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/Determinaticn of Fat Content

of'a Controlled Weight Reduction Diet///

A Paper
Submitted to the School of Graduate Studies
of Nicnigan State College of Agriculture and Applie
Science in partial fulfillment of the requirements
for the degree of
Master of Science

Department of Foods and Nutrition
1951

a. u! '
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THS

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Tetle of Contents

Cnapter
Introcuction
“3v1ew of Literatire . . . . . . . . . . . . . . . . . . . .
Experimental Procedure . . . . . . . . . . . . . . . . . . . h
Results and Discussion . . . . . . . . . . . . . . . . . . . 7
Sun'ery and Conclusion . . . . . . . . . . . . . . . . . . . 18
. . . . . . . . . . . . . . . . . . . . . . 19

Literature cited .

Intro suction

 

1; many laboratories conducting etzperiments involving the pr3para-

t ,significant di fVer:noes have been observed tetraen the

.9.
F]
C)
D
O
H
(D

(I)

caloric vale es as calculate 1 from tables of food composition and those
obtains d ty the arrlic ation of conversion factors to the values for car—
bohadr ates, protein and fat deternineo by proximate analysis.

In the research proj; ct concerning tne itilizu-lon of calories and
protein by overweight college women on a weiaht—re‘rot on diet, ard con—
‘ucted ty the Foods and Vutrition Desertnent of Hichigan Etete College,
discrepancies were otserv ed bet «seen the food calories as computed from the

tahles and as determined from the bomb calorimeter

U!

A the diets were characterized by a high fat content, a likely
'ariation in the actual fat con mte.t of the diet from tie values computed
from the food tables, w 5 considered an important factor in causing the
discrepancy. The purpose of this study therefore was to eterrine the

fat content of tne composite diets used in the atove-men‘woned research
project, by quantitative analysis. A con pa arisen of the valtes thus otters-
ed with the fat content as calculated from the food composition tables,

was also a matter of consideration.

It night be ad< led that sore of the fecal sarples of the subjects were

also analyzed for their fat content.

Review of Literature

 

Various methods have teen developed and used for the extrac tion of

at from focus, ranging fro; direct ertraction from the dried product by

F4)

solvents, as an1 'fi cation of t‘e fat followed by extraction of the libera a
fatty acids tyl ight ne troleun, to acid treatment followed by extraction
with mixed ethers (Eransby et al. l9h6).
The first method fails to extract the fat completely from many cooked
st.arohy foods, such as bread and biscuit. In the second method the calcu-

lation of the fat content re

U)

ts on the assumption that'fiiglycerides are

the only conround sof fattr ac1ds xresent in the material originally tdcen.
Besides, since light petroleun takes up sterols as Well as fatty acids
stars should have to be taken for their removal.

The prescerce of lower fatty acids in butter, mare arine and hydrogen-

ted coconut oil introduces errors since these acids are appreciably
volatile under the conditions re to drive off the ext ra cting solvent
and to dry the recovered fatty acids. Further, as tutyric is miscible
with water, and other acids are appreciably soluble in it, they are not
come ole tel y extracted by solvents.
The th 1rd method con1* )iSlS in treating the sample with hydrochloric
acid followed by alcohol and extracting the fat with ethyl ether and light
petroleum. ihe extract 0 tained by this method includes glycerides, phos-
phatids, sterols and free fatty acids.

A reliable method of fat-analysi s is by contirwuou extraction with
ether in a soxhlet apparatus, although it takes a long time. Extraction
determines crude fat, including neutral fats, phospholipids and other
substances in small amounts. Extraction method is convenient, and con—

sidered more suitable if the samp la is proparl1y grouni and drie d. (FiC', 19h?)

This method was adopted for the present investigation.

Fickelsen et al. (19h?) analyzed batches of three different low.
fat diets used in rotation over a period of six months as part of a large
eXperiment on semi—starvation. Results of analysis of different batches
of the same diet showed that the apparent fat content was subject to the
greatest day-to-day variation. Using the soxhlet method of extraction
with et er, they found that an addition of a detergent (.S% Duponol) to
the food collected for fat analysis resulted in a very considerable in—
crease in the weight of material extracted by ether. This was explained
as being due to the homogeneity produced in the sample by the surface active
agent. The difference amounted to as much as 33 per cent in a dry food
mixture atho as much as 330 per cent in a mixture analyzed in the moist
state.

