FISHERIES AND MARINE SERVICE Translation Series No. 3274 A study on cholesterol in foods (Report No. 5) on effects of lipids in foods of animal origin on the level of serum cholesterol by Yuko Koga, and Yurie Hijioka Original title: Sholuhin-Chu no koresteroru no Kenkyu (Dai 5 I-Io) Dobutsu-Sei Shokuhin Shishitsu no Kessei koresuteroru-ryo ni oyobosu eikyo ni tsuite From: Eiyo to Shokuryo 26 (7) . 441-456, 197.3 Translated by the Translation Bureau(KS /p S) Multilingual Services Division Department of the Secretary of State of Canada Department of the Environment Fisheries and Marine Service Halifax Laboratory Halifax,.N.S. 1974 42 pages typescript
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FISHERIES AND MARINE SERVICE
Translation Series No. 3274
A study on cholesterol in foods (Report No. 5) oneffects of lipids in foods of animal origin on the
level of serum cholesterol
by Yuko Koga, and Yurie Hijioka
Original title: Sholuhin-Chu no koresteroru no Kenkyu (Dai 5 I-Io)Dobutsu-Sei Shokuhin Shishitsu no Kessei koresuteroru-ryoni oyobosu eikyo ni tsuite
From: Eiyo to Shokuryo 26 (7) . 441-456, 197.3
Translated by the Translation Bureau(KS /p S)Multilingual Services Division
Department of the Secretary of State of Canada
Department of the EnvironmentFisheries and Marine Service
Halifax Laboratory
Halifax,.N.S.
1974
42 pages typescript
DEeARTMENT OF THE SECRETARY OF STATE
TRANSLATION BUREAU
MULTILINGUAL SERVICES
eLe
. *ePrt-re CANADA
DIVISION MULTILINGUES
SECRÉTARIAT D'ÉTAT
BUREAU DES TRADUCTIONS
DIVISION DES SERVICES
INTO - EN
Engl i sh
TRANSLATED FROM - TRADUCTION DE
. Japanese AUTHOR - AUTEUR
TRANSLATION BUREAU NO. NOTRE DOSSIER NO
TRANSLATOR (INITIA LS) TRADUCTEUR (INITIALES)
DEC - 5 19 74
d 07 4/
Yuko Koga and Yurie Hijioka TITLE IN ENGLISH - TITRE ANGLAIS
A Study on Cholesterol in Foods (R%xIct N. 5) On Effects of Lipids in Fcods of Animal Origin on the Level of Serum Cholesterol
TITLE IN FOREIGN LANGUAGE (TRANSLITERATE FOREIGN CHARACTERS) TITRE EN LANGUE ÉTRANGÈRE (TRANSCRIRE EN CARACTÉRES ROMAINS)
Shokuhin-Chu no koresteroru no Kenkyu (Dai 5 Ho) Dobutsu-Sei Shokuhin Shishitsu no Kessei koresuteroru-ryo ni oyobosu eikyo ni tsuite
REFERENCE IN FOREIGN LANGUAGE (NAME OF BOOK OR PUBLICATION) IN FULL. TRANSLITERATE FOREIGN CHARACTERS.
RÉFÉRENCE EN LANGUE ÉTRANGÉRE (NOM DU LIVRE OU PUBLICATION), AU COMPLET, TRANSCRIRE EN CARACTÉRES ROMAINS.
Eiyo to Shokuryo Vol.
