338 SepLember 2013, vol.

24 (8)
A c|ass|c examo|e o syneg|st|c ant|ox|-
oant |nteacnons can be obseveo between
-tocoo|eo| (v|tam|n L) ano ascob|c
ac|o (v|tam|n C) |n ce|| membanes. 1||s
syneg|sm |s oue to annox|oant egen-
eanon" v|a t|e ecyc||ng o a stonge
(o|may) ant|ox|oant w|t| a wea|e
(seconoay) annox|oant. uy t||s o|nc|-
o|e, t|e o|may annox|oant (-tocoo|-
eo|) scavenges ee ao|ca|s |n t|e ||o|o
o|ase ano |s t|en egeneateo by t|e
seconoay annox|oant (ascob|c ac|o).
lom a t|emooynam|c oo|nt o v|ew,
t|e eacnon |s avoab|e because t|e eouc-
non ootenna| o ascob|c ac|o |s muc| |owe
t|an -tocoo|eo|, |no|canng t|at ascob|c
ac|o oees to g|ve an e|ecton to ox|o|zeo
-tocoo|eo|. lom a o|ys|ca| oooety v|ew-
oo|nt, t||s eacnon |s a|so avoab|e because
o-tocoo|eo| |s |n t|e membane w|ee |t can
|nacnvate ||o|o ao|ca|s ano yet |s a|so at t|e
membane |nteace w|ee |t can eact w|t|
ascob|c ac|o.
1CCCnLkCL AND kCSMAkINIC
ACID LS1LkS IN CILINWA1Lk (C]W)
LMULSICNS
Whereas -tocopherol and ascorbic acid work synergistically in
biological membranes, this is not always the case in food systems
where lipids can be in the form of triacylglycerols and exist as
emulsion droplets. In addition, the range of antioxidant that could
1be cooceptool moJel sbowloq tbe lofoeoce of pbyslcol locouoos oo tbe oblllty of oouoxlJoots socb os olkyl estets of tosmo-
tlolc oclJ ooJ -tocopbetol to lotetoct lo oll-lo-wotet (O/w) emolsloos. losslblllues of lotetocuoos of oouoxlJoots wltb -tocopbetol
ot tbe O/w lotetfoce locloJe (lef) blqb polotlty, (ceotet) meJlom polotlty, ooJ (tlqbt) low polotlty oouoxlJoots.
Mechan|sms of annox|dant |nteracnons
|n o||-|n-wate emu|s|ons: t|e |nfuence o t|e|
o|ys|ca| |ocanons ano env|onmenta| ol
Ankorn anya, kennun k|mpongp|uaya,
M|cka| Laguerre, Chr|ste||e 8ayrasy,
Irme Lecomte, |erre V|||eneuve,
D. Iu||an McC|ements, and Lr|c A. Decker
S

