TitleANALYTIC CHEMICAL STUDIES ON FATMETABOLISM BY APPLICATION OF PAPERCHROMATOGRAPHY OF FATTY ACIDS

Author(s) TAN, NOBUTOSHI

Citation 日本外科宝函 (1959), 28(4): 1178-1196

Issue Date 1959-05-01

URL http://hdl.handle.net/2433/206851

Right

Type Departmental Bulletin Paper

Textversion publisher

Kyoto University

1178

ANALYTIC CHEMICAL STUDIES ON FAT METABOLISM BY APPLICATION OF PAPER

CHROMATOGRAPHY OF FATTY ACIDS

b~＇

N OBUTOSHI TAN

From the 2nd Surgical Didsion, Kyoto Uni,-ersity l¥Iedkal School (Director: Prof. Dr. YAsUMASA AoYAGIJ

<Received for publication Feb. 27, 1959)

I. INTRODUCTIO:¥

Recently there has been remarkable progress in the entire field of clinical

medicine. One of the contributing factors has been the improvement in nutritional

methods. Especially in surgery, consideration should be given to positive nutritional

supplements, because in most cases, patients cannot take oral！~－ enough nutriments

prior to and after surgical operations; beside日 surgicaltreatments inevitably accele-

rate the catabolic p1川 C日 ofmetabolism. In this case, most convenient is an intra-

venous supplement of nutriments such as glucose, amino acids, salts, vitamins, etc.

This method is currently in a practicable stage. However, intravenous administration

of fats still remains a very di出cultproblem. At our laboratory, the necessity for

parenteral administration of fats for supplementai‘v nutrition was earl）’ recognized,

and e汀ortshave been made to solve this problem. If fat勺 substancesare to be

administered intravenousb' at all, it is necessary that ample thought should be ac-

corded to the qualitative composition of fats --above all their acid components-

which a1℃ infused intravenous!¥・. The reason is this: in the digestive tract, some of

the fats which are orall>' taken, are in the non-h~·drob·zed form (triglycerides), some

in the partial!>・ hydrol>・zecl form (diglycerides & monoglycerides), and some in the

form of isolated free fatty acids, ancl th町 arereduced to corpuscles less than 0.5 11

in diameter lηbile salts and can be ail州 rbcdthrough the intestinal mucous mem-

brane. This absorption process is regulated lJ:, the species of fatt＞’ acids as to whether

they are absorbed direct!>・ into the portal blood or into the lymph of the thoracic

duct. The results of indirect examinations lηv isotopes IJy BwoM et al. indicate

that higher fatty acids can be absorbed mostlJア intothe b・mph of the thoracic duct,

while lower ones are absorbed into the portal blood. 人ccordingly, if we are to

infuse fattJア substancesdirectly into cutaneous veins, glyceride emulsions consisting

of higher fatty acids which are apt to be absorbed into thoracic duct lymph should

be inf・LI日cdinto cutaneous veins. This is because direct infusion of fat emulsion into

the portal ¥'ein cannot be considered.

In the abo,・e-mcntioncd experiments Iηv BwoM ct al. ,・ariou日 C''-labeled fatt）’

acids were u同 d,and the use of their radioactivit>・ enabled them to observe the

tran日portpathway of aiJ河川町lfatty acids. This is, so to speak, an indirect obser-

vation method. It is not l>y direct expじrimcntalvcri白cationthat lower fatty acids

ANAL YTff' CHEl¥TWAL只Tl'DIES 0 N F !¥ T 1¥TET A BO LIS'.¥T 1179

could actually he absorbed into the portal blood. i:yc11 if it is an admitted fact that

lower fatty acids arc absorbed through the intestinal mucり~！s membrane, it is IJ~・ no

means incor.ccivablc that th町’ mayundergo direct absorption into the portal vein in

yiew of the fact that almost all of them will not k、absot・bed into thoracic duct

］ ~·mph. The author intended to Yeri1下experimentallythe allegations by BwoM et

al. just by applying paper chromatograph~・ of fatt~’ acids, especially to determine

whether or not influx into the portal vein of lower fatty acids ever takes place.

It is theoretically the best method of supplementing fatt）ナ substancesto selectively

administer a triglyceride emulsion consisting of the commonest fatty acids which are

always found in animal organs or d句otfats. In this sense, the author resorted to

paper chromatographγof fatty acids with regard to the stud~’ of component fatty

acids in each organ or depot fats.

II. M人TERL¥LSAND METHODS

人. Experimental Materials

1)¥ Fat emulsions: 20 % sesame oil emulsion and 20 /o cod liver oil emulsion

were used. These contain 7 % glucose each. 2) Natural fats: Natural butter fat, cod livこroil, sesame oil, etc. ＇γere used.

3) S~＇nthesized simple glycerides: Tricaproin, tricaprylin, tricaprin, trilaurin, triolein, etc. were used.

4) Experimental animals : Healthy adult dogs weighing about lOkg were used.

In order to maintain as similar conditions as possible, each of them was fed a meas-

ured diet for more than日evendays, and then fasted for 24 hours so that experi-ments could be made in an immediate postabsorptive state.

B. Ex問rimentalMethods

1) Method of collecting chyle As in SttIROTANI’s method with cats, an incision approximately 6cm long was

made in the left supraclavicular fossa of an anesthetized clog. Then, ample exposure

of the ascending thoracic duct along the inside of the left jugular vein was made

and complete prevention of thoracic duct l>・mph influx into the blood was achieved

(Fig. 1). Next, a polycthγ！enc tube was inserted into the thoracic duct, and dripping

lymph was collected in a pre-arranged collecting flask. Subsequent to this, the

above-mentioned test fats ¥Vere administered into the stomach by stomach tube and

chyle, now cloud：＞ア asa result of fat absorption, was collected in a collecting司ask.

