nutrient content of the diet is calculated
with the aid of food composition tables. All calculated menus should
insure an adequate amount of all the essential nutrients. If the diet
does not fulfill the nutritional requirements, supplements in pill form
which contain the deficient nutrients should be given the subjects.

Food composition tables should only be used as a guide in planning the
diets (Widdowson and MCCance, 1943). Because of this factor Coons (1930)
emphasizes that_a11 diets require chemical analysis prior to the
beginning of the study.

Selection of the Menu

 

To determine the eating habits of the participants the researcher

should interview the subject. This information can aid in detemmining

 

15
the foods which are to be included in the menu (Presson, 1955). Sampson
and co-workers (1952) advocate the participantfs selection of the menu.
This avoids the continual encouragement that must be given to the subject
by the nutritionist when the subject does not desire the food. The
emotional disturbances created by forcefully eating an objectionable
food could contribute to unphysiological results. If the subject does
not consume the food, the results of the balance study would be affected.
Marble (1939) stresses that unconsumed food must be recorded. Subjects
with many food dislikes should be eliminated. This is only possible when
there is a large number of subjects from which to choose.

Caloric Requirement

 

The calories in the diet can be calculated according to the regular
dietary intake of the subject if the study is concerned with the main-
tenance of the subject's weight. Also, the caloric requirement of the
individual may be determined according to age and sex (Sampson, 25 al.,
1952).

.To fulfill the energy requirements of the participants whose needs
were not met by the calculated diet Lang 2; 31., (1965) served hard
candy and butter balls ad libitum. Their studies were to determine the
manganese metabolism of college men. To supply additional calories to
subjects on controlled caloric intake Pilcher (1961) administered sugar,
honey, butter, salad oil, jelly and jam.

Food Preparation

 

Johnston and McMillan (1951) in preparing mixed dishes weighed
the ingredients in individual casseroles prior to cooking. The
objective of their study was to investigate the effect of spinach on the

absorption of iron. Because the study was concerned with iron, all the

\

  
 

food was p1

Bauer
tance of v.
their clin:
a moisture :
food not co:
dish plus i:

by differentv

 

In addi'
dish, HaVks 5
its contents
the participg
Should not be
Where Vitati:
and Hummel S!
are not 1:901
treated in t1
jects. Furt]
tothe SUhje

bison...
in those Stuc

used, the ta'

the Study. 1

i “
DUKE of the”

16
food was prepared in pyrex or aluminum utensils.

Bauer and Aub (1927) and Gephart and DuBois (1915) stressed the impor-
tance of weighing, cooking and serving the food in the same dish. In
their clinical metabolic balance studies they recorded on the dish with
a moisture resistant enamel pencil the weight of the empty dish. Any
food not consumed by the participants was determined by reweighing the
dish plus its contents. The weight of the uneaten portion was obtained
by difference.

In addition to weighing, cooking and serving the food in the same
dish, Hawks 25.31., (1937) rinsed the dish with distilled water after
its contents had been consumed. This water solution is then consumed by
the participant. Hummel g£‘§1., (1942) stressed that the cooking water
should not be discarded in mineral balance studies or in those studies
where vitamins are to be determined. The importance of Hawks gt g1.,(1937)
and Hummel SE 31., (1942) suggestions may be questioned. These factors
are not important as long as the sample collected for analysis is
treated in the same manner as the diet which is consumed by the sub-
jects. Furthermore, the consumption of pot liquor might be objectionable
to the subjects.

Distifled water, according to Coons (1930), should only be employed
in those studies investigating minerals. If distilled water is not
used, the tap water should be analyzed for the minerals of importance in
the study. For studies involving mineral determinations the water
intake of the subjects should be recorded.

The mode of food preparation and cookery used for a balance study
is very important to assure uniformity and consistency of the product.

Since the cooked products should have no variable loss of nutrients,

steaming has been recommended for food preparation (Marble 1939).

17
Marble (1939) does not state the type of balance studies which would
require steaming. For mineral and vitamin studies this method might be
employed. However, if the diet sample for analysis is a duplicate of
the diet consumed by the subjects,the method of cooking need not be
limited to steaming. If the nutrient under investigation is not
destroyed by most cooking procedures, the choices for food preparation
are greatly increased.

In using Special recipes precautions are necessary to avoid
variations in content of water and solids. This is best achieved by
meticulous measurement of ingredients, standardization of cooking time
and temperature (Sampson £5 21., 1952). It was further suggested that
desserts, such as cake and pie, should not be used in balance studies
because of their inevitable variation in composition

In conducting a nitrogen balance study there is an increased
versatility inthe selection, in the type of storage, and in the cooking
procedures of the foods. This does not mean less control, for the
results of any balance study are only as reliable as the techniques
employed in conducting the study (Hawks 35‘31., 1937). If a vitamin,
such as ascorbic acid, is being investigated, food must be selected
which is not perishable and all cooking methods must insure minimal
variability in the losses of the vitamin.

Calculation of Mixed Dishes

If mixed dishes are to be employed in the study, the recipe should
be calculated. Accurate results cannot be attained by using for mixed
dishes the values listed by tables offimd crmposition (Patterson and

lflcHenry, 1941). This is due to the different proportions of the

18
ingredients in the recipe as presented by the tables and in those
designed for a balance study.

Weighing and Measuring the Food

 

Widdowson and McCance (1942) and Coons (1930) weighed their diets
on a Hanson spring balance. They report that the scale was accurate to
one gram. Contrary to their work, Sampson gt 31., (1952) and Lang and
co-workers (1964) employed a torsion balance for weighing all food items.
The torsion balance was accurate to 0.1 gram. Ceglarek 35 31., (1958)
measured liquids to the nearest ml.

Formula Diets

 

Because of the difficulty in controlling the variability when using
assorted natural foods, formula diets have been suggested by Ahrens 35 31.,
(1954). For Specialized metabolic studies, the formula diets may be
altered to give unusual proportions of fat, protein, and carbohydrate.

A formula diet can be devised to contain relatively few components of
definite composition which can be accurately formulated, homogenized,
and dispensed (Haust and Beveridge, 1958).

Many researchers using the formula diets have studied the effect
of various kinds and levels of fat upon plasma lipids. An example of
this type of formula diet is the one used by Beveridge and his co-
workers (1955). The ingredients of the basal diet were powdered skim
milk, corn oil, margarine and dextri-maltose. When other fats were
substituted, the sugar was used to adjust the calories to a constant
amount. Vitamins for the subjects were supplied in the form of capsules
and roughage was provided by the addition of celluflour to each formula.

Ahrens (1954) fed formula diets to adults for periods of two to

sixteen weeks. The formulas for all subjects proved to be entirely

 

 

19
acceptable, economical and simple to use. He suggests the use of formula
diets for those studies concerned with maintaining the weight of the
subjects. This can be obtained by increasing the caloric intake without
disturbing the composition of the diet. Formula diets can also be used
when the calories are to remain constant but the prOportions of carbohy-
drate, protein, and fat are to be varied or when the preportions of
these three are to remain constant.

Dietary Analysis

 

Reifenstein 35 31., (1945) and Bassett and Van Alstine (1935) report
three procedures for the analysis of the diets. They are: (1) each
foodstuff may be analyzed and the values obtained may be used in the
calculation of the diet, (2) one or more sample diets may be analyzed
.EE.EEE2 and the results obtained may be employed throughout the study
or (3) aliquots of the diet, as served each day, may be pooled and
analyzed for each collection period of the study. They report that it
is most accurate to sample and analyze the food from each collection
period, however, due to the increased labor and time requirements, they
suggest the second method. In the studies of Sampson st 31., (1952)
the diet is reanalyzed every time a new lot of any food is used.

Presson (1955) also advocates the occasional analysis of the total diet
or diets employed in a study. The values obtained are used to determine
the actual intake by the subjects. Lutwak g£.§1., (l964)determined the
composition of seven diets by collecting and analyzing a representative
sample every fourth week. Hunter 25.31., (1948) reports that the
assessment of diets by direct analysis immediately before ingestion is

the best means of determining the nutritive value. Assuming accuracy

in the collection of the diets, and also assuming reliability of the

analytical
analytical
Collecti:
The c:
constant me
that the C:
that the 11
She report:
torsion ba}
(1930) that
that it is
if)

even under
accurate Se
weigh each
50 that an

dish,

20
analytical techniques used for measuring the food constituents, the direct
analytical method can yield a high degree of accuracy (Hunter gt 31., 1948).
Collection of the Sample

The collection of the sample for analysis must be performed in a
constant manner throughout the study. It is reported by Coons (1930)
that the collection of the sample for analysis should occur at the time
that the item is served in order to avoid changes in moisture content.
She reports that weighing the sample which is to be collected requires a
torsion balance accurate to 0.1 gram. It is further reported by Coons
(1930) that the sample should be homogeneous. Hawks £5 31., (l937)state
that it is impossible to obtain a homogeneous sample of food for analysis
even under strictly controlled conditions. The principle to adopt for
accurate sampling as presented by Widdowson and McCance (1942) is to
weigh each ingredient separately or to make a mixture of the components
so that an average helping is likely to be representative of the whole
dish.

The amount of sample weighed for the analysis of the diet depends
upon the number and type of chemical determinations that are required.
Johnston and MeMillan. (1952) support the principle of collecting the
same amount of food that is served to the subjects. Bassett and
Van Alstine (1935) and Leichsenring gt 31., (1958) weigh one-fourth of
the prescribed weight of each item served.

Components of the Sample

The exact foods which will constitute the collected sample varies
‘With the chemical determinations employed in the study. High fat diets
and high sugar diets are often difficult to handle after drying (Coons,

1930). Leichsenring g£_31., (1958) and Reifenstein ££_§l., (1945)

 

O

21
suggest that butter, fat, sugar and other fats be excluded from the
collected diet sample and analyzed separately. If fat and calorie
determinations are required as a part of the study, the fatty foods can-
not be omitted (Leichsenring EE.§l°’ 1958). If only mineral and
nitrogenous components are being determined, Leichsenring £5 31., (1958)
advocates weighing a proportional aliquot of the fat items. These fatty
foods are heated with 50 to 100 ml. of distilled water until the fat is
melted. Upon cooling the layer of solidified fat is removed and the
remaining portion is then analyzed. The nutrients which are to be
investigated by the study must be considered prior to employing the
previous technique. For example, if the determination of phospholipids
c: phosphorus was an objective of the study, this method could not be
practiced.

The division of the diet into two portions is practiced by
Gutman and Low (1939). The samples containing the nutrients which are
subject to rapid deterioration are analyzed immediately, whereas, the
samples composed of stable nutrients are made into a slurry and frozen
in a low temperature refrigerator. No further description of the
refrigerator was presented.

Leichsenring 25 31., (1958) state that the number of analysis may
be reduced by preparing composites of several foods having similar
physical and chemical properties. In studying the protein requirements
of women on high cereal diets Bricker and co-workers (1949) employed four
<iivisions. These were: non-protein dessert which was cornstarch
cxaokies, low protein foods such as fruits and green vegetables, cereal

Plastein, and non-cereal protein such as potatoes, meats, and animal

products.
requirert
from diff
particula
£3333
Laic
square g]
collectir
London (1
home amp;
0f the d
women We

(1945),

22
products. The objectives of that study were to determine the protein
requirements of college women when consuming various nitrogen levels
from different sources. The samples were pooled as composites for the
particular length of the balance period.

Containers for the Collection

 

Leichsenring £5 31., (1958) report that wide-mouthed French-
square glass bottles with plastic screw caps are satisfactory for
collecting the diet because they can be compactly stored. Vought and
London (1964) in determining the iodine intake of subjects living at
home employed one gallon,wide-mouthed,polyethylene jars for the collection
of the diet. The liquid and solid components of the diets of lactating
women were collected individually in glass containers by Kaucher £5 21.,
(1945).

Weight of the Diet Sample

 

Upon placing the prescribed amount of food in the container, it
should be covered immediately and placed in the freezer (So. Coop. Series
Bull. No. 94, 1964). The weight of the container and the date of the
sample should be recorded on the container with a moisture resistant
pencil (Leichsenring gt 21., 1958). Bransby st 31., (1948) upon collect-
ing the entire diet, weighed the jar with its contents. The weight of
its contents was the difference between the two weights and was recorded
on the container. The weight of the diet was obtained after homogeniza-
tion in the diet studies on preadolescent children (So. Coop. Series Bull.
No. 94, 1964) .

Preparation for Analysis

Upon collection of the diet it is homogenized in a Waring blendor.

lflhenever needed, water is added to bring the mixture to the desired

consistency (Bransby gt 21., 1948). The amount of water added should

/\

be measurl
ing the sI
(Tharton 5
When
the honoge
portions 3
for the va
children a
energy, ni
For n
IEfrigerat.
1938), sp<
values or e
POUnds Suc'r
For light-5
and aliqum

1964),

23
be measured accurately and recorded. Other investigators suggest bring-
ing the slurry to a constant weight (McKay gt 31., 1942 ) or volume
(Wharton g£_§1., 1953).

When a composite is collected for a balance period, a portion of
the homogenate is saved each day until the period is completed. These
portions are then thoroughly homogenized and then apportioned in duplicate
for the various analytical determinations. In the studies on preadolescent
children a separate aliquot was removed for each of the following--
energy, nitrogen and fat (So. Coop. Series Bull. No. 94, 1964).

For nitrogen and mineral measurements the diet composites may be
refrigerated, frozen, or convened into digests (Leichsenring 25 31.,
1958). Special manipulation of samples is required where food energy
values or amino acid contents are to be determined or where labile com-
pounds such as thiamine, riboflavin or ascorbic acid are to be measured.
For light-sensitive nutrients the diet samples should be composited

and aliquoted in darkened laboratories (So. C00p. Series Bull. No. 94,

Storage of the Sample

The sample may be analyzed immediately following homogenization
or it may be stored for future analysis. If the latter is the case,
the sample must be stored with precautions so that no objectionable
chemical changes occur in the diet. Leichsenring and co-workers (1958)
suggest the use of pharmaceutical bottles or round glass bottles, wide
<3r narrow-mouthed. Amber colored bottles are employed for storing
Séunples when analysis for light-sensitive nutrients are to be made.
Pleastic screw caps are inert to most chemicals and can be used to

Ptflaserve samples, although in the presence of toluene and solvents of

like natur
1958). Le
metal scre
to be erpl
placed ove
protection
samle in
weeks with
for storin
The t
(50. C00p,
Young
ing tray it
a DeSivac
nitrOgEn W-
111 ch.-
homoge'JIZei
When Const

Hen Store

c
Lat, Prote

Basse
SOdi‘Jm, p0
analYSis d
PuiverizEd

DrYin

:1.) (11

(D
r?

/

24
like nature, a cardboard interliner may be used (Leichsenring st 31.,
1958). Leichsenring and co-workersIl958) also advise that because
metal screw caps easily rust, precautions should be taken if they are
to be employed. Generally, a piece of cellophane or aluminum foil
placed over the mouth of the jar prior to covering will provide sufficient
protection. McCay £5 21., (1945) stated that storing the homogenized
sample in a cellophane bag in a freezer protects the sample for several
weeks without decomposition. Pilcher (1961) utilized jars and cartons
for storing the homogenized samples in the freezer.

The temperature at which the samples should be stored is -200 C

(So. Coop. Series Bull. No. 94, 1964).
Drying of the Sample

Young EE.El°: (1953) placed the diet composites in a shallow dry-
ing tray in a freezer for twelve hours. The sample was then placed in
a Desivac for drying, then weighed and ground. Analyses of fat and
nitrogen were performed on the dry samples.

In the study conducted by Kaucher and co-workers (1945) the
homogenized samples were dried to a constant weight at 60°C in an oven.
When constant weight was achieved, the samples were weighed, ground, and
then stored in- a brown bottle in a dessicator. They determined vitamins,
fat, protein, and energy on these samples.

Bassett and Van Alstine (1935) in preparing their diets for nitrogen,
sodium, potassium, magnesium, calcium, phOSphorus, chlorine, and iron
analysis dried the foods at 90° to 95° C. The sample was then

pnalverized and stored.
Drying at a temperature of 700 C and higher according to Teague

E35.§1., (1942) may result in volatilization of essential oils, loss of

nitrogen ano

that a prote

 

weight may st
III. TM

The er

two distinctl
the individue
total diet, c
first concerr
has the poter
(Leverton _e_t
Levertor
values VOUId
when analyze-c
should not a;
different Va:
duct consume;
(1960) vai:
fOOdS On a Ye
Widdows<

the Organic ‘
do HOt give

i

25
nitrogen and decomposition of fats and carbohydrates. They further report
that a protein containing substance after having been dried to a constant
weight may still contain 2 to 3 per cent water.

111. Two Methods For Determining Food Composition

The energy and nutrient content of a diet can be determined by
two distinctly different methods: (1) laboratory analysis of either
the individual food items of a diet or the prepared aliquots of the
total diet, or (2) calculations from tables of food composition. The
first concerns controlled experimental investigations, while the latter
has the potential for adaptation to field epidemiological studies
(Leverton gt 31., 1960).

Leverton and co-workers (1960) reported that the calculated food
values would only present a range of that nutrient in which the food,
when analyzed, will fall. Thus food values obtained by the two methods
should not agree exactly. The figures in the tables are derived in
different ways and provide somewhat different information about the pro-
duct consumed. The calculated values as emphasized by Leverton st 21.,
(1960) provide averages of the nutritive values of the diet or similar
foods on a year-round basis.

Widdowson and McCance (1943) stated that composition tables present
the organic constituents in a food for comparison with other foods. They
do not give any information as to the exact amount of nutrients which a
particular item contains. Harris (1962) stated that food tables do not
present data with an accuracy of atomic weight determinations, neither

are they so unreliable as to be worthless.

26

Food Composition Tables

 

The constituents of each food as listed in the tables of food
composition represent the average values derived from chemical analysis.
The number of values which composes the mean varies for each nutrient.
Patterson and McHenry (1941) reported that occasionally only minimal
or maximal figures are presented. Leverton.§£.al., (1960) postulated
that if there is a vast amount of data for a particular item, the
average of the values can be considered fairly reliable, whereas, those
with the least number of values will be more in error when related to
the analyzed values. The termayear-round is considered in tabulating
the data for the tables. This takes into account variety, seasonal,
and geographical differences, the consequence of storage for periods
prior to marketing,and the effect of manufacturing and preparation
practices (Leverton.g£_§1., 1960). A factor not mentioned by Leverton
gt‘gl., (1960) is the error inherent in analytical techniques. Thomas
and co-workers (1950) pointed out that the analytical values do include
variability which is attributable to sampling, preservation, and
measuring as well as errors inherent in the analytical methodology.

Conversion Factors

 

Errors may also be prevalent in food tables because of the con-
version factors employed. The determination of conversion factors
for the calculation of food energy from carbohydrate, fat, and protein
and those for computing protein from.the determined nitrogen have been
greatly neglected (Mayer, 1952).

Calories

A distinction is required between the calories as derived from the

heat of combustion and those which represent the physiological value.

27
The former is the number of calories produced by burning a weighed
amount of food in a bomb calorimeter. The latter represents the food
energy that is available for the body. This value takes into considera-
tion the digestability coefficient and the energy lost to the body
through urea formation.

Atwater (1899) compiled from different groups of people residing
in the different areas of the U. S. one hundred eighty-five dietaries.
He calculated the per cent of protein, fat and carbohydrate in the diets
for the following food groups: meats, milk, cereals, sugar, vegetables
and fruits. Heat of combustion values were then determined for the
protein, fat and carbohydrate in each food group (See page 43 for
values). The physiological value was derived by multiplying the
digestion coeffiCEnt, which was determined for each division of foods,
by the amount of protein, fat and carbohydrate contributed by the
separate groups. For protein 1.25 calories per gram'were substracted
for the nitrogen compounds excreted in the urine. These values were
then adjusted according to the extent that each food group occurred in
the U. S. diet at that time (Maynard, 1944). The values thus secured
are: 4, 9, 4 calories per gram of protein, fat and carbohydrate,
respectively

The Committee on Calorie Conversion Factors and Food CompoSition
Tables (1947) found no evidence that Atwater's values, if prOperly used,
were not reliable. However, if the pr0portions of food are not similar
to his averaged mixed diets, the application of these values can
present inaccurate food energy values (Maynard, 1944). Mayer (1952)
stated that if the common figures are used in computing the physiological

value of a high bulk, low calorie diet the calculated values would be

28

too high. It is emphasized by Maynard (1944) that Atwater never intended
that the factors 4, 9, 4 should be applied without modification to all
diets. For mixed diets which deviate from the average U. S. diet the
calorie values may be determined by two methods. They are: (1) applying
the individual fuel values for protein, fat and carbohydrate, respectively,
to each food or food group and (2) readjusting the proportions in the
average mixed diet which is 60:40, anima1:vegetable, and determining new
average values for the protein, fat and carbohydrate components,
respectively.