Shannon (19h9) reported that the length of drying time after ex-
traction influenced the final percentage of fat obtained.

Iransby et al. (19L?) in their study of the comparison of the estima-
tion of the food intake for 3 days of a number of children obtained by
weighing and calculating, and by chemical analysis found that calculation
from tables over-estimated the fat content. In another study on the
methods of individual dietary survey Eransby et al. (lvhfi) found that for
individual diet differences between the values found by calculations and
by chemical analysis were in many instances so large as to throw doubt
on the usefulness of the individual results obtained by calculation.

Humnel and others (l9h2) found that the results obtained from the
analysis of foodstuffs varied widely from the standard figures. The few
foods other than butter and milk which contained fat in appreciable
amounts, were concentrated types of foodstuffs which are difficult to

sample accurately. In addition they were not uniform as purchased, though

1.9

every possible precaution was observed to control variations which might
arise at the source of supply. For instance, shoulder bee was recommended
as the most uniformly lean neat to be obtained, and all purchases were made
from one kitchen, but samples were found to vary as much as 1-15 per cent
in fat content. The energy values of teef confirmed the difference in com-
position. flunmel found that individual samples of a given food vary from
values reported in standard tables, but when analysis of composite diets
are compared with dietary figures calculated from the literature, there

is fairly good agreement in fat con,ent.

Thomas et a1. (1950) in a study of the nutritional status of children
evaluated the accuracy of calculated intakes of food components with respect
to analytical values. For fat and some other nutriments, 28 of 33 differences
showed smaller contents by analysis than were indicated by calculation.

The fat intakes estimated from food composition tatles were in no close agree-
ment with the actual amounts of fat ingested, emphasizing the great vari—
ations in the concentration of this constituent in foods. Thus a compari—

son of data obtained by analysis and by calculation from standard tables

of food composiqion showed that in diets eaten by two groups of children,

in fall and in spring, the results for fat were significantly different

in both the seasons. Consistent differences indicated that values selected
from the tables did not approximate the true values in either season.

The analyzed values for the fat content of diets containing large
amounts of complex recipes, could be different from values ottained by
calculation from tables. In calculating the diets of older women using the
tables of Donelscn and Leichsenring (l9h5), Ohlson (1950) found that the
calculated values for nitrogen, calcium and phosphorus did not correspond
to the analyzed values for these diets which contained a high proportion

of processed meats or complex cooking mixtures.

Egperimental Procedure

 

Experimental Packpround:

Q;

The iets analyzed for their fat content, were those used in the
metabolism study on weight reduction carried on six overweight college
women, by the Department of Foods and Nu rition of ‘iich iga an State College.
The general pattern of the weight reduction diet was: high protein,
high fat and low carhohydrate, with a total caloric intake of 1, €33 -
1,700 calories per day. (The National Research Council recommends an
ave ra._e Cf 2,090 calorie es daily for healthy, nodenmfly active young women)
The an ije sets were maintained on a self— selec ed diet for a two—week
observation period, prelizzinary to the peri.od of Wei ght loss. During the
second week a talance study was made to es tatlish calcium retention of the
subjects. After two weeks on the «eight reduction diet, arother balance
period was maintained for a week. Six talance periods were completed

during the entir study which lasted from January, 1951 to June, 1951.