2) Method of collecting chyle ancl rortal blood simultaneously

λ日 shownin Fig. 1, collection of chyle was【loneusing a pol>'ethylene tube

after oral administration of test fats. B>・ this procedure, complete interruption of

chyle flowing into the blood stream could be achieved.λt the same time, collection of

portal blood as it was after two hourぷ lap日 ofintragastric administration of test

fats was done. Chyle and portal blood were collected for fatty acid analysis.

3) Organs

The experimental animals were sacrificed by bleeding and the organs ＂℃ re

immediately removed, and pulverized finely by marine sand for analysing their

1180 日本外科宝酉第28巻第 i号

, V. juguhrl・Int orno slnlP •.rs

Due もu•

V, 1subclavla •1nl9tre

v, l;>racr.l♂rτcMl¥ce e!nlstra

Fig". 1 Method of Collecting Chyle

component fatt~， acids.

4) Paper chromatography of fatty acids

Paper chromatography was done by the NonA-HIRAYAMA’method, i.巴.the

development wa日 madeas the p-bromoph巴nacylester 2, 4-dinib可ophen~·lhycfatt~· acids.

(i) Filter paper：“T町ザ’ filterpaper No. 2 was used. A starting line was

penciled 3 cm above the upper border of the surface of the solvent.

(ii) Solvent s；，百tern日： λぉ themoving solvent, methanol『 glacialacetic acid-

petroleum h~·drocarbon (10: 2: 1. 2 by volume) was used and petroleum hydrocarbon

was L!Sed as the stationary solvent.

(iii) Procedure : The paper was spotted with samples an【l¥¥'as uniformly sp-

ra；，アeelwith the stationa1下 solvent,petroleu:m hJ・drocarbon. Then, the chromatogram was developed with the moving solvent by the ascending technique at 30°（司 The

development time required for a satisfactor；，’ separation was 5～6 hours b：－’ this method.

III. RESULTS

I) On the Tram;port Pathway of Absorbed Fats

In order to iclcntil¥・ the transport pathwaγof absorbed fats through the

intestinal mucous membrane, comparison was made between component fatty acids

contained in the test fats which were oralh・ administered to experimental animals and

component fatty acids contained in chJ・le (or portal serum) which ＂℃ I℃ collected

when the test fats were orally administered.

1) Absorption into thoracic duct

A.¥lALYTIC CHEMIC‘AL STUDIES ON FAT METABOLIS'.¥I 1181

In this experiment, the following materials ¥¥'ere used : cod liver oil emulsion containing a comparativel｝’larg℃ amount Gf high ！~， unsaturated fatty acids, eicosenoic acid, docosenoic acid, etc., butter fat containing lower fatt：－’ acids, and se悶 meoil .emulsion composed of higher saturated fatty acids, olcic acid, essential fatty acids and containing no lower fattγacids, eicoscnoic acid, docosenoic acid, highly unsaturated fattγacids.

The fatt~’ acids which are contained in the thoracic duct lymph of a dog in a postabsorptive state after 24 hours’fasting, as shown in Fig. 2, were m:-Tistic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. But no lower fatt；.’ acids than lauric acid and highly unsaturated fatty acid日 werefound.

A) Oral administration of cod liver oil emulsion

Ffg. 3 shows the analytic.: results of fatt；.’ acids contained in cod liver oil emulsion (F. I.) and fatt;.・ acids extracted from ch>・lc (L. F. I.) collected when 40g of cod liver oil emulsion was administered by stomach tube.

In cod liver oil emulsion, there are found myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, cicosenoic acid, clocosenoic acid, and high！＞’ unsaturated fatt；.’acids. Also in chyle collected when cod liver oil emulsion was orally administered, there were detected various kinds of fatt；.’ acids in the same p~rcentage as the above. It is apparent from this fact that all fatt;.・ acids contained in cod liver oil emulsion could well be absorbed into the thoracic duct.

Especially, it can be clearly seen that docosenoic acid, highly unsaturated fatty acids, eicoscnoic acid, etc. which were not detected in thoracic duct lymph in a postabsorptive state, have been well absorbed into the thoracic duct.

B) Oral administration of butter fat Fig. 4 shows the analytic results of fatt;.・ acids contained in natural butter

fat (B.) and fatty acids extracted from chyle (L. B.) collected after 40g of natural butter fat had been administered orally.

In natural butter fat are found butyric acid, caproic acid, capr~・lic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, and oleic acid. Especial!;.・ notcworth；.・ isthe fact that the percentage of both oleic and palmitic acids is high.

When the fatty acids extracted from chyle collected after natural butter fat had been orall;.• administered were examined by paper chromatography, it was found that fatty acids higher than myristic acid were alw匂’sin almost the same condition as the former, while fatty acids lower than !auric acid gradually showed spots which became thinner and fewer. And spots of fattj’acids lmYer than caproic acid were never seen.

In other words, it is considered that fatty’ acids higher than myristic acid could b巴absorbedinto thoracic duct lymph, but that fatt；ィ acidslower than lauric acid gradual!J’declined in the percentage of absorption into the thoracic duct in proportic川

to the decrease in number of carbon atoms.

C) Oral adminisJ.;ration of se泊 meoil emulsion Fig. 5 shows the analytic results of fatty acids contained in sesame oil

emulsion (F. I.) and fatty acids extracted from chyle (L. F. I.) when 40g of

1182 日本外科宝函第28巻 第4号

Table 1. :-Jculralisation and Iodine ¥'alues of Component Fatty Acids of Test Fats and Lipids （可。llectcdfrom Ch~ョ le

1 Cod Li、erOil Emulsion Sesame Oil Emulsion Butter Fat Acid

1 加fedT－孟yle瓦 r~－~ーヲ一両~··！加 fed I Chy le fat

::¥leutralisation Value Iodine Value

188.2

135.4

176.9 196.7 191.9

106.4 109.8 104.6

sesame oil emulsion was ~dministered IJ~’ stomach tube.