If the caloric value of a diet as determined by a bomb calorimeter

is to be compared with the calculated values, the values that Atwater

employed in calculating the conversion factors 4, 9, 4 should be used.
Erasers
Protein in a food is generally calculated from its nitrogen content

which is then multiplied by a conversion factor. These are the values

in food composition tables. Some food composition tables assume:

(1) all nitrogen in the food is protein and (2) all proteins contain

16 per cent nitrogen (Mayer, 1952). Jones (1931) reported that there

are few nitrogenous substances whose nitrogen is only in the form of
protein. Because of these gross assumptions the Committee on Calorie
Conversion Factors and Food Composition Tables (1947) classified the
common nitrogenous-foodstuffs into five groups and determined the con-
version factors for each division. They are: rice, 5.95; refined
wheat, 5.70; oilseeds and nuts, 5.30; meat and eggs, 6.25; milk, 6.38;
and fruits and vegetables,6.25. The latter factor for fruits and
vegetables is employed since there an no data to justify calculating a

special factor. If food composition tables determine protein values by

4"

\)

 

29
using the conversion factor 6.25, significant errors could result when
the research involves the comparison of the calculated and the analyzed
values.

There are many tables of food composition available. At best each
of them presents only approximate values for the nutrient content of
foods. According to Mayer (1952) there are two documents which take into
consideration the specific conversion factors as mentioned previously for
estimating calories and protein. The value presented for calories in
these tables is the physiological value. These two are the Food and
Agriculture Organization (United Nations) Food Composition Table for
International Use (Chatfield, 1949) and Agriculture Handbook No. 8 of
the United States Department of Agriculture Composition of Foods--Raw,
Processed, Prepared (Watt and Merrill, 1963). Mayer (1952) stated that
the U.S.D.A. publication may be considered as the authoritative document
for American foods.

Calculation versus Analysis

 

The constituents to be discussed in this comparison are protein,
fat, and food energy. The analytical methods employed for the chemical
analysis are as follows: protein, Kjeldahl; fat, ether extraction;
energy, bomb calorimeter.

Protein and Nitroggn

 

None of the investigators presented in the following section
indicated the conversion factors which were employed to compute the grams
of nitrogen from.the calculated protein value or the amount of protein

from.the analyzed nitrogen. These computations are necessary in order

to compare the calculated with the analyzed values for protein and

nitrogen. By implication it is possible to assume that the values used

30
were the same as those presented in the various tables of food
composition employed to calculate the diets.

Carroll 33 31., (1952) compared the calculated and analyzed protein
values for six diets from four cottages of a children's camp. Analysis
of variance revealed no significant differences between the two sets of
values. However, even though not significant the chemical analyses
resulted in higher values than the calculated in three of the four
cottages.

Wertz £5 31., (1952) found that the calculated values for protein
of twenty-one diets did not differ significantly from the determined
values. The statistical measure employed was the "'” test.

The nitrogen in the foods served in a hospital metabolic ward was
computed and analyzed by Bassett and co-workers (1931). They found close
agreement between the two methods. The statistical procedures used to
verify the authors' suggested close agreement‘une not reported.

Lutwak 35 31., (1964) in conducting a metabolic balance study on
children investigated the two methods for nitrogen. The authors report
an agreement that was within 10 per cent. Widdowson and McCance (1943)
found similar results.

Bransby 25 al., (1948) disagree with the preceding. They found a
statistically significant difference between the two sets of values
for protein. The statistics employed were not discussed. The average
analyzed value of the thirty-three composite diets was 76 grams of
protein per day, whereas, the average calculated value was 68 grams. The
composite represented the diets from three days of individuals living
at hOme. The standard deviation for the absolute difference in the

thirty-three composites was 5.6. Sixteen of the calculated values for

31
the diets revealed a difference of greater than 10 per cent of the
analyzed.

A consistent difference, even if slight, between the calculated
and analyzed values for nitrogen might be attributed to a variation in
the composition of foodstuffs produced under differing conditions of
atmosphere and soils. Donelson gt 21., (1931) reported the mean
determined value of nitrogen was 1.9 per cent higher than the calculated.
They encountered, on isolated days, marked divergences between the
calculated and determined values, whereas, on other days the two were
very close.

PE.

Diets of six subjects in which exact duplicates of the food eaten
were saved for analysis revealed that in five out of the six, the cal-
culated fat values were higher than the analyzed amounts. Two of the six
calculated values exceeded the analyzed values by more than 10 per cent.

Wertz gt $1., (1952) reported that the calculated values for fat
were significantly higher than the analyzed values. Of the twenty-one
diets eleven of the calculated values fell within 5 per cent of the
analyzed values. Significance was measured by the "t" test.

The calculated fat values of three moderate fat diets correlated
closely with the analyzed values (WOllaeger st 31., 1947). The analyzed
fat was 100.5 grams, whereas, the calculated value was 101.6. Lutwak
and co-workers (1946) found similar agreement. The analyzed values for
fat in their diets were within 10 per cent of the calculated values.

Fat and Calories

 

Fat and calories were determined by calculating and analyzing

the thirty-three composite diets in the studies conducted by Bransby

32
.gg'gl., (1948). The absolute and percentage difference between the
average nutrient values for all diets were not statistically significant
for calories or fat. For the individual diets the calculated values were
within 10 per cent of the analyzed in eighteen and twenty-seven of the
thirty-three composites for fat and calories, respectively. They suggest
that the better agreement for calories is due to the fact that an over-
estimate of the energy as derived from one of the three sources of food
energy, carbohydrate, protein and fat, is counterbalanced by an
underestimate of the other.

Protein, Fat and Calories

 

Mthy 25 31., (1945) conducted studies to evaluate the nutritive
value of the diets consumed at four large naval training stations. They
compared the values calculated for nitrogen, calories and fat with the
determined results. They reported that the calculated values were a
fair approximation of the determined values for nitrogen and calories,
however, the calculated value of fat was greater than the analyzed
amount. According to the authors the fat differences were probably due
to an over estimate of the amount of fat per cooked serving of meat. It
is difficult to understand why they reported that the analyzed food
energy is a fair approximation of the calculated value when the
calculated values for both diets were greater than the analyzed values
by more than 10 percent.

Similar trends are reported for thirty diets consumed by twelve
lactating women (Kaucher 55 31., 1946). The calculated value for fat
was fourteen per cent higher than the analyzed amount. The calculated
values for protein and calories were within 1.7 and 0.6 per cent,

respectively, of the analyzed values.

33
The authors reported that their analytical data represented food which
had been subjected to the hazards of marketing, storage, preparation,
cooking and serving, while the calculated values represented the fresh
foods.

Patterson and McHenry (1941) reported that for calories the calcu-
lated values are very close1:o the determined values for the same diet.
In the case of protein the calculated average is slightly lower, but is
still in good agreement. Their results are based on twenty different
lunches. The comparison of the two methods is reversed for fat, and
the large discrepancy leads to the opinion that the values for fat in
the food tables may be too great or that the loss of fat during food
preparation is fairly large. In their studies the deviation of
individual values from the mean is significant for protein and even
larger for fat. Such wide deviation indicates that the carbohydrate
value for foods, as listed in the food composition tables, is too low.
This assumes that the carbohydrate value was obtained by difference.

Leverton and Whiting (1960) collected approximately three hundred
cases in the literature which compared the two methods of determining
the nutritive composition of a food. The studies were performed in the
United States, Great Britain, and Canada. For protein and calories the
calculated values for more than 50 per cent of the cases fell within a
range of 10 per cent of the analyzed values. The calculated values for
fat were considerably higher than the analyzed determinations. In fact,
this factor was so consistent that if a chemically determined value for
fat was 140 grams then the calculated figure ranged from 130 to 185
grams. They concluded from their study that the data available in

in the literature support the fact that for epidemiological investiga-

tions the validity of food tables for giving a range of the actual intake

is questionable.

34
Individual Items versus Total Diet

Theoretically, the results of analyses of individual dietary
components should, on addition, provide the same value as that secured
from analySis of the total diet. To explore this possibility Hummel 35 31.,
(1942) analyzed twenty-two foods individually and as a composite diet.
These foods represented the foods consumed by normal children over a
period of several years. The results obtained from the individual
analysis of milk and banana more closely approached the values recorded
in food tables than for any other food analyzed. They reported that the
variability from time to time of the same foodstuffs is due to changes
in composition. The fat content of the analyzed foods showed the greatest
variation from the calculated figures. The individual values for the
same diet were within one standard deviation from the mean of the
analysis of the dietary composites for fat, nitrogen and food energy.

The composite diets showed a more constant composition than the components.
There was also better agreement between the analyzed and calculated

values for the composite than when the sums of the analysis of the
individual foods were compared with the calculated values.

Hunter and co-workers (1948) stated that the greater the number of
different food items in the diet being analyzed, the greater the chances
that the computed nutritive value will be correct. This statement
would not apply to an unstable nutrient like ascorbic acid. If the
analysis was not performed promptly on such diets or adequate means
adopted to prevent the destruction of the nutrient prior to analysis,
regardless of the number of food items, the analyzed results would not

approach the calculated values.

35
Individual Versus Average Diet Analysis

Toscani (1948) compared the analyzed and calculated values for
nitrogen in twelve diets. The diets were analyzed individually and
then as a composite. The individual diet analyses showed a large
variation when compared to the calculated nitrogen value. These
analyzed values ranged from 0.0 to 9.6 per cent below the calculated
values. The analyzed averages were only 3.7 per cent below the
calculated value.

Meyer and co-workers (1951) compared the analyzed values for the
nutrients in school lunches with the calculated values. Single lunches
were collected as well as the composites of the five lunches. There
was a wide range of difference between the calculated and analyzed values
for the single meals. However, for the composites the calculated values
were within 5 per cent of the analyzed values for calories, protein and
fat. Bransby 25 31., (1948) upon averaging the diet composites of thirty-
three subjects in their study found close agreement for protein and
calories. However, the values for the individual diets were so diffuse
that it was extremely doubtful, according to the authors, as to
whether it was worth the time and effort to calculate the individual
diets. The difference between the calculated and analyzed values for
calories was within 10 per cent in 82 per cent of the diets. For fat
and protein nearly 50 per cent of the analyzed values for the diets were
not within 10 per cent of the calculated values.

Variation in Analyses of Identical Diets

Hawks and co-workers (1937) hypothesized that there are variations

in the composition of identical diets when when experimental conditions

are carefully controlled. They analyzed eight and eighteen identical

36

diets, respectively, in two experiments for nitrogen and calories. The
correlation coefficient of variation ranged from 1.9 to 2.7 per cent for
calories. For nitrogen the spread was 1.6 to 3.0 per cent. The
coefficient of variation represents the extent to which the standard
deviation will vary from the mean. They presented three causes for
these variations in the composition of the same diet: (1) unavoidable
errors in chemical technique, (2) difficulty in obtaining a homogeneous
sample of food for analysis even under strictly controlled conditions
and (3) variation in the composition of food.

Bassett £5 21., (1931) prepared and analyzed the same diets for
nitrogen in different years and different seasons. Their average
analytical results agreed closely with the calculated values of all
the diets. They postulated that if a researcher throughout the course
of a study averaged the values obtained from analyzing d diet which
was replicated at ten times during the study the result would be the
actual composition of five of the ten diets.

Reifenstein 35 a1., (1945) in comparing the same diet prepared at
two different times and another diet prepared at three intervals found
a consistent 11 per cent deviation for the analyzed nitrogen values of
the same diet. Bassett and Van Alstine (1935) pointed out that the
fluctuations in the analyzed value for nitrogen of the same diets
represent the inherent variations in the concentration of the con-
stituents of the same food from time to time.

Differences in Analysis of Duplicate Samples
Hawks £5 31., (1937) reported that duplicate diets collected on the

same day and in the same manner varied as much as 4.8 per cent while

37
the average differences ranged from 1.0 to 2.0 per cent, respectively,
for calories and nitrogen. Again the variation in the composition of
food must be considered in these results. The differences observed in
the duplicate samples were less than those recorded for the same diet
analyzed on different days.

Upon analyzing duplicate samples, Thomas 25 31., (1950) found the
values for protein and food energy to be the most consistent and showed
little variation. However, the determined fat in the duplicate samples
varied immensely. These inconsistent differences, as hypothesized by
the researchers, are due to sampling. Many of the diets used in this
study contained mixed dishes. These were prepared in large receptacles.
It is impossible to control with any precision the amounts of the
individual conStituents of the dishes prepared in this manner. The
only way that a mixed dish can be prepared in a more exact duplication
is that the ingredients for each serving be weighed, cooked and served
in the same utensil. The authors concludelthat this procedure is
costly,yet in highly controlled studies it is more efficient to use
food preparation more amenable to exact sampling.

It is the purpose of this study to explain the dietary procedures
appropriate for a non-clinical metabolic balance study, to discuss the
calculated and analyzed values of the same diet, to present the
variability of composition of the same diet from different weeks of the
study, and to determine the time a sample should be removed for analysis

to assure that it represented the diet of the subjects consumed.

EXPERIMENTAL PROCEDURE

A metabolic balance study was devised to investigate bread as a
primary source of protein for man. The subjects were twelve male
college students. They were selected from forty volunteers who were
obtained by notifying several campus departments and through verbal
contact. The volunteers met with Dr. Olaf Mickelsen and the two
graduate students assisting with the study. At this time the purpose
of the study and the subject's responsibilities were explained. The
potential subjects were asked to record their age, height, weight,
religion and food likes and dislikes. Only those who were within
10 per cent of their weight as designated by standard height-weight
tables were chosen. Those subjects who were married or who did not live
close to the campus were eliminated. The subjects with food dislikes
which were not coherent with the objective of the study were rejected.
As a check on the subject's honesty, responsibility and adaptability to
the experimental controls, they were given the Minnesota Multiphasic
Personality Inventory Examination. The results of this and the assess-
ment of their health status by a medical examination served as the
final basis for the selection of the subjects. The subjects chosen were
three foreign and nine American students. Their ages ranged from
nineteen to twenty-seven years.

Each subject at the beginning of the study was given a notebook
in which he was to record daily activities, the amount consumed of the
pad libiggmpitems which were sucrose, tea, coffee and hard candies,
and the amount of daily rest. Any abnormal physical conditions were
also to be recorded. Also at this time each subject was assigned a

number.
38

39

The first part of the study consisted of twenty-one days during
which the subjects consumed the control diet. This was immediately
followed by a fifty day experimental diet. One week of the control
regimen followed the experimental period. The prime objective of the
study was for the diets to contain the same amount of protein in both
the control and experimental regimens.

Animal protein in small amounts was served during the control
period, whereas, throughout the experimental phase no animal protein
was allowed. The control regimen was calculated to supply an average
of 68.3 grams of protein, 115.6 grams of fat and 3,086 calories per
day (see Table 1). The diets (Table 2) contained only small amounts
of milk, eggs, meat and other animal protein. The experimental
regimen was calculated to provide an average of 63.6 and 89.2 grams of
protein and fat, reSpectively (Table 1). Wheat products accounted for
70 per cent of the calories and 90 to 95 per cent of the protein. The
remaining protein came from fruits and vegetables (Table 3).

The results obtained from.the subject during the control diet
were compared with the results of the same subject on the experimental
regimen.

The food was prepared by two cooks in the metabolic kitchen in
{the Home Economics Building. One cook worked between the hours of
5:30 a.m. and noon. The other from.noon to 6:30 p.m. Two students were
employed on a part-time basis to wash dishes and to clean the kitchen
and the dining areas.

All meals with the exception of the Sunday morning and Sunday
evening meals were served in the Home Economics Building. The Sunday
breakfast and supper were sack lunches. This was for economical

reasons and because of the subjects request for free Sunday mornings

40

and evenings. Only one cook was required to prepare Sunday's meals.
Meals were planned which could be packed and yet would provide the
same meticulous measurement as if the subjects received their meals
directly from the kitchen (Control Diet VII, Table 4 and Experimental
Diet VII, Table 5). The sweet rolls for Sunday breakfast during both
periods were baked on Saturday. The orange and sweet roll and the
bread, in the case of the experimental diet, were weighed and

packaged in cellophane sandwich bags. These were placed in paper sacks
which were labeled with the subject's number and were presented to the
subjects Saturday evening. The Sunday evening meal which consisted of
sandwiches, fresh fruit, and a baked dessert were weighed Sunday
morning, wrapped as previously described and given to the subjects
Sunday noon. On Monday through Saturday the subjects were given, at
dinner, a snack of either cookies, bread or crackers which was to be
consumed that evening (Tables 4 and 5). The snacks were packaged in
sandwich bags and labeled with the subject's number.

The control and eXperimental diets referred to the structure of
the diet designed for each period of the study. When used with the
Roman numerals,they designated a particular day of each regimen. Roman
numeral I through VII were the diets served Monday through Sunday,
reSpectively. The three meals, complements and snack composed one
day's menu or diet (Tables 4 and 5).

A seven day menu cycle was designed for each period (Tables 4 and 5).
The carbohydrate, protein and fat content of the menus and the recipes
(appendix) were calculated with the aid of the U.S.D.A. Handbook No. 8

(Watt and Merrill, 1950 and 1963) and Bowes and Church (1963).

.6»; .1
III ' b
4.1 «I \I.

l. /

\J

 

41

All recipes required by the menus were calculated on the following
basis. The ingredients were listed with the amount required in grams.
The carbohydrate, protein and fat in each ingredient were computed from
the food tables and totaled. If the product demanded a mode of cookery,
the weight prior to serving was determined. This weight’was obtained
when the recipes were standardized for time and temperature two months
before the study began. The grams of carbohydrate, protein and fat per
serving were then calculated in proportion to the prepared weight of
the recipe. Please refer to the appendix.

All foods employed in the study were purchased from the Michigan
State University Food Stores, Michigan State University Dairy Store
and a local grocery store. Whey-free butter was used for the experi-
mental phase of the study. It and the butter were produced by the
dairy department on the campus.

The frozen food items were purchased at the beginning of the
study in sufficient quantity for the entire study. The dry storage
items, such as flour and sugar, that could not be stored for the
duration of the study due to inadequate storage facilities were ordered
at the beginning of the study and stored until required at the University
food Stores. The perishable items such as fresh fruits and vegetables
were purchased twice a week and were of the same variety. The meat and
the eggs for the control period were purchased in sufficient quantity
for the length of this regimen. The meat was packaged in the amount
required by each day's menu. It was of the same variety and cut.

The fruits and vegetables used were frozen with the exception of
those listed as canned (Tables 2 and 3) or fresh. Apples, bananas and

oranges were the fresh fruits. The fresh vegetables were lettuce

42
radishes, green peppers, cucumbers, cabbage, tomatoes, celery and
carrots. All juices used were frozen and diluted with water according
to the specified directions on the container. The frozen strawberries
were sliced and sweetened.

The ingredients for each recipe and the food items served were
weighed on a Hanson spring scale and a Toledo scale. The Hanson scale
was employed for food items such as butter, jelly, sugar, salads, meats,
bread, crackers, and desserts; whereas, the Toledo scale was used for
weighing the ingredients of the mixed dishes and the foods served other
than those mentioned previously.

Upon weighing, the ingredients for the mixed dishes were mixed
thoroughly. In the preparation of the desserts and the bread, the
ingredients were mixed with the aid of an electric mixer. After the
mixed dishes were prepared, the amount specified per serving was
weighed into individual casseroles. The filled casserole dishes were
then covered and placed in a warming oven until they were served to the
subjects. The only mixed dishes not treated in this manner were the
dumpling and vegetarian stew in diet I (table 3) and the pizza dough
and sauce in diet VI (Table 3). The dumplings and the vegetarian stew
were cooked in a large receptacle and the individual servings were
weighed when the subjects arrived to be served. For the preparation of
the pizza, dough was weighed as wet dough per individual serving and a
designated amount of pizza sauce was placed on each individual pizza
tray.