REFERENCE IN ENGLISH - RÉFÉRENCE EN ANGLAIS
J. Jap. Soc. Food and Nutr., PUBLISHER- ÉDITEUR
DATE OF PUBLICATION DATE DE PUBLICATION
PAGE NUMBERS IN ORIGINAL NUMÉROS DES PAGES DANS
L'ORI GI NAL
441-456 YEAR
ANNÉE PLACE OF PUBLICATION LIEU DE PUBLICATION
ISSUE NO. NUMÉRO
VOLUME NUMBER OF TYPED PAGES
NOMBRE DE PAGES DACTYLOGRAPHIÉES
1973
Environment MI NISTÉRE-C LIENT
BRANCH OR DIVISION DIRECTION OU DIVISION Fisheries Service
PERSON REQUESTING Dr . R.G. Ackma.n DEMANDÉ PAR
YOUR NUMBER VOTRE DOSSIER N 0
DATE OF REQUEST DATE DE LA DEMANDE
3 05.20040.0 (REV. 2/08)
7 030-21.029-5333
26
REQUESTING DEPARTMENT
02-10-74
7
UNEDITED TRANSLATION For inkrination only
TRADUCTION WO N 12EVISEÊ interniâfkin sculernênt
42
676637
KS/PS
I
SECRÉTARIAT D'ÉTAT
BUREAU DES TRADUCTIONS
I ° DEPiIRTMENT OF THE SECRETARY OF STATE
TRANSLATION BUREAU
DIVISION DES SERVICES
MULTILINGUES
MULTILINGUAL SERVICES
DIVISION
CLIENTS NO. DEPARTMENT DIVISION/BRANCH CITY
N° DU CLIENT miNisTiRE misioN/DmEurm VILLE
Environment Fisheries Service Ottawa, Ont BUREAU NO. LANGUAGE TRANSLATOR (INITIALS)
el i) DU BUREAU LANGUE TRADUCTEUR (INITIALES)
676637 Japanese KS/Ps Nov./74
DEC - 5 1974
A STUDY ON CHOLESTEROL IN FOODSIRIEORT NO. 5) L441
ON THE INFLUENCE OF LIPIDS IN FOODS OF ANIMAL ORIGIN ON THE LEVEL
OF SERUM CHOLESTEROL
By Yuko Koga and Yurie Hijioka.
Department of Food and Nutrition, Nakamura Gakuen College.
Previously the authors measured the quantities of such sub- 1)2)
stances as fat and oil, cholesterol , coarse protein, and
essential amino acids, and made a report on the cholesterol-Coarse- 3) 4)
protein cOefficient and cholesterol-protein-value coefficient .
The present report deals with the measurements made on the compo-
sition of fatty acids in fat and oil, and on the increase in the
volute of serum cholesterol when they are consumed in their ori-
tu
ce 0 e z —r 0 0
a .-75« Z
n 32 0 111 C . c
•
E; o e LU Li. :r Z
et' • tfr
ginal states. 5)
Keys et al in 1956 denied the fact that exogenous choleste-
rol, i. e., the cholesterol in foods changed into serum choleste-
rol, and their work laid the basis for popular misunderstanding 6)
for a long time. However, soon after Beveridge et al , Connor 7) 8) 9)
et al , Steiner et al , Erickson et al all denied this 11)
hypothessis by Keys. Further Keys et al in 1962 reported the
SOS-200-10-91
finding that, other conditions being equal, there was a linear re-12)
lationship between the two. In 1965 they assumed that exo-
genous cholesterol was proportional to the square root of the
quantities administered, and as will be explained in a later sec-
tion, inserted a term related to cholesterol in the prediction
regression equation.
It is clear that endogenous cholesterol is formed with
acetyl coenzyme A(CH3-C0-S-CoA) as a source, and consequéntly it has
been widely conjectured that its formation started from the L442
products of intermediate metabolism of fatty acids, glucoside,
and amino acids. Of these by far the greater quantity is pro-
duced from lipids and this has been the min subject Of research.
It has been recognized by many scholars that the saturated
fatty acids in lipids were responsible for the increase in the
level of serum cholesterol. However, the effects due to their13) 14)
C-number are not uniform. Malmoros et al and Hegstedt et al
recognized that C12 and 014 saturated fatty acids raised the level
12)15)of serum cholesterol. Further Keys et al also recognized
that C121 C14 along with C16 fatty acids raised the level of serum
cholesterol. However, there is no such r-eaction associated with
C16 saturated fatty acid. Further it has been established that
saturated fatty acids lower than C10, and under certain condi-
tions those lower than C120 did not increase the level of serum
16)With reference to unsaturated fatty acids, Ahrens et al
maintain that there is a reverse correlation between the iodine
numbers of their fat and oil and their ability to reduce the level17)
of serum cholesterol. Gunning et al report that the square
cholesterol.
