J Sci Food A.gric 1991.

S4, 495-511
Review*
Recent Advances in Lipid Oxidation
Edwin N Frankel
6588
Department of Food Science & Technology, University of California. Davis.
California 95616. and Northern Regional Research Center. Agricultural Research Service.
US Department of Agriculture. Peoria. Illinois 61604. USA
(Received. 13 May 1990: revised version received 31 August 1990:
accepted 1 October 1990)
ABSTRACT
In a major parhway ofthe auto.r:idarion ofmethyl lino/enare. peroxyl radicals
of the inrernal hydroperoxides undergo rapid 1,J.<.:yciisarion to form
hydroperoxyepidioxides. Because linolenare h.vdroperoxides are relatively
UlfSraiJ/e. free radical amio:wianrs are much less effective in linolenate oils
than in linoleare oils. Tocopherols and carotenoids effectively inlrihi't
photosensitised oxidation of lJegetaiJ/e oils. Direct gas chromaiographic
analyses of malonaldeh.vde do nor con'elare wirh the iBA tesr. Model
fluorescence studies indicate that malona/dehyde ma.v not be so importanr
in cross/inking wirh DNA. In contrasr to oxidised methyllinoleare. oxidised
tn/inolein does not form dimers. A/though tri/inoletn oxidises with no
preference between the 1(3) and 2-triglyceride positions. the nJ double bond
of trilinolenin oxidises more in the 1(3) than in the 2-position. Synthetic
trig/ycerides oxidise in the following decreasing relative rates: LnLnL.
LnLLn. LLnL. LLLn (Ln =lino/enic and L = linoleic). io estimare the
flavour impact of 1J0iarile oxidarion products thetr relative threshold lJa/ues
musr be consideredtogether with their relative concentrarion in a givenfar.
Key words: Lipids. free radical autoxidation. hydroperoxides. photo-
sensitised oxidation. aldehydes. volatiles.linolenate. cyclisation. epidioxides.
tocopherol. carotene. antioxidants. malonaldehyde. dimerisation. tri
glycerides. trilinolein. trilinolenin. gas chromatography, stability, sensory,
flavour significance. flavour reversion. sensory assessment. aldehydes.
vegetable oils.
This revIew IS based on [he 1990 InternanonaJ Lecture addressed [0 [he SeT's Oils and Fats Grouo
In London. t 1 Apni 1990.
495
J SCI Fooa .-1qnc 0022-5142/91/$03.501.' 1991 SCI. Printed in Great Bntam
Supplied by U.S. Dept. of Agric.,
National Center for Agricultural
Utilization Research, Peoria, IL
496
INTRODUCTION
.V Frankel
Oxidation of polyunsaturated fatty acids is one of the most fundamental reactions
in lipid chemistry. Investigators working with polyunsaturated fatty acids and
lipids have to be seriously concerned with their oxidation as the products have
been implicated in so many vital biological reactions. The revival of the field of
lipid oxidation in the last 10-15 years can be attributed in large pan to the
accumulating evidence that free radicals and reactive oxygen species panicipate
in tissue injuries and in diseases. However, whether free radical species are the
cause or the effect of these diseases is a question that has been very difficult to
answer.
In the presence of initiatorS, unsaturated lipids (LH) form carbon-<:entred. alkyl
radicals (L) and peroxyl radicals (LOO-), which propagate in the presence of
oxygen by a free radical chain mechanism to form hydroperoxides (LOOH) as
the primary products of autoxidation (Franke! 1980).
LH - L
L +0% - LOa
LOO-+LH - LOOH+L
(1)
(2)
(3)
In the presence ofIig.h.t. unsaturated. fats can also form hydroperoxides by reacting
with singlet oxygen produced by sensitised photooxidation. which is a
non-free-radical process (Gollnick 1978).
Lipid hydroperoxides are readily decomposed into a wide range of carbonyl
compounds. hydrocarbons. ketones and other materials that contribute to flavour
deterioration of foods. Much work has been reported on the volatile oxidation
productS of unsaturated. lipids (Frankel 1982. 1985; Grosch (987) because they
cause rancidity in foods and cellular damage in the body. Different volatile
decomposition products are formed according to the relative thermal stabilities
of the lipid oxidation precursors and resulting carbonyl products. To evaluate the
oxidative and flavour stability of unsaturated edible oils. it is t:ssential to know
the structures of the oxidation products. how they decompose. the amounts of
volatile compounds produced. and the flavour significance of the volatiles.