For the preparation of the meats served during the control regimen,
please refer to the appendix.

The bread and all desserts in the menus were prepared daily. The

 

 

43
French bread, hot rolls and sweet rolls were made from the bread recipe
(appendix p.126 ). The amount of bread required for each meal was
weighed and packaged in ce110phane bags prior to serving.

The salads, fruits, and desserts were weighed and covered with
saran wrap prior to the serving periods.

The butter, jelly, honey and sugar were weighed in containers
labeled with the subject's number.

The method of cookery used for the hot cereals is listed in the
appendix under each specific cereal..

The diets supplied the recommended nutritive allowances with the
exception of calcium. Calcium was sufficient in the control diets,
however, during the experimental regimen the diets supplied an average
of 250 miflhgrams of calcium per day. In order to compensate for this
deficiency, calcium lactate pills which supplied 650 milligrams of
calcium were given to the subjects daily. A

The energy supplied by the diets was calculated by two methods.
The average physiological values of 4, 9, 4, for carbohydrate, fat and
protein, respectively, as derived by Atwater (1899) and the specific
heat of combustion factors for the particular groups of food. These are
the following:

Food Carbohydrate Protein Fat
(calqriés per gram)

milk 3.95 5.65 9.25
meat, fish 3.95 5.65 9.50
eggs 3.95 5.75 9.50
fruit 4.00 5.20 9.30
vegetables 4.20 5.00 9.30

1‘... hand]; . 211.11..
t

i. .

44

Food Carbohydrate Protein Fat
(Calories per gram)

wheat and

cereals 4.20 5.80 9.30
butter fat -- -- 9.25
animal fat -- -- 9.50
vegetable fat -- -- 9.30

The conversion factors employed to determine the amount of nitrogen
in the diet from the calculated protein were those of Jones (1931).
These factors for the reSpective food groups were milk, 6.38; meat,
eggs, fish, 6.25; fruits and vegetables, 6.25; wheat and cereals, 5.70;
and nuts and seeds, 5.30. The amount of nitrogen supplied by each food
group was determined by dividing the protein content of a food'group by its
specific conversion factor.

Since the objective of the study was to determine the effect of a
wheat diet on the nitrogen metabolic pattern in the subjects and,
furthermore, because weight loss or gain affects nitrogen balance, it
was most important to maintain the subjects' weight. During the control
period there was no change in weight (Bolourchi, personal communication
1965). However, during the experimental regimen some of the partici-
pants began either to loose or to gain weight. Additional calories
were supplied to those subjects losing weight in the form of whey-free
butter, jelly, hard candies, honey and sugar. These foods were allowed

.33 however, the amount consumed was weighed and recorded in

libitum;
the kitchen. For the subjects who were gaining weight, dietetic jelly
was allowed. Their whey-free butter and sugar intakes were rationed.

They received no hard candy or honey.

45

The items allowed 29 libitum to the subjects were coffee, tea,
salt and sugar. The sugar was not allowed ad libitum to those subjects
who were gaining weight. For this purpose instant coffee, tea bags
and sugar packs were employed. All items iSSued to the subjects were
recorded. The subjects were requested to record in their record books
the teaspoons of coffee, the number of tea bags and sugar packs that
were consumed each. day._ . The powdered coffee and tea were analyzed
for nitrogen. These results were employed to tabulate the additional
amount of nitrogen consumed by each subject.

A trial period utilizing five of the experimental menus was
employed prior to the balance study. The participants in the trial
period were not the same subjects who were chosen for the study. Both
males and females were in the trial group. The food was prepared and
served in the metabolic kitchen of the Home Economics Building. Food
samples which represented the same amount the subjects were to receive
were collected for analyses each day. The preliminary study occurred
one week prior to the actual study and was five days in length.

Because the balance period for the study was not the same length
as the number of menus, the diets were analyzed each day. The
determined values were totaled to represent each balance period. The
length of the balance period was five days.

During three weeks of the experimental period food samples of the
same amounts as served on the menus (Tables 4 and 5) were removed for
analyses when the first subject was served at zero hour. During the
fourth week of the experimental regimen the components of two diets

were collected. The first diet was removed as previously stated. The

second diet was removed when the last subject was served at one and

<44: .ul...!l.|.l.l 11:14.4

 

46
one-half hour after the serving time had begun.

To further evaluate the time a diet should be removed for analyses
to be assured of the subject's intake, two diets were prepared. The
meals of the diets were collected at zero, one-half, one, and one and one-
half hours after the serving time had begun. Individual food items of
three diets were collected separately for analyses. These samples were
weighed into Sealtest 16 ounce plastic-lined cartons. The total diets
for the same three days were also collected.

Upon collection the food items were placed in a tared #10 tin can.
The weight of the container was recorded on the can with a magic marker
pencil. The container was covered with a polyethylene cover and placed
in the freezer after the addition of each item. At the end of each day
the container plus the contents which included the three meals, comple-
ments and snack were weighed. The weight was recorded on the container
as well as the date and the diet number. This was stored overnight in
the freezer. The contents were then transferred with the aid of 710 m1.
of distilled water and a rubber Spatula into a Waring blendor. The diets
were homogenized for five minutes at medium speed and five minutes at
high Speed. Upon homogenization the slurries were put into 16 oz. Sealtest
plastic-lined cartons. The remainder of the slurry was discarded. The
cartons were labeled with the date and the diet number and placed in the
freezer until the analyses could be performed.

The wet slurries when removed from the freezer were thawed for
twelve hours. Then approximately 16 grams of the wet diet were placed
in a tared evaporating dish. All analytical weighings were performed

on a Mettler Balance. They were dried to constant weight at 60°C in

an oven with a vacuum of twenty-eight inches of Hg. Upon drying they

47
were ground in a Wiley Mill using a #20 mesh screen. The ground, dried
samples were placed in labeled bottles which were covered with plastic
screw tops. These were then placed in a dessicator.
The per cent moisture was determined by the difference in weight
between the wet sample and the dried sample. The actual per cent
moisture per diet was computed with the aid of the following formula:

(1) Wet weight of collected diet + water added x
(100 - avg. Z moisture) = dry matter.
(2) (Dry matter/gm. wet diet) x 100 = % dry matter.

(3) 100 - Z dry matter = Z actual moisture in original diet

The moisture was determined on the samples at two times. The time
intervals were Spring, 1964 and Spring, 1965. In 1964 the samples
were only dried for twenty-four hours at 60°C in an oven using a
vacuum of twenty-eight inches of Hg. In 1965 the samples were dried
as previously described.

One gram dried samples were weighed in duplicate for nitrogen
determination by the macro-Kjeldahl method. The formula used to
calculate the amount of nitrogen per sample was:

gm. of N in sample = ml. HCl (titration-blank x.§ of HCl
x 0.014 [milliequiv. wt. of NJ

Z'N (dry wt. basis) = gm. N/sample
sample wt.

 

gm. of N/diet =% N x dry wt. of diet

For fat determination duplicate 1 gram dried samples were extracted
with ethyl ether for three hours in the Goldfisch apparatus. The ether

was evaporated and then the beaker which contained the fat was weighed.
The weight of the dry beaker substracted from the former value was the
amount of fat per sample. The grams of fat in each diet were calculated

with the aid of the following formula:

48

gm. of Fat in diet = wt. fat extracted x gm. dry diet
wt. dried sample analyzed

 

One gram pellets were formed in duplicate from each dried diet
sample for the energy determinations. The instrument employed to
measure calories was the Parr Oxygen Bomb Calorimeter. Correction
factors were employed for the nitric acid produced and the unburned fuse
wire (Parr Instrument Company, 1960). The nitric acid was titrated with
0.0508 g NaOH. One ml of NaOH is equivalent to 0.801 calories and one
cm of fuse wire is equivalent to 2.3 calories. The formula employed to
calculate the heat produced in calories per gm of dry sample was:

,.

Gross heat in calories/gm = KE) (w) — c
S

t = temperature increase

w = water equivalent of the calorimeter = 1364 cal/degree F

c = calories of total correction from unburned wire and acid
formed

3 = sample weight

_ The water equivalent was determined by combusting standard benezoic acid.
Upon analyses, the determined values were compared with the
calculated values by the difference as per cent analyzed, standard
deviation, 3—way analysis of variance and the "t" test.
The standard deviation was determined with the aid of the formula

given in Dixon and Massey (1957).

N
s.d. =EZ, (xi " i)2 / (N ' U325
1 = 1

where N = the number of observations

xi- the value of each observation

i = the mean of the observations.

 

 

JR, “1.7

’1“. I... 4.3-... 3 .

 

 

49

A three-way analysis of variance was used to determine significance
between the calculated and analyzed values for the same diets. This also
determined the significance between the calculated and analyzed diets of
one week of the experimental regimen when compared with the other weeks
and the significance between the calculated and analyzed diets within
the same week. The calculated "F" values were compared with those listed
in the "F" distribution tables in Dixon and Massey (1957).

The "t" test was employed to further determine the reliability of

the food tables and differences in sampling time.

t = (M1 - M2) / (s.E.M.f + sang) $5
Where M1 = the mean of one set of observations
M2 = the mean of the second set of observations

SEMF Standard error of the mean of each observation
The standard error of the mean was determined by the method of Mantel
(1951). By this method the standard error is the range of values for
a set of observations divided by the number of observations. This
nethod is very accurate for samples of fifteen or less (Madal, 1951).
The "t" test was also used to determine the probability that the analyzed
values agreed with the values calculated from.the food tables.

One-way analysis of variance was applied to determine the signifi-
cance between the same diet replicated at four intervals during the
experimental period. The calculated "F" values were compared with those
listed in the "F" distribution tables in Dixon and Massey (1957, p. 388).

In order to evaluate the meals collected at the four time invervals

the maximum deviation was expressed as per cent of the mean.

50

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59

Table 4

Menus for the control period.

The Roman numerals refer to the different diets which are listed in

Table 2.

Complements are the items which the subjects could use with
one or all meals for that day.

 

 

 

 

 

 

Meal1 I gm. II gm.

Grapefruit Juice 150 Orange Juice 150

B Puffed Rice 14 Oatmeal 236

Toast 50 Toast 50

Milk 240 Milk 240

Spanish Rice 300 Onion Soup (canned) 200

French Bread 50 Saltine Crackers 25

L Relish Plate - Celery 25 Lettuce 50

Carrot 25 Tomato 50

Lime Sherbet 75 Bread 100

Ginger Cookies 50 Potato Chips 20

Cherry Kuchen 100

Roast Beef 50 Pork Chap 60

Oven Drowned Potatoes 100 Bread Dressing 150

D Green Beans (canned) 75 Carrots (canned) 75

Lettuce wedge 50 Shredded Cabbage/ 50
Pineapple Salad

Hot R0118 100 Hot Rolls 50

Peach Crumb Pie 135 Applesauce Cake 75

Butter 45 Butter 50

C Jelly 20 Jelly 20

S Refrigerator Cookies 60 Crunchy Cookies 50

 

 

 

1 The letters denote the following:

Complements, C; Snack, S.

Breakfast, B; Lunch, L; Dinner, D;

m. I. 'JIPIfsll. \IlLu .. a ‘L

I’ll.

 

60

Table 4 (Cont.)

 

1|

 

 

 

 

 

Meal III gm. IV gm.
Grapefruit Juice 185 Orange Juice 150
B Shredded Wheat Biscuit 22 Sweet Rolls 130
Toast 50 Rice Krispies 28
Milk 240 Milk 240
Beef Patty 4O Potatoes au Gratin 130
Bun 38 Bread Crumbs 20
Pickles -Dill 30 Asparagus Spears 50
Catsup 15 Mixed Salad

L Lettuce 20
Mustard 10 Green Pepper 10
French Fries 50 Tomato 20
Apricots (canned) 120 Bread 50
Cupcake 50 Melon Balls 100
White Icing 30 Molasses Cookies 40
Roast Chicken 40 Canadian Bacon 50
Steamed Rice 100 Harvard Beets 100
Stewed Tomatoes (canned) 100 Pineapple/Carrot Salad 50
D Lettuce 30 Hot Rolls 100
Green Pepper 20 Chocolate Cake 100

Hot Rolls 50

Lemon Pie/MEringue 160
C Butter 50 Butter 40
Jelly 20 Jelly 20
S Refrigerator Cookies 50 Ginger Cookies 50

 

 

 

1 The letters denote the following

Complements, C; Snack, S.

Breakfast, B; Lunch, L; Dinner, D;

61

Table 4 (Cont.)

 

 

 

 

 

 

 

 

 

1

Meal V gm. VI Aggm.
Sliced Banana 100 Grapefruit Sections 100
B Corn Flakes 25 Puffed Wheat 12
Toast 50 Toast 50
Milk 240 Milk 240
Vegetable Soup (canned) 200 Creole Spaghetti Sauce 100
Saltine Crackers 25 Spaghetti 100
Peanut Butter 20 French Bread 100
L Jelly 30 Pears (canned) 115
Bread 75 Refrigerator Cookies 60

Celery Sticks 25

Apple 230
Baked White fish(raw wt) 60 Veal Cutlet 4O
Butter 5 Boiled Potatoes 75
MaShed Potatoes 100 Fr. Cut Green Beans 100
D Peas 50 Fruit Salad 50
Lettuce Wedge 50 Lettuce 20
HOt R0113 75 Hot Rolls 75
Cherry Pie 135 Gingerbread lOO
Lemon Sauce 50
C Butter 40 Butter 50
Jelly 20 Jelly 30
S Crunchy Cookies 75 Mblasses Cookies 50

The letters denote the following: Breakfast, B; Lunch, L; Dinner, D;

Complements, D; Snack, 8.

62

Table 4 (Cont.)

 

 

 

 

 

MealIi VII gm.
B Orange 190
Sweet Roll 130

Cube Steak 50

Baked Potato 100

Whole Carrots (canned) 75

Lettuce 40

L Radish 10
Hot Rolls lOO

Milk 240

White Cake 100
Strawberries 100

Ham 40

Bread 50

D Potato Chips 20
Crunchy Cookies 75

Apple 230

C Butter 40
Jelly 10

 

 

The letters denote the following: Breakfast, B; Lunch, L;
Dinner, D; Complements, C.

63

Table 5

Menus for the experimental period.

The Roman numerals refer to the different diets which are listed in
Table 3. Complements are the items which the subjects could use with
one or all meals for that day.

 

 

 

 

 

 

MealIi . I II
Grapefruit Juice 120 Orange Sections 100
B Pettijohn 180 Farina 250
Toast 100 Toast 100
Dumpling (raw wt.) 120 Tomato Slices 50
Vegetable Stew 75 Lettuce 30
L Bread 100 Potato Salad 75
Pears (canned) 115 Bread 150
MolasSes Cookies 50 Gingerbread lOO
Tomato/Rice Casserole 245 Vegetable Chop Suey 350
Bread Crumbs/Butter 3O Chow Mein Noodles 38
D Lettuce 50 Lettuce/Green Pepper 40
Hot Rolls 200 Hot Rolls 200
Peach Crumb Pie 135 P' ple Upside Down Cake 75
C Butter (protein free) 40 Butter (protein free) 40
Jelly 10 Jelly 20
Bread 50 Crunchy Cookies 25
S
Jelly 15

 

 

 

The letters denote the following: Breakfast, B; Lunch, L; Dinner, D;
Complements, C; Snack, 8.

Table 5 (Cont.)

 

 

 

 

 

 

 

 

Meall III gm. IV gm.
Strawberries 100 Orange Juice 100
B Pettijohn 180 Cream of Wheat 235
Toast 100 Toast 125
Fruit Plate 227 Vegetable Soup 250
Bread 150 Bread 150

L
Refrigerator Cookies 50 Melon Balls 75
Molasses Cookies 50
Spaghetti 146 Creole Green Beans 170
Spaghetti Sauce 200 Lettuce wedge 25
D Sh. Cabbage/Pineapple 50 Hot Rolls 200
French Bread 150 Cherry Kuchen 100

Apple Pie 130

Butter (protein free) 40 Butter (protein free) 50

C
Jelly 15 Jelly 30
Bread 50 Ginger Cookies 75

S

Jelly 15

 

1 The letters denote the following:

Complements, C; Snack, S.

Breakfast, B; Lunch, L; Dinner, D;

65

Table 5 (Cont.)

 

MealI V

 

 

 

 

 

gm. VI gm.
Orange Juice 100 Grapefruit Sections 100
B Wheatena 270 Cream of Wheat 177
Toast 150 Toast 100
Tomato Slices 50 V-8 Juice 100
Cucumber Slices 25 Macaroni Salad 240
L Lettuce 30 Bread 100
Bread 150 Cherry Pie 130
Apricots (canned) 120
Crunchy Cookies 50
Boiled Dinner 150 Pizza Dough (raw wt.) 150
Fruit Salad 50 Pizza Sauce 175
D Lettuce 20 Lettuce 25
Hot Rolls 200 French Bread 100
Applesauce Cake 100 Fruit Ice 100
Refrigerator Cookies 25
C Butter (protein free) 40 Butter (protein free) 40
Jelly 20 Jelly 15
S Saltine Crackers 25 Bread 25
Jelly 15

 

 

 

1
The letters denote the following:

Complements, C; Snack, S.

Breakfast, B; Lunch, L; Dinner, D;

66

Table 5 (Cont.)

 

 

 

 

 

 

MealI VII gm.
Orange 100

B Sweet Rolls 200
Bread 50

Baked Potato 100

Fr. Cut Green Beans 100

Chef's Salad -Lettuce 30

L Radish 10
Hot Rolls 200
Shortcake 50
Strawberries 50
Cucumber Slices 25

Lettuce ' 25

D ‘Bread 150
Applesauce Cake 75

Apple 150

C Butter (protein free) 40
3 Ginger Cookies 25

 

 

l The letters denote the following: Breakfast, B; Lunch, L; Dinner, D;
Complements, C; Snack, S.

RESULTS AND DISCUSSION

Coons (1930) suggested that if the experimental diet is a drastic
change from the control diet a period should be allowed for the subjects
to establish equilibrium on the new regimen. No transitional period was
required by the "bread study" since the experimental diet contained approxi-
mately the same amount of amino acids as the control. Because of the low
level of nitrogen in the experimental diets, the control period was
employed so that the subjects could attain equilibrium. The objectives of
the study were to determine whether or not the subjects could maintain
nitrogen equilibrium on a diet in which 90 per cent of the protein was
from wheat flour. Reifenstein gt 31., (19455 warned against the
interpretation of a balance study when the subjects are placed on a diet
which is a change from the normal intake.

It has been pointed out that the length of a study is determined by
the objectives of the project. Donelson.g£.gl., (1931) stressed the
fallacies which a researcher can depict by interpreting a study that is
too short to reasonably fulfill its purpose. The interpretation of long
balance studies can also reveal results which are not possible (Walker,1962).
The twenty-one day control period allowed the boys to attain equilibrium
on the moderately low protein diet.

In the literature there are no statements concerning the relation-
ship between the personnel of a study and the subjects. It is feasible
to assume that a rapport, as discussed by Leichsenring 25.31,, (1958),
should be established so that the subject will admit any errors
committed. The errors in the "bread study" that were reported by the
subjects were eating food other than that allowed, collecting only some

of the excreta and not recording the ad libitum items. From the

67

68

observations of this study it can be hypothesized that the personnel

should deve10p rapport yet remain detached from the subjects. This was
apparent when breakfast was served. The serving time of breakfast was

from 7:15 to 8:30. There were several occasions when the morning cook
would tell one subject that it would be permissible if he came in later
than the set time. When this was discovered, stern comment was required

by the research staff in order to stress the importance of the meal serving
time. After several encounters the problem was improved and the subject
reported to breakfast within the set serving hours.

