1
Chemical and senso y e ec s o he eezing p ocess on he a oma 1
p o ile o black u les (Tube melanospo um) 2
3
Lau a Culle é (*)a, Vicen e Fe ei aa, Ma ía E. Ven u inib, Ped o Ma cob and Domingo 4
Blancob 5
a Labo a o y o A oma Analysis and Enology, A agón Ins i u e o Enginee ing 6
Resea ch (I3A), Depa men o Analy ical Chemis y, Facul y o Sciences, Uni e si y o 7
Za agoza, 50009 Za agoza, Spain 8
b Plan Foods Resea ch G oup. Depa men o Food Technology. Uni e si y o 9
Za agoza. Miguel Se e , 177. 50013. Za agoza, Spain 10
11
(*)To whom co espondence should be add essed 12
Phone: 34 976761000 Ex 3330 13
Fax: 34 976761292 14
Email: lcu[email p o ec ed] 15
16
2
ABSTRACT 17
The aim o his wo k is o e alua e he e ec o eezing black u les (Tube 18
melanospo um) on hei a oma bo h in senso y and chemical e ms. The u les we e 19
ozen a empe a u es o -20ºC o -80ºC. Senso y and chemical analyses we e ca ied 20
ou a e 24 hou s and a e 20, 40 and 60 days. The senso y analysis was bo h 21
desc ip i e and disc imina i e. Chemical analysis was done by headspace solid phase 22
mic oex ac ion ollowed by gas ch oma og aphy mass spec ome y analysis (HS-23
SPME-GC-MS). Fi een compounds wi h high a oma ic po en ial in u les we e 24
de e mined. Thei selec i e ion peak a eas we e calcula ed, summed and exp essed as 25
pe cen age o ac i e odo compound, in o de o moni o changes in odo p o ile. 26
The senso y s udy concluded ha he a oma o ozen u les di e ed signi ican ly 27
om he a oma o esh u les. This esul was ully con i med by ola ile composi ion 28
da a, which e eal ha T.melanospo um a oma ic p o ile is deeply modi ied as a 29
consequence o a eezing p ocess. Odo p o iles become en iched in compounds such 30
as diace yl, 1-oc en-3-one, 1-oc en-3-ol, 2-me hylisobo neol and dime hyl isu ide a e 31
he eezing pe iod, while isoamyl alcohol, me hane hiol and e hyl 3-me hylbu y a e 32
dec eased. These a oma ic changes could explain he loss o eshness obse ed in all 33
ozen u les. 34
On he o he hand, me hional and some phenols such as 3-e hyl-5-me hylphenol 35
inc eased a e 40 days which sugges ha hese compounds may be used as ma ke s o 36
eezing ime. In e es ingly, he pe cen age a ea o 1-oc en-3-one was cons an in ozen 37
samples and much highe han in esh ones, which sugges s ha i is a gene al ma ke 38
o eezing p ocess. 39
Keywo ds: eezing s o age; eezing ma ke s; eshness; a oma; Tube melanospo um; 40
1-oc en-3-one; black u le 41
42
3
In oduc ion 43
The black u le o Tube melanospo um is highly app ecia ed due o i s in ense and 44
dis inc i e a oma. I has been desc ibed as he queen o u les and is one o he mos 45
p ized oods wo ldwide. T u les ha e hei highes o ganolep ic alue when esh. 46
Howe e , like many o he ege able commodi ies, hey a e highly pe ishable mainly 47
due o bac e ial and mold g ow h and dehyd a ion which con ibu e o he apid loss o 48
o ganolep ic p ope ies such as ex u e, a oma, and as e (Ri e a, Blanco, Sal ado & 49
Ven u ini, 2010). Mo eo e , u les a e seasonal so long e m s o age me hods a e used 50
o ensu e hei a ailabili y h oughou he yea . F eezing is one o he mos impo an 51
me hods o e aining ood quali y du ing long- e m s o age. Fac o s such as a ie y, 52
ma u i y, g owing a ea and seasonal a ia ions in luence he ege able eezing p ocess 53
o an ex en ha may o e ide he posi i e e ec o a high eezing a e (Sk ede, 1996). 54
Fo many p ocesses i is impo an o know he ypical pigmen and ola ile compound 55
