CHAPTER 1

Definition and classification of lipids

The term ‘lipids’will be defined here as those substances which are (a)
insoluble in water; (b) soluble in organic solvents such as chloroform,
ether or benzene ; (c) contain long-chain hydrocarbon groups in their
molecules; and (d) are present in or derived from living organisms.
This definition covers a wide range of compounds and includes long-
chain hydrocarbons, alcohols, aldehydes, and fatty acids, and deriva-
tives such as glycerides, wax esters, phospholipids, glycolipids, and
sulfolipids. Also included are substances which are usually considered
as belonging to other classes of compounds, e.g., the ‘fat-soluble’
vitamins A, D, E, and K, and their derivatives, as well as carotenoids
and sterols and their fatty acid esters. They are included here because
they are usually associated with membranes and are obtained together
with the phospholipids and neutral lipids when cells or tissues are
extracted with ‘fat’ solvents. The sterols and carotenoids, however,
will be treated only briefly since several recent monographs or collec-
tions of articles dealing with these classes of compounds are available
e.g. Clayton’s (1969) Methods in Enzymology, Vol. 15 ; Marinetti’s
(1969) Lipid Chromatographic Analysis, Vol. 2 ; and Goodwin’s (1965)
Chemistry and Biochemistry of Plant Pigments.
The remainder of this chapter will be devoted to outlining the chemi-
cal structures of the various types of lipids usually encountered in
studies on cellular lipids. The classification given below is fairly
comprehensive, but is by no means all-inclusive. The reader should
refer to the monographs by Hanahan (1959) and Ansell and Hawthorne
279 Subjecf indexp. 601
280 TECHNIQUES OF LIPIDOLOGY

(1964) for basic background knowledge of lipid chemistry, and to the

following specialized books or articles for detailed structural infor-
mation on : bacterial lipids (Kates 1964b ; Asselineau 1966 ; O’Leary
1967); plant lipids (Mazliak 1970; Kdtes 1970); animal lipids (Van
Deenen 1965); fatty acids and glycerides in general (Gunstone 1967;
Sober 1968). The IUPAC-IUB Tentative Rules for biochemical
nomenclature (see Sober 1968)will be followed throughout this book.

1.I. Hydrocarbons
This class represents the simplest type of lipids ;they occur as normal,
branched and unsaturated chains of various lengths.

I .I .I. Normal, saturated paraffins

These have the general formula, CH3(CH,),CH3 (n= 6-36 or greater).
Some common hydrocarbons in this class are listed in table 1.1.

1.I .2. Monobranched-chain paraffins

These occur in two general types : the iso-series with general formula,
CH3
I
CH3CH(CH,),CH3 (n=4-32 or greater);
and the anteiso-series with general formula,
CH3
I
CH3CH,CH(CH2),CH3 (n= 3 - 31 or greater).
Table 1.1 lists some is0 and anteiso branched-chain hydrocarbons.

I .I .3. Multibranched parafins : saturated isoprenoids

The known members of this group are:
CH3
I
farnesane H-[CH,CHCH,CH2]3-H
(c15H32) 2,6,10-trimethyldodecane
Ch. 1 DEFINITION AND CLASSIFICATION 28 1

TABLE1.1
Some normal and branched-chain hydrocarbons.
____~ ~

Formula Normal paraftins [so-paraffins Anteiso-paraffins

Decane 2-methylnonane 3-methylnondne

Dodecane 2-methylundecane 3-methylundecane
Tetradecane 2methyltridecane 3-methyltridecane
Hexadecane (cetane) 2-meth y lpentadecane 3methylpentadecane
Heptadecane 2-meth ylhexadecane 3methylhexadecane
Octadecane 2-methylheptadecane 3-methylheptadecane
Eicosane 2-methylnonadecane 3-methylnonadecane
Heneicosane 2-methyleicosane 3methyleicosane
Docosane 2-meth ylheneicosane 3-methy 1heneicosane
Tetracosane 2-methyltricosane 3-methyltricosane
Pentacosane 2-meth yltetracosane 3-methyltetracosane
Hexacosane 2-methylpentacosane 3methylpentacosane
Octacosane 2-methylheptacosane 3-methylheptacosane
Triacontane 2-methylnonacontane 3-methylnonacontane
Dotriacontane 2-methylhentriacontane 3-methylhentriacontane
Tetratriacontane 2-meth yltritriacontane 3-rnethyltritriacontane
Hexatriacontane 2-methylpentatriacon- 3-methylpentatriacon-
tane tane

CH,
I
pristane H-[CHzCHCH2CHz]3-CHzCH(CH3)2
(c19H40) 2,6,10,14-tetramethylpentadecane

CH,
I
phytane H-[CHzCHCHzCH2]4-H
(C2oH42) 2,6,10,14-tetramethylhexadecane

and squalane or spinacane (C30H62)

CH, CH,
I 1
H-[CHzCHCHzCHJ 3-[CH,CHzCHCHJ3-H
2,6,10,15,19,23-hexamethyltetracosane
Subject m d e x p . 601
282 TECHNIQUES OF LIPIDOLOGY

I .I .4. Nonnul, mono-unsaturated para&

These have the general formula, CH3(CH2),CH=CH(CH2),CH3.The
odd-numbered members of this series may be derived by decarboxyla-
tion of the natural even-numbered monoenoic fatty acids (Oro et al.
1967): e.g., 10- or 8-heptadecene from vaccenic or oleic acid, respec-
tively ; 8-pentadecene from palmitoleic acid.

1.I .5. Monobranched, unsaturated paraffins

These are known to occur as monounsaturated iso- and anteiso-
branched hydrocarbons (Tornabene et al. 1967), of general formulae :
CH3
I
CH3CH (CH2),CH=CH(CH2),CH3 ( x + Y = 16-23)

and CH3
I
CHjCH2CH(CH2),CH=CH(CH,),CH3 (X +y = 15- 22),
respectively.

1.1.6. lsoprenoid polyenes

Some members of this group are: phytadiene (2,6, 10, 14-tetramethyl-
hexadecadiene ; C,,H3,), of which three positional isomers are so far
known (Blumer and Thomas 1965), namely,

CH3 CH3
I I
H-[CH2CHCH2CH2]3-CH=CCH=CH,
1,3-phytadiene (two geometric isomers)

CH3 CH3 CH3

I I I
H-[CH2CHCH2CH2],-CH,CHCH2CH=CHCH,
2,4-phytadiene (four geometric isomers)

and CH2
II
H- [CH ,CHCH CH ,] 3-CH ,CCH=CH
neophytadiene
 Ch. 1 DEFINITION AND CLASSIFICATION 283

squalene or spinacene (C30H50)

CH3 CH3
I I
H-[CH ,C=CHCH ,I3-[CH2CH=CCH 2 1 3-H
dehydrosqualene (bacterial ‘phytoene’;C,,H48) (Suzue et al. 1968)

CH3 CH3 CH3 CH3

I I I I
H [CH,C=CHCH,],CH,C=CHCH=CHCH=CCH,[CH,CH=CCH2] ,H

1.1.7. Carotenoids
This group consists of C,, isoprenoid polyene hydrocarbons with con-
jugated double bonds; they show strong light absorption and often are
brightly coloured. Only a few of the most common members of this
group are given here:

phytoene (C40H64 ; colourless)

Eycopene (C40H56 ; red)

y-carotene (C40H56;orange)

Subjecr index p. 601

284 TECHNIQUES OF LIPIDOLOGY

a-carotene (C40H56;yellow)

1.2. Alcohols
Aliphatic alcohols occur naturally in free form or more usually in
esterified or etherified form; they occur with normal, branched, or
unsaturated chains of various chain lengths and with primary,
secondary or (rarely) tertiary alcoholic function.

1.2.1. Normal, saturated alcohols

The primary alcohols are most common and have the general formula,
CH3-(CH2),-CH20H (n = 6 - 30 or greater) : see listing in table 1.2.
Two examples of secondary alcohols, found in bacteria, are 2-D-
octadecanol and 2-~-eicosanol.

1.2.2. Monobranched-chain alcohols

Two main types are known in this group : the iso-series, with general
formula,
CH,
I
CH,CH(CH,),CH,OH (n= 4-24) ;

and the anteiso-series, with general formula,

CH,
I
CH$H2CH(CHI),CH20H (n=3-21)
Some common branched-chain alcohols are listed in table 1.2.