The trial period that was carried out prior to this study did clarify
the working procedures of the cooks. However, because the participants
were not the same pe0ple who were employed in the study, the remaining
purposes of such a period were not fulfilled. These were a development
of rapport between the staff and the subjects and an adjustment by the
subjects to the low protein diet and to the routines of the study. Thus,
the trial period seemed to have little merit. The control period met the
objectives of the trial period. Because of this factor it seems advisable
to use part of the control period as the trial period. Leichsenring 35.21.,
(1958) reported that during the trial period analyses need not be
performed. This is not true, for it is important that the personnel
involved in the analytical laboratory develop reliable analytical
techniques and also adapt to the demands of the study. Also, if the con-
stituents of the diets are determined, the reliability of the computed

diets can be evaluated.
It has been pointed out that there is less supervision of the sub-
jects in a non-clinical study. Bauer and Aub (1927) stressed the

importance of control of the subjects throughout the study, whereas,

69

McKay 23 gl., (1942) stated that the environment plays an important role in
the results of the study. Therefore, the latter authors report that the
environment should be, as much as possible, the normal living situation

to which the particular subjects are accustomed. Throughout this study

the only supervision the twelve subjects received was during the meal serv-
ing periods and during the various tests to which they were subjected. It
was believed that the subjects were trustworthy and honest throughout the
study. To create a home-like atmosphere during the meal service the sub-
jects picked up their own hot food from the cooks in the kitchen and their
cold food dishes from the tables in the dining room.

According to Johnston and McMillan (1951) the only fresh fruit to be
employed in metabolic studies is citrus fruit. The other fruits and
vegetables should be frozen or canned. The research with which the
authors were concerned was the investigation of the absorption of iron
from spinach. The "bread study" was mainly concerned with nitrogen in-
take from.wheat products. Since the fruits and vegetables chosen for the
study contributed only a negligible amount of nitrogen, the use of only
frozen and canned fruits or vegetables was not followed.

(The food items (Tables 2 and 3) consisted of many mixed dishes and
desserts. They were used due to the length of the study, the subject's
emotional aspects, and the objectives of the study. Because no meat or
dairy products were allowed in the experimental diet, an entree was
impossible to prepare without the use of mixed dishes. If mixed dishes
had not been employed, the subjects would have received several separate
dishes of individual vegetables. Spaghetti and macaroni could have been
presented with the vegetables. This type of diet is bland and unattractive.

For fifty days the subject had to be presented food that was appealing and

[14:05. I |.|1'I. I. ..

i.

 

7O

appetizing. Most of the mixed dishes contributed only a small amount of
nitrogen to the total diet. The emotional disturbances which might be
produced by forcing the subject to eat undesired food could yield results
which would not be a direct effect of the diet but rather due to the sub-
ject's emotional conflicts. Schottstaedt and co-workers (1958) showed
that there were metabolic changes in the excretion of nitrogen when the
subjects were under stressful conditions.

Sampson 25 31., (1952) implied that desserts such as cake and pie
are not to be incorporated into metabolic balance study menus. The
researcher must avoid such a statement without the consideration of the
objectives of the study. During the experimental phase 90 to 95 per cent
of the protein was derived from wheat products. One loaf of bread per day was
consumed by each subject. They also received cake, pie, cookies, cereals,
and some entrees which contained wheat products. These foods were
required to attain the level of the type of protein required by the study.
Such foods could not have been used if the prime objective of this study
had been the determination of fat and calories. These were determined
but only for the purpose of knowing the amount consumed by the subjects.

In calculating the nitrogen intake of each subject from coffee and
tea several problems were encountered. Because the coffee was weighed
only at the beginning and end of the study, it was impossible to deter-
mine the amount consumed by each subject in a balance period. The total
weight as obtained from weighing the returned coffee at the end of the
study did not in 75 per cent of the cases equal the amount recorded by
the subjects. In these instances the amount recorded by the subjects
was too low. Since tea bags were employed, it was impossible to determine

the exact strength of tea consumed. As the strength of the tea

71

increases, the amount of nitrogen increases.1 In future nitrogen balance
studies the instant coffee and tea should be weighed at the beginning and
end of each balance period in order to determine the exact amount consumed
by the subject. With the coffee and tea,sugar was dispensed to the sub-
jects in individual packets. These were easy to handle and the amount
of sugar as listed on each packet allowed for accurate recording.

Maintaining the weights of the subjects during the experimental
regimen required excessive amounts of whey-free (protein-free) butter,
jelly, sugar and honey per day. The amounts of these complements
were as much as 150, 100, 50, and 35 grams of butter, jelly, honey and
sugar, respectively, per day. The whey-free butter contributed many cal-
ories to the diet of these individuals without increasing their nitrogen
intake. No weight gain was observed in the individuals losing weight
even when the intake was 4300 calories per day. In some subjects there
was a slight continual loss of weight even at this caloric level. The
weather at the beginning of the study was cold, however, after the
experimental regimen began the weather moderated. The subjects began to
increase their physical activity with this change of weather. Bricker
.g£.§l., (1949) suggested an experimental design in which there is a con-
trol group throughout the experiment. Because the facilities for such a
study were inadequate on this campus, this design could not be followed.
However, had it been employed more valid assumptions could be made. If
the weight loss had been observed in both the experimental and control
groups during the same weather conditions, the weight loss could be
attributed with more certainty to the change in weather which in turn

increased the physical activity. There were three subjects who showed

 

1 The analyses of the tea bags when placed in 170 ml. boiling water

"for one-half, one, and one and one-half minutes revealed 0-014:
0.024, and 0.027 gm. of N, respectively.

72

weight gains. One subject received only dietetic jelly and small amounts
of sugar after the first week of the experimental regimen in order to
keep his weight gain to a minimal.

When requesting food dislikes of an individual some times they cannot
all be recalled by the subject. Such a case occurred in this experiment.
One of the subjects disapproved of Harvard Beets. However, he did not
note this on the questionnaire. Harvard Beets were included in control
diet IV (Table 4). The subject's dislike for these was conveyed to the
other participants and aroused some emotional disturbance within the
group. This observation agrees with that of Marble (1939), Presson
(1955), and Sampson and co-workers (1952). The importance of the listing
should be stressed to the potential subjects of a balance study. Many
people think they can tolerate any food. However, when the subject is
placed on a control regimen and is required to eat each dish at least
once a week, the tolerance the subject stated that he had for the food,
lessens.

The Hanson spring scale according to Widdowson and McCance (1942)
and Coons (1930) is accurate to one gram. The accuracy of this scale
and the Toledo scale was determined by weighing a specified gram weight
eight successive times. Each time that the weight was removed the indica-
tor on the Hanson scale was returned to zero (Refer to Table 6).

It is true that the Hanson scale does weigh accurately to one gram.
However, the per cent error for the various weights must be considered.
The results show that there is an increase in the per cent of error for
the smaller weights for both scales. The Hanson scale shows an under-
estimation for all weights except for the 100 gram.weight. This weight

on the Hanson scale reveals less deviation than the same weight on the

 

 

 

 

73

Table 6

The validity of the scales employed in the metabolic kitchen.

 

Hanson spring scale

 

 

 

 

 

 

gm.‘weight
Weighing 10 20 50 100
1 9.0 19.0 49.5 100.0
2 10.0 18.5 50.0 99.5
3 9.0 19.0 50.0 100.0
4 10.0 19.5 50.0 100.5
5 10.0 19.0 50.0 100.0
6 9.5 19.0 50.0 100.0
7 9.5 19.0 49.5 100.0
8 9.5 19.0 49.0 100.0
Delta Mean Deviation 0.44 0.88 0.25 0.13
Per Cent Error 4.4 4.4 0.50 0.13
Toledo scale
gm. weight
Weighing 100 200 1200 3200
1 101.0 201.0 1200.5 3201.5
2 100.0 200.5 1200.5 3201.0
3 100.5 200.5 1200.5 3201.0
4 100.5 201.0 1200.0 3201.0
5 100.0 200.5 1200.5 3201.0
6 100.5 200.5 1200.0 3201.0
7 100.0 200.5 1200.5 3201.0
8 100.0 200.0 1200.5 3201.0
Delta Mean Deviation 0.31 0.56 0.38 1.06
Per Cent Error 0.31 0.28 0.03 0.03

 

74
.
Toledo scale. It is possible to say that for weighing small items
(less than 50 grams) where less than 5.0 per cent error is desired, the
Hanson scale should not be employed.

The food items served which weighed less than 50 grams and which were
weighed on the Hanson scale in both regimens were butter, jelly, sugar,
salads, saltine crackers, dry cereals, bread, meat and cookies. The meat
and dry cereals were served during the control diet. The remaining foods
listed are low in nitrogen or nitrogen-free with the exception of bread,
cookies and saltine crackers. The majority of these items were greater
than 50 grams and none of these were below 25 grams. The ingredients
of the recipes which weighed less than 100 grams were weighed on the
Hanson scale, whereas, for those greater than 100 grams the Toledo scale
was employed. Only in one of the recipes did the items that contributed
to the source of wheat protein weigh less than 40 grams and only in two
recipes were the weights of the wheat products less than 56 grams. Since
the nitrogen contributing items were greater than 50 grams throughout
most of the experiment, the error induced by the employment of these
scales was negligible. The scales were checked for accuracy periodically
throughout the study with weights.

Upon using food tables to calculate the nutritive value of the diets
an error caused by a misrepresentation of the protein value of flour
occurred. The amount of protein in all purpose, enriched flour as
listed in the U.S.D.A. Handbook No. 8 (Watt and Merrill, 1963) is 10.5
per cent. This figure was employed in calculating the diets. Analysis
of the flour used in the study revealed a value of 12.8 per cent protein.

This figure was based on the nitrogen analysis by the macro-Kjeldhal

-

7.5
method. The protein was calculated by multiplying the nitrogen by the
conversion factor, 5.70 (Jones, 1931). The "high protein" flour used
in the study was Gold Medal Kitchen Tested, enriched, all purpose,
brominated "Better for Bread." This was introduced by General Mills in
1953 (Celender, Personal Communication, 1964). It is packed only in 25,
50, and 100 pound bags and its distribution is primarily in areas where
there is home bread baking. Its protein content ranges from 12.0 to
13.0 per cent. When 12.8 per cent was employed to calculate the protein
in the flour, a closer correlation was observed between the calculated
and analyzed values for the diets. The average calculated values using
the higher protein figure for the flour revealed 68.4 and 63.7 grams of
protein for the control and experimental diets, respectively. The
analyzed average values for the seven diets were 64.4 and 63.3 grams of
protein, respectively, for the control and the experimental regimens.

The flour was calculated to contain the following by weight:

water 12.0 per cent
Protein 12.8* per cent
Carbohydrate 73.8 per cent
Fat 1.0 per cent
Ash 0.4 uper cent

* represents analysis from this laboratory

The error introduced by employing the protein value listed in the
food tables for flour demonstrates the inadequacy of such tables. They
only can be assumed to present the approximate nutritive value of the
items.

The results of the two methods employed in calculating the energy
content of the diets can be observed in Table 7. If the heat of combustion
values are employed, there is a closer agreement with the analyzed values.

It can be observed that if the 4, 9, 4 values, which represent the

76

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77

available physiological food energy per gram of carbohydrate, fat, and
protein, respectively are employed errors can result. Thus, to make a
more valid comparison, the correct heat of combustion factors must be
used.

Jones (1931) criticized the protein values of foods listed in the
food tables. If the food tables do not take into consideration the two
assumptions that are assumed in nitrogen determinations, the protein
value represented could be false. These assumptions are: (1) that all
protein contains 16 per cent nitrogen which accounts for the conversion
factor of 6.25, and (2) that all the nitrogen in the food is protein.

The U. S. D. A. food composition tables do consider these assump-
tions. The protein values reported therein (Mayer, 1952) represent those
calculated using the Specific factors for converting the analyzed nitrogen
to protein. The effect of using the wrong conversion factor to deter-
mine the amount of nitrogen in the control diet from the calculated
protein figure can be seen in Table 8. Since the control diets contained
only a minute amount of animal protein, the value of 6.25 which is
employed to determine the protein intake in most American diets, could
produce false values. However, when the specific factors for the food
groups are employed, a closer correlation between the calculated and
analyzed nitrogen is secured. In the experimental diets a different
picture is presented. The factors 5.70 and 6.25 were used to convert
calculated protein from the wheat products and fruit and vegetables,
reSpectively. Since such a small amount of protein came from foods other
than those containing wheat, ifi did not matter whether 5.70 was used

exclusively or the specific factors were employed. Both factors

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Comparison of dietary nitrogen calculated using various factors for converting

78

Table 8

the protein values listed in tables of food composition with the analyzed
nitrogen values.

 

 

 

 

 

Calcd. N (gm.) Analyzed

Diet Reg. 6. F. 1 Specific c. F. 2 I N (gm.)3
Control I 10.6 11.1 11.4
Control II 11.3 11.9 12.9
Control III 10.7 11.0 10.8
Control IV 10.8 11.4 11.0
Control V 11.0 11.6 10.7
Control VI 10.9 11.4 11.2
Control VII 19_9_ 1L2 11_l_+_
Mean 10.9 11.4 11.3
Exptl. I 11.5 11.4 11.8
Exptl. II 11.4 11.3 11.1
Exptl. III 10.8 10.7 10.7
Expfl. IV 11.3 11.2 11.2
Exptl. V 10.9 10.9 11.2
Exptl. VI 10.9 10.8 10.9
Exptl. VII 1112_ 11.2 1111
Mean 11.1 11.1 11.1

 

 

 

 

N

The regular conversion factor used for the control diets was 6.25; that for

the experimental diets was 5.70 which was prOposed by Jones (1931) for

wheat and cereal products.

The specific conversion factors proposed by Jones (1931) were applied to
each food group. (Refer to p. 44).

The control diet's nitrogen determination represents the average of the
analyses on the same diet collected at zero hour and one and one-half hour
after the food was prepared. The experimental diet's nitrogen determination
represents the average of the analyses on the same diet collected at zero
hour for each of four successive weeks.

Us.)

79

resulted in a close agreement between the calculated values and the
determined nitrogen content.

The reliability of food tables was demonstrated when the calculated
and analyzed values of the diets were compared. Table 9 shows that the
calculated nitrogen fell within 5 per cent of the analyzed values in five
out of the seven control diets. Diet V reveals that the calculated nitro-
gen is 8.4 per cent greater than the analyzed, whereas, diet II discloses
that the analyzed value is 7.8 per cent greater than the calculated. The
calculated fat value of diet I is within 3.5 per cent, diet II is within
5 per cent, and all the other diets (III-VII) reveal that the calculated
‘fat is greater than 10 per cent of the analyzed values. The determined
food energy is within 5 per cent for five of the seven calculated
values. Diets III and VI show calculated calories to be greater than 10
per cent of the analyzed values.

The nitrogen data is in agreement with Lutwak £1 31., (1964) and
Widdowson and MCCance (1943). In their studies they reported that the
calculated nitrogen was within 10 per cent of the analyzed. Kaucher
and co-workers (1945) found in their studies a close comparison between
the calculated and analyzed values for food energy. However, the
values for fat were 14 per cent higher than the analyzed. The calculated
fat values in the "bread study" are 14 per cent and 13 per cent higher
than the analyzed for the control and experimental diets, respectively.

In observing diet III of the control menu in Table 4, there are
two food items exclusive of butter which contained large amounts of fat.
These are lemon pie/meringue and refrigerator cookies. The fat probably
was not distributed homogeneously in these food items which would lead

to error. Hawks 35 31., (1937) and Thomas et a1., (1950) stated that it

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81

is impossible to collect a sample for analysis which is homogeneous.
The latter author further emphasized that no two servings of a food
which is prepared in one large receptacle can contain the same components.
Diet IV also contains two items which are high in fat content; they are
sweet rolls and chocolate cake. The milk and the ueats which were all
lean and trimmed of fat prior to cooking can be considered homogeneous
in their fat content when compared to the mixed dishes. The desserts in
diet V could also be subject to variation in the amount of fat dispersed
in each serving. Refrigerator cookies appear again in diet VI. They
were served in diet I but the difference produced in the analyzed versus
calculated fat was not affected. Diet VII food items which could con-
tribute to the differences in calculated versus analyzed fat are sweet
rolls, white cake, and crunchy cookies. The desserts and the bread which
could play a role in the fat differences were prepared with the aid of
an electric mixer in order to make the foods as homogeneous as possible.
The inability to disperse the fat evenly in the foods could also account
for the differences between the calculated and analyzed values for
calories. However, these differences are less than for fat, but the
trends are the same except for diets I and II where the calculated
values are higher than the analyzed.

Similar results are found for the experimental diets (Table 10).
All but four of the diets calculated are within 5 per cent of the

analyzed values for nitrogen. The diets not within this range are IA,

IIB, VID, and VIIB.1 According to the literature

 

The capital letters denote the four successive weeks of collected
diet samples: A, week 1; B, week 2; C, week 3; and D, week 4.

82

 

 

 

 

 

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86

(Leverton and Whiting, 1960) the accepted range for the calculated values
for protein, fat, and calories to represent the determined amount is 10 per
cent. Leverton and Whiting (1960) after reviewing three hundred cases in
the literature, which compared calculated diets with the same diets
analyzed, found 58 per cent and 54 per cent of the calculated values for
calories and protein, respectively, to be within 10 per cent of the analyzed
values; however, only 25 per cent of the calculated diets were within 10
per cent of the analyzed values for fat. The calculated nitrogen and
energy values are within 10 per cent of the analyzed for 86 per cent of
the experimental diets in the "bread study"; however, the calculated fat
is within 10 per cent in 50 per cent of the analyzed diets. Leverton and
Whiting (1960) found only 25 per cent of the calculated diets for fat to be
within 10 per cent of the analyzed values . About half the calculated
fat values were higher than the analyzed. In the "bread study" during
both regimens the calculated values are higher than the analyzed in 100
per cent of the cases. For the experimental diets 71 per cent of the
analyzed calories are below the calculated values. In reviewing the
sources of fat in the diets, it is possible that the heterogeneity of the
samples caused the deviation of the calculated fat and calories from the
analyzed values.

Upon applying 3-way analysis of variance, no significance is found
in the differences between the analyzed and calculated nitrogen values.
The values from diet to diet are also not significant for the seven diets.
The analyzed fat values show a significant difference from the calculated
values at the l per cent level. There is also a 1 per cent significance
among the seven analyzed diets when compared with the calculated values.

The differences between the calculated~and analyzed food energy are

87

significant at the 5 per cent level. There is also a significant differ-
ence among the seven experimental diets at the l per cent level in the
calculated versus the analyzed food energy. There is no significant
differences for nitrogen, fat, and energy for the diets served one week
to those in any other week. It can be assumed that the seven diets were
the same in composition from one week to another.

These results suggest that the food table values for fat are not
reliable. The calculated values for energy give slightly better agree-
ment with the analyzed values than the fat values.

The ”t" test for nitrogen in the control diets reveals a probability
that the analyzed value will approximate the calculated value 80 per cent
of the time (Table 9). For food energy there is only a 20 per cent proba-
bility that the analyzed value will approximate the calculated value. An
even lesser probability is found with fats there being only a 5 per cent
possibility that the determined value will represent the calculated figure
as obtained from food tables. In the experimental diets similar results
are found (Table 10). These are in agreement with the 3-way analysis
of variance. The calculated nitrogen will represent the analyzed values
nearly 100 per cent of the time. However, for fat, as well as food
energy, there is only a 1 per cent and 5 per cent probability,
reSpectively, that the determined value will represent the calculated
value.

Upon observing the "t" test for the individual diets differences '
can be observed. The nitrogen is below 0.6 in two cases. These are diets
I and V. This means that in some diets the calculated results are more
reliable than in other diets. In diet I it can be postulated that the

reason for a low probability is due to the dumpling and vegetarian stew

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88
served at lunch. The dumpling dough was weighed upon mixing and placed in
the stew. The subjects were then served one dumpling and 75 grams of the
stew. Since the dumpling contributed approximately 10 per cent of the
nitrogen, errors made by the kitchen personnel could have caused a differ-
ence in the amount of dumpling the sample received, thus causing signifi-
cant differences in the amount of nitrogen in the total diet. In future
studies entrees of this type should be eliminated. In diet V some varia-
tions could have been introduced by the boiled dinner served at dinner or
the cereal served at breakfast. However, there is a high probability of
the analyzed nitrogen approximating the calculated values in the remain-
ing experimental diets in which hot cereals and similar entrees were
served. Because the cereals and entrees were prepared in one receptacle,
the chances of obtaining a sample which is representative of the whole
is highly unlikely. This would account for the discrepancies in the
"t" test results for nitrogen.

