G asas y Acei es
Vol.
50. Fase. 2(1999), 114-121
E ec o he ex ac ion coodi io is o i gin oli e oil on he lipoxygenase cascade:
Chemical and senso y implica ions
By Ma ia T Mo ales (1), F anca Ange osa (2) and Ramó i Apa icio (1) *
(1) Ins i u o de la G asa. A da. Pad e Ga cía Teje o 4, 41012 Se illa, Spain.
(2) Is i u o Spe imen ale pe la Elaio ecnica, 65013 Ci á S. Angelo, Pesca a, I aly.
RESUMEN
E ec o de las condiciones de ex acción del acei e de
oli a i gen en la u a de la iipoxigenasa: implicaciones
químicas y senso iales.
Los compues os olá iles p oducidos a a és de la u a de la Iipo-
xigenasa
son
los esponsables de los a ibu os senso iales más impo -
an es del acei e de oli a i gen. El p esen e abajo es udia la
e olución de es os compues os según di e en es condiciones de
em-
pe a u a y iempo del p oceso de ex acción. Se ha de e minado la in-
luencia de es os pa áme os en la p oducción de compues os
olá iles
Ce
y Os así como el e ec o que ienen es os compues os en
los desc ip o es senso iales más des acados del acei e de oli a i gen
( e de, ama go-pican e, dulce e indeseable). Se dan alo es óp imos
de iempo y empe a u a pa a la p oducción de acei es de oli a í ge-
nes de al a calidad
senso ial.
Los a ibu os senso iales de es a ieda-
des,
la española Picual y las i alianas F an oio y Co a ina, ue on
e aluados y los compues os olá iles cuan i icados median e dos me-
odologías di e en es con el in de e i a que los esul ados pudie an
ci cunsc ibi se a una a iedad especí ica o a un único mé odo de
cuan i icación.
PALABRAS-CLAVE:
Acei e de oli a i gen - Compues os olá-
iles - Lipoxigenasa ( u a) - Sis emas de ex acción.
SUMMARY
E ec o he ex ac ion condi ions o i gin oli e oil on
he lipoxygenase cascade: Chemical and senso y implica ions.
The ola ile compounds p oduced h ough he lipoxygenase
cascade a e esponsible o he mos ema kable senso y a ibu es
o i gin oli e
oil.
The pape analyses
he
e olu ion o
hese
compounds
acco ding o di e en condi ions o empe a u e and ime o he
malaxing
p ocess.
The in luence o hese pa ame e s
on
he p oduc ion
o Ce and Cs ola ile compounds is s a ed oge he wi h he e ec
ha he amoun o hese compounds has on he mos ema kable
i gin oli e oil senso y desc ip o s (g een, bi e -pungen , swee and
undesi able). Op ima alues o empe a u e and ime a e gi en o
p oducing high senso y quali y i gin oli e
oils.
Th ee
cul i a s,
Spanish
Picual and I alian F an oio and Co a ina, we e cha ac e ised by
senso y a ibu es and ola ile compounds quan i ied by wo
di e en me hodologies in o de o a oid ha esul s could be
ci cumsc ibed o a speci ic cul i a o a quan i ica ion me hod.
KEY-WORDS: Ex ac ion sys ems - Lipoxygenase (cascade) -
Vi gin oli e oil -
Vola ile
compounds.
1.
INTRODUCTION
Vi gin oli e oil is he oil ex ac ed om he ui o
he oli e ee, Olea eu opea L. This ege able oil is
consumed wi hou a u he e ining p ocess so
e aining ola iles and o he mino compounds ha
gi e ise o a ag an and delica e la ou ha has
been app ecia ed by consume s since ancien imes.
In p e ious /o ks (Apa icio e al., 1994; Apa icio
and Mo ales 1995) di e en senso y no es esponsible
o i gin oli e oil la ou we e es ablished h ough
he consume app ecia ion, he mos ema kable
being g een, swee , ui y, ipe ui , ipe oli es,
undesi able and bi e -pungen . Mos o hese
senso y pe cep ions a e p oduced by ola ile
compounds (Fla h e ai, 1973; Gu ié ez e
al.,
1975;
Solinas
é al.,
1988; Mo ales e
a/.,
1995;
Apa icio
é al.,
1996b, Ange osa e al., 1996). F om all o hem, he
g een senso y pe cep ion is o g ea impo ance
because he ag an la ou o he i gin oli e oil is
p oduced by he balance be ween g een and ui y
no es (Mo ales e ai, 1996). I has been s a ed ha
alipha ic
Ce
compounds and he co esponding hexyl
es e s (Gu h and G osch, 1991; Gu h and G osch,
1993;
Mo ales e ai, 1996) a e he main con ibu o s
o he «un ipe» componen o he ui la ou .
Mo eo e , almos all ola iles esponsible o g een
senso y no es a e he majo componen s o he
i gin oli e oil headspace (Salinas e al, 1987;
Mo ales e al., 1994; Di Gio acchino e al., 1996;
Ange osa e al, 1997),
These compounds a e biosynîhesised om
Gi8-unsa u a ed a y acids in plan s oilowing he
lipoxygenase (LOX) pa hway (Vick and Zimme man,
1987;
Ha anaka e al, 1987). This pa hway in oi es
he ac ua ion o di e en enzymes gi ing ise o
di e en amoun s o aldehydes, alcohols and es e s
ha ing all o hem senso y p ope ies and so
con ibu ing o he o e all la ou . In he case o oli e
ui s,
i has been demons a ed (AIR, 1994) ha he
LOX pa hway p omo es he o ma ion o Ce ola ile
compounds agains Cg ola ile compounds and, in
consequence, a g ea amoun o hose ola ile
compounds esponsible o g een senso y no es
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Vol. 50. Fase. 2(1999) 115
(Mo ales e al., 1996) can be ound in esh and high
quali y i gin oli e oils.