3
roots of the iodine numbers are correlated negatively to the
levels of serum cholesterol. Of the unsaturated fatty acids,
which form the sources of the iodine numbers, it is believed that
those with a single value hardly contribute to the reduction in 12)13)
serum cholesterol (Keys et al )., With respect to the poly- 19)
unsaturated fatty acids, Kinsell et al recognized their contri-
bution in reducing the cholesterol level. However, there are 12)20) 16) 21)
opinions by Keys et al y Ahrens et al and Grande et al
with respect to their abilities in reducing the cholesterol level.
In order to study the effects of various fatty acids on serum
cholesterol as described above, and to predict the level of serum
cholesterol on the basis of the composition of fatty acids, Keys 20)
et al formulated the following regression equation (multiple
regression equation) by means of the least squares method:
AChol = 2.76 LIS -0.05 4M -1.35 A -1.63 • (1)
where AChol represents the level of serum cholesterol
mg/d1, and
1/s, 414, and AP represent, respectively, the percentages with respect to the total calories in glycerides of
saturated fatty acids, monounsaturated acids, and poly-
unsaturated acids in fat and oil.
In equation (1), since the numerical values of the coeffici-
ent of AM and of the constant term is small, we may neglect them and obtain the following equation:
AChol = 2.74 As - 1.31 41) (2)
It will be observed that these two equations contain no term 12)
associated with cholesterol. Later, Keys et al conducted
detailed investigations on the effects of cholesterol and made
h.
4
the following modification:
AChol = 2.7 AS - 1.3 AP + 1.5 AC (3),
or JChol = 1.35 (2 AS - AP) + 1.5 AC (31)..
iahere C represents the amount of daily cholesterol in-
take ( dg). 12)
They further modified the equation to read (Keys et al
QChol = 1.2 (2 AS - AP + 1.5 AZ (4)
where Z represents the square root of the amount of cho-
lesterol (mg) in one thousand (1,000) cal. of food.14)
Hegstedt et al obtained the following regression equation
after large scale investigations:
-,dChol = 3.27 S14 + 0.$9 S16 - 0.61 P + 5.$8 C - 6.7 (5)..
where 514, 516, and P represent, respectively,.myzistic
acid, palmitic acid, and polyunsaturated acids (main-
ly linolic acid), and C represents the amount of
cholesterol taken daily through foods (mg).1) .
Meanwhile, as has briefly been reported in a previous report
The U. S. Heart Association organized a Diet Heart Study group dur-
ing 1963-1965, and made concerted large scale investigations of
a type which had never been carried out before. That is to
say, experiments were carried out in six metropolitan areas such
as Baltimore: in the first experiment, which was carried out
over a period extending fifty-two (52) weeks, observations
were made on two thousand one hundred and seven (2,107) subjects,
and in the second experiment one thousand five hundred and ninety
(1,590) _subjects were examined over a period covering twenty-eight
(2$) to thirty-six (36) weeks, with a total of one hundred and
forty (140) odd medical men and biochemists taking part. These
5
investigations resulted in two sets of regression equations (pre- 22)
diction type) concerning the level of serum cholesterol
(mg/di):
Chol = 2.16 AS - 1.65 41) - 6.77 AC - 0.5 (6)
Ahol = 1.35 (2 LIS - LIP) ± 1.5A (7)
where the abbreviations have the identical meanings
as in equations (1) and (5), and
del% represents the square root of the amount of cho-
lesterol in foods mg/1,000 cal. (It should be noted
that equation (7) is similar to the aforementioned
equation (3) of Keys et al except for modifications
made on the basis of equation (4).)
It is the opinion of the authors , that equations (6) and (7)
are the most reliable as they are the results of efforts of a
large number of researchers , working with a large number of peo-
ple. Thus in the present report the authors calculate the ef-
fects of the composition of lipids on the level of serum choles-
terol by means of equation (7) with those materials, which they
had used in the previous reports, augmented by fresh data. As
was mentioned Dreviously, it has been reported that in addition 23) 24)
to glycocide and protein other substances such as phytoste-25)27) 26)
rol and pectin also affect the cholesterol level. How-
ever, the subject will be dealt with in later papers.
