Isolation of ethylene glycol from the lipids of beef lung

Vol. 13, No. 2, 1963

BIOCHEMICAL

AND BIOPHYSICAL

ISOLATION

OF ETHYLENE

FROM THE LIPIDS H. E.

Carter,

Division

P. Johnson,

Received

August

are

mammalian the

Noyes

studies

widely

and monoglycerides is

glyceride

the

only

of the During

the

beef

lung

of

and stearic were

in

three

solvent

however,

normally

sources.

(1962)

that

found

in

Recently,

reported

the which

and ethylene

of

extract

(Teets

complex

isolation on

glycol

readily

--et al.

identified

ester

band

Paper presence

had the systems.

hydrolyzed

hydroxide

yielded

chromatography of glycerol

same Rf as that

156

detailed

a

a fraction in

the

under

the

Hydrolysis

soluble

A more

from

1963)

(1946).

ether

lipids

et al.,

showed a strong

negative.

which

alcohol

isolation

as the

showed the

appears,

of di-

derivatives.

potassium acids

triglycerides

amounts

of a wax fraction

was not

Schmidt

that

It

glycerol

which

methanolic

substance

Chemistry

of glycerides

lesser

mammalian

benzoate

and which

fraction

with

yielded

conditions

tests

occurring

avium

was encountered

with

demonstrated

and Miguel

saponification

infrared

isolation

have

from

Mycobacterium

commercial

of the

polyhydroxy

fractions

by means

of

Urbana

respectively.

Damareau-Ginsburg from

and R. K. Yu

Laboratory

of Illinois,

tissue

most

glycerol

OF BEEF LUNG

23, 1963

Extensive from

GLYCOL

D. W. Teets

of Biochemistry, University

RESEARCH COMMUNICATIONS

of

mainly

this palmitic

constituents. of the plus

Sterol water-soluble

a second

of ethylene study

fraction

glycol of this

Vol. 13, No. 2, 1963

problem fresh

BIOCHEMICAL

was undertaken beef

lung

at

beef

ground

extracted

and for

four

lung

In

times

was then

re-extracted

combined

ether

of 14.5

with

extract g. of lipid

prepared

after

removal

methanol

and activation

machine acetone

with

evaporation,

the

and

tissue ether.

was loaded silicic

fines

--in

was eluted

The

vacua with

a in benzene

acid

by repeated

by heating

ice,

representing

of Mallinkrodt of

dry

by stirring

anhydrous

(213 mg. P),

The column

12 hours.

(430 g.)

suspension

(80 mm.)

benzene

at

in 140'

and 50 ml.

were collected. The first

three

of

in

The remaining

times

after

a column

mincing

24 hours. four

onto

fractions

extracts

transported

reagent-grade

for

solution

for

purpose

(30 lb.),

an electric

room temperature

total

this

RESEARCH COMMUNICATIONS

were prepared.

Fresh was finely

AND BIOPHYSICAL

1550 ml.

fractions:

appeared

A (l-12),

to consist

investigated

mainly

characterized

into Fraction

A

and was not

in

of spots

with

in benzene typical

in

B and to a lesser

the

extent

B and C were then

times

with

an excess

for

of Amberlite 157

di-

(iodine

and

tri-glyceride in

24 hours.

ether.

on

triglyceride

treated

to pH 1 with diethyl

were

chromatography

C showed in addition

were acidified

four

g.)

layer

B gave

KOH (90% MeOH) at 37'

hydrolysates

and C (3.01

5% methanol

Fraction

A series

Fractions

was stirred

C (18-31).

and by thin

and Fraction

were apparent

g.)

with

of spots).

monoglycerides.

extracted

B (13-l?),

B (0.87

G developed

properties

methanolic

were combined

of hydrocarbons

by infrared

Gel

detection

eluent

further. Fractions

Silica

of

region

C. with

1 N

The

5 N HCl and The aqueous

phase

MB-3 and allowed

Vol. 13, No. 2, 1963

to stand

BIOCHEMICAL

before

was reduced residue

filtering

Gel

ethanol

acid

periodate

resin.

The clear

at 40'

for were

RESEARCH COMMUNICATIONS

thin

(14 mm.)

layer

and the

chromatography

chromatographed

(65:25:10),

filtrate

in

and developed

on

chloroform:

by spraying

and benzidine.

Table fractions

in

the

evaporator

The plates

G.

methanol:formic with

from

on a rotary

dissolved

Silica

AND BIOPHYSICAL

I shows Rf values

B and C compared

of

with

compounds

obtained

from

standards.