On he o he hand, en a i e p e ious s udies
(Gio acchino and Se aiocco, 1995; Ranalli and
Se aiocco, 1996; Ranalli and Ange osa, 1996) ha e
demons a ed ha he condi ions o he malaxing
p ocess du ing oli e oil ex ac ion a ec o he
senso y quali y o i gin oli e oil. Thus, he mos
impo an goal o his esea ch is o es ablish an
index ela ed o he bes ex ac ion condi ions
h ough he in o ma ion o ola ile compounds
esponsible o he unique senso y a ibu es o i gin
oli e oil (Apa icio e
al.,
1996b).
This pape emphasises he impo ance o
empe a u e and ime du ing he malaxing p ocess
on he p oduc ion o Cs and Ce ola ile compounds.
Di e en me hodologies and a ie ies ha e been
used o a oid ha esul s could be ci cumsc ibed o
a speci ic cul i a o quan i ica ion me hod.
2.
MATERIALS AND METHODS
Samples
Oli e ui s om h ee di e en Eu opean cul i a s
-Picual (Spain), Co a ina and F an oio (I aly)- we e
used in his s udy. Vi gin oli e oil was p ope ly
ob ained om esh, heal hy ui s o good quali y
collec ed a ea ly g een s age o ipeness ha
co esponds o alue 2 acco ding o he scale
sugges ed by F ias e al., (1993). The ex ac ion o
he oil samples was ca ied ou a analy ical scale
using expe imen al oil mills. Tempe a u e and ime o
malaxing we e he pa ame e s s udied. Di e en
empe a u e alues we e selec ed, he minimum
being 25° G ha co esponds o he «cold» ex ac ion
condi ions while he maximum 35° G is in he neighbou
o he maximum sugges ed by expe s (Humanes,
1993). Fi e malaxing imes (15, 30, 45, 60, 90 min)
we e selec ed in o de o co e he ange o ime
habi ually used o ex ac he oils om he di e en
ypes o oli es selec ed. Thi y i gin oli e oil
samples we e cha ac e ised by hei ola ile con en
using dynamic headspace p ocedu es and senso y
desc ip o s.
Ano he expe imen was ca ied ou wi h in ac
and cu oli e ui s in o de o de ec i he LOX
pa hway is induced when oli e ui s a e damaged
(Gailla d,
1980). Twen y g ams o in ac ui s we e
analysed ollowing he me hodology desc ibed below
(DHS using Tenax TA aps). Gu oli es we e
ob ained placing wen y g ams o in ac ui s in he
ex ac ion essel and cu ing i e imes he pulp o
each
í uü.
Cu oli es we e analysed immedia ely
hey we e cu using he same me hodology.
Dynamic headspace-Gas Ch oma og aphy
Vola ile compounds o i gin oli e oil c Picual
we e analysed by a dynamic headspace echnique
p e iously epo ed (Mo ales e al., 1994; Apa icio
and Mo ales, 1994). Samples o 0.5 g we e hea ed a
40° G and swep wi h N2 (200 ml/min) o 15 min and
he ola iles adso bed on o a Tenax TA ap
(Ch ompack, Middlebu g, The Ne he lands) a oom
empe a u e. A Ch ompack he mal deso p ion cold
ap injec o (TCT) was employed o ca y ou he
he mal deso p ion o he apped ola iles by
hea ing a 220° G o 5 min. The ola iles we e hen
condensed on o a used silica ap cooled a -110° G
wi h liquid ni ogen o 5 min jus be o e injec ion ha
was ca ied ou by lash hea ing o he cold ap a
170° G whe e i was held o 5 min. The ola iles
we e ans e ed on o a used silica J&W (Folsom,
GA) DB-WAX column (60m x 0.25 mm i.d., 0.25 ^m
ilm hickness). The o en empe a u e was held a
40° G o 6 min and p og ammed o ise a 2° G/min
o a inal empe a u e o 200° G whe e i was held o
10 min. A Hewle -Packa d 5890 se ies II wi h a FID
de ec o was employed. Quan i ica ion was ca ied
ou using ¡sobu yl ace a e as in e nal s anda d.
The iden i ica ion o ola ile compounds was
ca ied ou by MS using iden ical condi ions o Gas
Ch oma og aphy. A Fisons Mass De ec o MD800
coupled o a GG 8000 se ies was employed. Masslab
1.3 was he so wa e used. Sample componen s
we e e i ied by compa ison o mass spec al da a
wi h hose o au hen ic e e ence compounds.
Vola ile compounds o i gin oli e oil c . Co a ina
and F an oio we e analysed by a dynamic headspace-
HRGC echnique p e iously de eloped (Ange osa
e al., 1990). Fi y g ams o oil was pu in o a 120 ml
D echsel gas washing bo le wi h a po ous dis ibu o
o which was added a known amoun o nonan-1-ol
(in e nal s anda d). Vola iles we e s ipped wi h
ni ogen (1.2 dm^ /min, a 37° G) o 2 h, apped on
50 mg o ac i a ed cha coal, and elu ed wi h
1
mL o
die hyl e he .