TABLE
1.2
Some normal and branched-chain primary alcohols
Formula Normal alcohols Iio-alcohols
1-hexanol 4-methyl-1-pentanol
1-octanol 6-methyl- 1-heptanol
1-decanol 8-methyl- 1-nonanol
1-dodecanol (lauryl alcohol) 10-methyl-I-hendecanol
(isolauryl alcohol)
c l 3 H 2 7 OH 1-tridecanol 1 1-methyl-1-dodecanol
C14H290H 1-tetradecanol (myristyl alcohol) 12-methyl-1-tridecanol
(isomyristyl alcohol)
1-pentadecanol 13-methyl-I-tetradecanol
1-hexadecanol (cetyl alcohol) 14-methyl-I-pentadecanol
(isopalmityl alcohol)
1-heptadecanol 15-methylhexadecanol
1-0ctadecanol (stearyl alcohol) 16-methyl-]-heptadecanol
(isostearyl alcohol)
1-nonadecanol 17-methyl-1-octadecanol
I-eicosanol 18-methyl-]-nonadecanol
I-docosanol 20-methyl-1-heneicosanol
I-tetracosanol 22-methyl-I-tricosanol
1-hexacosanol 24-methyl-1-pentacosanol
1-octacosanol 26-methyl-1-heptacosanol
1-triacontanol
0
P
Anteiso-alcohols
3-methyl-1-pentan01
5-methyl-1-heptanol
7-methyl-1-nonanol
9-methyl- 1 -hendecanol
(anteisolauryl alcohol)
10-methyl-1-dodecanol
1 1-methyl- 1-tridecanol
(anteisomyristyl alcohol)
12-methyl-1-tetradecanol
13-methyl-1-pentadecanol
(anteisopalmityl alcohol)
14-methyl-1-hexadecanol
15-methyl-1-pentadecanol
(anteisostearyl alcohol)
16-methyl-1-0ctadecano1
17-methyl-1-nonadecanol
19-methyl-I-heneicosanol
N
m
w!
286 TECHNIQUES OF LIPIDOLOGY

1.2.3. Multibranched alcohols : saturated isoprenoids

Some examples of this group are:

tetrahydrogeraniol H-[CH2CHCH2CH2I2-OH
(CI OH2201 3,7-dimethyl- I-octanol

CH,
I
farnesanol H-[CH,CHCH2CH2]3-OH
(C15H320) 3,7,11-trimethyl-l-dodecanol

TABLE
1.3
Some normal rnonoenoic alcohols and aldehydes.

No. of Alcohol Aldehyde

carbons

10 cis-3-decen-1-01 -
11 10-undecen-1-01 10-undecenal
12 cis-7-dodecen-1-01 -

14 cis-9-tetradecen-1-01(myristoleyl 'cis-9-tetradecenal (myristoleyl

alcohol) aldehyde)
16 cis-9-hexadecen-1-01(palmitoleyl cis-9-hexadecenal (palmitoleyl
alcohol) aldehyde)
18 cis-6-0ctadecen~1-01 (petroselenyl cisd-octadecenal (petroselenyl
alcohol) aldehyde)
18 cis-9-octadecen-1-01(oleyl alcohol) cis-9-octadecenal (oleyl aldehyde)
18 cis-11-octadecen-1-01(cts- cis-I 1-0ctadecenal (cis-vaccenyl
vaccenyl alcohol) aldehyde)
20 cis-9-eicosen-1-01(gadoleyl alcohol) cis-9-eicosenal(gadoleyl aldehyde)
22 cis-13-docosen-1-01 (erucyl alcohol) cis-13-docosenal (erucyl aldehyde)
24 cis-15-tetracosen-1-01(nervonyl cis-15-tetracosenal (nervonyl aldehyde)
alcohol)
Ch.1 DEFINITION AND CLASSIFICATION 287

CH3 CH,
I I
pristanol H-[CH,CHCH,CH,],-CH,CCH,OH
(CI9H400) 2,6.10,14-tetramethyl-1-pentadecanol

CH,
I
phytanol H-[CH2CHCH,CH2],-OH
(C20H420) 3,7,11,15-tetramethyl-l-hexadecanol

1.2.4. Unsaturated alcohols

Monoenoic alcohols usually are analogous to monoenoic fatty acids
(see tj 1.6.6). Their general formula is CH3(CH2),CH=CH(CH,),-
CHIOH; some individual alcohols in this class are listed in table 1.3.

1.2.5. Isoprenoid alcohols (terpenols or polyprenols) (Hemming 1969)

This series of alcohols has the general structure :

CH3
I
H-[CH,C=CHCH,],-OH (n=2- 13 or greater) .
Some individual members of the series (prenologues) are as follows :

n No. ofcarbons Name

2 10 Geraniol
3 15 Farnesol
4 20 Geranylgeraniol
9 45 Solanesol (all-truns)
10 50 Decaisopr en ol
11 55 Undecaisoprenol Castraprenols or
12 60 Dodecaisoprenol ficaprenols
13 65 Tridecaisoprenol (cis-trans)

Subjrcf index p . 601

288 TECHNIQUES OF LIPDOLOGY

Polyisoprenoid alcohols with one saturated isoprenoid unit

(dihydroprenols)are also known, e.g., bactoprenol ( C ,,) ;and dolichol
(C85 to CI 10).
A partially saturated terpenol which should be mentioned here is
phytol ( C 2 0 H 4 2 ) ,
CH, CH3
I I
H-[CH2CHCH2CH,]3-CH,C=CHCHzOH
3,7,11,15-tetramethylhexadec-2-en-l-ol

1.2.6. Sterols
These compounds are widely distributed in animals (zoosterols) and
in plants (phytosterols); only the most common members of this group
are mentioned here (see Sober 1968) :
cholesterol (CZ7H4,O)

HO J Y Fcholest-5-ene-3/?-01

ergosterol ( C ,,H4,0)

HO
ergosta-5,7,22-triene-3/?-01

&
stigmasrerol (C,,H4,0)

HO stigmasta-5,22-diene-3P-ol
Ch. 1 DEFINITION AND CLASSIFICATION 289

8
fl-sitosterol (CZ9H,,O)

HO stigmasta-5-ene-3P-01

1.2.7. Vitamin alcohols

The following fat-soluble vitamins are in this group :
vitamin A, (CzoH3oO)
CH20H

retinol

vitamin A, (CzoHzsO)
CH20H

dehydroretinol

ergocalciferol

cholecalciferol
Subject index p . 601
290 TECHNIQUES OF LIPIDOLOGY

vitamins E (tocopherols) - Four forms of this vitamin are known :

a-tocopherol
HO

5,7,8-trimethyltocol

8-tocopherol

5,8-dimethyltocol

y-tocopherol

CH3
7,8-dimethyltocol

&tocopherol

c H7
8-methyltocol

1.2.8. Long-chain polyols

Examples of this type of lipid are rare, but 1,14S-docosanediol has
recently been found as the disulfate ester in microorganisms (Mayers
et al. 1969), and 1,18-octadecanediol occurs in Spanish-broom. More
complex polyols, such as the ‘phthiocerols’, C34H7003 and C36H7403,
Ch. 1 DEFINITION AND CLASSIFICATION 29 1

have been isolated from Mycobacteriurn tuberculosis ; the C,,-phthio-

cerol has the structure :
OH OH OCH,
I I I
CH,(CH,),,CHCH,CH(CH,)4CHCHCH2CH3
I
CH3
3-methoxy-4-methyl-9,ll-tetratriacontanedioi

Other complex polyols found in Mycobacteria have been reviewed by

Asselineau (1966).

1.3. Long-chain amino alcohols

This class of lipids is essentially confined to derivatives or homologues
of sphingosine, a C ,,-aminodiol. These long chain bases occur largely
in complex form as ‘sphingolipids’ :ceramides, cerebrosides, sphingosyl
phosphatides, and gangliosides (see & 1.11 and 1.12). The most
common long chain bases are the following:
sphingosine CH,(CH,),,CH=CHCHCHCH20H
I I
H O NH,
4-sphingenine (2~-aminooctadec-truns-4-ene-1,3u-diol)

dihydrosphingosine CH, (CH,),,CHCHCH,OH

I I
HO NH,
sphinganine (2u-aminooctadecane-1,3o-diol)

C,,-dihydrosphingosine CH,(CH,),,CHCHCH,OH
I I
HO NH,
eicosasphinganine (2~-aminoeicosane-l,3~-diol)

phytosphingosine CH,(CH,), ,CHCHCHCH,OH

I l l
H O HO NH2
4-hydroxysphinganine (2u-aminooctadecane-1,3~,4u-triol)
Subject index p 601
292 TECHNIQUES OF ILIPIDOLOGY

C,,-phytosphingosine CH3(CH,),,CHCHCHCH20H
I l l
HO HO NH,
4-hydroxyeicosasphinganine(2o-aminoeicosane-l,3~,4~-triol)

dehydrophytosphingosine
CH3(CH,),CH=CH(CH,),CHCHCHCH20H
I l l
HO HO NH,
4-hydroxy-8-sphingenine(2~-aminooctadec-trans-8-ene-
1,3D,4D-trio])

1.4. Aldehydes
Lo& chain aldehydes occur in free form, as for example in essential
oils and insect pheromones, but also in the form of vinyl ether (alk-l-
enyl ether) analogs of glycerides and phosphatides (plasmalogens;
see & 1.10 and 1.11).

1.4.1. Normal, saturated aldehydes

These have the general formula CH,(CH,),CHO (n= 6-20 or greater);
see table 1.4 for a listing of individual aldehydes.