A better understanding of the mechanisms of oxidation of linoleic and linolenic
acids may lead to improved. methods for control of /lavour deterioration in
vegetable oils. Several reviews of the literature have appeared (Frankel 1980. 1985.
1988; Chan 1987; Grosch 1987; Gardner 1989). The mechanism of autoxidation
of linoleic acid and esters has rea:ived special attention (Porter 1986). This paper
summarises recent progress made in understanding the mechanism by which
polyunsaturated. edible oils can undergo oxidative and /lavour deterioration.
FREE RAOlCAL AUTOXIDAnON
Linolenate' esters
Since 1961. when the isolation of pure hydroperoxides of methyl linoienate was
first reported (Frankel ec ai 1961l. considerable advances have been made in
Lipid oxidation
0-0 OOH
rvrV-<
C
1"**......., " 1 1d.

F"II I. Free raciica1 autoxidation of methyl linolenate.
497
understanding linolenate autoxidation by the application of new powerful
separation and analytical tools. The fie] unsaturation of linolenate provides a key
mechanistic feature affecting the nature of its primary and secondary oxidation
products. In the presence of free radical initiators. such as heat. metals. irradiation
or light. hydrogen transfer occurs with a suitable radical acceptor X from the
two activated doubly allytic methylene groups on carbon- (1 and carbon-14 to
form two pentadienyl radicals (Fig i). Reaction with oxygen at the end carbon
positions produces a mixture of four peroxyl radicals leading to the corresponding
conjugated dienoic 9-, 12-, 13- and 16-hydroperoxides containing an isolated
double bond. The fact that the external 9- and 16-hydroperoxides are formed in
amounts significantly higher than the internal 12- and 13-hydroperoxides has been
known for a long time (Frankel er at 1961, 1977: Chan and Levett (977). Only
recently has it been possible to explain this uneven distribution of isomeric
hydropeToxides of methyl linolenate. The peroxyl radicals of internal 12- and
13-hydropeToxides undergo rapid 1.3-eyclisation (A - B) to form five-membered
hydroperoxyepidioxides (C. Fig 1) (Coxon er ai 1981; Neff er ai 1981). This rapid
cyciisation is a major pathway which accounts for the lower concentrations of the
internal 12- and 13-hydroperoxides (25%) relative to the external 9- and 16-
hydroperoxides 150%) (Frankel er ai 1961. 1977). By adding 5
1
% :C-l:Qcopherol as
a hydrogen donor. Peers er at (1981) showed that this cyciisation was completely
inhibited. methyl Iinolenate producing an even distribution of the 9-. 12-. 13- <lnd
16-hydroperoxide isomers. A mixture of dihydroperoxides (0 and E) is formed in
smaller concentrations than the hydroperoxyepidioxides. by a reaction competing
498
0-0
_

a
E .V Frankel
0-0 OOH
- '--J '--'-./- -

o 0
V
Q

Fig 2. Formation of bicycJoendopetOltides and malonaldehyd.e from oltidised. methyl linolena!e.

with cyclisation (Neff et al 1981). Coxon et al (1984) showed that in the presence
of 10% the 9.16-dihydroperoxide (E) was formed seiectiveiy during
oxidation of methyl linolenate.
The intenned.iate free radical (B) formed after cyclisation can either cyciise again
to form bicycioendoperoxides (F), structurally reiated to the prostagiandins. or
undergo cieavage to produce maionaJdehyde (G) and to give a positive
thiobarbituric acid (TBA) test (Fig 2) (Dahl et al 1962; Pryor et al 1976). In
contrast to methyl Iinoleate (Porter 1986; Chan 1987), the cis, trans-hydroperoxides
of methyl linolenate are not readily isomerised to the crans.trans configuration.
apparently because cyclisation is favoured much more than geometric isomerisation
(Porter et al 198 I). The bicycioendoperoxides from oxidised 1inolenate were shown
by O'Connor et al (1984) to have mainly cis substituents in contrast to the natural
trans stereochemistry of the enzymically derived prostaglandins. The physiological
importance of this structural difference has not been established.
Inhibirion
Free radical autoxidation may be interrupted by several kinds of antioxidants
which can react with either chain-carrying peroxyl radicals or the alkyl radical
intermediates (Scott 1985).