All sol en s, o o ganic esidual analysis, we e
pu chased om J.T Bake (De en e , Holland);
nonan-1-ol (in e nal s anda d) om Ald ich
(S einheim, Ge many); ac i a ed cha coal (0.5-0.85
mm;
20-35 mesh ASTM) om E. Me ck (Schucha d ,
Ge many). Cha coal was cleaned by ea men in a
soxhie appa a us wi h die hyl e he and es ed
be o e he analyses.
Gas ch oma og aphy was ca ied ou wi h a Ca lo
E ba Mega Se ies 5160 i ed wi h a No dion silica
capilla y Ca bowax 20 M column (50 m leng h; 0.32
mm i.d.; 0.5 mm ilm hickness), and equipped wi h
an on~column injec ion sys em, a GO2 c yogenic
accesso y o hold he o en a 25° G and a Flame
Ioniza ion De ec o (FID). The o en empe a u e
p og am was he
ollowing:
iso he m a 25° C o 7 min,
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om 25 o 33° C a 0.8° C/min, om 33 o 80° C a
2.4° C/ min and om 80 o 155° C a 3.7° C/min; inal
iso he m a 155° C o 20 minu es. The empe a u e
o he de ec o was held a 240° C; ca ie gas was H2
a 30
kPa.
Injec ion olume was 0.5
|iL.
The iden i ica ion
o ola ile compounds was ca ied ou by MS unde
iden ical condi ions o GC bu now using a
Hewle -Packa d Mass De ec o coupled o a GC
5890A model was used. Sample compounds we e
iden i ied by compa ison o mass spec al da a wi h
hose o au hen ic e e ence compounds and by hei
e en ion imes.
The au ho s ha e poin ed ou in p e ious s udies
ha he e is a good co ela ion be ween he echniques
jus desc ibed abo e (Mo ales and Apa icio, 1993).
Senso y p ope ies o ola ile compounds
Assesso s: The senso y p ope ies o ola ile
compounds (odou and as e) we e e alua ed by i e
assesso s wo wi h mo e han en yea s expe ience
and h ee who we e habi ual consume s o i gin
oli e oil.
HRGC-ol ac ome y: To assess he a oma no es
co esponding o oli e oil ola ile compounds, an
High Resolu bn
Gas
Ch oma og aphy (HRGC)-ol ac ome y
echnique was applied o i gin oli e oil samples
(Mo ales
e al.,
1994). The e luen o he GC column
was spli ed 1 o 10 o he de ec o and he sni ing
po , espec i ely. The desc ip ions o he odou -
ac i e egions o he elua e we e no ed on a o m
wi h a p e-p in ed ime scale, assesso s did no see
he ch oma og am. Assesso s basically ag eed on
he odou s o ola iles, al hough di e en seman ic
e ms whe e used o desc ibe some o hem. A
consensus-building discussion was held wi h
assesso s o decide he inal senso y desc ip o s.
Th eshold alues and
Tas ing
o
pu e
compounds:
Odou h eshold alues o ola ile compounds, in a
ma ix o deodo ised sun lowe oil, we e de e mined
by he au ho s using he me hod desc ibed by Gu h
and G osch (1993).
The assesso s also ca ied ou he smelling and
as ing,
in duplica e, a oom empe a u e o ou pu e
ola ile compounds ha p e iously had been
cha ac e ised as esponsible o senso y pe cep ions
o he han g een: hexan-1-ol (Me ck, Da ms ad ,
Ge many), (E)-2-hexenal, (E)-2-hexen-1-ol and
(E)-3-hexen-1-ol (Ald ich, Milwaukee, Wisconsin,
USA).
In ac , Apa icio e al., (1996b) ound ha
(E)-2-hexen-1-ol, (E)-3-hexen-1-ol and hexan-1-ol
we e be e cha ac e ised by he global senso y
pe cep ion o undesi able. This cha ac e isa ion
ag ees wi h Bedoukian (1971) who ound ha he
ans o ms o hexenols a e cha ac e ised by a a y
senso y pe cep ion while he c/s o ms a e sha p and
g een.
The ola ile compounds we e p e iously dilu ed
in wa e o pa a in oil, depending on hei solubili y,
o he same app oxima e concen a ion as ound in
i gin oli e oil samples.
Panel es
The senso y e alua ion o oli e oil is egula ed
inside The Eu opean Union (EC,1995). The
desc ip o s o he o icial panel we e se up o
desc ibing he mos ema kable senso y pe cep ions
o i gin oli e oil and no o di e en ia e sub le
changes in he «g een» pe cep ion. In o de o
dissec and desc ibe di e en la ou sensa ions
associa ed wi h g een pe cep ion, eigh ull ained
I alian assesso s we e eques ed o e alua e some
oils ob ained om un ipe ui s o di e en a ie ies.
In all o he aspec s, i was s ic ly ollowed he
egula ion o he Eu opean Union (EC, 1995).
Assesso s chose eely hei own desc ip o s by
means o associa ions o he sensa ions pe cei ed
wi h own o me expe iences. The desc ip o s wi h
enough ag eemen be ween assesso s we e
selec ed o he p o ile shee (Table
I).