TABLF: I Thin

Layer

Chromatography

on Silica

CHCLs:MeOH:HCOsH

(65:25:10)

Compounds

Ethylene

Fraction

Rf

Glycerol Glycol

B

0.59

0.59

C

0.43,

0.65

D-Glyceraldehyde

0.50

indicate

that

fraction

ethylene

glycol

while

fraction

C gave

ethylene

glycol,

with

glycerol

the

(based

on visual

by vapor

phase

according

to

of

standard

glycerol

and

both

predominant results

chromatography

of the

acetates

samples

--et al. of

(1957)

ethylene

respectively

when chromatographed and 75 ml./min.

times

were 2.2

The solid 158

component were

with

diacetate min.

the

behavior

and glycerol

and 14.3

on an 8 ft. support

confirmed

(prepared

compared

glycol

Retention

140'

only

These

Shriner

0.59

B yielded

inspection).

triacetate.

at

Rf

0.43

1,2-Propanediol

The data

Gel G in

min.

lO$ W-1

was 60/80

column

Vol. 13, No. 2, 1963

BIOCHEMICAL

Chromosorb

W which

disilazane. while in

the

had been

Fraction

Fraction

In blank through

other

than

pretreated

glycerol of 4:l.

runs

in which

entire

glycerol

hexamethyl

ethylene

glycol

and ethylene

ratio

the

RESEARCH COMMUNICATIONS

with

B showed only

C gave

approximate

carried

AND BIOPHYSICAL

glycol

glycerol

acetates

solutions

experimental

was detected

diacetate

were

procedure,

by thin

layer

no spot chromatography.

DISCUSSION The above beef

lung

of a combined

extracted

with

ethylene

from occurred

Furthermore, glycol

bacterial

lipids

ruminants

of bacteria found

is

in

extracting

the

which

escaped

rumen.

products

detection

that

due to its

in our

layers

standards.

silica

It

is 159

also

may be

(1956)

fractions other

than

contaminants. of

glycol method

gel

presence

Lovern

compound

to the

by the

of

from

it

glyceride

similarity

solutions.

glycol

hand,

class

may

aqueous

case due to the

to be non-lipid this

cannot

tissue.

and Miguel,

other

of

and ethylene the

of hydrolysis

laboratory

lung

of ethylene

Ethylene

behavior.

lipid

fractionation

fresh

of fish

is

amounts

lung

concentrating

On the

in

which

of glycerol

a special

possible

had an Rf on thin triglyceride

the

saponification

chromatographic synthesized

total

by Damareau-Ginsburg

the

glycol

partial

isolation

were assumed

entirely

since

in

existence

The relative in

losses

of the

water-soluble

glycerol It

data

constitute in

that

yielded

these

occurred

In view

of ethylene

lipid

differential

may have

the

fractions.

glycol-containing

have

that

demonstrate

form

glyceride

be estimated well

results

glycerides

in

dipalmitate of

Paquot

comparable

probable

has

that

(1947)

to various appreciable

Vol. 13, No. 2, 1963

BIOCHEMICAL

losses of ethylene procedures

glycol

AND BIOPHWCAL

would occur during

Further

or assigned

the extent

glycol

in beef lung and the nature

in the ester

to non-lipid

work is in progress

tively

of the occurrence

linkage. may be.

of esterified of the fatty

Other tissues (In single

glycol.)

The possible

quantitaethylene

acids involved

from beef and other how general

preliminary

and beef suet were found to contain ethylene

contaminants.

to determine

are being examined to determine

occurrence

concentration

and that small amounts of the compound remaining

may have been overlooked

animals

RESEARCH COMMUNlCATlONS

this

experiments

lard

very small amounts of

metabolic

significance

of such

compounds is also under investigation.

ACKNOWLEDGMENTS Part of this (E574)

Blindness,

work was supported

from the Institute TJ. S. Public

Welcome Trust

of Neurological Health

for a Travel

Service.

Scholarship

by a research

grant

Diseases and We wish to thank the

to one of us (P. J.)

REFERENCES Damareau-Ginsbur H. and Mi uel, biol., 48: 679 (1962 7

A. M., --Bull.

sot.

chim.

Jones, W. S. and Tamplin, W. S., in Glycol, Amer. Chem. .Soc. Monograph Series, Reinhold, New York (1952) PP. 30-56 Lebaron, F. N., Folch, J. and Rothleder, E., Federation -Proc., 16, 209 (1957) Lovern, J. A., Biochem. J., 63, 373 (1956) Paquot, M. C., --Bull. Schmidt, G., Benotti, J ., 2. --Biol.

sot. chim. France, 14, 926 (1947) J., Hershman, B. and Tannhauser, S. Chem., 166, 505 (1946) 160

Vol. 13, No. 2, 1963

Segur, Shriner,

Teets,

BIOCHEMICAL

J. B., in Glycerol, Series, Reinhold,

AND BIOPHYSICAL

RESEARCH COMMUNICATIONS

Amer. Chem. Sot. Monograph New York (1953) p. 254

R. L., Fuson, R. C. and Curtin, D. Y., The S stematic Identification of Organic Cornends, -5i-TJo n lley and Sons, Inc.,71957) p. 212 D. W., Johnson, Federation =.,

P.,

Tipton, 22, 414

161

C. L. and Carter, (19631

H. E.,