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Number 52 December 25, 1978

Citation ClassicsBligh E G & Dyer W J. A rapid method of total lipid extraction and

purification. Can. J. Biochem. Physiol. 37: 911-917, 1959.

The Bl igh and Dyer method has

cert ai nly been wi del y u sed , and i t i s

most gratifying to have it identif ied as a

Citation Classic. I t all began back in the

1950s at the Hal ifax Laboratory of the

F isheries Research Board o f Canada,

where Dr. Wil liam J. Dyer and his group

were investigating the deter ioration of

f ish musc le prote ins dur ing f rozen

s to rage. The group s p ioneer ing work

a tt ra ct ed i nt e rna ti ona l a tt en t ion and

st imulated a g reat deal o f research ins imilar laborator ies a round the wor ld .

They had found that protein denaturation

in f rozen f ish muscle cont r ibuted

substan tial ly to consumer rejec tion o f

f rozen seafoods and that i t was

a cc ompani ed b y l ip id d et er io ra ti on ,

par ticularly hydrolysis. When I returned

to the group in 1956, fol lowing graduate

work at McGil l Univers ity, my task was

to investigate the role of lipids in the

cont inu ing research program ondeterioration in frozen-stored fish.

On initiat ing this l ipid study, the well-

known need f or a gen tl e and r el iable

method for extraction and purif ication of

total lipid from biological tissues became

acute. We were work ing wi th cod

muscle, where the h ighly unsaturated

lipids rare ly exceeded 1 % wet we ight

and consisted primarily of protein-bound

phosphol ipid. Existing procedures were

unsatisfactory and attention was focused

on the use of mixtures of chloroform and

methanol to isolate the l ipids from moist

biological materials.

Examinat ion o f thechloroformmethanol-water phase diagram

led to the hypothesis that optimum lipid

extraction should result when the t issue

is homogenized wi th a mix tu re of

chloroform and methano l which , when

mixed with the water in the tissue, would

y ie ld a monophas ic so lut ion. The

r esul ti ng homogenat e c ou ld t hen be

diluted wi th water and/or chloroform to

produce a b iphas ic sys tem, the

chloroform layer of which should contain

the l ip ids and the methanolwater layer

t he n on li pi ds. T he h yp ot he si s wa s

readily confirmed by experimentation and

the me thod was pro ve n to be very

ef fec t ive. I t was enthus iast ica l l y

accepted by the scien ti fi c communi ty ,

and after 19 years, i t is sti ll being used

ext ensi ve l y i n r esea rch l abo ra t or ie s

through out th e wor ld, on a host o f

biological materials.

I wou ld ment ion that as a smal l

government research laboratory, we were

not encouraged to do fundamental

research but in the course of providing a

re sea rch and devel opment servi ce

related to the problems of the Canadian

sea food i ndus tr y, we were con ti nual y

faced wi th s i tuat ions where bas ic

scient i fi c know ledge was inadequate .

This is even more true today as the

world directs greater attention toward theoceans and their resources. Although Dr.

Dye r i s n ow re ti re d, h is c ol le ag ue s

con ti nue t o pur sue t he cha ll enges i n

f isher ies sc ience a t the Hal i fax

Laboratory where I am now Director.

Lipid decomposition studies in frozen fishLipid decomposition studies in frozen fishLipid decomposition studies in frozen fishLipid decomposition studies in frozen fishLipid decomposition studies in frozen fish

have led to the development of ahave led to the development of ahave led to the development of ahave led to the development of ahave led to the development of a simple,simple,simple,simple,simple,

rapid,rapid,rapid,rapid,rapid, and and and and and reproduciblereproduciblereproduciblereproduciblereproducible method for the method for the method for the method for the method for the

extraction and purif ication of l ipids fromextraction and purif ication of l ipids fromextraction and purif ication of l ipids fromextraction and purif ication of l ipids fromextraction and purif ication of l ipids from

biological materials. The method has beenbiological materials. The method has beenbiological materials. The method has beenbiological materials. The method has beenbiological materials. The method has been

applied to fish muscle and may easily beapplied to fish muscle and may easily beapplied to fish muscle and may easily beapplied to fish muscle and may easily beapplied to fish muscle and may easily be

adapted to use with other t issues. [Theadapted to use with other t issues. [Theadapted to use with other t issues. [Theadapted to use with other t issues. [Theadapted to use with other t issues. [The

SCISCISCISCISCI indicates that this paper was ci ted indicates that this paper was ci ted indicates that this paper was ci ted indicates that this paper was ci ted indicates that this paper was ci ted

2,554 times in the period 1961-1977.]2,554 times in the period 1961-1977.]2,554 times in the period 1961-1977.]2,554 times in the period 1961-1977.]2,554 times in the period 1961-1977.]

E. Graham Bligh

Canada Dept. of Fisheries & Environment

Fisheries & Marine Service, Halifax Lab.

1707 Lower Water Street

Halifax. Nova Scotia, Canada B3J 2S7

April 3, 1978