1.4.2. Unsaturated aldehydes (Goldfine 1964)

Normal monoenoic long-chain aldehydes usually encountered are
those analogous to the monoenoic fatty acids (see 9 1.6.6); they have
the general formula, CH3(CH2),-CH=CH(CH2),-CHO. Table 1.3
lists some common monoenoic aldehydes.

1.4.3. Cyclopropane fatty aldehydes

Aldehyde analogs of thecyclopropane fatty acids (see9 1.6.4)areknown
to occur in some bacteria (e.g., Clostridium ~ u ~ y r i c u as
m )constituents
of plasmalogens; for example C , ,-cyclopropane aldehyde (9, 10-
methylene hexadecanal) and C ,-cyclopropane aldehyde (11, 12-
methylene octadecanal).
Ch. 1 DEFINITION AND CLASSIFICATTON 293

TABLE1.4
Some normal aldehydes and ketones.

Formula Aldehydes Methyl ketones Symmetrical ketones

Octanal 2-octanone ~

Nonanal 2-nonanone Dibutyl ketone

Decanal (capricaldehyde) 2-decanone -

Hendecanal 2-hendecanone Dipentyl ketone

Dodecanal (lauraldehyde) 2-dodecanone -

Tridecanal 2-t ridecanone Dihexyl ketone

Tetradecanal 2-tetradecanone -

(myristaldehyde)
Pen tadecanal 2-pentadecanone Diheptyl ketone
(caprylone)
Hexadecanal 2-hexadecanone -

(palmitaldehyde)
Heptadecanal 2-heptadecanone Dioctyl ketone
Octadecanal 2-octadecanone -

(stearaldehyde)
Nonadecanal 2-nonadecanone Dinonyl ketone (caprone)
Eicosanal 2-eicosanone -

(arachidaldehyde)
Heneicosanal 2-heneicasanone Didecyl ketone
Docosanal 2-docosanone -

(behenaldehyde)
Tricosanal 2-tricosanone Diundecyl ketone
(laurone)
Didodecyl ketone
Ditridecyl ketone
(myristone)
Ditetradecyl ketone
Dipentadecyl ketone
(palmitone)
Dihexadecyl ketone
Diheptadecyl ketone
(stearone)

Subjecl index p . 601

294 TECHNIQUE3 OF LIPIDOLOGY

1.4.4. Isoprenoid aldehydes

Some of these aldehydes are the volatile odorous components of
plants, and are also insect pheromones (Regnier and Law 1968) e.g. :

CH3 CH3 CHO

I I /
CH,C=CH(CH,),C-CH

3,7-dirnethyl-cis-2,6-octadienal

CH3 CH3
I I
CH,C-TCH(CH,)~C=CH
I
CHO
3,7-dirnethyI-trans-2,6-octadienal

CH, CH3
I I
CH3C=CH(CH2),CHCH,CH0
3,7-dirnethyl-6-octenal

CH, CH3
I I
H-[ CH 2 C=CHCH2 1 2 -CH ,C=CHCH 0
3,7,1 l-trirnethyl-2,6,10-dodecatrienaI

CH, CH3
I I
geranylgeranial H-[CH2C=CHCH2]3-CHZC=CHCHO
(C20H320), 3,7,11,15-tetramethyl-2,6,10,14-hexadecatetraenal

1.5. Ketones and quinones

Long-chain ketones are widely distributed in nature where they occur
largely in free form.

1J.1. Methyl ketones

These have the general formula CH3(CH2),-CO-CH3 (n= 5-20 or
greater) ;see table 1.4for a listing of individual methyl ketones.
Ch. 1 DEFINITION AND CLASSIFICATION 295

1S.2. Symmetrical ketones

These ketones have the general formula [CH3(CH2),],C=0 (n= 3-20
or greater), and are of necessity odd-carbon-numbered; see table 1.4
for a listing of individual symmetrical ketones.

1.5.3. Unsaturated, branched, andlor cyclic ketones

Unsaturated symmetrical ketones are known to occur in bacteria
e.g. : A 7-palmitenone (7-hentriaconten-16-one ; C3 H,,O).
Some branched, unsaturated ketones act as insect pheromones, e.g. :

CH3
I
4-methylheptan-3-one CH,CHz-CO-CH(CH2)2CH3

CH3
I
2-methylheptan-4-one CH,CHCH,-CO-(CH,)2CH,

CH3
I
2-rnethyl-2-hepten-6-one CH3C-CH(CH2),-CO-CH3

Examples of cyclic ketones, some of which have sex pheromone

activity, are civetone (1-cycloheptadecen-10-one),and muskone (1-
methyl-cyclopentadecan-3-one).

1 S.4. Quinones with isoprenoid side chains (Sober, 1968)

The following are well-known members of this group :
uitamin K , or phylloquinone ( K )

*[4. CH3

0
2-methyl-3-phytyl-1 ,.l-oaphthoquinone
Subjccr mder p 601
296 TECHNIQUES OF LIPIDOLOGY

vitamin K , or menaquinone-n ( M K - n )

(n=6-10)

2-methyl-3-multiprenyl-
1,4-naphthoquinone
coenzyme Q , or ubiquinone-n (Q-n)

(n = 6-10)

-
1,4-benzoquinone
2,3-dimethoxy-5-methyl-6-multiprenyl-
plastoquinone-n (PQ-n)
<
c (n=6-9)

0
2,3-Dimethyl-6-multiprenyl-1,4-benzoquinone

1.6. Fatty acids

Long-chain carboxylic acids occur in many diverse forms, with varia-
tions in degree and kind of branching, number of double bonds, pre-
sence of other functional groups, and chain length. They are found
largely in esterified form, e.g. as waxes, glycerides, phosphatides, etc.

1.6.1. Normal, saturated acids

These have the general formula CH,(CH,),COOH ( n = 4 - 30 or
greater); see table 1.5 for listing of individual acids.

1 h.2. Monobranched saturated acids

Two main types are known in this group : the iso-series, with general
formula,
CH,
I
CH,CH(CH,),COOH (n= 1-20 or greater);
 TABLE1.5
Some normal and branched chain fatty acids.

Formula Normal acids Iso-acids Anteiso-acids

Pentanoic (valeric) 3-methylbutanoic 2-methylbutanoic

(isovaleric)
Hexanoic (caproic) 4-methylpentanoic 3-methylpentanoic
(isocaproic)
Heptanoic (enanthic) 5-methylhexanoic 4-methylhexanoic
Octanoic (caprylic) 6-methylheptanoic 5-methylheptanoic
(isocaprylic)
Nonanoic (pelargonic) 7-methyloctanoic 6-methyloctanoic
Decanoic (capric) 8-methylnonanoic 7-methylnonanoic
(isocapric)
Undecanoic (undecylic) 9-methyldecanoic 8-methyldecanoic
Dodecanoic (lauric) 10-methylundecanoic 9-methylundecanoic
(isolauric)
Tridecanoic 11-methyldodecanoic 10-methyl-
dodecanoic
Tetradecanoic 12-methyltridecanoic 1l-methyl-
(myristic) (isoinyristic) tridecanoic
Pentadecanoic 13-methyltetradecanoic 12-methyl-
tetradecanoic
(sarcinic)
Hexadecanoic (palmitic) 14-methylpentadecanoic 13-methyl-
(isopalmitic) pentadecanoic
Heptadecanoic 15-methylhexadecanoic 14-methyl-
(margaric) hexadecanoic
Octadecanoic (stearic) 16-methylheptadecanoic 15-methyl-
(isostearic) heptadecanoic
Nonadecanoic 17-methyloctadecanoic 16-methyl-
octadecanoic
Eicosanoic (arachidic) 18-methylnonadecanoic 17-methyl-
nonadecanoic
Docosanoic (behenic) 20-methylheneicosanoic 19-methyl-
heneicosanoic
Tetracasanoic 22-methyltricosanoic 21-methyltricosanoic
(lignoceric)
Hexacosanoic (cerotic) 24-methylpentacosanoic
Octacosanoic 26-methylheptacosanoic
(montanic)
Triacontanoic (melissic)
Dotriacontanoic (lacceroic)
298 TECHNIQUES OF LIPIDOLOGY

and the anteiso-series, with general formula,

CH3
I
CH3CH,CH(CH,),COOH (n=0-20 or greater);
see table 1.5 for listing of individual branched acids. It should be noted
that the anteiso acids have an asymmetric carbon atom and so can
exist in enantiomeric forms. The D-form is, however, usually encoun-
tered.
An example of a methyl-branched acid in which the methyl group
is centrally located in the chain is tuberculostearic acid (C, ,H3,Oz),
D- 10-methyloctadecanoic acid.