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SepLember 2013, vol. 24 (8) 339
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interact synergistically is limited because most natural antioxi-
dants have higher reduction potentials than -tocopherol. Tere-
fore, in foods it is not well understood if natural antioxidants can
regenerate - tocopherol or if - tocopherol is actually a second-
ary antioxidant that regenerates other antioxidants to produce
synergistic effects. In addition, it is not well understood how
physical locations of antioxidants in O/W emulsions infuence
antioxidant regeneration.
To address these issues, rosmarinic acid and its esters
provide an excellent tool for analyzing the infuence how physi-
cal locations can impact interactions with -tocopherol because
rosmarinic acid can be esterifed with faty acids to alter polarity
and thus physical location in O/W emulsions. Rosmarinic acid
and its esters have higher oxidation peak potentials (approxi-
mately 1115 mV) than -tocopherol (635 mV). Moreover, 90%
of rosmarinic acid (R0) partitions in the aqueous phase, whereas
butyl (R4), dodecyl (R12) and eicosyl (R20) rosmarinate esters
as well as -tocopherol are highly associated with the emulsion
droplets (approximately 29% in the aqueous phase). A concep-
tual model is shown in Figure 1 of how these diferences in physi-
cal location could impact interactions between -tocopherol and
rosmarinic acid and its alkyl esters used O/W emulsions.
SNLkGIS1IC, ADDI1IVL, Ck
AN1AGCNIS1IC LIILC1S?
In the absence of -tocopherol, rosmarinic acid and its alkyl esters
exhibited nonlinear antioxidant activity, the so-called cut-of efect
as shown in Figure 2 (page 540). Te combination of -tocopherol
(T) with R0 showed strong synergistic activity (synergistic index
= 5.25) as calculated from the induction period of hexanal forma-
tion in O/W emulsions. R4 and R12 showed slight synergistic
effects (synergistic index = 1.92.0). However, an antagonistic
efect was observed with combinations of -tocopherol with R20
(synergistic index = 0.3). Also, the synergistic interactions between
-tocopherol and the alkyl esters of rosmarinic acid were pH depen-
dent. Under acidic conditions (pH 35), synergistic interaction
between -tocopherol and rosmarinic acid decreased to an additive
efect. For example, the synergistic index of T + R0 at pH 3 was 1.2.
Tis highlights the problem that predicting synergistic interactions
between antioxidants in food systems is difcult and may involve
more than just electron-transfer mechanisms.
LVIDLNCL CI AN1ICkIDAN1
IN1LkAC1ICNS MLCnANISMS
Front-face fuorescence quenching and electron parametric reso-
nance techniques are two techniques that can be used to analyze
direct physical interactions and direct electron transfer among
the antioxidants. Figure 3 (page 540) shows that rosmarinic acid
(mostly dispersed in aqueous phase) highly interacted with and/
or regenerated -tocopherol at the interface of emulsion droplets
more than with the other rosmarinate alkyl esters. In addition,
the regeneration efficiency of rosmarinic acid decreased with
decreasing pH, which could explain why synergistic activity also
decreased under acidic conditions.
CCn1lnuLu Cn ACL 341
340 SepLember 2013, vol. 24 (8)
loJocuoo petloJs of tosmotlolc oclJ ooJ lts olkyl estets lo tbe pteseoce ot tbe obseoce of -tocopbetol.
llootesceoce poeocbloq coostoots (footesceoce stoJy) ooJ teqeoetouoo e[cleocles (electtoo potomoq-
oeuc stoJy, lk) of tbe copoclty of tosmotlolc oclJ ooJ lts olkyl estets to pbyslcolly lotetoct wltb ooJ to teqeoetote
-tocopbetol lo O/w emolsloos, tespecuvely. 1be losett fqote sbows cbooqes lo teqeoetouoo e[cleocles betweeo
-tocopbetol ooJ tosmotlolc oclJ ooJet Jl[eteot pn cooJluoos.
SepLember 2013, vol. 24 (8) 341
Another way to study antioxidant interactions is to look at
the kinetics of their decay during storage of O/W emulsions. In
this method, if one antioxidant is regenerating another then one
would expect to see one of the antioxidants decompose faster
than the other. Interestingly, a signifcant reduction of the oxi-
dation rate of -tocopherol was observed in the presence of
rosmarinic acid, but this did not seem to be only due to regener-
ation of -tocopherol, as rosmarinic acid degradation resulted in
the formation of cafeic acid (a part of the rosmarinic acid mol- the formation of caf the formation of caf
ecule). Since cafeic acid also has antioxidant activity in O/W
emulsions, it is possible that the formation of this second anti-
oxidant is responsible for the observed synergistic activity. Tus, oxidant is responsible for the observed synergistic activity. T oxidant is responsible for the observed synergistic activity. T
synergistic antioxidant interaction cannot be explained solely
by electron-transfer mechanism; the complexity of several other
mechanisms, antioxidants, and their oxidation products must be
considered as well. Overall, this study shows the importance of
understanding antioxidant interactions in complex food systems
so that beter predictions can be made on which antioxidant com-
bination could result in synergistic inhibition of lipid oxidation.
Aukoto looyo ls o teseotcbet ot tbe looJ 8lotecbooloqy lobo- Au Au
tototy, Nouoool ceotet fot Ceoeuc oqloeetloq ooJ 8lotecb- tototy, Nou tototy, Nou
ooloqy (8lO1c), 11J 1bollooJ 5cleoce lotk, kblooq Noeoq,
klooq loooq, lotbom 1bool, 1bollooJ, 12120. ne coo be coo-
tocteJ ot Aukoto.pooblotec.ot.tb. tocteJ ot Au tocteJ ot Au
keuooo klm keu keu pooqplu ooo klm ooo klm oyo ls o Joctotol stoJeot, u. Iolloo pooqplu pooqplu
Mcclemeots ls o ltofessot ooJ letqosoo clyJesJole oJoweJ
cbolt, ooJ tlc uecket ls ptofessot ooJ beoJ of tbe uepott-
meot of looJ 5cleoce, uolvetslty of Mossocbosetts, cbe-
oowetb lobotototy.
Mlckoel loqoette, cbtlstelle 8oytosy, Itme lecomte,
ooJ llette vllleoeove ote teseotcbets ot clkAu lo Mootpel-
llet, ltooce.