p o ile o a esh p oduc in o de o iden i y he colo and a oma changes p oduced 56
du ing ea men (de Ancos, Cano & González, 1999; Ibañez, López-Sebas ian, Ramos, 57
Tabe a & Regle o, 1998). Some s udies ha e been ca ied ou in o de o examine he 58
e ec o he eezing p ocess on he ola ile composi ion o se e al p oduc s. Fo 59
example, de Ancos, Ibañez, Regle o and Cano (2000) p ocess and long- e m ozen 60
s o age condi ions do no signi ican ly a ec he a oma o he aspbe y cul i a s hey 61
s udied. Howe e , a oma is usually a ec ed by eezing. Some changes in he a oma ic 62
p o ile o many o he p oduc s a e ozen s o age ha e been obse ed. Fo example, 63
Kjeldsen, Ch is ensen and Edelenbos (2003) de ec ed an o - la o , desc ibed as soapy 64
and pa a in-like, in unblanched ca o s du ing long- e m eezing. O he au ho s 65
concluded ha he a oma in unblanched leek slices changed a e ozen s o age o 12 66
mon hs as he amoun o sul u compounds dec eased (Nielsen & Poll, 2004). Some 67
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possible ma ke s we e p oposed o he di e en ia ion be ween esh and ozen- hawed 68
ish, as o example 1-oc en-3-ol o 1-pen en-3-ol (Iglesias, Medina, Bianchi, Ca e i, 69
Mangia & Musci, 2009). O he examples o changes be ween esh and ozen p oduc s 70
ha e been e alua ed in lamb mea (Bueno, Resconi, Campo, Cacho, Fe ei a & 71
Escude o, 2011), Tu kish Mo al cheese (Andic, Tunc u k & Ja idipou , 2011) o in 72
can aloupe melon (Ma e al., 2007). The la e pape demons a ed ha wi h he 73
p olonga ion o he eezing ime, an inc easing numbe o unsa u a ed alcohols and 74
aldehydes wi h 6 and 9 ca bons we e p oduced, he g een no es o ozen melon became 75
mo e and mo e in ense, and he es e ag ance became inc easingly less appa en . 76
I is he e o e impo an o e alua e he e ec s on a oma o he di e en me hods o 77
p ese a ion. Ve y ew p e ious wo ks (Al-Ruqaie, 2006; Jawo ska & Be nás, 2009) 78
ha e e alua ed he e ec o ozen s o age on u les and mush ooms. Al-Ruqaie 79
(2006) e ealed ha he eezing p ocess seems o be mo e e ec i e han d ying, a leas 80
in he wo species o u les s udied (T. cla e yi and T. hakizi), and concluded ha 81
blanching in a 4 % NaCl solu ion and s o age a -18 ºC p o ed o be he bes 82
p ese a ion me hod in e ms o quali y o he u les. The o he pape ci ed (Jawo ska 83
e al., 2009) p oposed a maximum s o age pe iod o ou mon hs o he ozen p oduc 84
in he case o unblanched mush ooms. 85
The goal o he p esen wo k is o s udy he in luence o eezing and long- e m ozen 86
s o age on he a oma composi ion o T. melanospo um, using senso y analyses and 87
analy ical echniques (HS-SPME coupled wi h GC-MS). 88
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90
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92
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2. Ma e ials and Me hods 93
2.1. T u le samples 94
The ascoca ps o T. melanospo um we e collec ed in cul i a ed u le-g ounds unde 95
holm oak ees (Que cus ilex subsp. balllo a) in Sa ión (Te uel, Spain), wi h he help o 96
a hun e dog. The u les we e ha es ed in Janua y-Feb ua y and we e in s age VI o 97
ma u a ion ( ully ma u e). Thei o ganolep ic p ope ies we e a hei maximum, 98
acco ding o he desc ip ion o ma u a ion s ages gi en in (Mon an , Kuli ak & Gleyze, 99
1983). The u les we e anspo ed o he labo a o y wi h co e ing soil in insula ed 100