Experimental diet:

0

seven eenus were planned for the weight—reduction diet, and these
Wi"c
were repeated each week. The meals crepared and serves in the dome Econo-
f.‘
mics Department. All foods eaten were weighed on a hansen dietetics

Tiling of food:

During the second geek of the self—selection diet, all servings were

5 :.J

13d and a weighed aliouots equal to one-fifth of the food eaten were
frozen, after each meal. i s‘mflar proeedare was followed for the weight

reduction riet. Liouids and solids w3re saved secarately for convenience

-

(U

in handling. The seven—day collection period was divid d into A.L—day

period and B. 3—day period for convenience. The sanples were thawed before

*1

blending at the completion of each week. The food was blended

in a Waring food blender for five minutes (until mixture became
homogenous), transferred to a 2-1iter volumetric flask and made to
volume with distilled water. After mixing the aflnrry, two sanples
were measured into 250 milliliter volumetric flasks, then transferr-
ed to weighed evaporating dishes and dried in an oven at 40 degrees
centigrade for 5 to 4 days until dry. The samples were stirred at
intervals to permit more even drying, and after drying to constant
weight, the sample was scraped from the dish, transferred to a
bottle and stored in a desiccator until analysed.

During the balance periods daily fecal collections_were made
in waxed containers. fit the end of each balance period the fecal
collectionswere transferred to a blender and blended for five minute
then made to volume in a two-liter volumetric flask. Three samples
were measured into 100 m1. volumetric flasks, transferred to weigh-
ed evaporating dishes and dried partially over a steam bath and
then under infra-red lamps. After drying to constant weight the
sample was scraped, ground and stored in bottles in a desiccator
until required for analysis.

The average weekly loss of weight was one kilogram per person.
Fat - extraction method:

Approximately tvo grains of the dried sample was weighed out
directly in an ether-extracted extraction thimble (weighed previous-
ly) in an analytical balance. The samples were then extracted in a
soxhlet apparatus for twelve hours.

The soxhlet apparatus consists of a wide glass tube with a
side siphon, connected at the bottom to a receiver flask and fixed
to a condenser at the top. The apparatus is fitted up on a water

bath and the condenser cooled with a running stream of water from

‘d

the tap.

After the water had been allowed to run for some time, ether
was poured into the receiver flask and the latter was fixed
tightly to the soxhlet. The steam was turned on the water bath.
The ether evaporated, was condensed in the condensor and allowed to
drop into the soxhlet tube with the extraction thimble and the
sample in it. The ether dissolved the fat in the sample, and when
enough of it collected, it was automatically siphoned back into
the receiver. Thus the extractior was continuous. After the 12-
hour extraction, the steam was turned off, the apparatus disconnect-
ed and the thimtles carefully removed with a pair of tongs, and left
in beakers to dry in the air for an hour. They were dried in the
disiccater afterwards for 18 hours before being weighed. The dif-
ference in weights gave the weight of fat extracted from the sample.
The percentage of fat in the given sample was calculated from this
and from the recorded weight of the dried material in the 250 ml.

7a., “raw-45¢ .4. Lt

aliquot of the blended compositefiwas obtained. Finally the grams
of fat in the weidht reduction diet for the given period per day
was calculated as shown in Table 1.

Fat extractions were-made of duplicate samples of both A
and B parts of the following weighed controlled periods:
ll, l2, 14, 15, 17,-20 and 22.

The conditions for drving were standardized in order to elimin-
ate any variation in moisture content.

‘ Fat extractiwns of the fecal samples of one of the subjects

'L' for the following balance periods were made:
2, 6, 13, and 2i.

The amount of fat in the feces per day was calculated as

shown in Table 2.

Results and Discussion

 

The average weights of fat in the diets as obtained from
analysis by ether-extraction are given in Table 3. The fat contents
of the diet as calculated from the food tables (U. S. Department
of Agriculture, 1950) are given in Table 4.

The daily average weight of fat in the weight-reduction diet
as ortained by extraction was 92.8 grams, while ttat calculated
from food composition tables was 100.2 grams as shown in Table
0. This result is in accordance with thst obtained by Thomas
et al. (1950) who also found that the fat content of the diets
was smaller when obtained by analysis than the values obtained
from food composition tables.