1.6.3. Multibranched acids ; saturated isoprenoids

Mycobacteria contain a series of multimethyl branched acids, for
example (see Asselineau 1966):

mycoceranic acid (C31H6202), 2,4,6,-trimethyloctacosanoic acid ;

mycocerosic acid (C32H6402)r 2,4,6.8-tetramethyloctacosanoic
acid ;
mycolipenic acid (phthienoic acid; C 2 7 H 5 2 0 2 ) r 2,4, &trimethyl-
trans-2-tetracosenoic acid.
Some examples of saturated or partially saturated isoprenoid acids
are as follows:
CH3 CH3
I I
farnesanoic acid H-[CH,CHCH,CH2],-CH,CHCHzCOOH
(cl SH3O02)3 acid
3,7,ll,-trimethyldodecanoic

CH3 CH3
I I
pristanic acid H-[CH2CHCH2CH2I3-CH2CHCOOH
(cl 9H3802)9 acid
2.6,10,14-tetramethylpentadecanoic

CH3 CH3
I I
phytanic acid H-[CH2CHCH2CH2],-CH,CHCHzCOOH
(CzOH4002)r 3,7,11,15-tetramethylhexadecanoicacid
Ch. I DEFINITION AND CLASSIFICATION 299

CH, CH,
I I
phytenic acid H-[CH,CHCH2CH2],-CH,C=CH-COOH
(C20H38 O2) 3,7,11,15-tetramethyl-2-hexadecenoicacid

1 d . 4 . Cyclopropane and cyclopropme fatty acids

Fatty acids containing a cyclopropane ring are found in some bac-
teria and plants ; they are biosynthesized by transfer of a methylene
group from S-adenosylmethionine across the double bond in a mono-
enoic acid. A few examples of cyclopropane and cyclopropene acids
are :
C17 cyclopropane CH3(CH2),CH-CH(CH,),COOH
acid (c17H3202) V
CH2
cis-9,lO-methylenehexadecanoicacid

dihydrostercu lic ucid CH, (CH,) CH-CH (CH,) C 00H

(cl 9H3602) V
CH,
cis-9,lO-methyleneoctadecanoicacid

sterculic acid CH,(CH,),C=C(CH,),COOH

(C19H3402) V
CH2
acid
cis-9,10-methylene-9-octadecenoic

lactobacillic or CH3(CH,),CH-CH(CH2)9COOH
phytomonic acid V
(c1g H 3 6 O2) CH2
c i s - I 1,12-methyleneoctadecanoicacid

1 h . 5 . Cyclopentenefatty acids
Fatty acids containing a cyclopentene ring also occur in plants, e.g. :
alepric acid (Cl4H2,O2) DCH,(CH,),COOH
acid
9-(2-cyclopentenyl)-nonano1c
Subject rndexp 601
 300 TECHNIQUES OF LlPlDOLOGY

hydnocarpic acid (C 6H280,) D C H , (CH,),COOH

11-(2-cyclopentenyl)-hendecanoicacid

chaulmoogric acid (C1 ,H3 2 0 2 ) D C H , (CH,), CO OH

13-(2-cyclopentenyl)-tridecanoicacid

gorlic acid (C18H3002) ~(CH,)6CH=CH(CH2)4COOH

acid
13-(2-cyclopentenyl)-6-tridecenoic

1 A.6. Normal, monoenoic acids

Mono-unsaturated normal acids are widespread among microorga-
nisms, plants and animals, where they occur mostly as the cis-isomer.
They have the general structure, CH3(CH2)xCH=CH(CH2)yCOOH.
Table 1.6 lists some common monoenoic acids in this series.

TABLE1.6
Some monoenoic fatty acids.
~ ~~

No. Structural formula Systematic name Common Occur-

of car- name rence
bons

H
4 CH,C=C-COOH trans-2-butenoic Crotonic Croton oil
H

6 CH,CH,CH=CHCH,COOH 3-hexenoic Dihydrosorbic

H H
10 CH,(CH2)5C=CCH2COOH cis-3-decenoic Bacteria

11 CH,=CH(CH,),COOH 10-undecenoic Undecylenic

H H
12 CH,(CH,),C=C(CH,),COOH cis-5-dodecenoic Denticetic Bacteria

H H
14 CH3(CH,),C=C(CH,),COOH cis-5-tetradecenoic Physeteric Bacteria

H H
14 CH3(CH2)3C=C(CH2)7COOH cis-9-tetradecenoic Myristoleic Bacteria,
plants
 Ch. 1 DEFINITION AND CLASSIFICATION 30 1

TABLE1.6 (continued)

No. Structural formula Systematic name Common Occur-

of car- name rence
bons
~

H
16 CH,(CH,), ,C=CCH,COOH trans-3-hexadecenoic Plants
H
H H
16 CH,(CH,),C=C(CHJ,COOH cis-9-hexadecenoic Palmi toleic Bacteria,
plants
H H
18 CH3(CH2)loC=C(CH2)4COOHcis-6-octadecenoic Petroselenic Plants
H H
18 CH, (CH &C=C(CH J,C OOH cis-9-octadecenoic Oleic Bacteria,
plants,
animals
H
18 CH,(CH2),C=C(CH,),COOH t runs-9-octadecenoic Elaidic
I H
H H
18 CH,(CH,),C=C(CH,),COOH cis- 1 I-octadecenoic cis-Vaccenic Bacteria
H H
20 CH,(CH,) C=C(CH,),COOH cis-9-eicosenoic Gadoleic

H H
22 CH,(CH2),C=C(CHZ),,COOH cis-13-docosenoic Erucic Plants
H
22 CH,(CH,),C=C(CH2), ,COOH trans-13-docosenoic Brassidic
H
H H
24 CH,(CH,),C=C(CH,),,COOH cis-15-tetracosenoic Nervonic Animals

1 h.7. Polyenoic acids

Normal unsaturated fatty acids with two or more double bonds are
widespread among plants and animals; they also occur widely among
microorganisms such as yeasts, fungi, molds, and algae, but are virtually
absent in bacteria. Table 1.7 lists some common poly-unsaturated fatty
acids.
Subject index p . 601
 TABLE
1.7
Some poly-unsaturated fatty acids.

No. of Structural formula Systematic name Common name

carbons

Dienoic Acids
6 CH,CH=CHCH=CH-COOH 2,4-hexadienoic acid Sorbic acid
H H
18 CH,(CH2)4[C=C-CHz]z-(CH,),COOH cis,ci~-9,l?-octadienolcacid
H H
18 CH,(CH2), [C=C-CH2],-(CH2),C0OH cis. cis-6,9-octadienoic acid
H H
7
U
20 CH,(CH2)4[C=C-CH2]z-(CH2),COOH cis, cis-l 1,14-eicosadienoic r

Trienoic Acids
0
H H 0
18 CH,CH, [C=CCH,],-(CH,),COOH all cis-9,12,15-octadecatrienoic a-Linolenic acid *

18 CH ,(CHz )4 [CH=CHCH2],-(CH2 ),COOH all cis-6,9,12-octadecatrienoicacid y-Linolenic acid

H H H
18 CHJCH,), [C=Cl2-C=C-(CHz),C0OH cis-9-trans-1 1-trans- 13- Eleostearic
H octadecatrienoic acid
H H
20 CH,(CH2)4[C=CCH,],-(CH2),COOH all cis-8,I 1,14-eicosatrienoic acid
H H
20 CH3CH2[C=C-CHZ],-(CH2),C00H all cis-11,14,17-eicosatrienoicacid
No. of
carbons
Structural formula Systematic name Commonname -
7

Tetra-, penta- and hexaenoic acids

H H
20 CH,(CH,),[C=CCH,],-(CH,),COOH All ci~-5,8,11,1Ceicosatetraenoic
acid Arachidonic acid

H H
20 CH3CH2[C=C-CH2],-(CH2)2CO0H All cis-5,8,11,14,I7-eicosapentaenoic
acid
H H
22 CH,CH2[C=C-CH2]5-(CH2)4COOH All cis-7,10,13,16,19-docosapentaenoic
acid
H H
22 CH,(CH,),[C=C-CH,],CH,COOH All cis-4,7,10,13,16-docosapentaenoic
acid
H H
22 CH,CH,[C=C-CH,.,-CH,COOH All cis-4,7,10,13,16,19-docosapentaenoic
acid

W
w
0
304 TECHNIQUES OF LIPDOLOGY

Isoprenoid or terpenoid fatty acids are also known, e.g.