boxes wi h ice packs. The samples we e b ushed wi h a we so b ush, insed wi h ap 101
wa e and o ced-ai d ied in a lamina cabine . Quali a i e selec ion o he ca popho es 102
was made by disca ding u les wi h so ex u e as well as hose pa asi ized o 103
damaged du ing ha es . 104
2.2. S anda ds 105
Fi een compounds we e chosen o his s udy due o hei po en ial a oma ic 106
impo ance in T.melanospo um. Table 1 p o ides use ul in o ma ion ela ing o hei gas 107
ch oma og aphic e en ion da a, he masses employed o he s udy and hei a oma ic 108
desc ip o s. 109
The s anda ds used o iden i ica ions we e supplied by Ald ich (S einheim, Ge many), 110
Fluka (Buchs, Swi ze land), PolyScience (Niles, USA), Lancas e (S asbou g, F ance), 111
and Al a Aesa (Ka ls uhe, Ge many). 112
2.3. Expe imen al design 113
Figu e 1 illus a es he expe imen al design o his wo k which e alua es possible 114
changes in T. melanospo um a oma p oduced by eezing. Two eezing empe a u es (-115
20ºC and -80ºC) and di e en ime pe iods (1, 20, 40 and 60 days) ha e been assessed. 116
The scheme shown in he igu e was ollowed in o de ha o educe he e ec s o he 117
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s udy should no be a ec ed by he in insic indi idual a ia ions o he u les. Fou 118
sphe ical esh u les o app oxima ely 20 g ams each we e used. A di e en u le 119
was used o each ime pe iod. The ou u les we e di ided in o h ee pieces o almos 120
iden ical size. One piece ac ed as he con ol sample. The o he wo we e acuum 121
packed in polye hylene bags (O ien ed Polyamide/Polyp opylene, 15/65, 80 µm (O ed, 122
Musile di Pia e, I aly)) wi h a VM-12 acuum seale (O ed). Then, one piece was 123
ozen a -20ºC in a Templow-S eeze (J.P. Selec a, Ba celona, Spain) and he o he a 124
-80ºC in MDF-U3286S eeze (Sanyo Elec ic Co., Japan). The empe a u e o bo h 125
eeze s was moni o ed h oughou he expe imen using a Tes o 177-14 Da a Logge 126
wi h a J ype p obe (Tes o Ins umen s, Ba celona, Spain). The empe a u es we e 127
main ained a - 20 ± 1 ºC and a -80 ± 1 ºC. 128
2.4. Senso y analysis 129
The ozen u les we e unpackaged, sliced (∼1 mm) and le a oom empe a u e o 2 130
h o wa m he p oduc . The samples we e hen p esen ed o he panelis s in closed 131
opaque con aine s in andom o de . 132
2.4.1. Senso y desc ip i e analysis 133
P io o he eezing expe imen , a panel o i e ained judges e alua ed he quali y 134
cha ac e is ics o black u les. Two speci ic 1 h aining sessions we e ca ied ou in 135
which he judges iden i ied desc ip i e e ms o de ine he di e en T. melanospo um 136
samples used in his s udy. As a esul o hese discussions, se en a oma a ibu es we e 137
chosen as he mos app op ia e o desc ibe he samples. This g oup o desc ip o s 138
comp ised: sul u , mush oom, mould, animal, boiled po a oes, bu e and inally cheese. 139
In he hi d aining session, he panelis s sco ed he in ensi y o each a ibu e using a 140
i e-poin scale (5 = e y in ense; 4 = in ense; 3 = mode a e; 2 = sligh and 1 = no 141
smell) and in he ou h session hey we e asked o sco e gene al pa ame e s such as 142
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in ensi y o cha ac e is ic a oma using a nine poin a ing scale (9 = ull ypical a oma; 7 143
= mode a ely ull a oma; 5 = mode a e a oma; 3 = sligh a oma and 1 =no ypical 144
smell). 145
2.4.2. Senso y disc imina i e analysis ( iangula es s) 146
The es panel ha ca ied ou he senso y disc imina i e analysis was composed o 8 147