With the exception of hens , eggs, foods of animal origin
6
which were available commercially in the city of Fukuoka, were
used as source materials. They consisted of seventy nine (79)1)2)
varieties, reported in two previous reports , augmented by
fresh materials (bought since Sept., 1969). The detailed break-
down of these materials-with respect to the time of their pur-
chase or of collection is shown in Tables 1-4. These materials
were cut-into various parts and served as foods in the form and1)2)
in the way identical to those of the previous reports
2. Experimental Method
1) Pfethod of General Analasis: The determinations of moisture,
coarse fatty acids, and cholesterol were made follow)the me-1)2
identical to those of the two previous reports . The
determination of coarse protein was made in a manner identical3)
to that of the third report
2 Measurements of the Composition of Fatty Acids:,. The methods
used for the analyses-of flesh of animals, birds, and whales,
hensT eggs, processed foods, and foods of marine origin were given1)
in an outline in a previous report . They were analyzed by L4432$)
the BF3 methanol method ; and most of marine foods were methy-
29)lated following the method employed by Shinma et al who
used a mixture of inethanol, benzol, and concentrated sulphuric
acid (123: 61•5: 1 v/v), and the resulting fatty acid methyl
ester was analyzed by means of gas chromatography. The equipment
used was a Hitachi gas chromatograph ( K-53 type; hydrogen flame
ionization detector). The conditions for analyses were identical30)
to those of a previous report . 0.2-0.5 /uL of the test
materials were used for analysis.
For the identification of the chromatograms, K-106, K-107,
and K-l08 of Applied Science Co. were used as the standards: 31
for other peaks the ECL values due to Hofstetter et al were
used.
3) Calculation of Abhol: As was explained in a previous sec-
tion, equation (7) was used to ascertain the relationship between
the composition of the fatty acids and the level of serum cho-
lesterol. For the purposes of this report the ratios with re-
spect to the total calories of the quantities of saturated fatty
acids (S) and of polyunsaturated fatty acids (P) coexisting•
in 100 g of coarse protein were calculated and used as As and AP, respectively. As the total caloric intake, two thousand
one hundred fifty (2,150) cal., a nutritional standard for Japa-
nese (Nutritional Enquiries Commission (Eiyo-Shingi-Kai)), was
used.
Experimental Results and Discussion
The results are shown in Tables 1-4.
The tables show, in addition to the . results of general ana-
lyses, quantities of fat, which coexist with one hundred (100) g
of coarse protein, the amounts of cholesterol in fat, and the
composition of fatty acids. Further, they show the amounts of
saturated fatty acids (S), polyunsaturated acids (P), and mono-
unsaturated acids (M), which have been calculated on the bases
of the composition of fatty acids (M). They also show the
amounts of serum cholesterol ( AChol) which have been calculated
on the bases of the total amounts of cholesterol, S, and P.
8
A great deal of the findings were reported in the previous 1)-3)
three (3) reports with reference to moisture content, coarse
fat, coarse protein, and cholesterol level in fat. Thus the pre-
sent report will deal mainly with fatty acids and2Chol. For
this purpose Fig. 1 has been added.
1. Flesh of Animals, Birds and Whales
With respect to the composition of fatty acids in the flesh 32)
of animals and birds, Hornstein et al reported that in the
case of beef the influence of age was recognizable; and Machlin et 33)
al reported that in the flesh of fowl the influence of feeds was
recognized. As the samples were obtained through commercial out-
lets, nothing is known of the effects of these factors in the
present report.
1) Beef: With respect to the composition of fatty acids 35)
in beef produced in Japan, there are reports by Otake and 36)
Morohoshi et al in addition to detailed investigations by Naka- 34)
nishi et al . It should be noted that, for the same parts, the
analytical values of C16:0 in"sirloinI ! and - fillet, and of C18:1
in nrunip" . which have been observed in the present experiments, 36)
are somewhat higher than those observed by Morohoshi et al .