The probability that the fat and food energy will approximate the
calculated values is low in all diets for fat and in five for calories
(Table 10). In diets V and VII, there is a probability of 0.6 and 0.9,
respectively, that the analyzed values for energy depict the values as’
calculated from the food tables. Since the fat revealed a low probability
for all diets, it can be said that this was due to the heterogeneity of
the sample. Since these diets were relatively low in fat (especially
when considered on a total caloric basis), any difference in fat content
would have a rather minor affect on caloric value of the total diet.

Upon making corrections for the calories according to the deviations
for nitrogen and fat these results agreed very closely with the differ-

ences in calories between the calculated and analyzed figures. These

89
calculations were based on the assumptions that the differences were
furnished by carbohydrate.

These statistical tests performed on the experimental and control
diets agree with those of Patterson and MCHenry (1941). These authors
reported that the values for fat in the food tables may be too great.
They obtained their values for calculation from the Canadian Tables of
Food Composition. However, it is possible to interject the statement that
the United States food composition tables are also too high for fat. If
this is true, the estimated carbohydrate in the diet is too low. This is
assuming that the carbohydrate is that which remains after subtracting
protein, fat, and minerals. Leverton and Whiting (1961) and Widdowson
and McCance (1942) reported that when the amount of variation between
the calculated versus analyzed fat is greater than 10 per cent, doubt can
be cast on the use of food tables for calculating fat for epidemiological
studies. FUrthermore, the tables should not be used with any validity
for the determination of the nutrient values of diets in metabolic
balance studies. Thomas _£_g1,, (1950) also found no close agreement be-
tween the calcu1ated and analyzed fat in their studies. They state that

this is due to the great variation in the concentration of fat in foods.

It is interesting to note that the analyzed.mean t Standard devia-

tion for fat in each diet is lower than the calculated values. This
only further points to the fact that the composition tables are not
reliable for calculating fat.

The argument for the unreliability of fat in regard to food tables
‘mmst take into consideration the weighing and preparation techniques
employed in the kitchen and the method used to determine fat. First are

the weighing errors in the kitchen. These include weighing the

”Gift!!!
xi.

 

9O

ingredients for each recipe and the amount to be served. The losses of
fat during cooking produces little if any volatilization of the fats. The
method of mixing the ingredients is the second source of error produced

in the kitchen which affects the dispersion of the fat in the food. For
this reason, Sampson EE.§l" (1952) warned against the use of mixed

dishes in a balance study. A dish consisting of many ingredients will be
less accurate in total fat composition than a dish with only one or a few
ingredients.

Many authors (Leichsenring 35-31., 1958, and Coons, 1930) did not
advocate placing fat in the diet collected for analysis. This is due to
the difficulty of preparing a homogeneous mixture when fatty substances
are included.

In the M.S.U. study the fats were included in the diet collection
to provide a measure of the total caloric intake of the subjects.

In the analytical methods there are several sources of errors. The
most important errors are those in the methods itself. The fat content
which was determined by the Goldfisch apparatus represents crude fat.
This method involves weighing a beaker of approximately sixty grams. A
one to two gram sample is extracted with diethyl ether. The fat which
remains in the beaker is determined by reweighing the beaker after evapo-
rating the solvent. This fat amounted to approximately 0.1200 grams in
the diet samples analyzed. This is a minute weight in comparison to the
weight of the beaker. In order to determine the reliability of this

method, severa1 samples were extracted on three occasions (Table 11).

91

Table 11

Per cent fat in three separate diets
analyzed on three different days.

 

Sample Trial1
a b c
1 9.13 8.92 8.91
2 9.48 9.51 9.53
3 9.67 9.74 9.78
1

All values represent the average of duplicate analysis.

These variations appear small when employing them in determining
the total fat per sample based on a dry weight basis; however, these
figures give some indication of the analytical variability. The collected
fat represents the fat that is ether extractable. Supposedly, the fat
listed in the food tables represents the fat that is ether extractable.
Harris (1962) reported that a more efficient and reliable method needs
to be devised for determining fat. The type of fat employed in both the
control and experimental regimens was butter, Wesson oil and "BlueiLabel
Kraft" all vegetable shortening. The fat intzhe butter and Wesson oil
was considered ether extractable. The Kraft Foods Company2 stated that
the shortening is entirely ether extractable.

Thomas 22021-3 (1950) reported that energy values are highly subject
to miscalculation. The heat of combustion factors were employed in the
M811. study for comparing the calculated energy as determined by a bomb
calorimeter. A non-homogeneous sample could be the factor producing the
differences in the calculated versus analyzed energy. Also, if the values

for fat in food tables are too'high, agreement betweenthe calculated and

analyzed results could not be expected.

The Kraft Foods Company (1962) Personal Communication.

92

The calculated nutrients in diets as reported in the literature
gave a closer correlation with the analyzed values when the entire
diet was analyzed rather than when the components of the same diet
were analyzed individually. Hummal'g£.§1., (1942) analyzed 22 in-
dividual foods for nitrogen, fat, and calories. The diet analyzed
13.1212 more closely approximated the calculated values than the sums
of the individual components. From Table 12 similar results can be
observed. In experimental diets III and IV there is essentially no
difference between the values for the entire diet when the analyzed
and calculated values are compared. There is also a closer agreement
between the calculated values and the diet analyzed in total than
when compared with the individual analyzed values in diet V. The accuracy
of determining a small amount of nitrogen is one of the errors en-
countered in this experiment. Since the analyzed values for nitrogen
are in close agreement with the calculated figures, it seems apparent
that the diet should be analyzed as a total composite. Bricker
21‘31., (1949) divided their food to be analyzed according to the type
of nitrogen in the products. The results of Table 12 cast doubt
on the reliability of analyzing minute amounts of food for nitrogen.

The literature does not clearly state the number of times that
a diet in a balance study should be analyzed so that one can be
assured that the analyzed results represent the composition of the
diet. Reifenstein 31 21., (1945) and Bassett and Van Alstine (1935)
advocated analyzing a diet several times throughout a study and thus,
employing the determined results for the length of the study. It

was stated by Reifenstein 31.31., (1945) that the most reliable

93

Table 12

Comparison of calculated versus analyzed values of individual foods that
were used in preparing three experimental diets.

 

 

Foodstuff E.P.3 N. (gm.) N. (gm.) N. (gm.)
gm. Anal. Calc}' Anal.2

EXPERIMENTAL DIET III

Strawberries 100 0.10 0.09

Fruit Plate 227 0.27 0.24

Spaghetti Sauce 200 0.40 0.32

Shredded Cabbage/Pineapple 50 0.17 0.11

Pettijohn 180 0.61 0.52

Spaghetti 146 1.26 1.30

Bread 450 6.53 7.03

Apple Pie 130 0.31 0.52
Refrigerator Cookies 50 9152. 9152'

10.18 10.72 10.7

EXPERIMENTAL DIET IV

Orange Juice 100 0.12 0.13

Melon Balls 75 0.08 0.07

Vegetable Soup 250 0.25 0.25

Creole Green Beans 170 0.34 0.49

Lettuce 25 0.04 0.05

Cream of Wheat 235 0.85 0.90

Bread 475 6.91 7.44

Molasses Cookies 50 0.44 0.49

Ginger Cookies 75 0.80 0.82

Cherry Kuchen 100 9169 9121

10.42 11.21 11.2

 

94

Table 12 (Cont.)

 

 

 

Foodstuff E-P. N. (gm.) N- (gm.) N. (gm.)
gm. Anal. Calcfl Anal.2
EXPERIMENTAL DIET V
Orange Juice 100 0.12 0.13
Fruit Salad 50 0.05 0.04
Apricots 120 0.10 0.10
Tomato 50 0.09 0.08
Lettuce 50 0.08 0.10
Cucumbers 25 0.02 0.03
Boiled Dinner 150 0.32 0.22
Wheatena. 270 0.86 0.87
Bread 500 7.26 7.81
Saltine Crackers 25 0.43 0.39
Crunchy Cookies 50 0.41 0.44
Applesauce Cake 100 0.59 9163_
10.33 10.84 11.2

 

1 Nitrogen represents protein content calculated from.the U.S.D.A. Hdbk.
No. 8 (watt and Merrill, 1950 and 1963) divided by conversion factors
for each food group as prescribed by Jones (1931) (Refer to p. 44).

Represents the average of duplicate analysis of the diet collected

during four successive weeks.

E,P, represents edible portion of the food items.

95

method-to be used is to analyze the diet daily or for each collection
period. However, this is laborious and costly for any experiment.
In this study the determined nitrogen values for the same diet prepared
in successive weeks are significantly different at the l per cent level
in all but two cases (Table 13). Diet VI shows significance at the 5
per cent level and the differences for diet IV are not significant.
That is to say, in all instances except one there is significant
variation in the dietary nitrogen of the same diet prepared at different
times. There are two diets of the seven which show significant dif-
ferences for energy; whereas, for fat all showed differences which
are significant, either at the l or 5 per cent level. Why this occurs
is difficult to understand, particularly when the diet was prepared
and analyzed each week in the same manner. Of course, both diet
preparation and analyses are subject to human error.

A closer agreement can be noted between the duplicate samples of
all the diets than for the values secured on different days (Table 13).
Thomas 31 31.,(1950) upon analyzing duplicate samples of a diet found
the values for protein and food energy to be the most consistent with
relatively small variations. However, the determined fat in the
duplicate samples varied greatly. This difference as postulated by
the authors was due to sampling. They further reported that kitchen
techniques include inherent variability of sampling, preservation and
measuring, as well as errors unavoidable in the best analytical
mathods. The variability of sampling in this study cannot be over-

looked. It is not feasible to assume that from a kettle of mixed

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food two identical samples can be removed. Thus when a dish of this
type is prepared at different intervals there is less possibility

that the samples removed will be the same in composition. Thus in

a study, such as the "bread study", where many mixed dishes and desserts
are required, the diet must be analyzed each day to assure the exact
content of the diets.

Coons (1930) advocated that the diet sample should be collected
for analysis at the time it is served. The serving period of a non-
clinical balance study cannot be accurately controlled. The subjects
in this study were informed to come within a two hour period, but due
to examinations, seminars, and other committments on campus, there
was not complete adherence to this policy. A time schedule, devised
to aid this problem, resulted in only a slight improvement.

The results of the sampling time for the control period are
portrayed in Table 14. The means of both serving times show only
slight differences in the smount of nitrogen, fat, or food energy
analyzed in the diets. The "t" test reveals that there is no dif-
ference between the samples collected at zero hour and one and
one-half hour. There is a high probability for each nutrient that
the analyzed values will be in close proximity regardless of the time
it is collected.

The results for the experimental diet (Table 15) agree with
the control diet for food energy and fat, but for nitrogen a dif-
ference with time is demonstrated. The standard deviation for nitrogen
is small which indicates that the nitrogen from diet to diet had small
variation. This factor is supported by the results of the 3-way

analysis of variance as mentioned previously.

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101
The results reveal that the nitrogen values for one and one-half hour
are greater in five of the seven diets than the values analyzed for zero
hour. Diets VI and VII show slight trends in the Opposite direction.
In viewing the constitutents which composed each diet (Tables 4 and 5)
it can be observed that both menus, control and experimental, contained
mixed entrees and desserts. Fewer mixed entrees were employed in the
control regimen. Diet VI contained a mixed dish entree for which the l
individual servings were weighed prior to cooking. This was performed 3
with pizza. This would produce a greater uniformity between the
individual servings. Macaroni salad in diet.VI was mixed in one dish
and then the individual portions were weighed. However, this had only
an infinitesimal amount of liquid. Thus, it can be said that because
of the compactness of this dish, a serving was likely to represent the
entire salad. Many of the entrees in the diet contained a fair amount
of liquid. This would produce more error in obtaining samples which
are representative of the whole dish. Because of the dryness of the
bread that was required during the experimental period the liquid
dishes made the meals more palatable. This was borne out by the fact
that these meals were easier to consume.

EXperimental diets I through V all contained hot cereal. It can
be postulated that there was a loss in moisture in the cereal upon
standing; thus the last serving was more concentrated in nitrogen.

The control diets contained hot cereal only in diet II. Table 14
shows that for nitrogen in control diet II there is the greatest
change between the two collection periods. However, this change is
opposite to the results revealed in the experimental diets. This

deviation in the results could be attributed to the bread dressing

102
served at dinner. Since two duplicate samples do not contain the
same components, the serving at one and one-half hour might have
contained less nitrogen than the sample at zero hour. Where mixed
entrees or cooked cereals are served from one receptacle, there are
possible changes in the nitrogen composition as the serving time is
extended. Because of the way the cereals were prepared, it is in-
evitable that there was a moisture loss with time. Since cereals
contributed notrogen to the experimental diets in a fairly large amount,

they should have been prepared on an individual basis. This method

m .‘-A--_ ._

(Presson, 1955) gives a high degree of accuracy. Unfortunately, this
could not have been done in the kitchen employed in the "bread study".
The analyzed values for nitrogen show a closer agreement than

the determined fat values in the individual meals (Table 16) col-
lected at different times during the meal service. It can be assumed
that if the values are within 10 per cent of the maximum deviation
there is no difference in the values with regard to sampling time.

The specific sampling time varies with the objectives of the
balance study. It is apparent that when the entire diet is Sampled.
nitrogen is the constituent which varies most over the one and one—
half hour collection period of the experimental regimen. The analyzed
nitrogen in the individual meals disclose only small differences.
Upon analysis for fat in the individual meals the maximum variations
from.the mean range from 5.7 to 39.4 per cent. This is in agreement
With the fat values discussed throughout this presentation. This
great deviation in fat is due to the difficulty of preparing and

obtaining food samples which are homogeneous in their fat content.

103

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104

Probably the diet sample for analysis should be collected when the
majority of the subjects are being served in order to be relatively
assured that the analyzed nutrients in the diet represent the diets
consumed by the subjects.

In the Spring of 1964, the diets were dried in an oven at 600 C.
for twenty-four hours using a vacuum of 28 inches of Hg. In the
Spring of 1965, the same procedure was used to dry the diets except I
that they were dried until constant weights were obtained. From. {
Figure 1 it can be observed that after 60 hours of drying time there '
is no appreciable change in moisture content. Thus it is accurate } ‘
to consider the sample dry after 60 hours of drying when using the
above specified conditions. Table 17 presents the per cent of moisture
obtained when the experimental diets were analyzed on two separate
occasions. A loss of moisture upon storage is shown for all of the
diet samples. A mean loss of 3.95 i 3.53 per cent occurs in the
experimental diets.

To further evaluate the moisture loss upon storage, two cartons
were filled with a measured amount of distilled water. This experiment
showed a 7.5 and a 10.0 per cent moisture loss after being stored in
a freezer for a two month and a three and one—half month period,
respectively. The loss pf moisture was measured volumetrically
and metrically. Since water was used, a greater loss is disclosed
than is seen in Table 17. This is due to the fact that the water is
maintained in the diet suspension and, therefore, less evaporation
can occur. This moisture loss could be attributed to several factors.
First, the cover did notifit tightly on all the cartons. Second. it

.is possible that some of the moisture was absorbed into the walls of

 

 

.30
.25
.20

z

I

z:

u .15

Is?

<1
.10
.05
.00

105

Figure 1. Effects on drying with time.

0

u

.- o

 

 

“51 J 1 r
24 6 48 6O 7 84
Hrs.

Where M is the average moisture (in Z) of 33 exptl. diets dried in duplicate
for the hrs. indicated on the abscissa and N is the average moisture (in%)
Of the same samples after they were dried to constant weight.

W- ‘wxw
D

 

 

 

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. I: -I I ,
y

 

106

Table 17

Moisture Determinations on diet samples.

Exper. 7. Moisture 1 7. Moisture2 A Aas ‘7. of
Diet Spring, 1964 Spring, 1965 Spring, 1965
IA1 N.A.3 70.04 -- --
IA4 N.A.3 69.97 -- --
13 70.21 69.35 0.86 1.24
IC 72.70 71.10 1.60 2.25
101 70.43 69.61 0.82 1.18
104 70.19 68.32 1.87 2.74
HA.1 N.A.3 71.28 -- --
IIAh N. A. 71.54 -- --
IIB 72.42 70.46 1.96 2.78
IIC 72.62 69.84 2.78 3.98
IIDl 71.54 70.82 0.72 1.02
IID4 70.80 68.68 2.12 3.09
IIIA 72.05 71.94 0.11 0.15
IIIB 72.42 66.31 6.11 9.21
IIIC 71.19 69.02 2.17 3.14.
IIID 72.04 67.72 4.32 6.38
1110: 70.09 68.10 1.99 2.92
IVA 71.51 71.19 0.32 0.45
IVB 70.86 68.34 2.52 3.69
IVC 70.12 67.32 2.80 4.16
IVDl 70.07 68.98 1.09 1.58
IVDA 69.58 69.20 0.38 0.55
VA 73.48 73.42 0.06 0.08
VB 69.98 66.76 3.22 4.82
VC 70.15 65.59 4.56 6.95
VDl 70.12 68.70 1.42 2.07
v04 68.56 67.16 1.40 2.08
VIA 72.38 71.94 0.44 0.61
VIB 74.66 70.92 3.74 5.27
VIC 71.58 66.34 5.24 7.90
v101 71.25 68.08 3.17 4.66
VID4 71.59 67.54 4.05 6.00
VIIA 68.15 67.58 0.57 0.84
VIIB 69.50 60.14 9.36 15.56
VIIC 66.29 62.20 4.09 6.58
VIID1 67.20 59.29 7.91 13.34
VIID4 66.76 64.67 2.09 3.23

Mean Per Cent = 3.95%

Delta Mean PerCent = 2.63%
3.0. = 3.53%

“‘Alfls I; ‘3. m_

107

Table 17 (Cont.)

Average of duplicate samples dried for 24 hours at 600C in a vacuum
oven.

2 Average of duplicate samples dried at 60°C. in a vacuum oven until
constant weight was obtained.

3 Diets not analyzed Spring, 1964.

Subscript 1 represents collection at zero hour.
Subscript 4 represents collection at one and one-half hour.

 

the carton. Third, some moisture loss could occur when the sample is

fi—‘m.aamm_
I

removed for analysis from the same carton at different times since this
involves thawing and refreezing the samples.

The literature does not discuss moisture losses due to storage. The
type of container and the amount of sample placed in each container should
be emphasized. The container employed to store the diet sample should
be air tight and non-porous. Furthermore, to avoid losses in moisture
which occur upon thawing and refreezing the sample, the amount to be
used at one time for analysis should be stored in individual containers.
Whenever more samples are required for analysis, a sample which has not

been opened previously could be removed from the freezer, thawed, and dried.

SUMMARY AND CONCLUSIONS

Twelve subjects participated in a study to determine whether
or not they could remain in positive nitrogen equilibrium when
consuming a diet which contained 90 to 95 per cent of their protein
intake from wheat products. This study was conducted in a non-clinical
situation. From this study various conclusions can be made concerning
the conduct and dietary management of such a study.

The selection of the subjects should be based on their age, sex,
health, dietary habits, integrity, and dependability. The health
status of the subjects should be evaluated by a medical physician.
Because this is such a broad statement, Specific instructions for the
type of evaluation desired should be given to the physician. Persons
with physical handicaps or not within 10 per cent of their ideal weight
should be eliminated in studies which attempt to evaluate normal
individuals. To eliminate those subjects who may have emotional
problems to the extent that they would be unreliable, irresponsible,
and dishonest, a psychological examination evaluating this aSpect
should be given.

To acquaint the personnel and subjects with the demands of the
study a trial period should be included. This trial period preferably
should be a part of the control period. It is during this time that
the subjects and personnel become adapted to the controls and pro-
cedures of the study.

Because the subjects were going to consume a low protein diet
during the experimental period, the control period provided a diet

which contained the same amount of amino acids as would be provided by

108

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L

 

mu. IL
>. .hleruTIa-

 

109

the experimental diet. This was done to permit the subjects to

establish nitrogen equilibrium prior to the initiation of the ex-

perimental diet. For this reason no transitional period was required

between the control and experimental periods.