These senso y
desc ip o s basically ag eed wi h hose desc ibing
he «g een» a oma pe cep ion, by ei he habi ual o
po en ial consume s (Apa icio e al., 1994; Apa icio
and Mo ales, 1995), al hough some o he desc ip o s,
mos ly ela ed o as ing (bi e and pungen ), we e
also selec ed o desc ibing he «g een» p o ile o
i gin oli e oil.
Table I
Senso y desc ip o s e alua ed in i gin oli e oil
samples (c . Co a ina. Picual and F an oio).
S uc u ed scale om 0 o 5
Senso y desc ip o s
Cu g een lawn
G een lea o wig
G een oli es
Wild lowe s
G een banana
G een oma o
Almond
A ichoke
Apple
Walnu husk
G een hay
Bi e
Pungen
Co a ina
0.7
1.5
2.0
—
0.1
0.6
0.1
0.7
—
1.1
—
2.8
2.5
Picual
1.0
0.8
1.8
0.1
0.2
0.8
0.6
0.3
0.1
0.5
0.1
1.8
1.9
F an oio
1.0
1.0
2.0
0.2
0.2
0.7
0.3
0.6
0.1
1.1
0.1
2.1
2.0
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Vol. 50. Fase. 2(1999) 117
S a is ical Analysis
STATISTICA (S a So , 1995) was he s a is ical
package employed. Response su aces we e used
o plo he i ed model o he selec ed chemical
compounds in a su ace along wi h hei obse ed
alues in he expe imen s a di e en ime and
empe a u e du ing he ex ac ion p ocess. A
quad a ic smoo hing algo i hm was applied o he
s anda dised alues o each ola ile compound.
Canonical Co ela ion (Tabachnick and Fidell,
1983) was applied o assessing he ela ionship
be ween senso y a ibu es and ola iles. This
algo i hm allows in es iga ing he edundancies
be ween bo h se o da a.
p esen in he i gin oli e oil headspace we e also
p oduced a e cu ing he oli es. This expe imen
s a es ha ola iles a e p oduced, h ough he
lipoxygenase pa hway, as soon as oli es a e cu o
damaged.
M JÀL.l JÂn
3. RESULTS AND DISCUSSION
The senso y e alua ion o samples by di e en
desc ip o s allowed knowing whe he hey we e
basically cha ac e ised by g een a ibu es. The
esul s o he senso y p o iles showed ha a ie ies
we e cha ac e ised by di e en pe cep ions o g een
senso y a ibu e. Oil om Picual oli es is usually
cha ac e ised by high alues o a ibu es ui y
( oma o) and pungen (Apa icio e al., 1994, 1996a,
1997) while his panel es (Table I) cha ac e ised he
Spanish a ie y as g een oli es, g een oma o and
cu g een lawn.
Co a ina oil is habi ually cha ac e ised by he
as ing a ibu es bi e and pungen (Apa icio e al.,
1994;
McEwan, 1994) bu , conce ning he g een
pe cep ion, he desc ip o s we e g een oli es, g een
lea o wig and walnu husk (Table I). High in ensi ies
o g een, ui y and pungen a ibu es cha ac e ised
F an oio oils (FLAIR, 1991) bu ocusing on g een
pe cep ions his oil shows high alues o a ibu es
g een oli es and cu g een lawn (Table I).
F om he la ge da abase o ola iles iden i ied in
i gin oli e oil (Mo ales e al., 1994) only a ew a e
esponsible o g een pe cep ions (Apa icio e al.,
1996b). These ola ile compounds a e p oduced
h ough LOX pa hway and hey a e usually known as
«g een ola ile compounds». In his pa hway he
p oduc ion o he ola iles is associa ed o cell
damage (Gailla d, 1980). Figu e
1
clea ly shows he
di e ences be ween wo ch oma og ams co esponding
o ola iles p oduced in in ac oli es and cu oli e
ui s
c .
Picual. I is no ewo hy he main p esence o
hyd oca bons in in ac ui s and he absence o
ola ile compounds esponsible o he inal i gin
oli e oil a oma, only aces o hexanal and
hexan-1 -ol
-bo h coming om linoleic
acid-
could be
de ec ed.
The ola ile p o ile o cu oli es shows a
conside able change due o he p oduc ion o g een
ola ile compounds, coming om bo h linoleic and
linolenic acids. O he ola ile compounds usually
Figu e 1
P oduc ion o some ola ile compounds in in ac (A) and cu (B)
oli es. Quan i ica ion by dynamic headspace
Table II shows he
Ce
ola ile compounds analysed
in samples. The HRGC-ol ac ome y showed g een
senso y no es co e ing he ange be ween
swee - ui y-g een o bi e -powe ul-g een. The
odou h eshold and odou ac i i y alue (OAV)
(G osch,
1994) a e also gi en. A ola ile compound
wi h an OAV g ea e han 1.0 con ibu es o he
g een senso y pe cep ion, while ola iles ha ing
lesse OAVs could con ibu e o i gin oli e oil la ou
h ough possible syne gism phenomena.