LOO-+AH - LOOH+A
L'+Q- - LQ
(4)
(S)
The first class of antioxidants (AH) includes hindered phenols such as butylated
hydroxyanisole. butylated hydroxytoluene and To be effective. these
compounds must compete with the unsaturated lipid substrate (reaction 31 which
is normally present in the highest concentration. The second ciass of antioxidants
(Q.) includes quinones such as ubiquinone and 'Z-cocopheroquinone which must
compete with 0: in the fast reaction (2). These compounds may therefore only
be active in biological systems where the oxygen pressure is relatively low.
In the presence of trace amounts of transition metals. hydroperoxides are readily
L.ipid oxidation
499
decomposed to form alkoxyi radicai intermediates (La) and (LOa), which can
effectively propagate the free radical chain:
LOOH + M" - La +OH- + Mnn.,.l (6)
LOOH+M,,+l - LOa' +H'" +M" (7)
The catalytic effect of metals will be greatly enhanced in methyl linoienate because
linoienate hydroperoxides are much more readily decomposed than linoleate
hydroperoxides (Frankei 1962). In the presence of metais, the activity of free
radical-acceptor antioxidants is also significantly diminished because their
reactivity toward LO' is oniy one order of magnitude higher than that of the
unsaturated lipids (Erben-Russ er ai 1987). For these reasons, phenolic and other
antioxidants are much less effective in inhibiting the oxidation of linolenate-
containing oils, such as soya bean and rapeseed oils, than that of
linoJeate-containing oils, such as sunflower and saffiower oils.
Metal chelators act as preventive antioxidants by complexing metai ions and
thus retarding free radical formation and hydroperoxide decomposition. Because
linolenate hydroperoxides are so readily decomposed in the presence of metai
catalysts. metal chelators are particularly effective in preventing linolenate
oxidation. Metal chelators are thus more effective than phenolic antioxidants in
controlling oxidative deterioration of soya bean oil that contains linoienate
(Frankel er ai 1959). Antioxidant synergism is a process by which the antioxidant
effect of multi-component systems is reinforced. Significant synergism is generally
observed between free radical acceptor antioxidants .and metal chelators.
Antioxidant synergism is particularly important between natural tocopherols found
in soya bean oil and metal chelators. such as citric acid. which are essential to
ensure oxidative stability (Frankel er ai 1959). Another type of antioxidant
synergism is produced by reducing agents such as ascorbic acid (Frankel 1989).
PHOTOSENSITISED OXIDAnON
Linolemue esters
Oxygen becomes excited. into the singiet state by an energy transfer mechanism
from a sensitiser (such as chlorophyU) that has been exposed to light energy (Foote
1968). The resulting singlet oxygen reacts with methyllinoleate at least 1500 times
faster than normal oxygen (Rawls and Van Santen 1970) to form hydroperoxides.
The breakdown of hydroperoxides produced by singlet oxygen may go on to
initiate normal free radical autoxidation (Rawls and Van Santen 1970). Each
carbon-Qrbon double bond of the fatty acids reacts directly with singiet oxygen
by a concerted. 'ene' addition to produce hydroperoxides with a double bond
shifted to an aliylic position and isomerised to the crans contiguration (Gollnick
1978: Frankel 1980, 1982). Methyllinolenate thus forms six Isomers. 9-, lO-, [2-,
13-, 15- and 16-hydroperoxides. by singiet oxygen addition at each unsaturated
carbon. According to the ene addition mechanism an even distribution of these
isomeric hydroperoxides would be expected. However, an uneven distribution was
observed (Frankel er al 1979). The internal 10-, 12-, 13- and 15-hydroperoxide
500
Fie 3. Formation of bis-etndioxicie:s from oxidised
methyl Iinoleftate.
EN Frankel
0-0

H 00'

J 0-0

0-0
, til a't: _
K
isomers of methyl linolenate were foimd in lower concentrations than the external
9- and 16-hydroperoxide isomers. The peroxyl radicals of these internal isomeric
hydroperoxides are readily cyclised in methyl linoleate and methyl linolenate
(Mihelich 1980; Frankel et ai 1982; Neff et ai 1982) into hydroperoxyepidioxides
because they have a unique homoallylic unsaturation similar to the peroxyl radicals
of the internal hydroperoxides in autoxidised methyllinolenate (Coxon et ai 1981:
Neff er ai 1981). Although singlet oxygen participates in the formation of the
hydroperoxides. the cyclisation is a facile free radical process occurring as a side
reaction that is not photosensitised lFrankel ItC ai (982). In methyl linolenate.
serial cyciisation (H - I) produced hydroperoxy-bis-epidioxides (I - J - K) and
hydroperoxybicycioendoperoxides (F) (Neff er ai (982) (Fig 3).