CH3 CH, COOH
I I /
geranoic acid CH3C=CH(CH2),C=CH
(clOHI6O2) acid
3,7-dimethyl-cis-2,6-octadienoic

CH3 CH2
I I
nerolic acid CH,C=CH (CH,),C=CH
I
(cl OH 16O2) COOH
3,7-dimethyl-trans-2,6-octadienoicacid

CH3 CH3
I I
farnesoic acid H-[CH ,C=CHCH 2] 2-CH ,C=CHC 0OH
(CISH24O2) 3,7,11-trimethyl-2,6,1O-dodecatrienoicacid

1 h.8. Acetylenic acids

Fatty acids containing a triple bond are known to occur in plants, for
example :
tariric acid (C18H3202) CH,(CH~)~,CEC(CH~),COOH
6-octadecynoic acid
8-octadecynoic acid (c18H320 2 ) C H ~ ( C H ~ ) ~ C(CH2)6CO
SC OH
stearoh acid (C18H3202) CH,(CH2)7CS(CH2)7COOH
9-octadecynoic acid

1.6.9. Hydroxy acids

Hydroxy fatty acids occur as various positional isomers, and in both
saturated and unsaturated forms. The a-hydroxy saturated acids are
constituents of cerebrosides,and form a series of even-carbon acidswith
general formula, CH,(CH,),-CH(OH)COOH (n=odd, 7 - 21). Exam-
ples in this series are : 2-hydroxylauric,2-hydroxymyristic,2-hydroxy-
palmitic, 2-hydroxystearic, 2-hydroxyarachidic, 2-hydroxybehenic,
and 2-hydroxylignoceric (cerebronic) acids.
Ch. 1 DEFINITION AND CLASSIFICATION 305

j-Hydroxy saturated acids are constituents of lipopolysaccharides

in the cell walls of Gram-negative bacteria ; they also occur in yeasts
as ester-bound or glycoside-bound extracellular lipids. They form a
series of even-carbon acids with general formula, CH3(CH,),CH(OH)-
CH,COOH (n=even, 6-14).Examples in this series are: 3-hydroxy-
capric, 3-hydroxylauric73-hydroxymyristic, 3-hydroxypalmitic, and
3-hydroxystearic acids. The D-3-hydroxy acids are intermediates in
fatty acid biosynthesis and the L-3-hydroxy acids are intermediates in
the j-oxidation pathway of fatty acids.
WHydroxy acids are intermediates in w-oxidation of fatty acids by
bacteria and other microorganisms and may accumulate as extra-
cellular glycosidically-bound glycolipids. Their general formula is,
HO-CH,(CH,),-COOH (n=8- 16 or greater); some examples are,
12-hydroxylauric (sabinic), 1Chydroxymyristic, 16-hydroxypalmitic
(juniperic), and 18-hydroxystearic acids.
Some examples of unsaturated hydroxy acids are : 12-hydroxyoleic
(ricinoleic),and 2-hydroxy-15-tetracosenoic(hydroxynervonic)acids.
Polyhydroxy normal fatty acids are also known, e.g., 15,16-dihy-
doxyhexadecanoic (ustilic A), 9,10-dihydroxyoctadecanoic (9,IO-di-
hydroxysteark) and 2,15,16-trihydroxyhexadecanoic(ustilic B) acids.
Complex hydroxy, branched-chain fatty acids (mycolic acids) occur
in bacteria such as Corynebacteria, Mycobacteria and Nocardia.
Some of these are:
OH COOH
I I
corynomyculic acid CH3(CH2)l4CH-CH(CH2)1 3CH3
(c3 2 H 6 4 O3)

a-smegmamycolic acid (C79H,5403)

CH, OH COOH
I I I
CH,(CH,), 7CH=CH(CH2)1,CH=CHCH(CH2)17CH-CH-C22H,,

A more detailed description of the mycolic acids and other complex

acids has been presented by Asselineau (1966).
Sirbject index p . 601
306 TECHNIQUE OF LIPIDOLOGY

1.6.10. Keto acids

Various positional isomers of keto acids are known ;the most common
ones may be classified into the following series:
a-keto acids CH3(CH,),-CO-COOH ( n = 7 - 2 0 or greater);
P-keto acids CH3(CH2),-CO-CH,COOH
( n = 6 - 20 or greater); and
(o- 1)-keto acids CH3-CO-(CH2),-COOH ( n = 7 - 20 or greater).
4-Keto and 6-keto fatty acids are also known. An unsaturated keto
acid which has insect pheromone activity is 9-keto-2-decenoic acid.

1.6.11. Dicarboxylic acids

These acids have the general formula, HOOC-(CH2),-COOH (n=
4 - 18 or greater). Some individual members of this series are :
~~

Formula Name

C6H 1 0°4 Hexanedioic acid (adipic)

C7Hl 2O4 Heptanedioic acid (pimelic)
C8H 14O4 Octanedioic acid (suberic)
C9H 1 6 0 4 Nonanedioic acid (azelaic)
C I O H 18O4 Decanedioic acid (sebacic)
c l 2 H 2 2 O4 Dodecanedioic acid
C13H2404 Tridecanedioic acid (brassylic)
C14H2604 Tetradecanedioic acid
C16H3004 Hexadecanedioic acid

1.6.12. Prostaglandins
This group of compounds includes unsaturated hydroxy or keto-
hydroxy derivatives of the parent Cz0 cyclopentane acid, prostanoic
acid [7-(2’-octylcyc1opentyl)-heptanoicacid]. These acids are present
in seminal plasma and in accessory genital glands and have smooth
Ch. 1 DEFINITION AND CLASSIFICATION 307

muscle and vasodepressor activity. A few examples of prostaglandins

are (see Ramwell and Daniels 1969):
prosfaglandin E , (PGE,) (C20H3405)
H H H
CH,(Cc( 214 CH ( 0H ) -5=HCI o \G '
y 6 C O O H

1 1a, 1 5(S)-dihydroxy-9-oxo-13-trans-prostenoic
acid

prostaglandin F , ( P G F ,a ) (C20H3 0,)

H H H
CH3(CH2)4CH(OH)-F=C I -&~U)~
,\COOH

HO-
9a, 1 la, 15(S)-trihydroxy-l3-trans-prostenoicacid

prostaglandin E , (PGE,) (C,,H,,O,)

'
H H H H H
I I
CH,(CH,), CH(OH)-$=C e : C H2-C =C-(CH,),COOH
H HO-
1 la, 1S(S)-dihydroxy-9-oxo-5-cis-
13-trans-prostadienoicacid

prostaglandin E , (C20H3,0,)

CH3CH&=k
H H
-CH2CH(OH)-C =C
H H
I +J-- H H H
H2-c I =C I -(cH,), COOH

HO-

1 la, 1 S(S)-dihydroxy-9-oxo-5-cis-l3-trans-
17-cis-prostatrienoicacid

1.7. Waxes
Waxes are defined as fatty acid esters of fatty alcohols. They occur in
the skin surface of animals, in the cuticle of leaves and in certain bacteria
such as the Mycobacteria and Corynebacteria. There are two main
types of waxes, simple and complex.

1.7.1. Simple waxes

These are monoesters of normal fatty acids and normal primary long-
Subjerr index p. 601
308 TECHNIQUES OF LIPIDOLOGY

chain alcohols, with general formula,

0
II
CH,(CH,),C-O-CH2(CH2),CH3 (n and m= 8 - 18 or greater).

Some examples are : dodecyl hexadecanoate (lauryl palmitate),

hexadecyl hexadecanoate (cetyl palmitate), tetradecyl octadecanoate
(myristyl stearate), octadecyl octadecanoate (stearyl stearate), etc.
Unsaturated waxes are also known, e.g., hexadecyl octadec-9-enoate
(cetyl oleate), 9-octadecenyl hexadecanoate (oleyl palmitate), etc.
Preen gland waxes of birds contain esters of normal alcohols with
mono- and multibranched fatty acids (Odham 1967).

1.7.2. Complex waxes

In this type either the fatty acid or the alcohol component or both may
have complex structures. Examples of this type are the ‘cerides’ or
waxes of Mycobacterium tuberculosis which are diesters of phthiocerols
(C33- C,, branched-chain diols) with mycocerosic acids (C29-C32
branched-chain acids). A detailed review of this class of lipids has been
presented by Asselineau (1966).
Animal skin surface lipids contain diester waxes of two types: (i) a
hydroxy acid in which the hydroxyl group and the carboxyl group
are esterified, respectively, with another fatty acid and a fatty alcohol ;
and (ii)an alkane cc,j?-diolboth hydroxyls of which are esterified with a
fatty acid (Nicolaides et al. 1970).

1.8. Esters of sterols and vitamin alcohols

Sterols and vitamin alcohols commonly occur as esters of fatty acids,
the range of chain lengths and degree of unsaturation being similar to
those of fatty acid components in glycerides.