217.0 192.2

34目。 66.I

Sesame oil emulsion contains myristic acid, palmitic acid, stearic acid, oleic acid,

linoleic acid and linolenic acid, especial!>' linoleic acid and oleic acid in large

quantities.

In contrast, various fatt? acid日 extract吋 fromch>'le after oral administration

of se泊 meoil were found in the same percentage as the former.

This indicates clearly that each of these fatty acids in se回 meoil emulsion

could九Nellbe absorbed into the thoracic duct.

Neutralization and iodine value of the component fatty acids contained in the

afore-mentioned three test fats and the component fatt~＇ acids in chyle collected

during oral administration of these test fats, are shown in Table 1.

According to the results mentioned above, it is roughly presumed that lower

fatty acids graduall~’ decline in percentage of absorption into thoracic duct lymph in proportion to the decrease in number of carbon atoms. To further clarify this

problem, simple gl~·ccrides such a日 tricapr~·lin, tricaprin, and trilaurin were synthe-

sized, and then a simple glyceride mixture containing equal amounts of them were

administered orallγto experimental animals in ordむ1・ to日tudYagain the absorption

of lower fatty acids into thoracic duct lγmph.

D) Oral administration of tricaprylin, tricapr加， andtrilaurin lOg of each of the above simple glycerides was mixed, and 30g of this

mixture was administered by tube into the stomachs of experimental animals.

Fig. 6 shows the anal~·tic results of fatty acids contained in the synthesized

simple g］ ~·ceri 〔le mixture (D) and fatt>・ acids extracted from chyle (L.) collected when the above mentioned可 nthesizedsimple glyceride mixture was administered

orally. It is apparent from these results that lower fatty acids such as !auric acid,

capric acid, and caprylic acid gradually decline in their percentage of absorption into

the thoracic duct in proportion to the decrease in number of carbon atoms.

In regard to the absorption of lower fatty acids into the thoracic duct, the

conclusion is that the fewer the carbon atoms, the lower is the absorption ratio.

In short, our direct examinations in which paper chromatography of fatt＞’ acids

was used to observe the absorption of fatty acids through the intestinal mucous

membrane when fats were oral ！~· administered, ha＼℃ gained the same I℃suits as the indirect verification method h>・ CHAIKOFF, REISER, BoLLMAN et al. using isotopes, and

the experiment H’ FERNANDES on a child with ch>・lothorax.

ANALYTIC CHEMICAL STUDIES ON FAT METABOLISM 1183

2) Absorption into the portal vein

From the results mentioned above, the percentage of absorption of lower

fatty acids into thoracic duct lymph is exceedingljァlowas compared with that of higher

fatty acids. However, no reports have yet appeared confirming the direct observation

of the absorption of lower fatty acids into the portal blood during the oral admini-

stration of fats.

The author, therefore, studied the absorption of caprylic acid and caproic acid

into thoracic duct lymph when large amounts of synthesized tricaprylin and tricap-roin were orally administered. In parallel, the author studied also the absorption of such fatty acids into the portal blood.

Fig. 7 shows the analytic results of fatty acids which are alwaγs found in the

portal serum of dogs in the postabsorptive state. The component fatty acids in

portal serum of a dog in postabsorptive state are myristic acid, palmitic acid, stearic

acid, oleic acid, eicosenoic acid, linoleic acid, linolenic acid, and highl~’ unsaturated

fattv acids. However, the author could detect no fattγacids lower than lauric acid.

A) Oral administration of tricap1下lin

Fig. 8 shows the anal~＇tic results of fatty acids extracted from chyle (L.) and

portal serum (B.) collected after oral administration of tricapQァlin. As shown in

Fig. 8, caprylic acid is generally absorbed into the portal blood through the intestinal

mucous membrane, but still some portion of it gets absorbed into thoracic duct lγmph.

B) Oral administration of tricaproin

Fig. 9 shows the analytic results of fatt>・ acids extracted from ch:yァleand

portal serum collected after oral administration of tricaproin. The comparison between

fatty acids in chyle (L.) collected after oral administration of 40g of tricaproin to experimental animals, and fatty acids extracted from portal serum (B.) suggests that just as in the experiment on administering tricaprylin, the portal serum con-

tains more caproic acid than chyle. That is, as caproic acid generally tends to be

absorbed into the portal blood, it is possible that certain portions of it get absorbed into thoracic duct lymph.

Thus, the absorption of orally administered fats through intestinal mucous mem-brane varies according to the type of fattγacid. Higher fatty acids than myristic

acid are generally transported into the blood stream by way of the thoracic duct,

while lower ones are absorbed directly into the portal blood mostly through the

intestinal mucous membrane. However, it is not necessarily proper to think that

absolutely none of the fatty acids lower than m円isticacid are absorbed into thoracic

duct lymph, since it was discovered that certain portions of them are absorbed into

thoracic duct lymph. It was also learned that lower fatty acids showed a gradual decline in the p2rcentage of absorption into thoracic duct h・mph,as the number of

their carbon atoms decreased. It is considered that the di百erencein the transport pathway of absorbed fatty acids based upon the number of carbon atoms constituting

such fattyァ acids,is closely associated with the degree of water-solubility and hydro-

lysis with lipase.

From these facts, the conclusion has been reached that for intravenous adminis-

第 4号

tration of fats, ti ・ig-J~·c~ridcs composed exclusively of as high fatty acids as possible

should be emulsified into globules less than 0.5;.L in diameter and be infu日eelas an

emulsion.