For psychological and emotional reasons rapport between the
researchers and the subjects must be established very soon after the
study begins. One factor that aids in this aspect is a "homey"
and pleasant atmosphere in which the subjects eat their meals.

The purchase and selection of food items is also a very important

a..- mt"

facet in the conduct of any metabolic balance study. All food used
1mmst be of the same brand and from the same lot. To purchase all

the food at the start of the study requires adequate and proper

storage facilities.

The type of menus used are limited by the purpose of the metabolic
study. Since this study was concerned with nitrogen balance, a
large variety of food items and methods of preparation could be used.
If the purpose of this study was to accurately determdne fat, food
energy, and various other nutrients, mixed dishes certainly could
not be used.

Coffee, tea, and sugar were allowed ad libitum. Since the first
‘two items contained nitrogen, the amount consumed by the subjects
The subjects were required to record all of these

Inust be known.

:items which they consumed. Ideally, instant coffee and tea should be

I’rovided in weighed packages that contain approximately enough for

One balance period. The amount consumed should be measured from

tlle weight of this material remaining at the end of each balance

Period.

110

If the subjects are of "normal" weight and the intention of the
study is to maintain such a body weight, prOper precautions should be
taken to accomplish it. For subjects who were losing weight non—
protein foods such as sugar, honey, hard candies, jelly, and whey-free
butter were administered. Those individuals gaining weight were
restricted in their use of whey-free butter, jelly, and sugar.

The weight loss which was observed in the majority of the subjects P
during the experimental regimen was hypothesized to be due to the

increased physical activity which was brought about with advent of E

 

summer-like weather. TherefOre, if the purpose of a balance study is

to maintain the weights of the subjects, it should occur during the
time of year when there is the least change in weather conditions.
For a metabolic balance study of this nature the Hanson spring
scale and the Toledo scale are sufficiently accurate providing they
are checked at frequent intervals during the study.
To determine the amount of protein, fat, carbohydrate, and other
essential nutrients in the diets tables of food composition must
be employed. However, complete confidence in such tables is not
'valid as was shown from the comparisons of the analyzed values of the
(iiets with the calculated values of the same diets.

The factors used to convert the calculated protein to nitrogen
eand the calculated carbohydrate, fat, and protein to food energy are
Ilunerous and must be used with discretion. Jones' factors used
t4) calculate the nitrogen from.the calculated protein values were:
nmilk,6.38; meat, eggs, fish, 6.25; wheat and cereals, 5.70; and nuts

and seeds, 5.30. These factors are more Specific for each respective

111

food group than is 6.25. The factors used to convert grams of car-
bohydrate, fat, and protein to calories were 4, 9, 4, respectively,
and the specific heat of combustion factors for each food group. Heat
of combustion factors must be used if a comparison is to be made with
the determined values obtained from an oxygen bomb calorimeter.

There is no statistically significant difference between the
calculated and analyzed values for nitrogen in the experimental
diets as shown by the 3-way analysis of variance test. The values
for fat and calories were significantly different. The levels of
significance for fat and food energy are one per cent and five per cent,
respectively. All of the analyzed values for fat were lower than the

calculated values. Five of the seven control diets showed a deviation

greater than 10 per cent. Upon comparing the calculated versus analyzed

‘values for fat in the experimental diets 50 per cent showed a deviation
greater than 10 per cent. All but two of the analyzed values for food
energy in the control diets were lower than the calculated values. Two
()f these diets showed a deviation greater than 10 per cent. Eight out
()f the twenty-eight experimental diets had larger calculated values
than the analyzed values for food energy. Only one of the experimental
ciiets showed a deviation greater than 10 per cent. In comparing the
CLalculated versus analyzed values for fat and calories there were fewer
dciets in this study that had a deviation greater than 10 per cent when
cOmpared to the values presented in the literature .

A closer correlation is observed when the diet is analyzed in Eggg
and compared with the calculated values than when the individual
(“anonents of the same diet are analyzed and summed.

Various reports state that analysis of diets needs to be performed

only occasionally and the results obtained can be used as the actual

 

112

composition of the diet throughout a balance study. Analyses performed
on diets replicated during four successive weeks showed that it is
important to analyze the diets each time the diet is prepared.

No significance was observed between the chemically determined
nitrogen, fat, or calories when the meals were collected at two
different times during the meal serving period. However, for the
nitrogen in the experimental diets the "t" test showed that the

analysis does depend upon time. Contrarily, when individual meals of

 

the experimental diets were collected at four intervals during the meal

 

service, no significant differences were observed for nitrogen. There
‘was a significant difference for fat. Therefore, it appears that the
analyses of the diets does depend somewhat upon the time the diet sample
is collected. The most accurate time is probably when most of the
subjects are being served.

It appears that the drying time required to completely dry a diet
sample is sixty hours using a temperature of 60° C in an oven with a
“vacuum of 28 inches of Hg.

The container used to store the diet samples for analyses should
13a non-porous. Because moisture loss occurs when a sample is frozen
aand rethawed, only enough sample which can be used for the determinations
crn.one occasion should be stored in a container. The most suitable
Guantainer is a glass jar with a tight fitting cover or lid.

Good results can be obtained from a non-clinical metabolic

tMilance study if the proper precautions and techniques are employed.

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Processed, Prepared". U. S. D. A. Agr. Handbook No. 8, Agr.
Research Service, Washington, D.C.

'Wertz, A. W., M. E. Lojkin, E. H. Morse, G. C. Hogan, and P. S. VanHorn.
1952. "A Comparison of Determined and Calculated Amounts of Eight
Nutrients in One Day's Food Intake of Twenty-two Subjects."

Univ. Mass. Agr. Expt. Stat., Amherst, Mass., Bull. No. 469:79.

 

 

120

Wharton, M. A., D. Tyrrell, and M. B. Patton. 1953. Amino acid intake
and nitrogen balance of college women. J. Am. Dietet. Assoc.,29:573.

Widdowson, E. M. and R. A. McCance. 1942. Mineral metabolism of
healthy adults on white and brown bread dietaries. J. Physiol.,
181:44.

Widdowson, E. M., and R. A. McCance. 1943. Food tables-~their scope
and limitation. The Lancet 1:230.

Wollaeger, E., M. Comfort, and A. Osterberg. 1947. Total solids, fat
and nitrogen in feces. Gastroenterology 8:272.

Young, C. M., I. Ringler, and B. J. Greer. 1953. Reducing and
post-reducing maintenance on the moderate fat diet. J. Am. Diet.
Assoc.,I88:89O.

‘Young, C. M. 1964. Personal Communication.

 

APPENDIX

121

‘w
,_

 

THE RECIPES

Special recipes were devised for the "bread study" in order to
fulfill the requirement for the type of protein in the diets. These
recipes were formulated with the aid of the following references:

1. "Better Homes and Gardens New Cook Bookf'1953.

lst ed., Meredith Publishing Co., New York.
2. "Betty Crocker's Picture Cook Book." 1956.
2nd ed., McGraw Hill Book Co., New York.
3. Farmer, F. M. 1951. "The Boston Cooking-School
Cook Book." 9th ed., Little Brown and Co., Boston.
4. Fowler, S. and B. West. 1950 "Food for Fifty."
John Wiley and Sons, Inc., New York.

5. Givens, M. "Modern Encyclopedia of Cooking!’ 1959.

Rev. ed., J. G. Ferguson Publ. Co., Chicago. Vol. I and II.

The recipes which follow are listed in alphabetical order.

122

 

123

.Apple Pie
Yield: Two 9" pies
Temp.: 325° F

Time : 55 minutes

Gram
Ingredient Weight
Apples
(sliced, water packed) 1550
Sugar 300
Cinnamon _ 4
Butter (protein free) 30
Pie Crust (raw weight) 842
Prepared Weight: 2606
Serving: (one) 130

Instructions: Make double pie crust (see pie crust). Weigh 421 grams
of crust for each pie. Use approximately 210 grams for each crust.

Roll out crusts between two sheets of wax paper.

324.8

 

795.3

39.7

Protein

gm.

3.4

56.1

59.5

3.0

Fat

24.0

274.4

299.7

 

15.0

Slit pastry for

toP crusts in several places. Place bottom crust in 9" pie pan.

Place 775 grams apples on bottom crust.

and 2 grams of cinnamon. Sprinkle over apples.

Combine 150 grams of sugar

butter. Cover with top crust. Seal edges securely.

Dot with 15 grams

 

 

 

 

124

Applesauce Cake
Yield: Four 8" x 8" cakes
Temp.: 375°F

Time : 20 minutes

 

Gram CHO Protein Fat

Ingredient weight gm. gm. gm.
Flour 400 295.2 51.0 4.0
Baking soda 7 --- --- -_-
Salt 8 —-- -_- ---
Cinnamon 11 --- --- ---
Cloves 2 --— --- _--
Allspice 7 --- --- ---
Nutmeg 7 --- --- ---
Shortening 230 --- --- 230.0
Sugar 460 460.0 --- ---
Applesauce 550 114.6 1.4 0.9
(canned, sweetened) ‘———— '-—-
Prepared weight: 1453 869.8 52.4 234.9
Serving (one) : 100 59.9 3.6 16.2

Instructions: Line four 8" x 8" pans with wax paper. Weigh and
combine flour, baking soda, salt, Cinnamon, cloves, allspice and
nutmeg. Cream shortening and sugar until light and fluffy in

electric mixer bowl. Add sifted dry ingredients alternately with
tflie applesauce, beating well after each addition. Pour 420 grams

iJJto a wax paper lined 8" x 8" pan.

 

125

Boiled Dinner

 

Ingredient Gram CHO Protein Fat
weight gm. gm- gm.
Carrots (canned, drained) 670 42.9 4.5 3.6
Onions (raw, diced) 110 11.3 1.5 0.2
Potatoes (raw, diced) 700 133.7 14.0 0.7
Cabbage (raw, shredded) 340 18.4 4.8 0.7
Celery (raw, diced) 300 11.1 3.9 0.6
Vegetable bouillon cubes 4
(1 cube) '-- 0.5 "“
Water 1500 —— _._._. __
Prepared Weight: 3234 217:4— ‘29T2__ 5.8
Serving (one) : 150 10.1 1.4 0.3

Instructions: Cook raw vegetables in weighed amount of water with
vegetable boullon cube in 8 quart aluminum kettle. When done add
carrots. Heat thoroughly. Serve 150 grams in individual casserole

dishes. Cover and place in warming oven.

 

126

 

Bread

Gram. CHO Protein Fat

Ingredient weight gm. gm. gm.
Flour 4790 3535.5 611.2 47.9
Shortening 200 --- --- 200.0

Sugar 150 150.0 --- ---

Salt 85 --- --- ---
Yeast, dry (7 pkg.) 49 19.3 18.3 0.8

Water 2880 --- --- ---
Prepared Weight: 7047 3704.8 629.5 248.7
Serving per 100: 52.6 8.9 3.5

 

Instructions: Place into a 2 quart mixing bowl yeast, 6 teaspoons of
weighed sugar and 360 grams of lukewarm water. water should be 43°C.
Please test with thermometer. Set bowl in proofing area. weigh 2520
grams of water and pour in a large Hobart mixing bowl. Add 150 grams
shortening, remaining sugar and 85 grams salt. Mix 2 minutes at low
speed. Add 10 cups of flour. Mix 3 minutes at low speed. At this
time be sure dough is lukewarm (43°C).

If dough is correct temperature, add yeast mixture. Add the
remaining flour very slowly. Be sure to sop mixer for each addition.
After all flour and ingredients are mixed, knead at low speed for
l4'minutes.

Grease sides of mixing bowl and tOp of dough lightly with
shortening allowed for handling dough. Cover dough with wax paper

and a cloth towel. Place in proofing area. Mark time.

3150 grams in recipe and 50 grams for handling dough.

127

Let rise 1% hours. Punch down and knead at low speed for 2
minutes. Brush top of dough with shortening as above. Cover and
return to proofing area for 45 minutes. Mark time. While dough is
rising --

Grease lightly 12 bread pans from the shortening allowed for

2 ”I
handling. ‘1

'Weigh approximately 500 grams of dough and form into a loaf.

‘q. "i..- t‘ '-

Place in greased pans. Cover as above. Return to proofing area.

 

o V
Mark time. Let rise 1 hour. Preheat oven 350 F. Bake 35 minutes. i

FEJr rolls, weigh the amount requested and use 50 grams for each roll.
1lice in greased 9" x 13" pan. Bake 20 minutes at 425°F.

F331? French bread, shape 500 grams dough into oblong loaf. Place on
greased cooking sheet. Bake at 350°F for 35 minutes.

128

Bread Dressing
0
Temp.: 350 F

Time : 60 minutes

 

Ingredient ngzgt 2:? Prziein :3?
Bread Crumbs 840 441.6 75.0 2.9
Butter, melted 175 1.4 1.5 140.9
Salt 17 --— --- ---
Pepper 2 -—- --- -_-
Onion, minced 70 7.0 0.7 ---
Celery, minced 350 13.0 4.6 0.7
Water 1850 ‘_;;;__ --- ---

Prepared Weight: 2643 463.0 81.8 144.5

Serving (one) : 150 26.3 4.6 8.2

Instructions: Combine all ingredients except water. Mix thoroughly.

Add water and mix. Place in 9" x 13" pan and bake.

Carrot/Pineapple Salad

Gram CHO Protein Fat

Ingredient weight gm. gm. gm.
Carrots (raw) 420 39.1 5.0 1.3
Pineapple slices 280 59.0 1.2 0.2
(sirup packed) ‘—‘—— "‘“ “"
Prepared Weight: 700 98.1 6.2 1.5
Serving (one): 50 7.0 0.4 0.1

Instructions: Grate carrots. Drain pineapple. Cut in chunks. Stir

in pineapple. Weigh 50 grams for each subject.

 

129

Cherry Kuchen (Control)
Yield: Three 8" x 8" Kuchens
Temp.: 3500F

Time : 45 minutes

 

 

Gram. CHO Protein Fat
Ingredient weight gm. gm. gm.
water 270 --— --- ---
Sugar 450 450.0 --- ---
Flour1 450 --- --- ---
Baking Powder 14 --- --- ---
Salt 11 -—- --- ---
Water 270 --— --- _--
Butter (melted) ’ 280 --- --- ___
Vanilla 12 --- _-- ---
Topping:
Flour 1 85 --- --- --_
Brown Sugar 110 105.0 --- ---
Butter2 56 --- --- ---
Baking Powder 4 --- --- ---
Cherries 460 47.6 .81£1_ 1.2
(water packed, drained)
1 Total Flour Values 535 394.9 68.3 5.3
2 Total Butter Values 336 2.7 1.9 270.0

3 15 grams of butter is to be employed for lightly greasing the pans.

Prepared Weight: 2233 1000.2 73.4 276.5
100 44.8 3.3 12.4

Serving (one)
Instructions: Lightly grease three 8" x 8" pans with the allotted
butter. Blend 270 grams water and sugar. Add sifted flour, baking
powder, and salt.‘ Add remaining water, melted butter, and vanilla.
Combine and mix all t0pping ingredients.‘ Into each pan place 500 grams
of batter, 130 grams of drained cherries, and sprinkle with 70 grams

of topping.

\‘._ “~‘ *t -. . '
‘ . r"
I

 

130

Cherry Kuchen (Experimental)
Yield: Three 8")c8" Kuchens
Temp.: 350°F

Time : 45 minutes

Gram CHO Protein Fat
Ingredient Weight gm. gm. gm.
Water 270 --- --- ---
Sugar 450 450.0 --- ---
Flour1 450 --- --- ---
Baking Powder 14 --- --- -_-
Salt 11 —-- --- ---
water 270 --- --- ---
Butter, melted 2’:3 280 ___ ___ -__
(protein free)
Vanilla 12 ——- --- ---
Topping:
Flour]' 85 --- --- --_
Brown Sugar 110 105.0 --- ---
Butter 2 56 mm -..- --..
Baking Powder 4 --- --- ---
C1123: packed, drained) 46° —4—7-=§— —3-& —1—-—§-
1 Total Flour Value 535 394.9 68.3 5.3
2 Total Butter Value 336 --- --- 268.8

3 15 grams of butter is to be employed for lightly greasing pan.

Prepared weight: 2188 997.5 71.5 275.3

Serving (one) 100 45.6 3.3 12.6

Instructions: See Control.

 

131

Cherry Pie (Control)
Yield: Two 9" pies

Temp. and Time: 4250F for 10 minutes
3500F for 40 minutes

 

 

 

Gram CHO Protein Fat
Ingredient weight gm. gm. gm.
Filling:
Cherries 760 78.6 5.3 2.0
(water packed, drained)

Sugar 300 300.0 --- ---
Flour 56 41.3 7.2 0.5
Juice from cherries 240 trace trace trace
Butter 56 0.4 0.3 45.1

Pie Crust 840 324.8 56.1 274.4
Prepared Weight: (Filling & Crust) 2038 745.2 68.9 322.0
Serving (one) : 135 49.4 4.6 21.3

Instructions: Make double pie crust (See pie crust). Weigh 421 grams
of crust for each pie. Use approximately 210 grams for each crust.
Roll out crusts between two sheets of wax paper. Slit pastry for

top crust in several places. Place bottom crust in 9"_pie pan.

Drain cherries. Save juice. 'Mix sugar and flour in 3 quart sauce
pan. Add 240 grams of the juice. Bring to boiling point. Remove

and stir in drained cherries. Fill each pie crust with 756 grams

of filling.

132

Cherry Pie (Experimental)
Yield: Two 9" pies
Temp. and Time: 4250F for 10 minutes

350°F for 40 minutes

 

. Gram CHO
Ingredient Weight gm.
Filling:
‘Cherries 760 78.6
(water packed, drained)
Sugar 300 300.0
Flour 56 41.3
Juice from cherries 240 trace
Butter (protein-free) 56 ---
Pie Crust 840 324.8
Prepared Weight: (Crust & Filling) 2087 744. 7
Serving (one): 130 46.4

Instructions: See Cherry Pie (Control).

Protein
gm.

5.3

7.2

trace

 

68.6

4.3

Fat

0.5
trace
44.8

22.841.
321.7

20.0

 

 

133

Chocolate Cake

 

 

 

 

 

 

Yield: Four 8” x 8" layers
0
Temp.: 375 F
Time : 30 minutes
. Gram CHO Protein Fat
Ingredient Weight gm. gm. gm.
Shortening 200 --- --- 200.0
Vanilla 8 --- --- ---
Sugar 500 500.0 --- ---
Chocolate Squares 100 29.2 5.5 52.9
Flour 400 295.2 51.0 4.0
Salt 5 --- --- ---
Baking Powder 16 --- --- ---
Water 480 --- --- ---
Prepared Weight: 1482 824.4 56.5 256.9
Serving (one) : 100 55.6 3.8 17.3

Instructions:
chocolate.

and grated chocolate.

Line four 8"x:8" cake pans with wax paper.

Grate
Cream shortening, add vanilla and beat in (by hand) sugar

Combine flour with salt and baking powder.

Add flour mixture alternately with water to shortening mixture.

Pour 428 grams into each 8"128" layer pan.

134

Cream of Wheat

 

 

 

 

 

Ingredient Gram CHO Protein Fat
weight gm. gm- gm.

Water 3636 --- --- ---
Salt 10. --- --- ---
Cream of Wheat (Quick) 684 188888 8818 7.2
Prepared Weight: 3643 1022.2 79.2 7.2
Serving (one) : 235 65.9 5.1 0.5
Water 2376 --- --- ---
Salt 8. --- --- ---
Cream of Wheat (Quick) 456 681.4 8818_ 4.8
Prepared Weight: 2387 681.4 52.8 4.8
Serving (one) : 177 50.5 3.9 0.4

Instructions: Bring water and salt to rapid boil. Slowly sprinkle
in Cream of Wheat stirring constantly while the mixture thickens.
‘Lower heat and cook 5 minutes. Place over pan of hot water to keep

‘warm for serving.

135

Creole Green Beans

 

Ingredient WSEZEt 2:? Prgiein :3?
Onion (raw, chopped) 50 10.3 1.4 0.2
Salad oil 60 --- --- 60.0
Chili Sauce 360 178.6 18.0 2.2
Salt 13 --- --- ---
Green Beans (canned, drained) 1200 122.9 8288. .521

Prepared Weight: 3132 311.8 56.9 66.5

Serving (one) : 170 16.9 3.1 3.6

Instructions: Brown onion in oil in 8 quart aluminum kettle. Add
chili sauce. Add green beans. Heat thoroughly. Place 170 grams in

individual casseroles. Place in warming oven until served.