1.8.1. Sterol esters

The following are some examples of this group :
Ch. 1 DEFINITION AND CLASSIFICATION 309

cholesteryl esters

RCO=CI2-C2,, saturated and

unsaturated acyl groups
RCO-0

&
ergosteryl esters

RCO=C,2-C20,saturated and
unsaturated acyl groups

KO-0

3-0-acyl-ergosta-5,7,22-triene-3fl-ol

&
stigmasteryl esters

RCO=CI2-Czo,saturated and
unsaturated acyl groups

3-O-acyl-stigmasta-5,22-diene-3~-ol
RCO-0

&
B-sitosteryl esters

RCO=CI2-C,,, saturated and

unsaturated acyl groups

RCO-0 3-O-acyl-stigmast-5-ene-3~-ol

Subject index p . 601

310 TECHNIQUES OF LIPIDOLOGY

1.8.2. Vitamin alcohol esters

The following are some examples of this group :

vitamin A esters
CHz0CO-R RCO = C,, - C,,, saturated or
unsaturated acyl groups
acyl retinols

vitamin D esters

RCO=C,2-C20,saturated or
unsaturated acyl groups

RCO-0
acyl cholecalciferols

vitamin E esters
5H3
RCO=CI2-C,,, saturated or
RcO-O@[d]
H3C 3
/ unsaturated acyl groups
CH3
acyl a-tocopherols

1.9. Glycerides
Glycerides (neutral fats) are fatty acid esters of glycerol ;they occur in
three main types.*

* For asymmetrically substituted glycerol derivatives, the stereospecific numbering

(sn)system of nomenclature will be used to distinguish between the two primary car-
bin01 groups (see Sober 1968): If the secondary hydroxyl group in glycerol is oriented to
the left of carbon-2 in the Fischer projection, the carbon atom above carbon-2 is desig-
nated carbon-1, and the one below is designated carbon-3. The use of this stereospecific
numbering is indicated by the prefix 'sn' before the stem-name of the compound.
Ch. I DEFINITION AND CLASSIFICATION 311

1.9.1. Monoglycerides
These are fatty acid monoesters of glycerol and exist in two isomeric
forms :
u 1 CH2-0-CO-R CH20H
I I
2 HO-C-H CH-0-CO-R
I I
cx‘ 3 CH2-OH CH,-OH
sn-1- or a-isomer 2- or /I-isomeI
R =saturated or unsaturated hydrocarbon chains
1.9.2. Diglycerides
These are fatty acid diesters of glycerol and occur in two isomeric
forms :
CH2O-CO-R 3 CHZOH
I I R and R‘ =saturated
HO-C-H CH-o-Co-R
I
I or unsaturated chains
CH-0-CO-R’ 1 CH,-O-CO-R’
sn-1,3- or a.a’-isomer sn-1,2-or a,/?-isomer

1.9.3. Triglycerides
These are fatty acid triesters of glycerol; the fatty acids may be all
alike, two alike or all different. Natural triglycerides with at least two
different fatty acids groups have been found to be asymmetrically sub-
stituted.
3 CH2-0-CO-R
I
2 H-C-0-CO-R’ R, R’, and R ’= saturated
I
or unsaturated chains
1 CHZ-0-CO-R”
sn-f,2,3-tri-O-acyl glycerol

I .I 0. Glycerol ethers
Two general classes of glycerol ethers are known: aikyl ethers and
alk-1-enyl ethers (plasmalogens); see Snyder (1969a) for a detailed
review of this field.
Subjecr index p. 601
312 TECHNIQUES OF LIPIDOLOGY

1 J0.1.Alkyl ethers of glycerol

The following types of alkyl ethers may occur :
monoulkyl ethers, CH2-0-R CH2-OH
I I
HO-C-H CH-0-R
I I
CH2-OH CH2-OH
sn-1- or a-isomer 2- or j3-isomer
The a-isomers usually are found to have the D- or sn-l-configura-
tion, e.g., sn-1-0-hexadecyl glycerol (chimyl alcohol) ; sn-1-0-octa-
decyl glycerol (batyl alcohol) ;and sn- 1-0-octadec-cis-9-eny1 glycerol
(selachyl alcohol). However there is one example of an a-monoether
with the L- or sn-3 configuration, namely, sn-3-0-tetramethylhexa-
decyl glycerol (a-monophytanyl glycerol). The 2-0-tetramethyl-
hexadecyl glycerol (/3-monophytanyl glycerol) isomer also exists (Joo
et al. 1968).
Dialkyl ethers, CH2-0-R CH2-0-R
I I
R’-0-C-H CH-OH
I I
CH2-OH CH,-O-R‘
sn- 1,2- or GI,
j3-isomer 1,3- or a,cc’-isomer

The only naturally-occurring diether known so far is sn-2,3-di-0-

tetramethylhexadecyl glycerol (diphytanyl glycerol). Diethers in
which the R and R groups are alike or different, normal, saturated or
unsaturated chains have been synthesized in both 1,2- and 1,3-forms.
Trialkyl ethers, CH,O-R
I
CH-0-R R, R‘, and R” are saturated
I or unsaturated chains
CH20-R”
Triethers of glycerol have been synthesized but so far have not been
found in nature.

1J0.2. Alk-1-enyl ethers of glycerol

These aldehydo enol ethers occur naturally with the alk-1-enyl
Ch. I DEFINITION AND CLASSIFICATION 313

linkage in the cis-configuration. The monoalk-1-enyl ethers predomi-

nate but there is indirect evidence for the occurrence also of dialk-l-
enyl ethers ; tri-enol ethers have not been detected.
Mono-alk- 1-enyl ethers,
CH2-O-CH=CH-R CH2-OH
I I
HO-C-H CH-0-CH=CH-R
I I
CH,-OH CH,OH
sn- 1- or a-isomer 2- or /?-isomer
Only the sn-1-isomer has been found in nature. The aldehydo
chains are generally palmityl, stearyl, or oleyl.
Di-alk-1-enyl ethers,
CH2-O-CH=CH-R CH2-O-CH=CH-R
I I
R-HGCH-0-C-H CH--OH
I I
CH,-OH CH2-O-CH=CH-R
sn-1.2- or a, 8-isomer 1,3- or a, a‘-isomer

1 .I 0.3. Acylated alkyl ethers of glycerol

These occur as mono- and diacyl derivatives of alkyl glycerols :
CH 2- 0 - R CH2-0-R R, R’, and R”
I I
R’-CO-0-C-H R’-CO-0-C-H are saturated or
I I unsaturated
CH2-OH CHZ-0-CO-R” ,-.hains
sn-2-acyl- I-alkyl glycerol sn-2,3-diacyl-l-alkyl glycerol

1.10.4. Acylated alk-1-enyl ethers of glycerol (neutral plasmalogens)

The mono- and diacyl derivatives of alk-1-enyl ethers are known :
CH 2- 0-CH=CH -R CH -0-C H=C H-R
I I
R’-CO-O-CH R’-CO-0-CH
I I
CH2-OH CH2-0-CO-R”
1-alk-1’-enyl-2-acyl-sn-glycerol 1 -ak- 1 ’-enyl-2,3-diacyl-sn-glycerol

R, R’, and R” are saturated or unsaturated chains

Subject index p . 601
314 TECHNIQUES OF LIPIDOLOCY

1.1 1 . Phospholipids (phosphatides)

These lipids may be classified into two general groups : glycerophos-
phatides and sphingosylphosphatides.

1.I 1 .I. Glycerophosphatides

This group consists of derivatives of the parent compound 1,a-diacyl-
sn-glycerol-3-phosphate (phosphatidic acid), having the general
structure :
R'-CO-0-CH2 R', and R" are saturated and
I unsaturated chains, respectively
R"-C 0-0-CH
I
H2C-0-PO-0-R
I
0-
The R group represents amino alcohol (nitrogenous base) or
polyol residues ; some nitrogenous base-containing glycerophospha-
tides are listed in table 1.8 with their abbreviations.

TABLE 1.8
Nitrogenous base-containing glycerophosphatides.

R-group Name of phosphatide Abbre-

viation

-H Phosphatidic acid PA
-CH2CH(NH2)-COOH Phosphatidyl serine PS
(serine)
-CH2CH2NH2 Phosphatidyl ethanolamine (cephalin) PE
(ethanolamine)
-CH2CH2NH(CH3) Phosphatid y 1-N-methylethanolamine PE-Me
-CH2CH2N(CH3), Phosphatidyl-N,N-dimethylethanolamine PE-diMe
-CH,CH,N(CH,), Phosphatidyl choline (lecithin) PC
(choline)'
Ch. 1 DEFINITION AND CLASSIFICATION 315

The polyol-containing phosphatides fall into two groups :

(a) The polyglycerolphosphatides

phosphatidyl glycerol (PG)

II
O
R-C-0-CH
I I
HC- OH

1
H2C-O-
9
Y-O-CH,
1
I
-0 K f
3-sn-phosphatidyl-1'-sn-glycerol

phosphatidyl-0-aminoacyl glycerol (PG-AA)

0 NH2
I
II
R-C-O-CH2 H,C-0-co -C H-R"

II
O I
R -C-0-C H HC-OH

I
-0 K+
3-sn-phosphatidyl-l'-(3'-0-aminoacyl)-sn-g~ycerol

The aminoacyl R" groups are usually those corresponding to lysine,

ornithine, arginine or alanine.
phosphatidyl glycerophosphate (PGP)
0 0-K+
II
R-C-0-CH, H,C-OP
(I/

II
R-C-0-CH
O I
I
HC-OH
'0-K+

i i
H2C-0 -P-0 -C H
l
I
-0 K+
3-sn-phosphatidyl-1'-sn-glycerol-3'-phosphate
Sub/ecr indexp. 601
316 TECHNIQUES OF LIPIDOLOGY

diphosphatidyl glycerol (di-PG)

0 -0 K+
II 1
R-C-0-CHp H2C-O-P-O-CH2

II
R-C-0-CH
O I 1
HC-OH 8 1
H -0-C-R 8
I
H2C-O-P-O-CH2
d I R
H2C-0-c--R
I
-0'K
cardiolipin
l', 3'-di-0-(3-sn-phosphatidyl)-sn-gIycerol

(b) The phosphoinositides (for nomenclature, see Klyashchitskii et al.