II) Component Fatt：γAcids in Various Organs As mentioned earlier in this report, it is a matter of course that the most

appropriate method for parenteral administration of fats is a glyceride emulsion

consisting of the commonest fatty acids which are alw何日 foundin the body.

1) Component fatty acids in dog’s organs

The component fatt:vア acids in the cerebrum, lh℃r, kidney, spleen, muscle,

large omentum, subcutaneous adipose tissue, bile, serum, l;vアmph, etc. of a healthy

adult dog in the postabsorptive state, were analysed b~· paper chromatography.

The arnilytic results are shown in Figs. 10 to 17. The commonest fatt>・ acids

in all organs were m≫r i円ticacid, palmitic acid, stearic acid, oleic acid, linoleic acid,

and linolenic acid.

In regard to the neuturalization values subcutaneous adipose tissue and the large

omentum were highest with values of 202 to 2D4, containing palmitic and oleic acid

in great quantities. Characteristic, however, is the fact that the＞’ contain lauric

acid, but almost no highly unsaturated fatty acids.

Gen巴rall：－ァ speaking,large amounts of highly unsaturated fatty acids, docosenoic

acid, cicosenoic acid, etc. are found in liver and bile which give higher iodine values.

Although a certain amount of highly unsaturated fatty acids can alwa>・s be detected

in s巴rumtoo, their pres巴ncecan hardl＞ア becertified in thoracic duct lymph, when in

the postabsorptive state.

第28巻日本外科宝函1184

＋

＋

側

＋

土 l ー

＋

剛

士

tit

＋

Table 2. Fatty Acid Composition of Lipids in Dog’s Organs

I Liver i Kidr

一ト；！ = I = I = I = i -I_ I -

一

＋

州

＋

＋川市川廿

＋川市件

一

＋

十

州

＋

ぃ

士

一

＋

附

＋

＋川市川廿士

一＋川市十士

僻

土

＋仲

＋

掛

＋

掛

＋

tit

＋

掛

＋

＋

側

士

川

甘

＋

州＋川市＋

十？？十

＋

ti十

＋

一

制

＋

怖

＋

＋川相H

廿土

－－－~！ Brain Butyric -Caproic Caprylic Capric Laurie

》alrnitic

Stearic Arachidic

l:;・ I Hexa巾ccnoicl

rZ I Oleic !

τ I Eicosrnoic ! v I I

~ I Linoleic I

さ丘ILinolenic I g;·~ Highly I Lイ Unsaturated1

Iodine Value I 121.0 j

＋山mH廿

回一＝

υ44ン】パコ山F

何回一〕’ぷ吋H

目玉同庁山

川附＋品川十

川市＋川計十

土

122.3 J 111.2 : 73.9

＋ 川廿土土

l

uu 氏U

＋

90.7

＋

ワt

F同

ul

l

＋

s6.2 I

＋＋＋

141.0

＋

ANALYTIC CHEMICAL STUDIES ON FAT METABOLISM

Table 3. Neutralisation and Iodine Values of Component Fatty Acids in Human Organs

1185

恥 id I Serum I Omentum j Subcutaneous '

Neutralisation Value I 193.6 I 204目4 I 208.9

Iodine Value I 111.3 I 68.1 i 71.5

Table 2 tabulates the species and iodine value of the component fatty acids of

various organs. 2) Component fatt~ァ acids in human subcutaneous adipose tissue, large omentum,

and serum

The analytic results on .component fatty acids in subcutaneous adipose tissue,

large omentum, and serum of an adult, are shown in Table 3 and Fig. 18. The

experimental results are al.most the回 meas in the dog.

The analytic results on fatty acids in each organ, d句otfat, serum, and thoracic

duct lymph of dogs and humans indicate that the泊 festparenteral method of supp・lementing fats is to administer a gl:yァcerideemulsion consisting of various fatt:yァacids

such as myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic

acid which are invariably present in all organs almost in the same ratio. Therefore,

as shown in Fig. 19, sesame oil is considered very appropriate.

IV. DISCUSSION

In our laboratory, the necessit:yァ forparenteral administration of fats has been well recognized, and e町ortshave been made during the past several years to prepare

intravenously infusible fat emulsion. However, the problem still remains as to what

fats should be chosen for such an emulsion.

In order to solve this problem, the analysis of component fatty acids in chyle

and portal blood collected after various test fats had been orally administered to

experimental animals, was carried out by paper chromatography. The results were

in agreement with those of BwoM et al. in which isotopes were used ; higher fatty

acids than myristic acid are generally absorbed into the thoracic duct through the

intestinal mucous membrane, while lower ones gradually decline in the percentage

of absorption into the thoracic duct in proportion to the decrease in number of

carbon atoms, and are absorトedmain！~’ into the portal blood. Thus, it follows that

in infusing fats directly into the cut!lneous vein, triglycerides composed substantiall:yγ

of higher fatty acids than myristic acid shoulcl be emulsified into globules less than

0.5μ in diameter and be infused as an emulsion.

In this case, attention is called to the following fact. Even low℃r fatty acids do not necessarily resist absorption into the thoracic duct, but rather some portions

may be absorbed into the thoracic duct.

AsADA and IzuKURA in our laboratory have used sesame oil emulsion, cod liver

oil emulsion, and synthetic ti句leinemulsion for oral or intravenous羽ministration

in experimental animals and have carried out histochemical studies of various organs.

1186 日本外科宝函第28巻第4号

When corl li¥'cr oil c011bi11i11広 alarg2 amount of high!;,・ uns:ituratcd fatt>・ acids,

clDco~cnoic acid, and cicoscnllic acid, or butter，「ontaini11glower fatty acids, were

given oral!>・ or intravenous!>・ as an emulsion, much larger amounts of phospholipids

were demon .;trated in the pg,renc~ァmatou日 cells of the liver, in comparison with cases

in which r泊 meoil emulsion was used, containing nothing but higher saturated

fatty acids, oleic acid and the essential fatt）γacids. And Kuy AMA found the same

results as when he studied the phosrholipid content of various organs biochemically.