136

Creole Spaghetti Sauce

 

 

Ingredient Gram CHO Protein Fat
Weight gm. gm- gm.

Sauce:
Onion (raw, minced) 160 16.0 1.6 ---
Green Pepper (raw, chopped) 160 9.2 1.6 0.4
Vegetable Oil 56 --- --- 56.0
Ground Beef (cooked weight) 200 --- 51.3 34.1
Salt 15 --— --- ---
Sugar 1 6 6.0 --- ---
“mam“ (canned) 1700 66. 3 17 .0 3.4

(juice and solids) "“‘

Prepared Weight: ‘ 2068 97.5 71.5 93.9
Serving (one) : 100 4.7 3.4 4.5

Instructions: Cook ground beef in frying pan. Brown onion and green
pepper in oil in aluminum 8 quart kettle. Stir in ground beef and
any fat from beef, salt, sugar, and tomatoes. Simmer for 30 minutes.

Pour 100 grams of sauce over 100 grams cooked spaghetti in individual

casserole dishes. Cover with casserole lid and place in warming oven.

 

137

Crunchy Cookies

0
Temp.: 350 F

Time : 10 minutes

 

 

Gram CHO Protein Fat
Ingredient Weight gm. gm. gm.
Shortening 400 --- --- 400.0
Brown Sugar 440 420.0 --- ---
White Sugar 400 400.0 --- ---
Water 120 --- --- ---
Vanilla 15 -—- --- ---
Salt 8 --- --- ---
Baking Powder 10 --- —_- ---
Flour 600 442.9 76.6 6.0
Wheaties 260 §9§;2. _8888_ .888
Prepared Weight: 2049 1471.8 102.6 411.6
Serving (one) : 25 17.9 1.2 5.0

Instructions: Thoroughly cream shortening and sugars in Hobart mixing

bowl. Add water and vanilla. Beat well. Combine dry ingredients.

Add to previous mixture. Add Wheaties. Mix 1 minute(low speed).

Bake on ungreased cooky sheet. Remove from baking sheet and place

on wax paper. When cool, weigh and store in plastic bags in the walk- 1
in refrigerator. Be sure to label the bag with the contents, weight,

and the date prepared.

138

Cupcakes
Yield: 18 cupcakes
Temp.: 3750F

Time : 25 minutes

 

 

 

 

Ingredient Gram CHO Protein Fat
Weight gm. gm- gm.
Shortening 134 --- --- 134.0
Flour 400 295.2 51.0 4.0
Sugar 400 400.0 --- ---
Baking Powder 12 --- --- ---
Salt 7 —-— --- ---
Milk 365 18.6 13.5 12.7
Egg (Approximately 2) 108 0.6 13.8 11.0
Vanilla - 8 --- --- ---
Prepared weight: 1130 714.4 78.3 161.7
Serving (one) : 50 31.6 3.5 7.2

Instructions: Mix shortening to soften in electric mixer bowl. Sift
in dry ingredients. Add one-half of milk and the eggs. Mix only to
dampen flour mixture. Add remaining milk and vanilla. Beat for one
minute at low speed. Line the muffin tin with the paper muffin cups.

Fill each with 60 grams of dough.

139

 

 

 

Dumplings

Gram CHO Protein Fat

Ingredient Weight gm. gm. gm.
Flour 990 730.7 126.3 9.9
Baking Powder 53 --- --- ---
Salt 15 --- --- ---
Butter (Protein free) 60 --- --- 48.0
water 810 --- --- ---
Prepared Weight: 1939 730.7 126.3 57.9
Serving (one) : 120 1 45.2 7.8 3.6

Instructions: Combine all ingredients and mix well. Weigh out 120

 

grams raw weight per subject. Place the dumpling in the vegetarian

stew. Cook 20 minutes.

 

 

Farina

Ingredient Gram CHO Protein Fat
weight gm. gm- gm.

Farina 608 473.6 60.8 4.8
water 3232 --- --- ---
Salt 10 --- --- ---
Prepared weight: 3500 473.6 60.8 4.8
Serving (one) : 250 33.8 4.3 0.3

Instructions: Bring water to vigorous boil in 8 quart aluminum kettle.
Add salt, then add Farina very slowly (to avoid lumping), stirring
constantly. Return to boiling point. Lower heat half-way and cook

2% minutes stirring frequently. Place pan with cooked cereal in

receptacle of hot water to keep warm for serving.

1‘ 120 grams wet weight - yielded 150 grams cooked dumpling.

140 .

Fruit Ice

 

Ingredient Gram CHO Protein Fat
Weight gm. gm. gmo
Water 300 -- -- '-
Sugar 400 400.0 -- --
Salt 2 '- -- '-
Orange Juice
(frozen, diluted 1:3) 240 26.2 1.9 '-
Lemon Juice (Real Lemon) 160 12.3 0.6 0.3
Banana (fresh) 450 £88 8_._4_ 8:2
Prepared Whight: 1505 542.0 7.9 1.2
Serving: (one) 100 36.0 0.5 0.1

Instructions: Place bananas in electric mixing bowl: mix at low
speed. Add juices, sugar, and water to the banana. Mix well. Pour
into ice cube trays. Place in freezer of refrigerator. Freeze

24 hours before serving.

141 ,

Fruit Plate (Individual Plate)

1

 

 

Ingredient WSIZEt :2? Prggein :3?
Pear Half 50 9.2 0.1 ---
Pineapple Slicesl' 65 --- --- ---
Banana 75 17.2 0.9 0.2 F}
Pineapple Juice 15 --- --- --- l ,
Lettuce 20 0.6 0.2 --- i
Maraschino Cherry 2 0.6 ._;:;_ ‘_:;;_ ij
,6»
Total values for pineapple 80 16.9 0.3 0.1
Prepared Weight: 227 44.5 1.5 0.3
Serving (one) : 227 44.5 1.5 0.3

Instructions: Drain all fruit before weighing. Arrange fruit on
lettuce. Pineapple juice is to be placed over the banana to avoid

darkening.

142

Fruit Salad

 

 

Ingredient Gram CHO Protein Fat
Weight gm. gm. gm.

Pineapple slices (canned) 200 42.2 0.8 0.2
Banana (slices) 250 46.0 2.4 0.4
Apples (unpeeled, cubed) ‘ 250 .8118 _818 .888
Prepared Weight: 700 115.7 3.7 0.8
Serving (one): 50 8.2 0.3 0.1

Instructions: Drain pineapple and cut slices into dices. Prepare
bananas and apples as stated above. Toss all fruits together. Serve

50 grams on 20 grams of lettuce to each subject.

 

143 ‘

Ginger Cookies
0
Temp.: 375 F

Time : 15 minutes

 

 

Ingredient Gram CHO Protein Fat

weight gm. gm. gm.
Shortening1 225 -- -- 225.0 lr}
Sugar 225 225.0 -- -- "
Molasses (medium) 370 240.5 -- -- .
Vinegar 20 -- -- -- ‘J
Flour 745 549 . 9 95 . 1 7 . 4 “w
Soda 3 -- -- --
Cinnamon 3 -- -- --
Ginger 3 -- -- -_
Salt 3 -- -- --
Water 125 -- _;:; --
Prepared Weight: 1517 1015.4 95.1 232.4
Serving: (one) 25 16.7 1.6 3.8

Instructions: Bring shortening, sugar, molasses, vinegar to a

. boil in 8-quart aluminum kettle. Cool. Add water. Combine dry
ingredients. Add to liquid ingredients and mix well. Shape in a
roll, wrap in wax paper and chill. Cut into slices and bake on
lightly greased cookie sheet at 3750 for 15 minutes. Remove from
baking sheet and place on wax paper. ‘When cool, weigh and store in
plastic bags in the refrigerated walk-in. Be sure to label the

bag with the contents, weight and the date prepared.

1ten grams of the shortening is to be employed to lightly grease
the baking sheets.

144

Gingerbread
Yield: Four 8" x 8" pans
Temp.: 3750 F

Time : 20 minutes

 

Ingredient Gram CHO Protein Fat
Weight gm. gm. gm.
Flour 492 363.1 62.8 4.9
Baking Powder 15 --- --- ---
Salt 5 —-- --- ---
Molasses 517 336.0 --- _-_
Sugar 170 170.0 --- ---
Shortening 110 "' "’ 110.0
Soda 15 --- --- ---
Ginger 2 --- --- ---
Cinnamon 7 -—- --- ---
Cloves 2 --— --- ---
Allspice 8 --- --- ---
Boiling Water 400 --- --- ---
Prepared weight: 1504 869.2 62.8 114.9
Serving: (one) 100 57.8 4.2 7.6

Instructions: Line four 8" x 8" pans with wax paper. Combine
flour with baking powder and salt. Mix sugar and molasses in
electric mixing bowl. Add shortening and soda to sugar and
molasses. Add flour mixture. Mix spices and boiling water. Add
this spice mixture to the previously mixed ingredients. Pour

435 grams in each pan.

145 .

Iiarvard Beets

 

 

 

Ingredient Gram CHO Protein Fat
Weight gm. gm. gm.
Sugar 270 270.0 --- ---
Cornstarch 14 10.5 "' ---
Vinegar 145 --- --- ---
Water 145 --- -—— --- ’
Beets (drain) 1200 117.1 11.6 1.4 é
Butter 75 0.6 0.4 60.4 7
Salt 3 --- --- -—- i;
Pepper 1 “- 2.:— _-_-_-_
Prepared Weight: 1663 398.2 12.0 61.8
Serving: (one) 100 24.0 0.7 3.7

Instructions: Mix sugar and cornstarch in 3-quart sauce pan. Add
vinegar and water. Boil 5 minutes. Add beets. Let simmer 30 minutes.
Add butter, salt and pepper. Remove from heat and place in a pan of

hot water to keep warm for serving.

146

Lemon Pie with Meringue

“Yield: Two 9" pies

 

 

Ingredient GFam CHO Protein Fat
Weight gm gm. gm.

Cornstarch 80 70.0 --- ---
Flour 60 44.9 7.7 0.6
Salt 6 --- --- ---
Sugar 740 740.0 --- ---
Boiling water A 900 --- --- ---
Butter 35 0.4 0.3 28.5
Lemon Juice (Real Lemon) 205 15.8 0.8 0.4
Egg Yolks (slightly beaten) 170 1.0 28.0 54.0

Meringue:

Egg Whites 155 1.0 16.5 ---
Sugar 120 120.0 --- ---
Vanilla 6 --- --- ---
Pie Crust 421 162.4 881 137.2
Prepared Weight: (1 pie) 1200 577.7 40.7 110.4
Serving (one) : 160 77.0 5.4 14.7

Instructions: Mix cornstarch, flour, salt and sugar. Add boiling water.
Stir constantly over heat until the mixture boils. Cook until thickened.
Add butter and lemon juice. Add half of the hot mixture to slightly
beaten egg yolks. Mix. Add this egg mixture to the remaining hot

mixture. Cook and stir until thick. Cool.

For the meringue beat the egg whites until stiff, then gradually beat
in sugar until meringue is stiff and shiny. Add vanilla. Place 940
grams filling into 9" baked pastry shell (see Pie Crust). Top with

125 grams meringue. Bake at 3500F for 12 minutes.

Reference: Farmer, F. M. 1951. The Boston CookingeSchool Cook Book
9th ed. Little Brown and Co., Boston. p. 637.

 

1P? 5;. -.—._
.1
'1' J

 

147

Lemon Sauce

 

 

 

 

 

Gram CHO Protein Fat

Ingredient weight gm. gm. gm. |
Sugar 200 200.0 --- ---
Flour 28 21.4 2.8 0.3
Water 480 --- --- ---
Butter 55 --- --- 44.6
Lemon Juice 15 1.2 0.1 ---
Nutmeg 5 --- --- ---
Prepared weight: 720 222.6 2.9 44.9
Serving: (one) 50 15.8 0.2 3.2

Instructions: Combine sugar and water in 2-quart sauce pan. Stir
in flour to make a paste. Bring to a boil. Remove from heat. Stir

in butter, lemon juice, and nutmeg.

148 «

, 1
Lime Sherbet

 

 

Ingredient Gram CHO Protein Fat
Weight gm. gm. gm.
Milk (3.5% Fat) 19,505 991.2 719.4 679.4
Water 12,928 --- --- ---
Sugar- (cane) 9,072 9072.0 --- —--
Sugar- (corn) 3,629 3276.5 --- --- '
Stabilizer 226.8 "‘ --- ---
Prepared WEight: ‘ 45,361 1333977 .719Z4 679.4
Serving: (one) 75 22.6 1.2 1.1.

Prepared at the Michigan State University Dairy Department.
Calculation based on information obtained from the Dairy Department.

149

Macaroni Salad

 

 

 

 

 

Ingredient ngzgt :2? Prgiein :3?
Macaroni 23:33 383 634.5 106.5 12.0
Onion (raw, chopped) 80 8.3 1.1 0.2
Celery (raw, diced) 320 11.8 4.2 0.6
Lettuce (torn) 200 5.8 2.4 0.4
Pickle Relish 120 31.4 0.9 0.9
Pimento (minced) 30 1.4 0.2 0.1
Celery seed 21 --- --- ---

Dressing:
Salad Oil 220 —-- --- 220.0
Vinegar 240 --- --- ---
water 120 --- --- ---
Dry Mustard 1 --- --- ---
Celery Salt 2 —-— --- ---
Black Pepper dash -—— --- ---
Paprika 2 --— --- -_-
Caraway Seed 4 --— --- ---
Salt 5 --- --- ---
Sugar 15 15.0 --- ---
Thyme pinch --- --- ---
Prepared weight: 3490 708.2 115.3 234.2
Serving (one) : I 240 48.7 7.9 16.1

Instructions: Cook macaroni in 7 quarts salted (2 Tbsp.) boiling water
for 15 minutes. Drain. Rinse thoroughly with hot water. Drain. Add

remaining ingredients. Mix. Cover and marinate 1 hour in refrigerator.

Mashed Potatoes

, Gram CHO Protein Fat

Ingredient ,
Weight gm. gm. gm.

Potatoes

(cooked, well drained) 1800 343'8 36'0 1'8
Butter 50 0.4 0.3 40.2
Milk 100 5.1 3.7 3.5
Salt 12 --- --- ---
Pepper 1 --- --- ---
Prepared weight: 1950 349.3 40.0 45.5
Serving (one): 100 18.3 2.1 2.4

150

 

Instructions: Peel potatoes. Cut in halves. Cover with water and
cook 30 minutes. Remove and drain. Mash potatoes with the use of
the electric mixer. Combine milk and butter in 1 quart sauce pan.
HEAT. Do not scald or boil. Add the milk mixture and the seasonings
to the potatoes. Mix at low speed for 1 minute, then at high speed
until light and fluffy. Place the mashed potatoes in a container.

Place container in hot water until all the subjects are served.

151 .

Meats

The protein and fat composition of the meat prepared was obtained from

the tables of food composition.

Beef (choice grade, bottom, round roast): Season with salt and pepper.
Place with fat side up in roaster. Bake at 3250F. Allow 40 minutes
for each pound of roast. Trim off fat and serve.

Beef Patties (choice grade, ground beef): Form 75 grams of ground
beef into a patty. Cook in electric frying pan. When done place
on paper towels to absorb fat.

Canadian Bacon (roll): Trim and slice into 50 gram portions. Place

 

on baking sheet. warm in oven at 200°F for 30 minutes.

Chicken (roasting): Season with salt and pepper. Place in roaster
and bake at 325°F allowing 40 minutes per pound of chicken. Prior
to serving, remove fat and bones. Serve a combination of white
and dark meat to each subject.

Cube Steak (choice grade): Season with salt and pepper. Pan broil.
When done place on paper towels to absorb any excess fat.

Fish (frozen, White): Thaw. Weigh 65 gram portions and place on
baking sheet. Top each portion with 5 grams butter. (Season with
salt and pepper. Bake at 3250F for 45 minutes.

Ham (canned): Trim excess fat, slice and weigh individual servings.

Pork Chops (choice grade, center cut): Trim fat. Season with salt
and pepper. Place on baking sheet. Bake at 3250F for 50 minutes.

Veal cutlets (choice grade): Season with salt and pepper. Place

on baking sheet. Bake at 325°F for 40 minutes.

152 '

Molasses Cookies
0
Temp.: 350 F

'Time : 10 minutes

Instructions:

hot water to molasses.
shortening and sugar.

Speed for 2 minutes.

cookie sheet.

 

Ingredient Gram CHO Protein Fat
Weight gm gm, gm.
Shortening 200 --- --- 200.0
Brown Sugar 220 210.0 --- ---
Hot Water 240 --- --- ---
Molasses (medium) 320 192,0 --- ---
Flour 500 369.0 63.8 5 0
Salt 5 ---- --- ---
Soda 2 --- --- ---
Baking Powder 8 --- --_ ---
Ginger 5 --- --- _--
Cloves (ground) 3 --- --- -_-
Cinnamon 7 --- --- --_
Grapenut Flakes‘ 120 2ggg 11.6 ._LLZ
Prepared weight: 1341 869.6 75.4 206.7
Serving (one) : 50 32.4 2.8 7.7

Cream shortening and sugar in Hobart mixing bowl. Add
Add the water and molasses mixture to the
Combine dry ingredients and addgmixing at low

Drop dough from teaspoon onto a lightly greased

Remove from baking sheet and place on wax paper. When

cool, weigh and store in plastic bags in the refrigerated walk-in.

Be sure to label the bag with the contents, weight and the date prepared.

 

 

 

 

 

Oatmeal

Ingredient

Oatmeal (dry weight)
Water
Salt

Prepared Weight:

Serving (one) :

Instructions: Stir oats into boiling salted water.

stirring occasionally.

153

Gram
Weight

600

2900

10

3100

236

 

409.2

31.2

of hot water until all subjects are served.

Protein

6.5

 

44.4

3.4

Cook 5 minutes,

Cover pan, remove from heat and place in a pan

v Jugs-fir (. .c' n -7
..-

‘F"‘”
wua.

 

Peach Crumb Pie
Yield: Two 9" pies
Temp.: 3750F
Time : 45 minutes
Ingredient
Filling:

Sugar
Flour

Salt

154

Gram
Weight

400

1
56

2

Peaches(frozen, sliced, sweetened) 1000

Crust:
Salt
Flour
Shortening
Water
Topping:
Flour
Brown Sugar (light)

Butter (control)

(Exper. - protein free)

Baking Powder

1 Combined values for flour
(Control)

Prepared Weight: (Exper.)

. (Control)
Serv1ng (one): (Exper.)

Instructions: Drain peaches.

thoroughly with peaches.
Fill each crust with 715 grams of filling.

and sprinkle 125 grams over the filling of each pie.

220

135
60

85
110

56
56

361

1981
1980

135
135

Combine dry filling ingredients.

226.2

 

266.4

998.0
997.6

68.0
68.0

Protein

46.1

50.5
50.2

3.4
3.4

Fat

 

3.6

184.3
184.2

12.6
12.6

Mix

For preparation of crust, see Pie Crust.

Blend topping ingredients

 

Pettijohn Cereal

155

 

Gram CHO Protein Fat

Ingredient weight gm. gm. gm.

Pettijohn (dry) 448 339.2 41.6 9.6

water 2432 --- --- ---
Salt 8 --- --- :::_'

Prepared Weight: 2500 339.2 41.6 9.6

Serving (one): 180 24.2 3.0 0.7

Instructions: Bring salted water to boil. Sprinkle in pettijohn. Cook
5 minutes stirring occasionally. Remove from heat. Cover and place

in receptacle of hot water to keep warm until all subjects are served.

Pie Crust

Yield: Two double crust pies
Temp.: 4OOOF

Time : 12 minutes

 

 

Ingredients nggfit :2? Prggein :3?
Salt 12 --- --- ---
Flour 440 324.8 56.1 4.4
Shortening 270 --- --- 270.0
Water (cold) 120 --- --- __;::;_

Prepared Weight: (raw weight) 842 324.8 56.1 274.4

Serving (one): Calculated with each total pie recipe
Instructions: Cut shortening into flour,to which salt has been added,
with a pastry blender. .Add cold water. Mix lightly with a fork.