1969).
phosphatidyl inositol (monophosphoinositide, PI)
0
I1
R-C-O-CHB
I

II
O I
R- C-0-C H

-0
+K

3-sn-phosphatidyl-sn-l'-rnyo-inositol
phosphatidyl inositolrnonophosphate (diphosphoinositide, DPI)
0
II
R-C-0-CH,
I

R-C-
8 0 -C H
1

3-sn-phosphatidyl-sn-1 '-rnyo-inositol-4-phosphate
Ch. 1 DEFtNITION AND CLASSIFICATION 317

phosphatidyl inositoldiphosphate (triphosphoinositide, TPI)

0
II
R-C-0-CH,
+K K+

I1
-0
\!/
O 0-

R-C-0-FH
O I I

3-sn-phosphatidyl-sn- l'-myo-inositol-45'-diphosphate

Glycerophosphatides occur ubiquitously in the fatty acyl diester

form, but variations of this form are also fairly widespread. These
variations include structures such as plasmalogens (mono-alk-1-enyl
monoacyl), monoacyl alkyl monoether, alkyl diether, monoacyl
(lyso), alkyl monoether, and phosphono forms (table 1.9).

TABLE 1.9
Variations in structures of glycerolphosphatides.

Structure* Structural form Common

phosphatides

0-
H~C-O-PO-OR Diacyl ester form All
I
HC-0-CO-R'
I
H,C-0-CO-R

0-
I
H,C-0-PO-OR Monoacyl monoalk- PS, PE, PC
I -I-enyl ether form
HC-0-CO-R' (plasmalogens)
I
H,C-0-CH=CH-R"
(continued)
Subject index p. 691
318 TECHNIQUES OF LIPIDOLOGY

TABLE1.9 (continued)
Structure* Structural form Common
phosphatides

0-
I
H,C-0-PO-OR Monoacyl monoether PE, PC
I form
HC-0-C 0-R’
I
H2C-0-CH2R”

0-
I
H2C-0-PO-OR Diether form PG, PGP
I
HC-O-CH2-R’
I
H2C-0-CH2-R’

0- 0-
I I
H2C-0-PO-OR H2C-0- P-OR Monoacyl (lyso) PS, PE, PC
I I form
HCO-OH HC-0-CO-R
I I
H2C-0-CO-R‘ HZC-OH
a-isomer p-isomer

0-
I
H2C-0-PO-R Phosphono form** PE**
I
HC- 0-CO-R’
I
H,C-0-CO-R”
~~

* R=nitrogenous base or polyol residues. R’ and R”=saturated or unsaturated fatty

acid chains.
** Liang and Rosenberg (1966).

1.I 1.2. Sphingosyl phosphatides

The commonest lipid in this class is sphingomyelin, the phosphoryl-
choline ester of an N-acyl-sphingosine (ceramide) :
Ch. 1 DEFINITION AND CLASSIFICATION 319

H H H 0-
I I 1 I -+
CH,(CH,), ,C=C-C-C-CH2O-PO-0CH2CH2+N(CH3),
I I I
H OHNH
I
CO-R
1-phosphoryIcholine
N-acyl-trans-4-sphingenine-

The corresponding derivative of dihydrosphingosine is also known :

H H 0-
I I I
CH, (CH2)l~C-C-CH20-PO-O-CH2CH2+fi(CH3)3
I I
OH NH
I
CO-R
N-acyl-sphingenine- 1-phosphorylcholine
(N-acyl-o-erythro-2-aminooctadecane- 1,3-diol-l-O-phosphorylcholine)

Recently, phosphono analogs of sphingosyl phosphatides have been

found in various invertebrates :
0-
I
CH3(CH2),2CH=CH-CH-CH-CH2-O-P-CH,CH2NH,
I 1 II
OH NH 0
I
CO-R
ceramide aminoethyl phosphonate (ceramide ciliatine)

0-
I
CH3 (CH,), 2CH=CH-CH-CH-CH2-O-P-CH,CH2NH-CH3
I I II
OH NH 0
CO-R
ceramide N-methylaminoethyl phosphonate
Subject indexp 601
320 TECHNIQUES OF LIPIDOLOGY

1.12. Glycolipids
This class of lipids includes various types of long-chain derivatives of
sugars which may be classified into the following six groups :
1.12.1. Glycosyl digEycerides
These consist of mono-, di-, or trisaccharides linked glycosidically
to the hydroxyl group of a diglyceride, e.g.:
monogaluctosyl diglyceride (MGD) (plants ; Carter et al. 1956)

3-0-~-o-galactopyranosyl-sn-
1,2-diacyl-glycerol

digalactosyl diglyceride (DGD) (plants ; Carter et al. 1956)

Hoe- O ’ 0-CH,
I
HC- 0- CO -R
I
OH OH HZC-0-CO-R

G:oL
3-O-[-cr-~-galactopyranosyl-(l’
+6’)-O-~-~-galactopyranosyl]-sn-1,2-diacyl-glycerol

gulactosyl glucosyl diglyceride (bacteria)

H o < ~ o ~ o

HO
0- C H,
OH
I
HC-0-CO-R
I
H,C- 0-CO-R
3-O-[-a-~-galactopyranosyt-( 2-diacyl-glycerol
~’~2’)-O-cc-D-glucopytanosyl]-sn-l,
Ch. 1 DEFINITION AND CLASSIFICATION 32 1

Other examples of bacterial glycosyl diglycerides are given in table

1.10.
TABLE 1.10
Some bacterial sn-3-0-glycosyl diglycerides*.

Name Structure of glycoside moiety Occurrence

Monoglucosyl diglyceride a-o-glucopyranoside Pneumococcus,

Diglucosyl diglyceride
Diglucosyl diglyceride
Dirnannosyl diglyceride
Galactofuranosy l
diglyceride
Galactosylglucosyl
dig1yceride
Glucosylgalactosyl-
glucosyl diglyceride
Mycoplasma
/h-glucopyranosyl-( 1 46)-0- Staphylococcus
8-D-glucopyranoside
a-u-glucopyranosyl-( 1-+ 2)-0- Mycoplasma,
a-o-glucopyranoside Streptococcus
a-D-rnannopyranosyl-(1+3)-0- Micrococcus
-o-mannopyranoside lysodeikticus
b-1,-galactofuranoside Mycoplasma,
Bacteroides
a-u-galactopyranosyl-( 1+2)-0- Lactobacillus
a-D-gf ucopyranoside
a-D-glucopyranosyL(1-r6)-0- Lactobacillus
a-o-galactopyranosyl-( 1-2)-
0-a-o-glucopyranoside

* Compiled from data of Brundish et al. (1966) and Shaw (1970). An alkyl glycerol
diether analog of glycosyl diglycerides has been detected in extremely halophilic bac-
teria (e.g..Halobacterium cutirubrurn); its tentative structure is sn-2,3-di-O-phytanyl- 1-0-
[galactosyl-(l+6)-O-mannosyl-l (1+2)-O-glucosyl]-glycerol (Kates et al. 1967; Kates
1972).

1 . 1 2.2. Glycosides of hydroxy fatty acids

The following are examples of this type of glycolipid :
rhamnolipid (P. aeruginosa)

0 -CHCH2 C O - 0 - CHCH, COOH

O OH OH
D OH
! I I
C7H15 C7H15
2-O-a-~-rharnnopyranosyl-a-~-rhamnopyranosyl-o-3-hydroxydecanoyl-o-3-
hydroxydecanoic acid
Subject rndex p. 601
322

C2o-y
TECHNIQUES OF LIPIDOLOGY

sophorolipid (yeasts) CH,OH

- w2 ) , , c o o H

<y
HO CH3

CH20H

HO

OH
2-O-~-o-glucopyranosyl-~-o-glucopyranosyl-~-17-hydroxyoctadecanoate

Mycosides (Mycobacteria). Glycosides of methylated sugars with a

complex aromatic long-chain glycol (‘phenoglycol B’) esterified with
mycocerosic and palmitic acids (see Asselineau 1966).
1.12.3. Fatty acid esters of sugars
Some examples of this type of glycolipid are :
triacyl glucose (Mycoplasma)
CH2O-CO- R

<-:?
R-CO =mainly oleoyl (+ linoleoyl)
<>OH OCO-R
R-CO-0

OH
3,4,6-triacyl-~-o-glucopyranose
6,6-diacyl trehaloses (Mycobacteria)
HOCER

HO
Cord factor
OH OH

R-CO = palrnitoyl, C,,H,,O, in 6,6‘-dipalmitoyl-a-~-trehalose

R-CO = mycolanoyl,C,, H I 7JOz, in 6.6’-dimycolanoyl-a-n-trehalose
R-CO = corynomycolanoyl,C 3 2 H 6 3 0 2 , in 6,6’-di-(2’-tetradecyl-3”-hydroxyocta-
decanoy1)-a-o-trehalose
Ch. 1 DEFINITION AND CLASSIFICATION 323