According to the results mentioned above, high!>・ unsaturated fatty acids, lower fatty

acids, eicosenoic acid, dorosenoic acitl etc. are shifted onlJマ to the parenchymatous

cells of the liver ; and higher saturated fatt>・ acid日， oleicaεid, and essential fatty

acids arc shifted not onlγto the parench>・matous cells of the liver but also to extra-

hepatic tissues where they will b2 disposed of further.

It is natural that in my anal>・sis of component fatty acids of various organs,

much larger amounts of high!)・ unsaturated fatty acid日， docosenoicacid, eicosenoic

acid etc. were nntained in liver than in extrahepatic tissues. Accordingly, the

nutritional e百ectof fats should b2 cl巴cidedin consideration of these two metabolic

processcι （Fig. 20). That Iヘfatt)・ acids of the former group are a burden to the

liver, markedlyァ increaseketone bod;,・ production and maγcause ketosis, while fatt>・

acids of the latter group lighten the burden of the liver and produce ketone bodies

in smaller quantities than those of the former group. The author decided that in

view of the transport pathw町 ofabsorb2cl fats, triglycerides consisting mereb・ of

higher fatty acids have only to b3 infused intravenously in the form of emulsified

fat. Yet such a definition is still insufficient, so the following addition should be

made: Trigly・cerid2s which contain none of the highly unsaturated fatt;; acids, cico・

senoic acid, clocosenoic acid, but merely higher fatt)・ acids such as myristic acid,

palmitic acid, stearic acid, oleic acid, linol::ic acid, and linolcnic acid, should be used

as materials for a fat emulsion.

Lower Fatty Acids

Highly Unsaturated Fatty Acids, Eicosenoic Acid, Docosenoic Acid

Higher Saturated Fatty Acids, Oleic Acid, Essential Fatty Acid~

Fig. 20. 』 HepaticParenchymatous C0lls

／ク、！＼

＼

＼ 、、＼ 、、 Exlrahcpatic Tissues

V. SUMMARY

Using paper chromatography of fatty acid日， anattempt was made to pursue the

transport path way of absorbed fats through the intestinal mucous membrane when

fats are orally administerccl, and simultancou-;I~， to anal:v；；：、 componentfatt~’ acids in various organs, serum, bile, l~·mph, and depot fat etc.

ANALYTIC CHEMICAL STUDIES ON FAT METABOLISM 1187

The following conclusions were reached :

1) There are two trans1〕::lrtpathways of absorbed fats through the intestinal

mucous membrane : absorption into the thoracic duct and absorption into the portal

vcm.

2) Higher fatty acids than m~Tistic acid are generally absorbed into the

thoracic duct, while lower ones are absorbed mainlyア intothe portal vein directly.

3) In regard to lower fatty acids, the percentage of absorption into the tho-

racic duct gradually declines in proportion to the decrease in number of carbon atoms.

Even all the lower fatty acids such as caproic, caprylic acids, do not necessarily get

absorbed into the portal blood, but some are absorbed into the thoracic duct l~ ・mph

too.

4) In the postabsorptive state, lower fatty acids than lauric acid cannot be

found in any organ. And it was found that the fatty acids present in every normal

organ, and whose percentage in every organ is almost constant, are : mγristic acid,

palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid.

5) Depot fat contains lauric acid. However, highly unsaturated fatty acids,

docosenoic acid, eicosenoic acid etc. are present in the liver, bile, and blood in con-

siderable amounts ; those present in extrahcpatic tissues are far le田 inquantity than

in the liver. The origin of highly unsaturated fatty acids, docosenoic acid, eicosenoic

acid etc. contained in organs is attributable to alien fats coming from nutrients.

In closing, my hear勺rappreciation is extended to Dr. YoRINORI HrKASA, the instructor who

favored me with his tireless guidance and encouragement in this research as well as Prof.

MANJIRO NooA, an<l Mr. 0<AMIJ HIRAYAM¥, Assistant of the Biochemical Laboratory, Saikyo Univ-

ersity, Kyoto who directed and encouraged me in paper chromatography.

EXPLANATION OF FIGURES

The spot identi白cationsof paper chromatogram are as follows : 4: Butyric Acid, 6: Caproic Acid, 8: Caprylic Acid, IO: Capric Acid, 12: Lauric acid, 14: My-

ristic Acid, 16: Palmitic Acid, 18: Stearic Acid, 20: Arachidic Acid, OL: Oleic Acid, El: Eico-

senoic Acid, LE: Linoleic Acid, LN: Linolenic Acid, DO : Docosenoic Acid, HX: Hexadesenoic

Acid, HU: Highly Unsaturated Fatty Acids, S: Mixture of the Derivatives of Saturated

Acids from C4 to Czo (Standard), S': Mercurated Derivative of Oleic Acid (or Oleic and Eicosenoic Acids), S11: Mixture of Nonmercurated and Mercurated Derivatives of Oleic Acid (or Oleic and

Eicosenoic Acids), (Hg・) : Mercuric Compounds of the Ester Derivatives.

Fig. 2 Chromatogram of Component Fatty Acids in the Thoracic Duct Lymph Collected in

Postabsorptive State (Dog). Fig. 3 Chromatogram of Component Fatty Acids in Chyle Collected after Oral Administration

of Cod Liver Oil Emulsion (Dog). F. I.: Component Fatty Acids in Cod Liver Oil Emulsion. F. I. L.: Component Fatty Acids in Chyle.