IDhiDition
Tocopherol is highly reactive toward singlet oxygen and inhibits photosensitised
oxidation by both physica.Uy quenching singlet oxygen (ie by preventing activation
of oxygen into singlet oxygen) and by reacting with it to form stable products.
Other natural quenchers sucb as carotenoids protect lipids against photosensitised
oxidation by an energy transfer mechanism (Foote er ai 1970). Carotenoids can
also react with the triplet state of tbe excited sensitisers by a similar energy transfer
mechanism (Fujimori and Livingston 1957: Krinsky 1979).
In many foods carotenoids are bleached during processing. In distilled soya
bean oil esters cHocopherol was found to be more efficient than ,lJ-earotene in
inhibiting oxidation photosensitised by chlorophyll (Frankel er ai 1979). This
greater activity was attributed to the- dual effect of tocopherol in quenching and
reacting with singlet oxygen. With distilled soya bean oil esters a protective effect
for ,lJ-earotene was shown at a concentration of I g kg - l _ Later studies showed
Lipid oxidation
501
soya bean oil that contains natural tocopherols and citric acid to be adequately
protected against light oxidation by ,B-carotene at concentrations < 20 mg kg - l
(Warner and Frankel 1987). However. when soya bean oil was stored in the dark.
,B-carotene promoted peroxide development. At concentrations > 20 mg kg - l
carotenoids can produce objectionable colour and flavour. and can form secondary
products that initiate and promote free radical autoxidation.
DECOMPOSmON OF MONOHYDROPEROXlDES
Mechanism
Fragmentation of hydroperoxides occurs by homolytic and heterolytic cleavage
mechanisms (Fraiucel 1982). Homolytic ,B-scission produces alkoxyl radical
intermediates (L and M. Fig 4) that undergo further carbon-earbon splitting.
Homolytic cleavage a on one side of the aikoxy carbon forms pentane plus methyl
13-oxo-9,11-tridecadienoate from the 13-hydroperoxide of methyl linoleate. and
methyl oetanoate pius 2.4-decadiena! from the 9-hydropcroxide of methyl linoleate
(Fig 4). Homolytic cleavage b forms hexana! and methyl 9-oxononanoate from
the respective 13- and 9-hydropcroxides of methyllinoleate. Under acid conditions,
heterolysis produces ether carbocation intermediates (N and O. Fig 4) which cleave
selectively to form the same products as those of the homolytic pathway b. namely
hcxana1 and methyl 9-oxononanoate (Frankel er aJ 1984) (Fig 4).
The literature is not clear on the effect of antioxidants on the decomposition of
hydroperoxides. In one study. ::-tocopherol and butylated hydroxyanisole changed.
the carbonyl products formed from the 9-hydroperoxide of linoleic acid
decomposed with copper but not from the corresponding 13-hydroperoxide isomer
(Grosch er at 1981). In another study, ,;-tocopherol promoted the formation of
Fig "'- Homolytic and heterolytic scission
mechanisms for the decomposition of hydro.
perOXides.
502 E .V Frankel
..... 9-OiOIU...1I
, ODH
v==v=v;+1R
'"Me OctllllMa
HOG
Fie S. Main volatile decomposition productS of linolenate
hyciroperoxides. Pr..-I
dienals that produce fishy flavours in the copper-atalysed oxidation of butterfat
(Swoboda and Peers 1971). Recently a;-tocopherol and were
investigated to determine how they ati'ect the relative amounts of thermal
decomposition products formed from linoleate hydroperoxides (Frankel and
Gardner 1989). These hydrogen-donor compounds diminished the relative
percentages of pentane and methyl octanoate and increased the relative percentages
of hexanal and methyl 9-oxononanoate. This effect of 2-tocopherol and
1.4-cyclohexadiene was explained by their inhibition of homolytic .B-scission of an
alkoxyl radical intermediate (cleavage a. Fig 4). and promotion of heterolytic
cleavage (Fig 4).