The differences in the cholesterol levels in fat (hereinafter
referred to as the cholesterol levels) in the four test materials,
except for those of the liver, were not significant. The level
of S was the lowest in the outer shanks, and high in "runp"
Consequently the cholesterol level was high in the latter -- 2.6
times that of the outer shanks.
8a
Beef
l Shoulder roast
2 Chuck
3 Shank
4 Rib
5 Plate (Bara)
6 Brisket
7 Loin
$ Fillet
9 Flank
10 Sirloin
11 Round
12 Runnp
9^•
2) Pork: Of the three ( 3) samples, consisting of L446
fillet, shoulder roast, and "ribs", thé cholesterol level in
fillet was by no means low; its level of P was the highest and
,QChol was the lowest. ^/Chol in the shoulder roast was 2.7
times that of fillet. Thus, as in the case of beef, there was a
considerable difference in 2Chol due to the position of the meat.
3) Mutton: Among the flesh of animals and birds, C13:0 in
the thigh and fillet showed extremely high values. As in the37)
case of work by Otake C20:4 and a minute quantity of C15:0
were found.
The level of S was tohe second highest next to that of the
skin, which will be dealt with in a later section. However, the
cholesterol level was low and consequently _ZChol was low. This
observation also leads us to the belief that the cholesterol
level is a factor which greatly influences Chol.
4) Chicken: The levels of C13:2 in the three samples of
flesh of various types.were high and somewhat similar to those
of pork. As shown in Fig. 1, the level of _ZChol was the low-
est among the flesh of animals and birds. Of the three samples,
the one, which had shown the lowest value, was the breast.
The level of C16:0 in the thigh was higher by 44.3% in
comparison to those of the flesh; the level of S was also the
highest among all the samples. Howevér, the level of cholesterol
was low and the level of f^Chol was not too high.
The liver (beef, pork, and fowl): In all the three (3)
samples, the levels of C16:0 were lower by 35-4$% than those of the
corresponding flesh, C1$:0 were twice as high as those in the flesh,
and C20:3and C20:4 were detected in all the three samples. In
10
comparison to those of the corresponding flesh, //Choi's were
not low.
6) Whale Meat: During the course of the analyses of fat-
ty acids, two types of unidentifiable substances (x1 and x2 )
were found in two positions. These were not included in the cal-
culations ofAillChol. Since the quantities involved were extremely
small, the authors felt that the exclusion of these substances
should not influence in any way the values of AChol.
On the other hand, the levels of M, being 72.9 and 79.4%,
were extremely high in all the samples; the levels of S were low.
However, as a result of the influence of the level of cholesterol
in the red meat, AChol was the second highest following that
of pork.
2. Henst Eggs
There was hardly any difference in the levels of S and P
in yolks A and B. However 2/Chol was somewhat higher in A in
comparison to that of B. As is evident in Fig. 1, these values
were the highest among all the samples.
In comparison to those of the yolk, whites A and B contained
56.9 and 60.6% of C13:2 , respectively. C14:0 was higher by --
a factor of 4.7 and 2.3, 013:0 by 2.7 and 3.0 and C204 by
6.1 and 7 -*.' 020:3 was also detected. The level of /1Uol was
low and followed those of the ordinary meat of omaesou Saurids
undosquelis(RICHARDSON), and skipjack MIsmonmsmlamis (LINNE).
The findings concerning the latter will be dealt with in later
sections. 38)
At the present time Japan is reported to be the second
10
comparison to those of the corresponding flesh, _ACholts were
not low.
6) Whale Meat: During the course of the analyses of fat-
ty acids, two types of unidentifiable substances (x1 and x2 )
were found in two positions. These were not included in the cal-
culations of2Chol. Since the quantities involved were extremely
small, the authors felt that the exclusion of these substances
should not influence in any way the values of 4Chol.
On the other hand, the levels of MI, being 72.9 and 79.4%,
were extremely high in all the samples; the levels of S were low.