Table II
Senso y desc ip ion, odou h esliolds and odou
ac i i y alues o he g een ola ile compounds
Vola ile Compounds
Hexanal
(Z)-3-hexenal
(E)-2-hexenal
Hexyl ace a e
(Z)-3-hexenyl ace a e
Hexan-1
-ol
(E)-3-hexen-1-ol
(Z)-3-hexen-1-ol
(E)-2-hexen-1
-ol
Odo desc ip o s
G een,
apple, g een ui
G een,
cu g ass
G een,
ui y, almonds
F ui y, swee
F ui y, g een lea es
F ui , banana, so
G een
G ass, banana
G een,
g assy, swee
Odo
h esholds"
60
3.0
1200
1040
730
400
1500
5800
8000
OAV"
8.91
184.67
9.76
0.04
0.07
0.13
0.01
0.07
0.01
Legend:
^ Calcula ed in ^g/kg
^ OAV = odou ac i i y alue.
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The high alue o he canonical co ela ion coe icien
(R=0.98) be ween he Ce g een ola ile compounds
(Table II) and he senso y a ibu es (Table I)
demons a es ha he o me a e esponsible o he
la e . Howe e , he ola iles only explained 66% o
he senso y a ibu es ( edundancies be ween da a
se s) and his could mean ha he whole in o ma ion
o g een senso y a ibu es is no only explained by
he odou o he ola iles bu also by he as ing o
some o hese ola iles as assesso s e alua ed he
samples by hei ol ac o y-gus a o y- ac ile no es
(EC,
1995).
P e ious s udies using SSW (Apa icio e al.,
1994,
1996b) demons a ed ha some o hese
compounds mainly con ibu e o he g een senso y
pe cep ion i.e. (Z)-3-hexenal, hexyl
ace a e,
(Z)-3-hexenyl
ace a e, and (Z)-3-hexen-1-ol. Howe e , o he se o
hem could also con ibu e o o he senso y
pe cep ions i.e. (E)-2-hexenal, hexan-1-ol, (E)-3-hexen-1-ol,
and (E)-2-hexen-1-ol. The as ing o he la e
ola iles allowed hei cha ac e isa ion as as ingen
and bi e ((E)-2-hexen-1-ol and (E)-3-hexen-1-ol),
ough (hexan-1-ol) and sha p, bi e , as ingen
((E)-2-hexenal).
The e is good ag eemen among consume s in
ela ion o he senso y pe cep ions. They dislike high
in ensi ies o as ing pe cep ions bi e and pungen ,
while hey like almos all a oma desc ip o s quali ied
wi h he adjec i e g een (McEwan, 1994; Paglia ini
e al., 1994). The e o e, he p esence o ola iles
esponsible o pleasan senso y pe cep ions should
be p omo ed. Howe e i gin oli e oil mus be
ex ac ed solely using physical means, and hen he
a io be ween ola iles in he inal p oduc can only
be a ied by changing he empe a u e and ime o
he malaxing p ocess. Depending on empe a u e
and ime o malaxing, each ola ile compound
showed di e en beha iou .
Figu e 2 shows a scheme o he LOX pa hway
wi h i s h ee di e en b anches o he p oduc ion o
ola iles. The b anch coming om linoleic acid gi e
ise o hexanal, hexan-1-ol and hexyl ace a e, being
he o me and he la e esponsible o desi able
pe cep ions. This b anch could be seen as he
g een-swee aspec o he global g een la ou . The
second b anch could be seen as he esponsible o
he main g een pe cep ion, being cons i u ed by
(Z)-3-hexenal, (Z)-3-hexenol and (Z)-3-hexenyl
ace a e. The hi d b anch gi ing ise o (E)-2-hexenal
and (E)-2-hexen-1-ol could be conside ed as he
bi e -as ingen aspec o he g een senso y
pe cep ions.
Figu e 2
Response su aces o Ce ola ile compounds p oduced h ough he lipoxygenase cascade. The su ace has been i ed o he da a by a
2 o de polynomial unc ion and ola ile compounds we e p e iously no malised by Z-sco e. The igu es o ola ile compounds ha e
been o a ed (clokwise o coun e -clokwise) in o de o expose he esul s be e .
Hexanal,
esponsible o g een, apple, swee
pe cep ions, has an OAV = 8.91 (Table II), he e o e
con ibu ing o pleasan senso y pe cep ions. The
p oduc ion o hexanal is mo e in luenced by he ime
o malaxing han by he empe a u e, hexanal is
p oduced du ing he whole p ocess bu he highes
concen a ions we e ound a e 45 minu es o
malaxing.
A di e en beha iou showed ola ile
(E)-2-hexen-1-ol ha is cha ac e ised by a g een
smelling and an as ingen -bi e as ing, he la e
being an undesi able senso y pe cep ion o po en ial
consume s (McEwan, 1994). The p oduc ion o
(E)-2-hexen-1-ol is mainly a ec ed by empe a u e
since he highes concen a ions we e measu ed a
he highes empe a u e o malaxing (35° C). Hexyl
ace a e is also a ec ed by empe a u e bu , in his
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Vol.
50. Fase. 2(1999) 119
case,
he highes concen a ion o his es e was
measu ed a he lowes ennpe a u e (25° C).