Significant differences were found between the composition of products from
Iinolenate hydroperoxides decomposed thermaHy at 150C and catalytically with
ferric chloride and ascorbic acid (Frankel er ai 198Th). Figure 5 shows the main
volatile compounds expected from the 9-. 12-. 13- and 16-hydroperoxide isomers
of methyl 1inolenate. Thermal decomposition produced more methyl octanoate
and 2.4,7-decatrienal. and less 2.4-heptadienai, methyl 9-oxononanoate and
propanal.. than catalytic decomposition. Aithough these products represent a: small
portion of the total decomposition materials (7'4% by thermal decomposition and
2'1 % by catalytic decomposition). they have an important impact on the ITavour
a.nd biological effects of lipid oxidation (Frankel [982. (988).
Malonajdebyde fonnation
Malonaldehyde (G. Fig 2) has been assumed to be an important lipid oxidation
product in foods and biological systems but many studies in the literature have
been based on the non-specific TBA test. To determine maionaldehyde more
detinitively, a GC procedure was developed based on the stable acetal derivatives
formed under mild acid conditions (Frankel and Neff (983). Tn dilute
Lipid oxidation 503
HCl/methanol. hydroperoxides are readily cleaved to the diacetal derivatives and
maionaidehyde is converted to the tetramethyl acetals. This acid decomposition-
acetaiation procedure was used to study how much malonaldehyde is formed from
various primary and secondary lipid oxidation products.
As expected. the five-membered hydroperoxyepidioxides of methyl linolenate
(compound C. Fig 2) provided rich sources of maionaidehyde (Frankel and Neff
1983). The bicycloendoperoxides of methyl linolenate (compound F. Fig 2) were
also good sources of maionaidehyde. as predicted in the literature (Dahl ec ai
1962; Pryor et ai (916). The bis-epidioxides of methyl linolenate (compound K.
Fig 3) and the mono-epidioxides of methyl linoleate. oxidised with singlet oxygen.
were better sources of maionaidehyde than the bicycloendoperoxides of methyl
linolenate. There was. however. no correlation between the TBA values and the
amounts of malonaidehyde found by the GC procedure. The 10.11- and
13.15-dihydroperoxides and 9.12- and 13.16-<iihydroperoxides. from methyl
linolenate oxidised with singlet oxygen. were important precursors of
maionaidehyde. As expected. the 9.16- and lO.16-dihydroperoxides did not form
any maionaidehyde as measured by the GC method. On the other hand. high
values were obtained by the TBA test for ail the dihydroperoxides. From the lack
of correlation between the direct GC anaiyses for maionaldehyde and the TBA
test. Frankel and Neff (1983) concluded that the importance of malonaldehyde
may have been exaggerated in the literature.
The interactions between lipid oxidation products. DNA. metals and reducing
agents were investigated by determining the fluorescence formed in a model system
(Fujimoto et ai 1984). Hydroperoxyepidioxides (e. Fig 1l. hydroperoxy-
bicycloendoperoxides (F. Fig 2). dihydroperoxides (D and E. Fig [) and
hydroperoxy-bis-epidioxides (K. Fig 3) from oxidised methyl linolenate were all
rich sources of DNA fluorescence in the presence of iron and ascorbic acid.
Unsaturated aldehydes were much less active than their corresponding precursors
methyl linolenate hydroperoxides in forming DNA fluorescence in the presence
of iron and ascorbic acid (Frankel ItC ai 1981al. In the presence of DNA. metals
and reducing agents. maionaldehyde produced very little or no fluorescence and
the TBA test did not correlate with fluorescence formation. Therefore.
malonaldehyde may not be so important in its crosslinking properties with DNA.
A rapid headspace capillary GC method was recently developed to determine
hexanal as an important volatile product of n-6 polyunsaturated lipid oxidation
in rat liver samples (Frankel Itt ai 1989). Total volatiles were also determined by
this method as a measure of total lipid oxidation. This rapid and convenient
method is a more direct measure of lipid oxidation than the TBA test. which is
non-specific and subject to interference by many substances (Slater 1984l.
OIMERISATION OF HYDROPEROXIDES
Peroxide-linked dimers were identified during the initial autoxidation of methyl
linoleate at room temperature (Miyashita ec ai [98a.b I. Peroxide or ether dimers
isolated from methyllinoleate hydroperoxides were composed of unsaturated fatty
504
F"II 6. Thermal decomposition of methyl
linolenate dialers.