- uecker, L.A., 8. Chen, A. anya, and 8.!. Lllas, under-
sLandlng anuoxldanL mechanlsms ln prevenu sLandlng anu sLandlng anu ng oxl- oxldanL mechanlsms ln prevenu oxldanL mechanlsms ln prevenu
dauon ln foods ln dau dau OxlJouoo lo looJs ooJ 8evetoqes OxlJou OxlJou
ooJ AouoxlJoot Appllcou ooJ Aou ooJ Aou oos oxlJoot Appllcou oxlJoot Appllcou , edlLed by L.A. uecker,
8.!. Lllas, and u.!. McClemenLs, Woodhead ubllsh-
lng, CreaL AblngLon, Cambrldge, unlLed klngdom,
2010, pp.223-248.
- anya, A., M. Laguerre, C. 8ayrasy, !. LecomLe, .
vllleneuve, u.!. McClemenLs, and L.A. uecker, An
lnvesugauon of Lhe versau lnvesu lnvesu le anu gauon of Lhe versau gauon of Lhe versau oxldanL mecha- le anu le anu
nlsms of acuon of rosmarlnaLe alkyl esLers ln oll-ln-
waLer emulslons, I. Aqtlc. looJ cbem. 60:2692-2700
(2012).
- anya, A., k. klLLlpongplLLaya, M. Laguerre, C.
8ayrasy, !. LecomLe, . vllleneuve, u.!. McClemenLs,
and L.A. uecker, lnLeracuons beLween o-Locopherol lnLeracu lnLeracu
and rosmarlnlc acld and lLs alkyl esLers ln emulslons:
synerglsuc, addlu synerglsu synerglsu ve or anLagonlsu c, addlu c, addlu c e ve or anLagonlsu ve or anLagonlsu ecL? c e c e I. Aqtlc.
looJ cbem. 60:10320-10330 (2012).
342 SepLember 2013, vol. 24 (8)

1be fotmouoo of llplJ
byJtopetoxlJes (block
lloe) ooJ tbe Jepleuoo of
o-toc (o-tocopbetol, bloe
lloe) lo bolk olqol oll lo
tbe pteseoce of J00 M
oscotblc oclJ, 500 M
o-tocopbetol, ot oouoxl-
Joots combloeJ (J00 M
oscotblc oclJ - 500 M
o-tocopbetol) ot 45c.

lnteest |as been gow|ng |n consum|ng |ong-

c|a|n oo|yunsatuateo aty ac|os (LC-lblA)
ow|ng to t|e| oom|s|ng |ea|t| benets, suc|
as t|e |n||b|t|on o cao|ovascu|a o|sease,
cance, ano |nfammatoy o|sease (see lofotm
24:/-11, 2013). L|cosaoentaeno|c ac|o (LlA)
ano oocosa|exaeno|c ac|o (LlA), t|e two
o|nc|oa| LC-lblA account|ng o t|e outa-
nve benets, |ave been w|oe|y useo o ooo
ano beveage ot||cat|on. Ma|ne |s| ano
a|gae ae t|e two mao souces o LC-lblA.
Among t|em, a|ga| o|| |s |nceas|ng|y o s|gn|-
|cant commec|a| |nteest |n t|at |t exc|us|ve|y
conta|ns a |age amount o LlA.
However, LC-PUFA are vulnerable to oxidation during oil
extraction, food processing, and storage. Tis generates oxidation
products that negatively afect the favor, color, nutrient value, and
functionality of the fortifed foods and food components. In addi-
tion, some lipid oxidation products are detrimental to the health
of consumers. Tus, the benefts of LC-PUFA can be realized only
when no oxidation occurs before consumption.
Te incorporation of antioxidants is the most successful way to
achieve that goal and has been broadly applied in the food industry.
But the traditional natural antioxidants that are applied to prevent
common food oil oxidation are inefcient in inhibiting LC-PUFA
oxidation. Te obstacles impeding the oxidative stability of foods
having LC-PUFA are the higher susceptibility of LC-PUFA to
oxidation and the relatively lower efcacy of antioxidants. Hence,
special strategies should be developed to maximize the oxidative
stability of oil or food product containing LC-PUFA.
Our previous study suggested the existence of nanoreactors
in bulk heterogeneous vegetable oil that form by self-assembly
of endogenous surface-active minor compounds and trace water
(~200 ppm). We also found that those reactors could potentially
afect the activity of antioxidants of diferent polarity in bulk veg-
etable oil by altering their locations (Inform 21:577578, 2010).
Annox|dant regeneranon: a ranona|
stategy to |moove ox|oanve stab|||ty |n a|ga| o||
8|ngcan Chen, Iu||an McC|ements,
and Lr|c Decker
CCn1lnuLu Cn ACL 344