However, as a result of the influence of the level of cholesterol
in the red meat, AChol was the second highest following that
of pork.
2. Hens' Eggs
There was hardly any difference in the levels of S and P
in yolks A and B. However 4Chol was somewhat higher in A in
comparison to that of B. As is evident in Fig. 1, these values
were the highest among all the samples.
In comparison to those of the yolk, whites A and B contained
56.9 and 60.6% of 013:2 , respectively. 014 . 0 was higher by
a factor of 4.7 and 2.3, 013:0 by 2.7 and 3.0 and C204 by
6.1 and 5 . 7 . 020:3 was also detected. The level of AChol was
low and followed those of the ordinary meat of umaeso" Saurids
undosquamisiqICHARDSON), and skipjack Katsuwonus pelamis (LINNE).
The findings concerning the latter will be dealt with in later
sections. 38)
At the present time Japan is reported to be the second
11
largest consumer of eggs following the U. S. Eggs are used not
only in side-dishes but also in desserts. As their use is ex-
tremely varied, we must be conscious of the amount of their in-
take.
Further, eggs are processed in many ways -- for instanCe, they
can be frozen, powdered, condensed, or treated with enzymes. It
appears as though analyses on the processed materials will be
necessary in the future.
3. Aquatic Food (excludes whale meat)
39)40) There have been a number of reports on fatty acids in
fish in this country. Researches are being conducted over a con-
siderably wide range of topics from the establishment of the func-41)
tion of fatty acid amido to the study of properties of lipids 42) 43)
in fish meal . Further, there is a research by Shinma et al
on shellfish.
The materials, which were used in the present report, were
purchased through retail outlets. Thus the authors feel that they
bore factors, which were likely to contribute to the change in
the nature of constituent polyunsatil;ated fatty acids such as
the conditions under which the commodities had been frozen or
thawed out, or the storage conditions in the shops. Further, in
the case of fresh water fish, it is known that among the species
which change their habitats, the body fat changes from the fresh
water type to sea-water type at a certain stage of their develop-
ment. At the present time fish is being reared intensively.
Under these varying circumstances we can perceive the possibi-
lity that the analysis of fatty acids in fish may involve factors
p447
12
complicated above those of flesh of animals and birds. Whatever
the case may be, the results of the present investigations are
shown in Tables 3-1 to 3-5-
1) Fish Nleat: The fact that the level of P was high has been
well known. This is also noted in Tables 3-1 to 3-2, in which
the values for fish are higher by a factor of 2.0 to 5.5 in com-
parison to those in the flesh of animals and birds. A comparison
between fourteen (14) samples of ordinary meat of white, blue, and
fresh water fish and five samples ôf dark meat showed that the levels
of P in the latter, being 43.6-47.9%. were very much higher.
On the other hand, as shown in Table 3-2, comparisons between
the ordinary meat and dark meat from the same so-called blue fish
showed that C22:5 was found in very minute quantities in ordinary
meat, and that C22:6 was commonly found in quantities larger by
a factor of 2.0-10.$ in dark meat in comparison to those in ordi-
nary meat. Further in fresh water species ( ordinary meat) C1$
acid-contents were found to be as high as approximately 44-5$%,
of which Cl$:3 represented as much as 12.9-17.5%, C20 and C22
acid contents were low. These findings were similar to those of40)
Shinma et al .
With respect to AChol, it was found that although the le-
vel of P was considerably higher than those in the flesh of ani-
mals, birds, and whales, since the levels of S and cholesterol in
many samples were somewhat high, its levels were not always low.
Of the twenty-one ( 21) samples excluding t'maunagi" Anuuilla...
japonica TEMMTNCK et SCHLEGEL, and t 'maanago" Conger myriaster
( BREVOORT ), ZChol, being 4.1$, was the lowest,in «maeso" Saurids
13
undosguamis (RICHARDSON) (ordinary, meat) . This was followed,: :-
in increasing order, by skipjack Katsuwonus pelamis (LINNE)
(ordinary meat), flat fish (ordinary meat), albacore Thunnus