.V Frankel
ester units containing hydroperoxy, hydroxy and oxo groups (Miyashita et ai
1985). By gel permeation chromatography analyses before and after sodium
borohydride reduction. peroxide dimers were identified as main produet5 from
methyllinoleate and methyllinolenate autoxidised at 40C (Neff et ai 1988). The
dimers formed at 1S0C were entirely ether 0' carbon-carbon linked. Dipters
formed in the presence of ferric chloride and ascorbic acid consisted of both types
oflin.lcage. Other dimers from hydroperoxyepidioxides and dihydroperoxides were
mainly peroxidic in nature.
Significant differences were found between the volatile produet5 from thermal
and catalytic decomposition of monomers and corresponding dimers from oxidised
linolenate (Frankel et ai 1988). Major volatile decomposition products expected
from dimer structures P and Q are shown in Fig 6. Cleavage between the peroxide
link and the olefinic side of the 9- and 16-hydroperoxide groups produces methyl
9-oxononanoate, which is the most substantial thermal volatile decomposition
product. C1eavages on the opposite side of the peroxide links form methyl octanoate
on one side and propanal on the other side of the first monomer unit of dimer P
(Fig 6). Dimer Q undergoes cleavage on the right to produce methyl
9-oxononanoate and methyl octanoate and cleavage on the left to produce
propanal.
TRIGLYCERIDE AUTOXIDATION
Trilinoiein and trilinoienin were used as models for oxidation studies of vegetable
oil trig.lycerides (Frankel et ai 1990: Neff et ai (990). The main autoxidation
products from trilinolein were identified as mono-, bis- and tris-hydroperoxides
which are formed by sequential oxygen addition. The mono-hydroperoxides were
further oxidised to prodUce a mixture of I.J- and 1.2-bis-hydroperoxides. which
were also oxidised to tris-hydroperoxides (Fig 7). The hydroperoxides were
composed of a mixture of cis,trans- and crans,rrans-9- and -13-isomers. The
Lipid o:cidalion 505
2-Monoo
-
l.{l.OOH . l.{l.
l. 1.00101
l-llnO- 3-MI:lrIo-

1
0
2
,

1.2-811- 1.3-811-
I"",,*OIWCIiiidIe
F"11 1. Mcchani.sm of trilinolein autoxidation.



I.,......,. ..
1.2... 1.3-811-
IIjOQIJWCilCIIII
10
2
,
--{:
:m..
1olydI0iM' _
F"11 8. Mechanism of trilinolenin autoxidation.
triglyceride position of monohydroperoxides was determined by HPLC and by
pancreatic lipolysis. The ratios of the 9- and I3-1inoleate hydroperoxides in the
1(3)- relative to the 2-triglyceride position averaged a value of 2. Therefore. the
oxidation of trilinolein had no positional preference between the 1(3). and
2-triglyceride positions (Neff er a1 1990).
Trilinoienin produced. on autoxidation. 1(3)- and 2-monohydroperoxides.
1.3- and 1.2-bis-hydroperoxides and tris-hydroperoxides by sequential oxidation
(Fig 8) (Frankel er at 1990). However. in addition to hydroperoxidcs. trilinolcnin
produced significant amounts ofhydroperoxyepidioxidcs formed by 1.3-cyclisation
(Fig 1). The isomeric composition was the same as that of methyl linolenate
(Frankel 1980), 9-. 12-. 13- and 16-hydroperoxidcs. The cyclic peroxides were
mixtures of 9- and 16-hydroperoxyepidioxidcs. By HPLC the ratio of the
cis,trans 16-linoienate hydroperoxide in the 1(3)- relative to the 2-triglyceride
position was found to be higher (2'3) than that for the corresponding cis.lrans
9-linoienate hydroperoxides (l8). This evidence supports the small preferential
oxygen attack of the n-3 double bond oflinolenate in the 1(3 )-triglyceride positions.
rn contraSt co methyl linoleate and its hydroperoxides. which form SIgnificant
amounts of dimers (Miyashita e! ai 1982a.b. 1984. 1985). no evidence was found
for dimer formation in highly oxidised trilinoiein (NetT e! at 1990). Also. no dimer
formation was found when che purified monohydroperoxides of crilinolein were
506 tv Frankel
further oxidised. Dimerisation is evidently significant only in the methyl esters of
unsaturated fatty acids because intermolecular condensations of peroxyl radicals
are favoured. On the other hand. further oxidation of the monohydroperoxides
of trilinolein to bis- and tris-hydroperoxides is apparently the preferred reaction.
fntramolecular hydrogen abstraction from the linoleoyl residues can evidently
occur more favourably than intermolecular condensation of the peroxyl radicals to
form dimers. No evidence was found for dimerisation of tris-hydroperoxides. This
work therefore demonstrates that simple esters of unsaturated fatty acids do not
aecessarily provide valid models for the oxidative dimerisation of unsaturated
triglycerides.