The fat content of the diets analyzed for the seven different
periods varied fran 82.2 to 101.5 grams per day (Table b). The
actn 1 fat content of meat, and rather intricate y prepared recipes
containing high fat, has been found to vary and fluctuate con-
siderably from the values given in good composition tables. Althoug
conditions of preparation, the source and cuts of the meat (as
all other foodstuffs used in the diets) were standardized as far
as possible, it is likely that some factors of variables could
not be eliminated. Furthermore, there is no proof that the fat
extraction values are absolutely correct. Peoefited extractions d7
duplicate sump as need to be done in order to get appreciably
accurate results. The moisture content is an important factor in
causing variation in results. Although the conditions for moisture
content were standardized, in the present experiment, it cannot
be claimed that they were the same as ttcse conducted in other

laboratories.

In the weight reduction diets, an average of 1,650 calories
per day was supplied, of which about 360 calories were provided
from approximately 90 grams of protein, 460 calories from about
115 grams of carbohydrate and the remaining 840 calories hv about
90 grams of fat.

The average daily weight of fat in the diets obtained by
analysis was actually 92.9 arrms which provided 855 calories
which is nearly 49 per cent of the total calories (averaae
for the weight control periods analyzed) in the day's diet.

The latter determined by the bomb calori eter, was 1,708 calories.

The fecal samples of Subject L from the four balance periods
analyzed gave the results tabulated in Table 6. The fat ex-
tracted from the feces ranged from 2.65 to 3.65 grams per day
for periods II, VI, XIII. The validity of the results obtained
for peribd XXI is questionable since the Sgnginaal fecel sample
was too small,for convenient preparation of the dry sample. .It

was found thrt 51.5 to 46.5 per cent of the total calorie output

in the feces was provided by the fat content of the feces.

Table I

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Table II

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_ 11 _

Table III

Fat content of the diets as attained by analysis.

 

 

period Sample Sarule

 

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(1'3" ‘. . .' €("c,.\ [Lu 3'“?- ')
Teal ax'eraére flit. Of fat. per day fro-mJ'TQI‘iOdS 11 — 22 = 92.5

 

-12-

Table IV

Fat content (calculated from tables) of foods included in menus during
seven day period of weight reduction diet.

 

 

 

 

 

 

 

 

MONDAY
Food Quantity. wt. in gms. Gms. fat
Grapefruit and orange juice 100 0.1
Eggs — 2 12h 11.5
Tread 29 006
Butter 10 8.0
Milk hSJ 17.5
Pork Chop 125 32.5
Broccoli , 100 0.2
Hamburg 125 37.5
Creamed potatoes » 100 2.0
Total fat content of the day's diet . . . . . . . . . . . . . . . 109.9
TUESDAY

Grange juice 100 0.2
Eggs 12h , 11.5
Eread ‘ 20 0.6
Butter 10 8.0
Milk L53 17.:
Lamb Patties 12, 35.0
Pears, drained 110 .

Lettuce 15 -

Roast veal 125 15.0
Peas 75 0‘3
Butter 10 8.0
éggle saace 133 0.2
Total fat content . . . . . . . . . . . . . . . . . . . . . . . . . 96.h

’3

-13..

Table IV - continued

 

Food

Quantity. wt. in gm. Gm.

 

Orange juice
Eggs - 2
Butter

Lread

"ilk

Feat Loaf
Cottaye cheese

res“ tomatoes

Roast teef

to

Cl‘

toiled pot
a:

Sweet ch 'ries

100 0.2

12h 11.5

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80 O.h

 

Total fat content of the day's diet . . . . .'. . . . . . . . . . . 102.5

 

 

 

TflUpSDAY

Grapefruit juice 100 3.1
Eggs - 2 122. 11.5
Putter 15 12.0
{read 2’1 0.‘
T'ilk LSl 17.,
Swiss steak, with 125 16.2

fat for broiling 5 5’,L
CauliflCWSP 139 3.2
anet pork 125 32.5
Beets 130 0.1
Arricots 1‘0 0.1
Total fat content of the uav's diet . . . . . . . . . . . . . . . . 95.8

,7

................

Table IV - continaed

 

 

FRIDAY
Food Quantity wt. in gm. Sm.

Tomato juice 190 0.2
TEES 121: 11 .5

Ewtter 33 2L.0

Tilk [53 17.5
T J c /
LTDCC. (-q a.