Fig-. .t Chromatogram of Component Fatty Acids in Chyle Coll巴ctcdafter Oral Administration

of Butter Fat mo且・）． B. : Component Fatty Acids in Butter Fat. L.B. : Component

Fatty Acids in Chyle. Fig-. 5 Chromatogram of Component Fatty Acids in Chyle Collected after Oral Administration

of Sesame Oil Emulsion (Dog). F. I.: Component Fatty Acids in Sesame Oil Emulsion.

F'. I. L. : Component Fatty Acids in （句hyle.Fig・. 6 ChrnmatogTam of C'Lmponcnt Fatty Acids in Chyle Collected after Oral Administration

1188 日本外科宝！J:f 第28巻第L号

of Synth巴sizedSimple Glyceride Mixture <Dog). D: Component Fatty Acids in the

Mixture of Simple Glycerides (Equal Amount of Trilaurin, Tricaprin, and Tricaprylin).

L: Compon巴ntFatty Acids in Chyle.

Fig・. 7 Chromatogram of Component Fatty Acids in Portal Serum Collected in the Postabsorp-

tive State (Dog). Fig・. 8 C'hromatogram of Component Fatty Acids in Chyl巴 andPortal Serum C冶Ilectedafter

Oral Administration of Synthesized Tricaprylin (Dog). L: Component Fatty Acids

in Chyle. B: Component Fatty Acids in Portal Serum. Fig'. 9 Chromatogram of Component Fatty Acids in Chyle and Portal Serum Collected after

Oral Administration of Synthesized Tricaproin (Dog). L: Component Fatty Acids in

Chyle. B :Component Fatty Acids in Portal Serum.

Fig. IO Chromatogram of Component Fatty Acids in Liver (Dog).

Fig・.11 Chromatogram of Component Fatty Acids in Muscle (Dog).

Fig.12 Chromatogram of C》mponentFatty Acids in Kidney (Dog).

Fig.13 Chromatogram of Component Fatty Acids in Spleen iDogJ. Fig.14 Chromatogram of Component Fatty Acids in Brain (Dog).

Fig.15 Chromatogram of Component Fatty Acids in Bile (Dog).

Fig.16 Chromatogram of Component Fatty Acids in Subcutaneous Adipos巴 Tissue(Do宮）．

Fig.17 Chromatogram of C'ornponent Fatty Acids in Omentum (Dog).

Fig・. 18 Chromato且Tamof Component Fatty Acids in Serum, Omentum majus and Subcutaneous Adipose Tissu巴（Human).

Fig・. 19 Chromatogram of Component Fatty Acids in S巴sameOil and Cod LiYer Oil.

S. 0.: Component Fatty Acids in Sesame Oil. C. L. 0.: Component Fatty Acids in Cod

Li＂巴 rOil.

REFERENCES

1 , Artom, C & （’ornatzer, W. E: The Action of Choline and Fat on Lipid Phosphorylation in the

Li¥'cr. J. Biol. Chem., 171, 799, 1947.

2J Asada, S.: Histochemical Studies on the Intravenously Infused Fat Emulsion. Arch. Jap目

Chir., 22, 77, 1954.

3) Bauersfelcl, E. H.: Traumatic Chylothorax from Ruptured Thoracic Duct Treated by Intrave-nous Injection of Aspirated Chyle. J. A. M.A. 109, 16, 1937.

4J Berry, I. M. & Ivy, A. C.: Th巴 Toleranceof Dog’s to Intravenously Administered Fatty Chyle and Synthetic Fat Emulsion. Federation Proc., 7, 7, 1948.

5) Bloom, B., Chaikoff, I. L., Reinhardt, W. 0., Entenman, C&Dauben, IY. G.: The Quantitative Significance of the Lymphatic Pathway in Transport of Absorbed Fatty Acids. J. Biol. Chem., 184, 1, 1950.

6) Bloom, B., Chaiko汀， I.L., Reinhardt, W. 0. & Dauben, W. G. : Participation of Phospholipids

in Lymphatic Transport of Absorbed Fatty Acids. J. Biol. Chem., 189, 261, 1951.

7) Bloom, B., Chaiko庁， I.L. & R巴inhardt,W. 0.: Intestinal Lymph as Pathway for Transport

of Absorb巴dFatty Acids of Different Chain Lengths. Am. J. Physiol., 166, 451. 1951. 8) Bloor, W.R. . Th巴 OxidativeDetermination of Phospholipid (Lecithine & CephalinJ in Blood

and Tissues. J. Biol. Chem., 82, 273, 1929.

9) Bergstrカm,S. Bergstrom, B. Tryding, N. & Westiiii, G.: Intestinal Absorption and Metabolism

of 2: 2・DimethylstearicAcid in the Rat. Bioch. Journal, 58, 604, 1954. 10) Collins. H, S., Kraft, L. :¥I., Kinnet, T. D., Davidson, C. S., Young, J. & Stare, F. J.: Studies

on the Tolerance of Dogs to Intravenous Administration of Fat Emulsion. J. Lab. and Clin. :¥Iecl., 3.3, 143, 1948.

11) Davis, R. E.: The l¥Ielabolism of Tributyrin. J. Biol. Chem., 88, 67, 1930.

12) Eckstein, H. C.: Fat Absorption through Channels other than the Left Thoracic Duct. J. Biol. Chem., 62, 737, 1923.

13) Eckskin, H. ('.: The Influence of Diet on th巴 BodyFat of the White Rat. J. Biol.「hem.• 81, 613, 1929.

14) Frazer, A. C.: Fat Absorption and its Relationship to Fat l¥letabolism. Physiol. Rev., 20, 361.