membe s o he labo a o y s a ( h ee women and i e men, anging om 23 o 50 148
yea s o age). All o hem pa icipa e egula ly in senso y es s. 149
To assess whe he he di e en eezing condi ions ha e a signi ican e ec on he 150
a oma o T.melanospo um, iangula es s we e ca ied ou . The samples examined in 151
each es we e esh (con ol) and ozen (bo h om he same u le). The esh con ol 152
samples used on he di e en analysis days we e ob ained om he same cul i a ed 153
u le-g ounds and we e a he same s age o ma u a ion as hose p e iously subjec ed 154
o he eezing p ocess. Th ee closed con aine s we e p esen ed o each judge, who had 155
o decide which sample was di e en om he o he wo. The numbe o co ec 156
answe s was compa ed wi h abula ed alues o decide i signi ican di e ences exis ed. 157
In he case o a di e ence being de ec ed, he judges we e asked o no e he desc ip o s 158
which caused he di e ence. 159
2.5. Headspace Solid-Phase Mic o Ex ac ion (HS-SPME) 160
An SPME holde (Supelco, Belle on e, PA, USA) was used o pe o m hese 161
expe imen s. A ibe o medium pola i y was needed o a oid disc imina ion owa ds 162
e y non pola and pola ola ile compounds. A used silica ibe coa ed wi h a 50/30 163
µm laye o di inylbenzene/ca boxen/polydime hylsiloxane (Supelco) was chosen o 164
ex ac he i een a oma ic compounds. 165
The me hod applied o his analysis was desc ibed in (Culle é, Fe ei a, Ven u ini, 166
Ma co & Blanco, 2012). This is based on a me hodology designed and published in 167
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di e en pape s (Díaz, Ibáñez, Seño áns & Regle o, 2003; Diaz, Seño ans, Regle o & 168
Ibañez, 2002), bu wi h some changes, as o example he mass o u le analyzed. 169
App oxima ely 2 g ams o sample was placed in a 20 mL ial closed wi h a plas ic ilm. 170
Once he desi ed empe a u e (53ºC) was eached, he ial was allowed o condi ion o 171
he equilib ium ime (5 min). A e his ime, he ibe was in oduced in o he ial and 172
exposed o he headspace o he sample du ing 13.6 minu es. These ex ac ion 173
condi ions we e op imized in a p e ious wo k (Diaz e al., 2002). 174
Th ee uni s o u les o each ea men we e cu in o hin slices using a sha p kni e. A 175
o al o h ee eplica es o each u le species we e analyzed. 176
2.6. Gas Ch oma og aphy-Mass Spec ome y (GC-MS) 177
The analyses we e pe o med wi h a CP-3800 ch oma og aph coupled o a Sa u n 2200 178
ion ap mass spec ome ic de ec ion sys em supplied by Va ian (Sunny ale, CA, 179
USA). A DB-WAXETR capilla y column (J&W Scien i ic, Folsom, CA, USA) o 60 m 180
x 0.25 mm I.D., a ilm hickness o 0.25 µm, and p eceded by a 3 m x 0.25 mm 181
uncoa ed (deac i a e, in e media e pola i y) p ecolumn om Supelco (Belle on e, PA, 182
USA) was used. Helium was he ca ie gas a a low a e o 1 mL min-1. The o en 183
empe a u e was ini ially 40 ºC du ing 5 minu es, hen aised by 4 ºC min-1 o 140 ºC, 184
ollowed by a a e o 10 ºC min-1 o 220 ºC and inally held a his empe a u e o 10 185
minu es. The MS pa ame e s we e an MS ans e line and chambe ioniza ion 186
empe a u e o 200 ºC, and a ap emission cu en o 80 µA. The global un ime was 187
eco ded in ull scan mode (45-250 m/z mass ange). The injec ion was in Spli less 188
mode (spli less ime 5 min) a a empe a u e o 200 ºC. A deso p ion ime o 15 minu es 189
was used. The ch oma og aphic da a we e analyzed by Va ian Sa u n GC-MS Ve sion 190