The o he compounds esponsible o g een
a oma desc ip o s ((Z)-3-hexenal, (Z)-3-hexen-1-ol,
and (Z)-3-hexenyl ace a e) also showed he highe
concen a ions a low empe a u e, Figu e 2. The
in luence o malaxing ime was low o (Z)-3-hexenal,
medium o (Z)-3-hexen-1-ol and highe o
(Z)-3-hexenyl ace a e. This means ha he
p oduc ion o (Z)-3-hexenal is almos immedia e
while he p oduc ion o (Z)-3-hexenyl ace a e needs
mo e ime. This explana ion ag ees wi h he
sequence o p oduc ion o hese ola iles h ough he
LOX pa hway. The i s s ep o his biochemical
pa hway is he o ma ion o he aldehyde, hen i s
ans o ma ion o alcohol -by he ac ion o alcohol
dehyd ogenase enzyme (ADH)- and la e he
o ma ion o he es e , by means o he alcohol acyl
ans e ase enzyme (AAT). I is no ewo hy ha
(Z)-3-hexen-1-ol is ep esen ed by a sha p saddle
pic u e. I has wo maximum peaks -a low
empe a u e and low ime and a high empe a u e
and high ime- so sugges ing ha he la e condi ion
(high empe a u e and high ime o malaxing) makes
easie he o ma ion o his compound in de imen o
(Z)-3-hexenyl ace a e while he o me condi ions
enables i s ans o ma ion on (Z)-3-hexenyl ace a e,
bo h being cha ac e ised as g een ola iles.
Conce ning he ola iles esponsible o he
undesi able senso y pe cep ions ((E)-2-hexenal,
hexan-1-ol,
(E)-3-hexen-1-ol), hei concen a ions
a e highe when he empe a u e o he malaxing is
he highes . Figu e 2. In his case, he ime o
malaxing had no in luence on he o ma ion o he
alcohols bu i showed a g ea in luence on he
o ma ion o he aldehyde. The concen a ion o he
aldehyde was lowe a high and low malaxing imes
and highe a medium alues o ime o he Picual
a ie y hough ce ain di e ences we e ound om
he esul s o Co a ina and F an oio a ie ies. The
in e media e ola ile compound (E)-2-hexenal is
ep esen ed by a ypical saddle pic u e whe e, once
again,
he e a e wo maximum peaks a low and high
empe a u es unde medium ime o malaxing.
Acco ding o he senso y cha ac e isa ion o
(E)-2-hexenal and (E)-2-hexen-1-ol he low
empe a u e gi es he bi e as ing o i gin oli e oil
bu when empe a u e eaches high alues han his
senso y pe cep ion in i gin oli e oil is accompanied
by an undesi able pe cep ion ha can make he oil o
be ejec ed.
In gene al, he highes concen a ions o aldehydes
we e measu ed when he ime o malaxing was
sho e , he p oduc ion o alcohols was p omo ed a
high empe a u e (35° C) and he concen a ion o
es e s was highe a low empe a u e (25° C). This
means ha he malaxing p ocess a high empe a u e
(T > 35° C) p omo e he o ma ion o g een ola iles
esponsible o undesi able senso y pe cep ions
while,
on he con a y, low empe a u es (T > 25° C)
a ou ed he p oduc ion o desi able g een senso y
pe cep ions. The empe a u e is hen he main ac o
cha ac e ising he malaxing p ocess.
Recen ly Ange osa e
al.,
(1998) ha e sugges ed
a b anch o he lipoxygenase cascade o Cs ola ile
compounds. An alkoxy adical would be p oduced
om 13-hyd ope oxide by he ac ion o he
lipoxygenase, la e by p-scission would p oduce
1,3-pen ene adical and an unsa u a ed aldehyde. A
se ies o pen ene dime s would be p oduced by
dime iza ion o 1,3-pen ene while he hyd oxila ion
would p oduce 2-pen en-1-ol and 1-pen en-3-ol ha
will hen p oduce 1-pen en-3-one and 2-pen enal by
he ac ion o he alcohol dehyd ogenase. Cs ola ile
compounds ha e a senso y beha iou qui e simila
o Ce ola ile compounds, he e a e con ibu o s o
g een,
(Z)-2-pen en-1-ol and (E)-2-pen enal, swee ,
1-pen en-3-one, pungen , 1-pen en-3-ol, and
undesi able, (Z)-2-pen enal and 1-pen en-3-ol,
senso y pe cep ions (Apa icio e
al.,
1996b).
(Z)-2-pen en-1-ol, esponsible o g een banana,
is p omo ed a low alues o empe a u e (25° C) and
ime (15 min) while i s lowes alues a e a he
highes alues o hese a iables, so showing a
simila beha iou
o Ce
ola ile compounds esponsible
o g een pe cep ions. Figu e 3. The nex ola ile in
his lipoxygenase cascade, 1-pen en-3-ol, has i s
highes alues immedia ely he malaxing begins and
i s concen a ion diminishes when malaxing goes on
ime;
he empe a u e seems o ha e poo in luence
on i s p oduc ion. Figu e 3. This beha iou can be
in e p e ed as hese compounds a e p oduced in
pa allel p ocess and he p oduc ion o hese ola ile
compounds begins immedia ely he oli es a e
c ushed;
hese compounds al eady appea ed in he
ch oma og am o cu oli es. Figu e 1. (Z)-2-pen enal
is p oduced om he p e ious compounds and
Figu e 3 shows ha he ola ile eaches i s op imum
a e a ce ain delay; in ac , i does appea s a ace
le els in Figu e 1. The p oduc ion o 1-pen en-3-one
is inc eased wi h empe a u e, so sugges ing i is
p oduced h ough an oxida ion p ocess. Figu e 3
shows ha i s maximum is eached wi h a delay
simila o ha o (Z)-2-pen enal.