ANALYTIC CHEMICAL STUDIES ON FAT METABOLISM 1189

1940. 15) Fern旦ndes,J.: Fat AbsつrptionStudied in a Child with Chylothorax. Thesis, Leiden. 16) Futter, J. H. and Shorland, F. B.: Studies on the Composition of Lipids of the Rabbit. Bioch.

Journal, 65, 689, 1957. 17) Hashino, H.: Experimental Studies of Fat Metabolism from th巴 View-pointof Ketone Body

Formation. Arch Jap. Chir., 24, 488, 1955.

18) Hika百九 Y,. Ishigami, K., Asada, S., Zaitsu, A., Tsui王ada.A., and Nakata, K. . Studies on the

Intravenous Administration of Fat Emulsion. J. J. S.S., 52, 298, 1951.

19) Hikasa, Y., Asada, S., Zaitsu, A., Tsukada, A., and Nakata, K.: Studies on the Intravenous

Administration o.f Fat Emulsion. Arch. Jap. Chir., 21, l, 1952.

20) Hikasa, Y.: Studies on the Intravenous Administration of Fat Emulsion Prepared by High

Pressure Jet. Rev. Phys. Chem. Jap., 12, 82, 1952.

21) Hikasa, Y., Hashino, H., Osa, H. and Takeda, S.: The Role of Watersoluble Vitamins in Fat

Metabolism. Nipponrinsho., 13, 1225, 1955.

22) Hikasa, Y., Shir叫 ani,H., Shigenaga, M., Kuyama, T., Tan, N. and Shimada, Y. : Pa'rentcral

administration of Fats. I. Fat Metabolism in Vivo, Studied with Fat Emulsion. Arch. Jap. Chir., 27, 396, 1958.

23) Hikasa, Y., Kuyama, SηShi且・enaga,M., Chi Chien Hsii, Hanabusa, S., Tamaki, A叫 Matsuda,

S., Onishi, H., and ’Tobe, T. : Par巴nteralAdministration of Fats. ll. Clinical Application of

Fat Emulsion. Arch. Jap. Chir., 27, 736, 1958.

24) Hirayama, 0., and Noda, M.: Metallic Addition Compounds o.f Unsaturated Fatty Acids. ll. Microanalysis of Unsaturated Fatty Acid Esters by Mcrcuration Method. Scientific Reports

of the Saikyo UniYcrsity, Agricultur巴， 6,86, 1954.

25) Hirayama, 0. and Noda, M. : Paper-chromatography of Fatty Acids as their Acetol Ester Derivatives. Scientific Reports of th巴 SaikyoUniversity, Agriculture., 7, 43, 1955.

26) Hu広hes,R. H. and Wimmer, E.T.: The Absorption of Soluble, Volatile Fatty Acids. J. Biol.℃hem., 108, 141, 1935.

27) Ikeda, H. : Exp巴rimental Studies on Fat Metabolism with a Blocked Reticuloendothelial System. Arch. Jap. Chir., 26, 215, 1957.

28) Inoue, Y., Hirayama, 0. and Noda, M. : Separation and Identification of Fatty Acids. XX. 2 4-Dinitrophenylhydrazones o.f p-Bromophenacyl Esters as Derivatives for Characterization

of Unsaturated Fatty Acids. Bull. Agr. Chem. Soc. Jap., 20, 197, 1956. 29) Inoue, Y., Hirayama, 0. and Noda, M. : Separation an Identification of Fatty A~ids. XX!.

Paper-chromatography of Fatty Acids as their p-Bromophenacyl Ester Derivatives. Bull.

Agr. Chem. Soc. Jap., 20, 200, 1956.

30) Izukura, T.: Histochemical Studies on Intravenously Administered Fat Emulsion. Arch. Jap. Chir., 26, 215, 1957.

31) Kosaki, N.: Studies on Fat Metabolism after Total Pancreatectomy. Experiments in Dogs. Arch. Jap. Chir., 25, 13丹， 1956.

32) Kiyasu, J. Y., Bloom, B. and Chaikoff I. L.: The Portal Transport of Absorbed Fatty Acids.

J. Biol. Chem., 199, 415, 1952.

33) Ku:vama, T.: Clinical Studies on the Nutritional Effects of Intravenous Administration of

Fat Emulsion. Arch. Jap. Chir., 27, 64, 1958. 34) Lerner, S. R., Chaikoff, I. L., Entenman, C. and Dauben，羽T. G. : The Fate of C14-Labeled

Palmitic Acid Administered Intravenously as a Tripalmitin Emulsion. Proc. Soc. Exper.

Biol. and Med., 70, 381, 1949.

35) Little, J. 1¥1., Harriおりn,（‘ and Blalock, A.: Chylothorax and Chyloperitoneum. E汀ectsof

Reintroduction of Aspirated Chyle. Survery., 11, 392, 1942. 36) l¥laki. Y.: unpublished.

37) Manabe, K. and Kameda, I: Homolatenlity of the Portal Blood Flow in the Liver. Sogoigaku, 11, 255, 1954.

38) I阻むng,H.「 and Freeman, S.: Experimental Studies on the Intra¥・enous Injection of a Fat

Emulsion into Do且’s.J. Lab. and Clin. Med., 33, 698, 1948.

39) Nishino, T.: Laboratory Studies on the Intravenous Administration of Fat Emulsion in the

1190 日本外科宝函第28巻第4号

Light of Tissue Metabolism. Arch. Jap. Chir., 23, 607, 1954.

40) Oeken : Ein Fall von Zerreissun阜、 desDuctus thoracicus infolge Brustquetschung. Miinchen.

med. Wochenshr., 55, 1182, 1908. 41) Ono, K: Fat i¥Ictabolism. Saishin-Igaku, 10, 52, 1955.

42) Osa, H.: Experimental Studies on the Intravenous Admimstration of a Fat Emulsion for Nutritional Purposes. Arch. Jap. Chir., 25, 151. 1956.