5.2 so wa e. The iden i y o he odo an s was de e mined by a compa ison o hei 191
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ch oma og aphic e en ion index and MS spec a wi h hose o pu e e e ence 192
compounds. 193
The da a is exp essed in a ea pe cen ages. Howe e , gi en ha some o he selec ed 194
analy es do no gi e a signal in TIC o else appea oge he wi h peaks o in e e ence 195
ha make i di icul o in eg a e hem (as is he case wi h 2,3-bu anedione and 3-e hyl-196
5-me hylphenol), i was decided o wo k wi h a ea pe cen ages bu also conside ing 197
selec i e mass. In his way, 100 % cons i u ed he sum o he selec i e ion peak a eas o 198
all he compounds o in e es . 199
2.7. S a is ical analyses 200
Fo he da a ob ained om he senso y desc ip i e analyses, one- ac o ( ime) and wo-201
ac o ( ime and empe a u e) analyses o a iance (ANOVA) we e ca ied ou . A - es 202
analysis was also employed in o de o check possible changes be ween esh and 24 203
hou - ozen samples. 204
Simila one- ac o and wo- ac o ANOVA es s we e also ca ied ou in he case o he 205
chemical quan i a i e da a in o de o e alua e he exis ence o signi ican di e ences in 206
he ola ile composi ion be ween he con ol samples ( esh u les) and he samples 207
subjec ed o he di e en eezing condi ions. Di e ences be ween he a e ages in bo h 208
se s o da a we e s udied wi h Duncan’s es and signi ican di e ences we e es ablished 209
a p < 0.05. 210
All hese analyses we e pe o med using SPSS so wa e ( e sion 15.0) om SPSS Inc. 211
(Chicago. IL). 212
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Re e ences 364
Al-Ruqaie, I. M. (2006). E ec o di e en ea men p ocesses and p ese a ion 365
me hods on he quali y o u les: I. Con en ional me hods (d ying/ eezing). Jou nal 366
o Food P ocessing and P ese a ion, 30(3), 335-351. 367
Andic, S., Tunc u k, Y., & Ja idipou , I. (2011). E ec s o ozen s o age and acuum 368
packaging on ee a y acid and ola ile composi ion o Tu kish Mo al cheese. Food 369
Science and Technology In e na ional, 17(4), 375-394. 370
Bueno, M., Resconi, V. C., Campo, M. M., Cacho, J., Fe ei a, V., & Escude o, A. 371
(2011). Gas ch oma og aphic-ol ac ome ic cha ac e isa ion o headspace and 372
mou hspace key a oma compounds in esh and ozen lamb mea . Food Chemis y, 373
129(4), 1909-1918. 374
Culle é, L., Fe ei a, V., Che e , B., Ven u ini, M. E., Sánchez-Gimeno, A. C., & 375
Blanco, D. (2010). Cha ac e iza ion o a oma ac i e compounds in black u les (Tube 376
melanospo um) and summe u les (Tube aes i um) by gas ch oma og aphy-377
ol ac ome y. Food Chemis y, 122, 300-306. 378
Culle é, L., Fe ei a, V., Ven u ini, M. E., Ma co, P., & Blanco, D. (2012). E alua ion 379
o gamma and elec on-beam i adia ion on he a oma ic p o ile o black u le (Tube 380
melanospo um) and summe u le (Tube aes i um). Inno a i e Food Science and 381
Eme ging Technologies, 13, 151-157. 382
Da ie , P., Pons, M., Lamy, S., & Dubou dieu, D. (2000). Iden i ica ion and 383
quan i ica ion o geosmin, an ea hy odo an con amina ing wines. Jou nal o 384
Ag icul u al and Food Chemis y, 48(10), 4835-4838. 385
de Ancos, B., Cano, M. P., & González, E. M. (1999). Di e en ia ion o aspbe y 386
a ie ies acco ding o an hocyanin composi ion. Z. Lebensm.-Un e s.-Fo sch, 208, 33-387
38. 388
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de Ancos, B., Ibañez, E., Regle o, G., & Cano, M. P. (2000). F ozen s o age e ec s on 389
an hocyanins and ola ile compounds o aspbe y ui . Jou nal o Ag icul u al and 390