The main conclusion o he p esen s udy is ha low
empe a u e (T > 25° C) and medium imes (35-45 min.)
a e he bes ex ac ion condi ions o p omo e he
o ma ion o g een ola ile compounds esponsible
o desi able senso y pe cep ions. High empe a u e
(T > 35° C) wi h minimum alues o ime ( < 35 min.)
could also be use ul as a al e na i e way o ob ain
pleasan g een i gin oli e oils.
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120 G asas y Acei es
-^ 1.2:
Figu e 3
Response su aces o Cs ola ile compounds a di e en empe a u es and imes o malaxing. The su ace has been i ed o he
da a by a 2"^ o de polynomial unc ion. Values o ola ile compounds a e gi en in 10*ppm.
All he esul s, coming om ei he ola ile
compounds o senso y a ibu es in o ma ion, poin
ou ha low empe a u e du ing malaxing (an
indus ial example is pe cola ion) is de e minan o
ob ain pleasan g een i gin oli e oils in any o he
s udied a ie ies.
ACKNOWLEDGEMENTS
The au ho s would like o acknowledge hei
indeb edness o assesso s who ca ied ou he
senso y e alua ion o he samples. This wo k has
been suppo ed by AIR3-CT94-1967.
BIBLIOGRAPHY
AIR (1994).—«Eu opean P ojec AIR3-CT94-1967. Oli e
oil la ou and a oma: biochemis y and chemis y o
senso y ac o s a ec ing consume app ecia ion and
hei analysis by a i icial in elligence, The Commission
o he Eu opean Communi ies».
Ange osa, F., Di Giacin o, L and Solinas, M. (1990).—
«In luence o oli e ui mass s o age on he a oma o
esul ing oils. E alua ion o « us y» de ec by HPLC
and GLC analyses o ola ile compounds».—Ri .
Me ceol.,
29, 275-294.
Ange osa, R, Di Giacin o, L., Wo, R. and
Cumi ini,
S. (1996).—
«Senso y e alua ion o i gin oli e oils by a i icial
neu al ne wo k p ocessing o dynamic head-space gas
ch oma og aphic da a».—J. Sci. Food Ag ia, 72,
323-328.
Ange osa, F., Di Giacin o, L and D'Alessand o, N. (1997).—
«Quan i a ion o some la o componen s esponsible
o he "g een" a ibu es in i gin oli e oils».—J. High
Resol.
Ch oma og ., 20, 507-510.
Ange osa, F., Came a, L, d'Alessand o, N. and Melle io,
G.
(1998).—«Cha ac e iza ion
o se en
new hyd oca bons
compounds p esen in he a oma o i gin oli e
oils».—J.
Ag ie. Food Chem., 46, 648-653.
Apa icio, R. and Mo ales, M.T. (1994).—«Op imiza ion o a
dynamic headspace echnique o quan i ying i gin
oli e oil ola iles. Rela ionships among senso y
a ibu es and ola ile peaks».—Food Qual. and
P e .,
5, 109-114.
Apa icio, R. and Mo ales, M.T. (1995).—«Senso y wheels:
a s a is ical echnique o compa ing QDA panels.
Applica ion o i gin oli e
oil».—J.
Sci. Food Ag ie, 67,
247-257.
Apa icio, R., Alonso, M.V., Mo ales, M.T. and Cál en e, J.J.
(1994).—«Rela ionship be ween COI es and o he
senso y p o iles by s a is ical p ocedu es».—G asas
Acei es, 45,
26-41.
Apa icio, R., Cál en e, J.J. and Mo ales, M.T. (1996a).—
«Senso y au hen ica ion o Eu opean ex a- i gin oli e
oil a ie ies by ma hema ical p ocedu es».—J. Sci.
Food Ag ie, 72, 435-447.
Apa icio, R., Mo ales, M.T. and Alonso, V. (1996b).—
«Rela ionship be ween ola ile compounds and
senso y a ibu es o oli e oils by he senso y
wheel».—J.
Am. Oil Chem. Soc, 73,1253-1264.
Apa icio, R., Mo ales, M.T. and Alonso, V. (1997).—
«Au hen ica ion o Eu opean ex a- i gin oli e oils by
hei chemical compounds, senso y a ibu es and
consume s a i udes».—J. Ag ie. Food Chem., 45,
1076-1083.
Bedoukian, P.Z. (1971).—«The se en p ima y hexenols
and hei ol ac o y cha ac e is ics».—^J. Ag ie. Food
Chem.,
19,1111-1114.
Di Gio acchino, L., Ange osa, F. and Di Giacin o, L.
(1996).—«E ec o mixing lea es wi h oli es on
o ganolep ic quali y o oil ob ained by cen i uga ion».—
J.
Am. Oil Chem Soc, 73, 371-374.
Di Gio acchino, L. and Se aiocco, A. (1995).—«In luenza
dei sis emi di la o azione delle oli e sulla composizioni
dello spazio di es a degli oli».—Ri .
I al.
Sos anze
G asse, 72, 443-450.
EC.
(1995).—O icial Jou nal o The Commission o The
Eu opean Communi ies. Ma ch
28 h,
3-4 Regula ion
n.°
1
656/95.
FLAIR (1991).—Eu opean P ojec FLAIR-CT91-0046. The
s udy o senso y and nu i ional quali y o i gin oli e oil in
ela ion o a ie y, ipeness and ex ac ion echnology,
The Commission o he Eu opean Communi ies. Fi s
Yea Repo .