43) Otani, S. : On Mechanism of Adverse Reactions Caused by Intravenous Administration of

Fat Emulsion. Arch. Jap. Chir., 25, 172, 1956. 44) Reiser, R., Bryson, M. J.唱 Carr,M. J. and Kuiken, K. A. : The Intestinal Absorption of

Triglycerid巴s.J. Biol. Chem., 194, 131, 1952. 」5) Seno, A.: A Study of Fat Jictabolism in the Isolated Perfused Liver. Arch. Jap. Chir., 24,

179, 1955.

46) Shigenaga, M. : Experimental Studies on Fat Metabolism with Determination of Respiratory Quotient and Ketone Body Production of Tissues. Arch. Jap. Chir., 27, 91, 1958.

47) Shirotani, H.: Histochemical Studies on Fat Metabolism by Intravenous Administration of

Fatty Chyle. Arch. Jap. Chir., 26, 38, 1957. 48) Sinclair. Robert Gordon: The Metabolism of the Phospholipids. V. The Relationship between

the Amount of Fat In且・estedand the Degree of Unsaturation of the Phospholipids and

Neutral Fat in the Tissues of the Rat. J. Biol. Chem., 96, 103, 1932.

49) Takahashi, K. : Experimental Studies on the Absorption of Fat from the Intestine. Tohoku-

lgaku, 33, 239, 1943.

50) Tamaki, A. : unpublished 51〕 Tatsumi,W.: Uber <lie intraveniise Infusion des Fettes. Arch. Jap. Chir., 26, 1, 1957.

52) Tobe, T. : unpublished 53) Yasuda, M .. Lipid Metabolism. J. Biol. Chem. Jap., 17, 1, 1942.

54) Yasuda, l¥L: Lipid Metabolism. Nisshin-Igaku, 32, 501, 1943.

55) Yasuda, M.: Lipids in Tissues. Igakunoayumi, 13, 201, 1952. 56) Zilversmit, D. B., Chaikoff, I. L., and Entenman, C.: Are Phospholipids Obligatory Participants

in Fat Transport across the Intestinal Wall? J. Biol. Chem., 172, 637, 1948. 57) Zilversmit, D. B., Entenman, C. and Chaikoff, I. L. : The Measurement of Turnover of the

Various Phospholipids in Liver and Plasma of the Do且、 andits Application to the Mechanism of Action of Choline. J. Biol. Chem., 176, 193, 1948.

和文抄録

脂酸ペーパークロマトグラフィーを応用した

脂質代謝の分析化学的研究

京都大学医学部外科学教室第 2 講座 rJtr~ 青-WI皮誠教授）

丹 敏

脂酸ペーパークロマトグラフィ一法を駆使，応用す

ることによってF 経口的に摂取された脂質の腸粘膜か

らの吸収経路についーて追究すると同時にp 生体内各種

臓器中の含有脂酸p 更には血清p 胆汁p リンパp 貯蔵

脂質中の含有JJ旨般の分析を行し、，次のような結論に到

J主した．

Ill 経口的に摂取された脂質の腸粘膜からの吸収経

路は 2つに大別される．即ち胸管リンパ中に吸収Zれ

るものと，門脈血中へ直接吸収されるものとの2つが

ある．

(2) この何れの吸収経路をとるかはp 脂酸の鎖の長

短によって規定されるものでy ミリスチン酸以上の高

ANALYTIC CHEMICAL STUDIES ON FAT MF:T ABOLISM 1191

級1旨酸は専ら胸管リンパ中へ吸収されるのに反して， リセライドをその原料として選ぶべきである．

それ以下の低級脂酸は主として門脈血中へ直接吸収さ (5) そのような意味では，われわれが斯る目的に使

れる． 用しつつあるゴマ油は，その原料脂質として合理的な

13！併し，更に低級脂酸についてみると，低級脂酸 ものということが出来る．

の中でも，その炭素原子数が減ずるに従って門脈血中 16) 各臓器を通じてみる時y ラウリン酸以下の低級

への吸収率は漸次増大する．併し炭素原子数が 6個あ 脂酸は Postabsorptivestateに於ける限仏何れの

るいは8個というような低級飽和脂酸（カプロ ン酸， 臓器組織内にも認められない．そして生体内各臓器の

カプリル酸）であってもP その全てが門脈血中に吸収 何れにも存在しP 而jも各臓器が常に略々一致した含有

されるわけではなしなおその幾許かは胸管リ ンパ中 比率を示している脂酸はp ミリスチン酸，パルミチン

へも吸収される． 酸，ステアリ ン酸p オレイン酸， りノール酸p リノレ

il I 従って脂質を乳化態として直接皮下静脈内 4 、；.j:_ ン酸であった．

入するに当つては，出来得る限り，高級脂駿のみから 171 貯蔵胞買はラウリ ン酸を比絞的多く含有してい

なるトリ・ ゲリセライ ドをその原料として選ぶべきで るためP その中平日価が高い．これに反して高度不飽和

ある．併し教室先人の行った組織顕微化学的倹索成績 脂酸は肝臓，胆汗・，血清中に比較的多く含有されて居

あるいは生化学的検索成績とを併せ考えると，高級脂 り，肝外組織中のそれが含有量は肝臓等に較べれば問

酸とはいうものの，高度不飽和脂酸，エイコセン酸， 題にならない程少量である．而して高度不飽和脂酸は

鯨油酸等を全く合有していてはならないもので，結局 生体内で合成されることはあり得ないから，少なくと

ミリスチン酸，パルミチン酸，ステアリン酸，オレイ もそれら臓器中に含有される高度不飽和脂酸の由来は

ン酸，リノール酸，リノレン畿のみからなるトリ・グ 外来脂質に求めざるを得ないー

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