1 .I 2.4. Sugar-phosphatide derivatives

Examples of this type of glycolipid are :
phosphutidyl glucosuminyl glycerol (in Bacillus megaterium)

"
(Op den Kamp et al. 1969)
3-sn-phosphatidyl-l'-(2-~-glucosaminyl)-sn-glycerol
phosphatidvl inositol polpmannosides (in Mycobacteria)

[a -. r . -,,"-
3-sn-phosphatidyl-sn-1 1 -.+ 6)-poly-1>-
'-myo-inositol-2'-/l-~-mannopyranoside-6'-~-(
mannopyranoside (Lee and Ballou 1964)

(n= 1) phosphatidyl inositol dimannoside

(n= 3) phosphatidyl inositol tetramannoside
( n=4) phosphatidyl inositol pentamannoside

1.12.5. Phytoglycolipid (plants)

This substance is an N-acyl-phytosphingosyl phosphorylinositol
linked glycosidically to a complex oligosaccharide (Carter et al. 1969):
0-
I
CHJCH,), ,CH-CH-CH-CH2-O-PO-O-1'-inositol-2'-O-mannose
I l l I
OH OH NH 1"+6'
I I
CO-R glucuronic acid
I
glucosamine

galactose, arabinose, fucose

Suhlect index p. 601
324 TECHNIQUES OF I.IPIDOLOGY

1 .I 2.6. Lipopolysaccharides (Bacteria)

These are high molecular weight, polysaccharide-lipid complexes that
occur in the cell walls of Gram-negative bacteria. They consist of a
polysaccharide linked to a glycolipid (‘lipid A’) through a 3-deoxyoctu-
losonate residue. Lipid A appears to be a poly-N-acyl-glucosamine
esterified with ethanolamine phosphate and with fatty acids ; the
following is a tentative structure for Escherichia coli lipid A (see Burton
and Carter 1964; Adams et al. 1968; Adams and Singh 1970):

CHZO-PO-OCH,CHZNH, CH,O-PO-OCH,CH,NHZ

---0‘
---
-
YH
C0.R’
YH
C0.R’ 2

where R-CO- and R’CO- are mainly acetic, lauric, hydroxylauric,

myristic, hydroxymyristic, and palmitic acid chains.

1.1 2.7. Steryl ylycosides

These are 3-O-P-~-glucopyranosidederivatives of sterols, most com-
monly cholesterol, sitosterol or stigmasterol, found in plants ; the 6‘-
palmitoyl-P-u-glucoside of p-sitosterol is also known (Lepage 1964).
1-Sitosterol ylucosides

0< po&=H or C,,H,,CO-

OH

1.I 2.8. Cerebrosides (ceremide hexosides)

These compounds are glycosides of N-acyl long chain bases (cera-
mides), which occur in animals and plants.
Ch. I DEFINITION AND CLASSIFICATION 325

Brain cerebrosides

N-acyl-sphingosine-
1-P-D-galactopyranoside

where
R-CO = lignoceryl (C,,H,,O) in kerasin
cerebronyl(2-hydroxylignoceryl)in phrenosin
nervonyl (C24H450) in nervone
2-hydrox ynervonyl in oxynervone
In addition to sphingosine, dihydrosphingosine has also been found
in brain cerebrosides ; glucose-containing animal cerebrosides are
also known.
Plant cerebrosides (Kates 1970)
H H H

(--j
I l
CH3(CH2),, 7l
- C - C - - CH2-0 OH
b H b H NH
I
c=o
I O d O H
R
N-(a-hydroxy)acyl-phytosphingosine-
1-0-8-glucopyranoside

where R-CO = 2-hydroxypalmitic, 2-hydroxystearic, 2-hydroxy-

behenic, and 2-hydroxylignoceric acid chains.
In addition to phytosphingosine, other long chain bases found in
plant cerebrosides are dihydrosphingosine, and dehydrophytosphin-
gosine.
Ceramide polyhexosides. These compounds are cerebrosides in
which the glycoside portion is an oligosaccharide with 2 to 6 sugar units.
An example of a ceramide dihexoside is :
Subject index p. 601
326 TECHNIQUES OF LIPlDOLOGY

Ceramide lactoside
CH,(CH2),2
\

H/ c =c ‘CH-CH-CH,-
I 1

OH NH
I
R-C=O

I\.qf
1-0..[/-o-galactopyranosyl-( 1-+4)-O-fi-D-glucopyranosyl] N-acyl-sphingosine

1.12.9. Gangliosides (see Svennerholm 1964; Ledeen, 1966)

These are complex cerebrosides in which the ceramide residue is
linked to a glucose-galactosamine-sialic acid containing carbohy-
drate moiety ;sialic acid (P-D-N-acetylneuraminic acid) has the follow-
ing structure :
H,C-OH
Sialic acid I
H-C-OH
I
H-$-OH

Because of the variety in the carbohydrate moiety, many types of

gangliosides exist. An example of a ‘parent’ ganglioside (GMlor GI),
a monosialoganglioside, is as follows :
Gal(l-3)-GalN-acetyl(l-4)-Gal(l-4)-Glu(l- 1’)-ceramide
I
2-3
I
N-acetyl
neuraminate
Gangliosides occur in high concentrations in brain, but are also
found in other organs and in erythrocyte membranes.
Ch. 1 DEFINITION AND CLASSIFICATION 327

1.13. Sulfur-containing lipids (Haines 1971)

Three types of sulfur-containing lipids are known :

1.13.1. Long-chain alcohol sulfates (alkyl sulfates)

Only one example of this type of sulfolipid is so far known, a disulfate
ester of a long-chain diol found in microorganisms (Mayerset al. 1969):

oso;
I
CH3(CH2),-CH-(CH2)12CH2-OSO;
1,14S-docosanediol-
1,14-disulfate(8)

1 . 1 3.2. Cerebroside sulfates (sulfatides)

These compounds are usually found in brain and consist of the galac-
tosyl-3-sulfate esters of cerebrosides, e.g.,

CH3(CH2),,CH-CH-CH20-~-~-galactopyranosyl-3-sulfate
I I
OH NH
I
CO-R

1.13.3. GIycoIipid sulfates

An unusual glycolipid sulfate ester has been found in extremely
halophilic bacteria; it has the following structure (Kates et al. 1967;
Kates 1972):

- 03SO-3-Gal(1+6)-Man(1-+2)-Glu(1+ 1’)-0-CH,
I
R-0-C-H
I
R-0-CH2

Another unusual glycolipid sulfate, 2,3,6,6‘-tetra-acetyl-a,a’-treha-

lose-2’-sulfate, occurs in Mycobacteriurn tuberculosis (Goren 1970).
Subject index p. 601
328 TECHNIQUES OF LIPIDOLOGY

1.13.4. Sulfoglycosyl diglyceride (SQD) (Benson 1963)

Only one example of this type of sulfonic acid derivative is known, the
plant sulfoquinovosyl diglyceride:

H,CO -CO --R

6-sulfo-cc-~-quinovopyranosyl-(
1+3’)-1’,2’-diacyl-sn-glycerol

1 .I 4. Amino acid-containing lipids

Apart from the phosphatidyl 0-aminoacyl glycerols (see 9 1.11.1)
several classes of amino acid containing non-phospholipids are known.

1 .I 4.1. N-acyl amino acids and ester derivatives

Some examples of these are:
Serratamic acid (Cartwright 1957)
COOH
I
CO-NH-C-H
I I
CH, CH,OH
I
H-C-OH
I
(CHZ),CH 3
N-(~-3-hydroxydecanoyl)-~-serine

Siolipin A (Kawanami et al. 1968)

RlCO-NH-CH-CO-OR2
Esters of N”-acyl-L-lysine

R ,C OOH =normal and branched acids, and P-hydroxy-branched

acids ;
Ch. 1 DEFINITION AND CLASSIFICATION 329

R,-OH = long chain polyalcohols

Siolipin B refers to the series of analogous esters of N*-acyl-L-ornithine

1.1 4.2. 0-acyl carnitines

COOH
I RCOOH = n-C,,, to
CH,
I n-C,, fatty acids
RCO-O-CH-CH,-y(CH,),

1.I 4.3. Peptidolipids

These lipids occur in Mycobacteria and Nocardia species of bacteria
and have the basic structure of N-acyl oligopeptides. This class of
lipids often occurs in the form of glycoside derivatives, as for example
the glyco-petidolipid in Mycobacteria (Laneele and Asselineau 1968):
C25HS -CH-OH-CH ,CO-Phe-allo-Thr-Ala-alaninol-0-Me,rhamnose
I
0-(Ac2-6-deoxytallose)

Subjecr indexp. 601