Roll on wax paper.1 For double crust, weigh out 421 grams per pie.

For single crust, use 210 grams per pie. Bake at 400°F for 12 minutes.2

1 NO extra flour is to be used in rolling the crust.
Only bake shell when required by recipe.

 

 

156

Pineapple Upside Down Cake

1

Yield: Three 8” x 8" cakes
Temp.: 350°F

Time : 30 minutes

 

Ingredient W21::t :2? Prggein :2?
Butter (protein free) 70 --- --- 56.0
Brown Sugar 275 262.6 --- ---
Pineapple slices (drained)1 488 184.1 3.6 0.7
Flour 338 249.5 43.1 3.4
Sugar 250 250.0 --- ---
Baking Powder 20 --- --- ---
Salt 7 --- --- ---
Shortening 163 --- --- 163.0
Pineapple juice1 385 ‘_;;;__ --- ---

Prepared Weight: 1715 946.2 46.7 223.1

Serving (one): 75 41.4 2.0 9.8

Instructions: Melt butter in 1 quart sauce pan. Add brown sugar.

Place 175 grams of the sugar and butter mixture in each pan. Arrange
162 grams of drained pineapple in this mixture. Combine dry ingredients
and soften shortening in electric mixer bowl. Add pineapple juice.

Beat 2 minutes. Weigh and divide into thirds (approximately 540 grams).
Place this amount in each pan. When done, let stand 5 minutes. Then

invert on wax paper.

Pineapple calculated values represent 488 grams of slices and
385 grams of juice.

 

1!?“-

1
‘.TF'

-._:~

157

 

Pizza Dough
Ingredient Gram CHO Protein Fat
Weight gm. gm. gm.
Sugar 30 30.0 --- ---
Salt 16 --- --- ---
Shortening 35 --- --- 35.0
Boiling Water 300 --- --- ---
Lukewarm Water 450 --- --- ---
Yeast (dry) 18 7.0 6.6 0.3
Flour 1130 834.1 133;g_ 11.3
Prepared Weight: 1979 871.1 150.8 46.6
Serving (one) : (wet weight) 150 65.5 11.3 3.5

Instructions: Combine yeast with 150 grams of lukewarm water in a

1 quart mixing bowl. Set aside in warm area. Place salt, shortening
and boiling water in Hobart mixing bowl. Beat until shortening is
dissolved. Add 300 grams lukewarm water. Add sugar and yeast mixture.
Add three cups of flour and beat until smooth. Addremaining flour.
Knead at low speed for 5 minutes. Into each 8" aluminum foil pie
plate, place 150 grams dough. Flatten the dough so that it covers

the bottom of the pan and comes up on the sides 1/2 inch.

 

1r-

'- ‘11.

158

Pizza Sauce
0
Temp.: 425 F

Time : 20 minutes

 

 

 

Ingredient WStiram CHO Protein Fat
ght gm. gm. gm.

Canned tomatoes (drained) 2760 107.6 27.6 5.5
Onion (fresh, chopped) 130 13.4 1.8 0.3
Green Pepper (fresh, chopped) 325 18.6 3.9 0.9
Sweet Basil 25 --- --- ---
Oregano 5 --- ---- ---
Salt 5 --- --- ---
Bouillon Cubes 8 ___ 1 0 ___

(2 vegetable cubes) °

Protein free butter 28 --- --- 22.4
Prepared weight: 2994 139.6 34.3 29.1
Serving (one) : 175 8.2 2.0 1.7

Instructions: Combine all ingredients in an 8 quart aluminum kettle.
Cook 45 minutes. Cover each individual serving of pizza dough with

' o
150 grams of sauce. Bake at 425 F for 20 minutes. After baking,

turn oven off -- leave pizzas in oven until all subjects are served.

 

‘1

{HAL-1-

159

Potato Salad

 

 

Ingredient WEEZEt :2? Prgiein :3?
Cold boiled potatoes
(peeled) 900 171.9 18.0 0.9
Onion (raw, chopped) 40 4.1 0.6 0.1
Parsley (dry) 7 0.1 0.1 ---
Pickle Relish 50 13.1 0.4 0.4
Vinegar 20 --- --- ---
Salad Oil 85 --- --- 85.0
Salt 8 --- --- __;;;_
Prepared Weight: 1110 189.2 19.1 86.4
Serving (one) : 75 12.8 1.3 5.8

Instructions: Scrub potatoes with skins and place in 8 quart kettle.
Cover with water and cook 45 minutes. Drain. Peel and slice potatoes
into large bowl. Add onions, parsley, and pickle relish. Combine
vinegar, salad oil and salt. Pour this mixture over the potatoes.
Toss lightly. Keep in cool place until time of serving. Turn once

before serving.

Potatoes au Gratin

Ingredient

Potatoes (cooked)
Butter

Flour

Milk

Salt

Pepper

Parsley (dried)

Prepared Weight:

Serving (one) :

160

Gram

Weight

1400
75
40

1350

dash

5

2650

130

CHO

267.4

0.6

29.5

68.6

0.3

366.4

18.2

Instructions: Peel 2000 grams raw potatoes.

butter in 8 quart aluminum kettle.

milk gradually. Simmer 15 minutes.

potatoes. Then add parsley.

Protein

gm.
28.0
0.5
5.2

49.0

0.1

 

82.8

4.0

Stir flour into butter.

Add salt and pepper.

6

4

Fat

1.4

0.4

0.4

9.8

0.1

 

11

Cook until done.

Add

Add

casserole. Top with 20 grams bread crumbs and 5 grams butter.

in warming oven until all subjects are served.

2.1

5.2

Melt

Place 130 grams potato mixture into a

Place

 

161 .

Refrigerator Cookies (Control Period)

0
Temp.: 350 F

Time : 8 minutes

 

 

Ingredient WGram CHO Protein Fat
eight gm. gm. gm.
Butter 6701 5.4 4.0 539.4 ,
Sugar 300 300.0 --- --- §
Flour 825 608.9 105.3 8.2 3
Lemon or Vanilla Extract 20 ._;;;__ __:::_- --_ J
Prepared Weight: 1571 914.3 109.3 547.6 L; ‘
Serving (one) : 50 29.1 3.5 17.4

Instructions: Cream sugar and butter until fluffy in Hobart mixing
bowl. Add flour and flavoring. Mix at low speed for 3 minutes.
Form.oblong roll and wrap in wax paper. Chill in refrigerator.
Slice crosswise. Bake on lightly greased baking sheet. Remove
from.baking sheet and place on wax paper. When cool, weigh and
store in plastic bag in the refrigerated walk-in. Be sure to label

bag with the contents, weight and the date prepared.

Reference: Swedish Food, 1946, 5th ed. Esselte, Gothenburg, Sweden.
p. 132.

 

10 grams of butter is to be used to grease the cookie baking sheets.

162

Refrigerator Cookies (Experimental Period)
0
Temp.: 350 F

Time : 8 minutes

 

 

Gram CHO Protein Fat
Ingredient Weight gm. gm. gm.
Butter (protein free) 670 1 --- --- 536.0 {‘3
Sugar 300 300.0 --- ~-- I
Flour 825 608.9 105.3 8.2
Lemon Extract 20 --- --- --- t;‘
(or Vanilla Extract J
Prepared Weight: 1570 908.9 105.3 544.2 k"
Serving (one) : 50 29.0 3.4 17.3

Instructions: Cream sugar and butter until fluffy in Hobart mixing
bowl. Add flour and flavoring. Mix at low speed for 3 minutes.
Form oblong roll and wrap in wax paper. Chill in refrigerator.
Slice crosswise. Bake on lightly greased baking sheet. Remove
from baking sheet and place on wax paper. When cool, weigh and
store in plastic bag in the refrigerated walk-in. Be sure to label

bag with the contents, weight and the date prepared.

10 grams of butter is to be used to grease the cookie baking sheet.

163

Rice (Boiled)

 

 

 

Gram CHO Protein Fat

Ingredient weight ng gm. gm.
Rice (dry) 418 332.2 31.9 1.1
Water 1922 --- -—- ---
Salt 10 --- --- ---
Prepared Weight: 1300 332.2 31.9 1.1
Serving (one) : 100 26.2 2.5 0.1

Instructions: Bring water and salt to boil. Add rice and stir.
Cook for 30 minutes using medium heat. Remove cover, place container

in pan of hot water to keep warm until all subjectsare served.

Shortcake
Temp.: 4OOOF

Time : 15 minutes

- mt..--

 

Spa.“
:1
.uw: '?‘_._

 

 

Gram CHO Protein Fat

Ingredient weight gm. gm. gm.
Flour 550 406.0 70.2 5.5
Sugar 60 60.0 --- ---
Baking Powder 3O --- --- ---
Salt 20 --- --- ---
Shortening 165 --- --- 165.0
water 505 --- --- ---
Prepared Weight: (raw weight) 1330 466.0 70.2 170.5
Serving (one): (raw weight) 65 22.8 3.4 8.3

Instructions: Combine dry ingredients. Cut in shortening with a

pastry blender. Add water. Stir only to mix. ‘Weigh out 65 grams

raw weight per subbct. Bake on ungreased baking sheet.

164

Shredded Cabbage with Pineapple Salad

Ingredient ngggt :fi? Przifiin :3?
Cabbage (shredded) 420 22.7 5.9 0.8
Pineapple Slices (canned) 280 52;9_ 1;2;_ 9;;

Prepared Weight: 700 81.7 7.1 1.0

Serving (one) : 50 2.7 0.7 0.1

Instructions: Drain pineapple and cut into chunks.

Combine these two and toss lightly.

 

Spaghetti
In redient Gram CHO Protein Fat
g weight gm. gm. gm.
Control
Spaghett? (eanChEd’ 530 407.9 68.1 7.4
dry weight)
Water 1580 --- --- ---
Salt 10 --- --- ---
Experimental
Spaghetti (enriched,
dry weight) 800 611.9 102.1 11.1
Water 2370 --- --- ---
Salt 15 --- --- ---
Prepared Weight: (Control) 1350 407.9 68.1 7.4
(Exper.) 2025 611.9 102.1 11.1
Servin (one)- (Control) 100 30.2 5.1 0.6
8 ° (Exper.) 146 44.1 7.4 0.9

 

Shred cabbage.

 

Serve 50 grams to each subject.

a“ -
——..

 

Instructions: Place water and salt in 8 quart alwminum kettle.
Bring to boil. Add spaghetti. Cook 20 minutes at high speed. Drain.
Rinse with hot water. Place the amount specified for the individual

serving in the casserole dishes.

165

Spaghetti Sauce

 

Ingredient WSEZEt 2E?
Onion (raw, diced) 250 25.8
Green Pepper (raw, diced) 250 14.3
Vegetable Oil 70 ---
Salt 10 ---
Sugar 9 9.0
Tomatoes (canned) 2400 93.6

Prepared Weight: 3000 142.7

Serving (one) : 200 9.6

Protein
gm.

3.5

3.0

24.0

 

30.5

2.0

Fat

0.5

0.7

70.0

 

76.0

5.0

Instructions: Sautee onion and green pepper in vegetable oil in

8 quart aluminum kettle. Add seasonings. Add tomatoes and heat

thoroughly (30 minutes). Must total 3000 grams.

cooked spaghetti with 200 grams of this sauce in individual casserole.

Place in oven prior to serving.

Cover 146 grams of

 

 

Spanish Rice

Ingredient

Bacon, raw
Onion (raw, chopped)
Green Pepper (raw, chopped)
Tomato Soup (Condensed)
Rice (dry weight)
Water
Whole Cloves
Bay leaves
Salt
Prepared Weight:

Serving (one) :

166

Gram
Weight

84
600
180

2700
450
675

18

4
12
4550

300

 

CHO Protein Fat
gm. gm. gm.
0.6 6.9 48.6
61.8 8.4 1.2
10.3 2.2 0.5
329.8 43.5 54.4
357.6 34.3 1.2
760.1 95.3 105.9
50.1 6.3 7.0

Instructions: Cut bacon into small pieces. Fry until crisp in

8 quart aluminum kettle. Add onion and green pepper. Fry until

golden. Add remaining ingredients.

Cover and cook slowly 50 minutes.

Remove cloves and bay leaves. Serve each subject 300 grams in

individual casserole. Place casseroles in warming oven until all

subjects are served.

 

Sweet Rolls
0
Temp.: 350 F
Time : 35 minutes

Ingredient

Control

Bread Dough
Sugar
Butter

Cinnamon

Prepared Weight:
Serving (one) :
Experimental

Bread Dough

Sugar

Protein free Butter

Cinnamon

Prepared Weight:
Serving (one) :

Instructions: Roll dough to 1/2 inch thickness.

sugar and cinnamon.

167

Gram
Weight

1700
200
200

12
1807

130

3000
350

1
350
22

3186
200

CHO

772.2
200.0

1.6

 

973.8

70.1

1363.0
350 .0

1713.0
107.5

Spread evenly over rolled out dough.

Protein

1.2

 

132.5

9.5

231.6
14.5

 

371.5
23.3

Combine butter,

Roll dough

into a roll. Slice 1/2 inch thick and place on greased baking sheet.

Cover and let rise one hour.

15 grams of the weighed butter for the Control sweet rolls is to be
used to grease the baking sheet.

30 grams of the weighed butter for the experimental sweet rolls is

to be used to grease the baking sheet.

 

168

Tomato and Rice Casserole

 

Ingredient Gram CHO Protein Fat
Weight gm. gm. gm.

Rice (converted, dry weight) 260 206.6 19.8 0.7
Salt 7 --- --- ---
Water 810 -—- --- ---
Onions, raw (minced) 160 16.5 2.2 0.3
Celery, raw (diced) 160 5.9 2.1 0.3
Green Pepper, raw (diced) 160 9.2 1.9 0.4
Vegetable Shortening 50 --- --- 50.0
Tomatoes (canned) 2000 78.0 20.0 4.0
Celery Salt 14 --- --- ---
Sugar 8 8.0 --- ---
Salt 28 --- --- ---
Pepper dash _-_-_-___ _._:_.__ ---
Prepared Weight: 3555 324.2 46.0 55.7
Serving (one) : 245 22.3 3.2 3.8

Instructions: Cook rice in boiling salted water until tender. Cook
onions, celery, green pepper in vegetable shortening in 8 quart
aluminum kettle. Add tomatoes and remaining seasonings. Add cooked
rice to this mixture. Serve 245 grams of tomato/rice in individual
casseroles. This is to be topped with 25 grams of bread crumbs and

5 grams butter (protein free). Place casseroles in warming oven until

all subjects are served.

 

Tim- 4...
1 .

169 '

Vegetable Chop Suey

 

 

 

Ingredient ngggt :3? Prggein :3?
Vegetable Bouillon Cubes (2) 8 --- 1,0 ---
Water 3760 --- -_- ---
Flour 240 177.1 30.6 2.4
Salt 10 --- ---- --- f4
Pepper 1 --- --- --_
Wercestershire Sauce 40 7.2 0.8 ---
Soy Sauce 40 4.0 trace 0.5 ‘
Celery (fresh, diced) 1000 37.0 13.0 2.0 PVT
Onions (fresh, diced) 240 24.7 3.4 0.5
Soy Bean Sprouts (canned, 135 11.8 11.8 1.7
drained) _______
Prepared Weight: 4884 261.8 60.6 7.1
Serving (one) : 350 18.8 4.3 0.5

Instructions: Combine bouillon cubes and 3000 grams of water in an
8 quart aluminum kettle. Bring to boil. Make smooth paste from 760
grams of water and flour. Add this paste to the bouillon water.

Add remaining seasonings. Cook celery and onions in small amount

of water. Drain and add to previous mixture. Add bean sprouts.
Check total weight which should be 4884 gramsE Serve 350 grams of
vegetable chop suey in each casserole. Place casseroles in oven to

keep warm.

1 Add water if required to meet this weight.

170

Vegetable Soup

 

 

 

Ingredient Gram. CHO Protein Fat
Weight gm. gm. gm.

Potatoes (raw, diced) 300 57.3 6.0 0.3
Carrots (canned, drained) 725 46.4 4.8 3.9
Celery (raw, diced) 350 13.0 4.6 0.7
Butter (protein free) 50 --- --- 40.0
Flour 40 29.5 5.1 0.4
Vegetable Bouillon Cubes (5) 20 --- 2.5 ---
Boiling Water 2700 --- --- ---
Salt 9 --- --- ---
Pepper 1 --- --- ---
Prepared Weight: 3724 146.2 23.0 45.3
Serving (one) : 250 9.8 1.5 3.0

Instructions: Cook celery and potatoes in small amount of water until
done. Drain. ‘Melt butter in 8cpart aluminum kettle. Stir in flour
until well blended. Slowly add boiling water. Add vegetable bouillon
cubes. Add salt and pepper. Add vegetables. Cook slowly 10 minutes
stirring constantly. Place 250 grams in each casserole, cover, and

place in warming oven.

. 171

Vegetarian Stew

 

 

Ingredient ngzgt 2E? Prgiein :3?

Cagrots (canned, drained, 370 23.7 2.5 2.0
iced)

Potatoes (raw, diced) 180 34.4 3.6 0.2
Celery (raw, diced) 150 5.5 1.9 0.3
Wax Beans (canned, drained) 150 11.6 3.6 0.3
water 960 --- --- ---
Vegetable Bouillon Cubes (2) 8 --- 1.0 ---
Flour 40 29.5 5.1 0.4
Salt 3 --- --- ---
Pepper dash --- --- ---
Butter (protein free) 50 --- --- _4Q;Q_

Prepared Weight: 975 104.7 17.7 43.2

Serving (one) : 75 8.0 1.4 3.3

Instructions: Cook potatoes and celery. Drain well. ‘Melt butter in

8 quart aluminum kettle. Stir flour into it. Add hot water, vegetable
bouillon cubes and seasonings. Add all the vegetables to this
thickened sauce. Add dumplings (See dumpling recipe). Cook 20
minutes. Serve one dumpling and 75 grams vegetable stew in individual

casseroles.

 

172

Wheatena
Gram
Ingredient Weight
Wheatena 672
water 3648
Salt 8
Prepared weight: 3780
Serving (one): 270

Instructions: Bring water to a boil.

slowly, so water never st0ps boiling.

 

CHO Protein Fat
gm. gm. gm
520.8 69.6 14.4
520.8 69.6 14.4
37.2 5.0 1.0

Add salt. Sprinkle in Wheatena

Cook 10 minutes. Remove from

heat. Place receptacle in pan of hot water to keep warm for serving.

”'1

 

"1'4.“ ;

N'W‘

173

White Cake

Yield: Three 8" x 8" cakes
Temp. : 325°}?

Time : 30 minutes

Gram
Ingredient Weight
Eggs 166
Sugar 525
Butter 196
Milk 420
Salt 4
Flour 490
Baking Powder 24
Vanilla 8
Prepared weight: 1585
Serving (one) : 100

Instructions: Line three 8")c8" cake pans with wax paper.

CHO
gm.

0.9
525.0

 

910.5
57.4

all dry ingredients in electric mixing bowl.

vanilla, and two tablespoons of the milk.

speed. Add egg and remaining milk.

Place 611 grams in each cake pan.

Protein

m.
21.2

 

100.4
6.3

Fat
gm.

16.9

157.8
14.6

 

 

4.9

 

194.2
12.2

Combine
Add the softened butter,

Beat 1 minute at medium

Beat 3 minutes at high speed.

Reference: Farmer, F. M. 1951. The Boston Cooking.School Cook Book
9th ed. Little Brown and Co., Boston, p. 701.

White Icing (for cupcakes)

Ingredient

Butter

Vanilla

Confectioner's sugar
Prepared Weight:

Serving (one) :

Instructions: Melt butter in 1 quart sauce pan.

Beat in vanilla and sugar.

174

Gram
Weight

75

340
415

30

Frost cupcakes using 30 grams per subject.

338.3

338.9

24.5

Protein
gm.

0.4

Fat

60.4

60.4

4.5

Remove from heat.

 

15g“

—.-

HICHIGQN STRTE UNIV. LIBRQRIES

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