Eaked white fish (Haddock) 125 7.3
Sauce 15- 2.3
Leked sweet notatoes 100 0.9

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Green beans 100 0.2

¢c+al fat content of the dag's diet 91.1
Saturday

Crange é grapefruit juice 130 3.1

1 rd

“6.1/1.8 1&1; 110E,

I-I“fiéid 20 O. 6

Vilk L511 17 . 5

danburg gq 37.§

.ossed salad 103 0.2
French dressing 5 1.3
”Fr“ roast / 32.g

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-15..

Tatle 1V continued

iUUDAY

 

Quantity wt. in gm.

Gm fat

 

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L

Lettuce
“ranch dressing
Cold meat loaf

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'T' J'.- 4. ~ 1- +‘..- ‘4‘ '
lotal 1a. content of chi day's diet

Table Showing the daily intak
seven—day period, as calcuiet

Da‘

Daily average

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e

71111113 V

of
d.fr 3 food composition tables.

Gm. fai/dav

 

109.9
96.1.

102.5

fat on the weight reduction diet in a

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-18;

Summarg*and Conclusion

 

The fat contents of dried samples of controlled weight_
reduction diets for seven periods were determined by extraction
with ether in a soxnlet apparatus, under standardised conditions.
An average daily fat content of the diet was calcu‘ated from the
seven—day menu using food composition tables. The average amount
of fat in the diet as obtained by ether extraction was found to
be 92.8 grams per day which was lower than the amount estimated
by calculation from food tables -- this being 100.2 grams per day.
AS the diet contained a daily average of 1,650 calories, tle fat
provided nearly 49 per cent of these calories. F

The discrepancy between the fat values of the diet obtained
by the two methods could be due to various factors already cited
in the review of literature. However, it must be noted that this
difference is not too great.

The dried fecal samples of one of the subjects L on the fat
reduction diet from four balance periods were also ether - ex-
tracted to determine the fat content of the feces. Approximately
5 grams of fat were excreted in the feces daily, and this formed

50 per cent of the total caloric output.

109 0804
38735

40

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10.

”I9 _

Literature Cited

Bransby, E. R. Daubney, C. G., King, J. 1948. Comparison of
results obtained by different methods of individual dietary
survey. British J. of Nutr. vol. 2, p. 89.

Bransby, E. R., Daubney, C. G., Ving. J. 1948 Methods of
individual dietary survey. Dr. J. of Nutr. vol. 2, p. 232.

Donelson, . G., J. M. Leichsenting. 1945. Food compostion
table for hort method of dietary analysis. J. Amer. Diet.
Assoc., vol. 21, p. 440.

..J (a [.‘3

Food and Agriculture Organization of the United Nations. 1947
Energy yblding components of food and computation of calorie
values.

Hummel, F. 0., M. L. Shepard, H. Galbraith, H. H. Williams,
G. Macy. 1942.. Chemical composition of twentyétwo common
foods and comparison.of analytical with calculated values of
diets. J. of Nutr. vol 24,up 41.

Mickelson, 0., E. 0. Miller, A. Keys and H. H. Mitchel 1947.
The determination of fat and its relation to the calculated
calorie values of diets. J. of Diet. Assoc. vol. 23, p. 952.

Ohlson M. A., L. Jackson, J. Bebk, D. C. Cederquist, W.D.
Brownr, E. G. Brown. 1950. Nutrition and dietary habits of
aging women. Amer. J. Pub. Health. Vol. 40., p. 1101.

Shannon, A. F. 1949. Refractive index and ether extraction
methods for oil and avocados. Chem. Abst. vol. 45, p. 9285 h.

Thomas, R. U., M. N. Rutledge, E. F. Beach, E. Z. Moyer,

M. C. Drummond, S. Milder, A. R. Robinson, 0. N. Filler, N. N.
Ceryell, I. G. Macy. 1950. Nutritional Status of Children.
XIII. Accuracy of calculated intakes of food components

with respect to analytical values. J. Amer. Diet. Assoc.

vol. 26, p. 889.

U. S. Department of Agriculture. 1950. Agriculture Handbood
No. 8. Composition of foods.

 

93 02237 2365