(c) Consejo Supe io de In es igaciones Cien í icas
Licencia C ea i e Commons 3.0 España (by-nc)
h p://g asasyacei es. e is as.csic.es
Vol.
50. Faso. 2 (1999) 121
Fla h,
R.A., Fo ey, R.R. and Guadagni, D.G. (1973).—
«A oma componen s o oli e
oil».—J.
Ag ie. Food
Ghem,
21,
948-952.
F ías,
L, Ga cia-O iz, A., He moso, M., Jiménez, A.,
Lla e o, M.R, Mo ales, J., Ruano, M.T. and Uceda, M.
(1993).—Analis as de Labo a o io de Almaza a,
Andalusian Minis y o Ag icul u e, Se ille (Spain).
Gallia d,
T. (1980).—The Biochemis y o Plan s, Vol. 4,
Academic P ess, London.
G osch,
W.(1994).—«De e mina ion o po en odou an s in
oods by a oma ex ac dilu ion analysis (AEDA) and
calcula ion o odou ac i i y alues (OAVs)».—Fla ou
F ag .J.,9,147-158.
Gu h,
H. and G osch, W. (1991).—«A compa a i e s udy o
he po en odo an s o di e en i gin oli e oils».—Fa .
Sci.
Technol., 93, 335-339.
Gu h,
H. and G osch, W. (1993).—«Quan i a ion o po en
odo an s o i gin oli e oil by s able so ope dilu ion
assays».—J.
Am. Oil Ghem. Soc, 70, 513-518.
Gu ié ez, R., Olías, J.M., Gu ié ez, F., Cab e a, J. and del
Ba io,
A.
(1975).—«The ch oma og aphic
and
o ganolep ic
me hods in he e alua ion o he a oma ic
cha ac e is ics o i gin oli e oil».—G asas y Acei es,
26,21-31.
Ha anaka, A., Kajiwa a, T. and Sekiya, J. (1987).—
«Biosyn he ic pa hway o Ce-aldehydes o ma ion
om linolenic acid in g een lea es».—Ghem. Phys.
Lipids, 44,
341-361.
Humanes, J. (1993).—P oducción de Acei e de Oli a de
Calidad:
In luencia del cul i o, Jun a de Andalucía,
Conseje ía de Ag icul u a y Pesca, Se ille (Spain).
McEwan,
J. A. (1994).—«Consume a i udes and oli e oil
accep ance: he po en ial consume ».—G asas y
Acei es, 45, 9-15.
Mo ales, M.T. and Apa icio, R. (1993).—«Op imiza ion by
ma hema ical p ocedu es o wo dynamic headspace
echniques o quan i ying i gin oli e oil
ola iles».—Anal. Chim. Ac a, 282,
423-431.
Mo ales, M.T., Apa icio, R. and Ríos, J.J. (1994).—
«Dynamic headspace gas ch oma og aphic me hod
o de e mining ola iles in i gin oli e
oil».—J.
Ch oma og . A, 668, 455-462.
Mo ales, M.T, Cál en e, J.J. and Apa icio, R. (1996).—
«In luence o oli e ipeness on he concen a ion o
g een a oma compounds in i gin oli e oil».—Fla ou
F ag . J.. 11,171-178.
Mo ales, M.T,
Alonso,
M.V,
Ríos,
J.J.
and
Apa icio,
R.
(1995).—
«Vi gin oli e oil a oma: ela ionship be ween ola ile
compounds and senso y a ibu es by chemome ics».—
J.
Ag ie. Food Chem., 43,
2925-2931.
Paglia ini,
E., Be uccioli, M. and Abba, S. (1994).—
«Consume a i udes and oli e oil accep ance: he
adi ional consume ».—G asas y Acei es, 45,16-19.
Ranalli,
A. and Ange osa, F. (Î996).—«In eg al cen i uges
o oli e oil ex ac ion. The quali a i e cha ac e is ics o
p oduc s».—J.
Am Oil Chem. Soc, 73, 417-422.
Ranalli,
A. and Se aiocco, A. (1996).—«E alua ion o
cha ac e is ics o oli e oil p oduced by inno a i e and
adi ional p ocessing echnologies».—Ri .
I al.
Sos anze G asse, 73, 303-315.
Solinas, M., Ange osa, F. and Ma silio, V. (1988).—«
Indagine su alcuni componen i dell'a oma degli oli
e gini di oli a in elazione alia a ie à delle
oli e».—Ri .
I al.
Sos anze G asse, 45, 361-368.
Solinas, M., Ma silio, V. and Ange osa, F.
(1987).—«E oluzione di alcuni componen i dell'a oma
degli oli e gini di oli a in elazione al g ado di
ma u azione delle oli e».—Ri .
I al.
Sos anze G asse,
44,
475-480.
S a So , (1995).—STATISTICA elease 5, Tulsa, USA.
Tabachnick, B.G. and Fidell, LS. (1983).—Using
Mul i a ia e S a is ics, Happe and Row Publishe s,
New Yo k.
Vick, B.A. and Zimme man, D.C.(1987).—«Oxida i e
sys ems o modi ica ion o a y acids: he
lipoxygenase pa hway».—In The Biochemis y o
Plan s: A Comp ehensi e T ea ise, Vol. 9, p. 53.
S ump , P.K. and E.E. Conn, (Eds.).—Academic P ess
Inc., New Yo k.
Recibido: Junio 1998
Acep ado: Sep iemb e 1998
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