Autoxidation of synthetic triglycerides containing linoleate and linolenate in
different known positions formed monohydroperoxides and hydroperoxy-
epidioxides as the main products (Miyashita er ai 1990). By reversed phase HPLC
the linolenate triglyceride components were found to be oxidised twice as much as
the linoleate components. However. the relative triglyceride positions of the
linolenate components had no influence on the rates of cyclisation of their internal
12- and 13-monohydroperoxides. LaLaL oxidised faster than LaLLa (L =
linoleate. La = linolenate) and LLaL oxidised faster than the corresponding LLLa.
The easier interactions between the two linolenoyl residues in LaLnL may explain
its lower oxidative stability than LaLLa. On the other hand. the easier interactions
between linolenoyl and linoleoyl residues in LLaL may explain its lower oxidative
stability than LLLn.
FLAVOUR SIGNIFICANCE OF VOLATILES
The genesis of volatile lipid oxidation products. their flavour and their biological
significance were reviewed previously (Frankel 1980. 1982). The types of flavour
imparted by lipid oxidation in foods is extremely difficult to assess because there
is wide variation in the sensory impact of different volatile products. in the methods
used for their determination and in the vocabulary used by taste or odour panels
to describe their defects.
Gas clII'omatographic: methods
Three commonly used capillary GC methods were compared. to determine volatile
oxidation compounds in vegetable oils (Snyder er ai 1988). Each method produced
different volatile profiles with oxidised soya bean oil. The weighted percentages
of each volatile were calculated in Table 1 on the basis of l-octen-3-ol which has
the lowest threshold value (Forss 1972) (defined as the lowest concentration of a
compound that a pane! can detect). By the direct injection method.
crans.cis-2.4-decadienal was the most flavour significant followed by crans.crans-
2.4-<iecadienal. l-octen-3-o1. crans.crans-?.4-heptadienal. hexanal and crans.cis-2A-
heptadienal. 2-Pentylfuran ranked tenth in importance. and pentane had the least
t1avour significance. By the dynamic headspace method. crans.cis-2.4-<iecadienal
was also the most t1avour significant. followed by crans.rrans-2.4-<iecadienai.
crans.cis-2.4-<iecadienal. l-octen-3-o1. hexanal and crans.cis-2A-heptadienai. By the
Lipid o.:cUUuiol'l
507
TABLE 1
Flavour significance of volatiles in oxidised soya bean oil
G
Major volatiles TH Rei % Weighted %b Relative order
values
Df DHS SHS Df DHS SHS Df DHS SHS
t,l-2.4-Decadicna1 O'lO 46-9 4O'S 0'3 47 4'[ 0-03 2 2 7
t.c- 2.4-Decadicna1 0-02 238 21'S
[-()
[,1'9 to8
(}S
I I 3
t,l-2.4-Hcptadicna1
(}04
65 133
2-{) . ['6
3'3
(}5 3 3 3
t-2-Hcptcna1 0'20 31 67 8'3 () 16 (}33 0'4 7 7 S
t.c-2.4- Hcptadienal O'lO H 5'4 25 (}31 (}S4
0'25 6 6 6
,,-Hexanal 0-{)8 69 5'4 247 0'86 (}68 3-[ 5 5 2
,,-Pentane 340 48 37 38'6
(}[4
C
(}II
C
I'[C [ 1 lO 10
t-2-Pentenal 100 19 1'4 12 0-02 0-01 0-01 9 8 8
I-O<:ten-3-o1 0-01 14 1'1 0'3 14 11 0'3
4 4 4
2-Pentylfuran 2-00 12
I-{) o-S
60QC 60QC 25
c
10 9 9
,,-Pt'opanal
(}{)6
05 20'6 0-08 3-4 8 I
GTH::athre:shold values (Forss 1972). DI=direa injection. DHS=dynamic headspace.
SHS::&static headspac:. t,l-::a t7'altSPaltS-. t.c - t7'altS.cis-.
b Calculated on the basis of I-octen-3-o1 which has the lowest threshold value.
C x lO-.o.
static headspac: method.. propanal was the most important flavour volatile
followed by hexanal. crallS.cis-2.4-decadienal and cl'aIIS.cis-2.4-heptadicnal.
Therefore. the amounts of each volatile compound found varied according to the
method. used. To estimate the flavour impact of volatile oxidation products. not
only their relative concentration in a given fat must be known. but also their
relative threshold values.
A GC sniffing procedure was recently employed by Ullrich and Grosch (1987.
1988a.b) and Guth and Grosch (1989) to assess the flavour impact of volatiles in
oxidised fatty acids. esters and soya bean oil by an aroma extract dilution analysis.
The most potent flavour volatiles found in oxidised linoleic acid included hexanal.
cis-2-octenal. cl'aIIS-2-nonenal. l-octen-3-o1 and l-octen-3-one (Ullrich and Grosch
1987). The relative contribution of these volatiles depended on the level of
oxidation. with crallS-Z-nonenal being most potent after 24 h oxidation. and
hexanal. Z.4-nonadienal and cis-2-octenal being produced in greater amounts after
48 and 72 h oxidation. The most significant volatile compounds found in oxidised
methyl linolenate included crallS.cis-2.6-nonadienal. l.cis-S-oetadien-3-one.
crallS.cis-3.'s-oetadien-Z-one and cis-3-hexenal (Ullrich and Grosch 1988a).
'Reverted' soya bean oil is defined as having a characteristic flavour defect occurring
at low oxidation levels. usually below a peroxide value of 10 (Frankel 1980). The
most flavour potent volatiles found in a .reverted' soya bean oil induded
cis-3-hexena!. octana!. l-octen-J-one. l.cis-5-octadien-J-one. nonana!. crans-Z-
nonena!. cis-2-nonena!. cis-J-nonena! and crans-2.cis-6-nonadienal (Ullrich and
Grosch 1988bI. In this study the "reverted' soya bean oil was prepared by storage
at room temperature under diffused daylight and the volatiles were concentrated
by distillation at 50C prior to capillary GC and sniffing at the GC exit port. fn
508
N Frankel
a later study by the same group, nonan-2.4-dione and 3-methyl-nonan-2.4-dione
were identified in a 'reverted' soya bean oil that had been stored at 21-23"C under
a northern light exposure (Guth and Grosch 1989), Although these studies provide
important qualitative data on the flavour impact of certain volatile compounds
in unsaturated fats. they are difficult to compare with other studies in the literature
because of the complexity of flavour formation in different unsaturated oils oxidised
under different conditions and analysed by different methods. Under the conditions
of direct injeaion (Snyder et al 1988) and dynamic headspace (Selke and Frankel
1987) capillary GC, the volatile profiles included only four of the potent compounds
reponed by Ullrich and Grosch (1988b) (2-/3-hexenal. octanal. nonanal and
2-nonenal) in soya bean oil stored at room temperature in the dark. However,
these results on major volatiles that can be readily determined quantitatively by
capillary GC cannot be related to the results of UHrich and Grosch (1987, 1988a.b)
and Guth and Grosch (1989. 1990) until an estimate of the concentration of the
flavour-intensive volatile compounds found in soya bean oil can be made.
Sensory assessment
Because of the subjeaive nature of panel testing there is much variation in the
vocabulary used in the literature by different workers to describe a given volatile
compound. The conditions used for storage are also critical in the assessment of
the impact of tlavour compounds formed in vegetable oils. In a recent study Warner
et a1 (1989) compared the flavour stability of different vegetable oils. Soya bean
oil after storage iIi the dark at 60C was described by a taste panel as grassy and
beany. and low-erucic rapeseed oil as characteristic of cabbage and sulphur
flavours; both oils after exposure to intense light were described as grassy, sour,
metallic or buttery. In a similar study by Guth and Grosch (1990) soya bean oil
after storage for 30 days at room temperature in daylight was described as strawy,
lard-like. beany, green, hay-like. buttery and fatty, and rapeseed oil as green.
strawy and fatty.
The diversity of sensory vocabulary used by different investigators to describe
the same flavour defect in an edible oil has led to controversy as to what individual
product or mixture of volatile oxidation products causes the so-<;ajled 'reverted'
flavour in soya bean oil. Clearly, a greater understanding of flavour development
in oxidised lipids is needed. Future progress in this area will require for the
analytical chemist to work more closely with the sensory investigators to correlate
qualitative and quantitative flavour analyses with p r o v ~ taste panel techniques
using commonly agreed terms to describe flavours and odours from oils that have
been stored under the same conditions.
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