Food Chemis y, 48(3), 873-879. 391
Díaz, P., Ibáñez, E., Seño áns, F. J., & Regle o, G. (2003). T u le a oma 392
cha ac e iza ion by headspace solid-phase mic oex ac ion. Jou nal o Ch oma og aphy 393
A, 1017 (1-2), 207. 394
Díaz, P., Seño ans, F. J., Regle o, G., & Ibañez, E. (2002). T u le a oma analysis by 395
headspace solid phase mic oex ac ion. Jou nal o Ag icul u al and Food Chemis y, 50 396
(22), 6468-6472. 397
Escude o, A., Cacho, J., & Fe ei a, V. (2000). Isola ion and iden i ica ion o odo an s 398
gene a ed in wine du ing i s oxida ion: a gas ch oma og aphy-ol ac ome ic s udy. 399
Eu opean Food Resea ch and Technology, 211(2), 105-110. 400
Ibañez, E., Lopez-Sebas ian, S., Ramos, E., Tabe a, J., & Regle o, G. (1998). Analysis 401
o ola ile ui componen s by headspace solid-phase mic oex ac ion. Food Chemis y, 402
63, 281-286. 403
Iglesias, J., Medina, I., Bianchi, F., Ca e i, M., Mangia, A., & Musci, M. (2009). S udy 404
o he ola ile compounds use ul o he cha ac e isa ion o esh and ozen- hawed 405
cul u ed gil head sea b eam ish by solid-phase mic oex ac ion gas ch oma og aphy-406
mass spec ome y. Food Chemis y, 115(4), 1473-1478. 407
Jawo ska, G., & Be nás, E. (2009). The e ec o p elimina y p ocessing and pe iod o 408
s o age on he quali y o ozen Bole us edulis (Bull: F .) mush ooms. Food Chemis y, 409
113, 936-943. 410
Kjeldsen, F., Ch is ensen, L. P., & Edelenbos, M. (2003). Changes in ola ile 411
compounds o ca o s (Daucus ca o a L.) du ing e ige a ed and ozen s o age. Jou nal 412
o Ag icul u al and Food Chemis y, 51(18), 5400-5407. 413
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La Gue che, S., Dauphin, B., Pons, M., Blanca d, D., & Da ie , P. (2006). 414
Cha ac e iza ion o some mush oom and ea hy o -odo s mic obially induced by he 415
de elopmen o o on g apes. Jou nal o Ag icul u al and Food Chemis y, 54(24), 416
9193-9200. 417
Ma, Y. K., Hu, X. S., Chen, J., Chen, F., Wu, J. H., Zhao, G. H., Liao, X. J., & Wang, 418
Z. F. (2007). The e ec o eezing modes and ozen s o age on a oma, enzyme and 419
mic o-o ganism in Hami melon. Food Science and Technology In e na ional, 13(4), 420
259-267. 421
Mon an , C., Kuli ak, M., & Gleyze, R. (1983). No e su la écol e de jeunes ascoca pes 422
du T. melanospo um Vi . (T u e noi e du Pé igo d) e leu é olu ion. Con es Rendus 423
del'Academie des Sciences, Pa is, 296, (pp. 463-468). 424
Nielsen, G. S., & Poll, L. (2004). De e mina ion o odo ac i e a oma compounds in 425
eshly cu leek (Allium ampelop asum Va . Bulga) and in long- e m s o ed ozen 426
unblanched and blanched leek slices by gas ch oma og aphy ol ac ome y analysis. 427
Jou nal o Ag icul u al and Food Chemis y, 52(6), 1642-1646. 428
Ri e a, C. S., Blanco, D., Sal ado , M. L., & Ven u ini, M. E. (2010). Shel -Li e 429
ex ension o esh Tube aes i um and Tube melanospo um u les by modi ied 430
a mosphe e packaging wi h mic ope o a ed ilms. Jou nal o Food Science, 75(4), 431
E225-E233. 432
Schiebe le, P., & Bue ne , A. (2001). In luence o he chain lengh on he a oma 433
p ope ies o homologous epoxy-aldehydes, ke ones and alcohols. In: A. S. S. Eds., 434
Ame ican Chemical Socie y, In a oma ac i e compounds in oods: chemis y and 435
senso y p ope ies (pp. 109-118). Washing on, DC. 436
Sk ede, G. (1996). F ui s, in eezing e ec on ood quali y, edi ed by Les e , E.J. (pp. 437
183-245), Ma cel-Dekke , Inc., New Yo k. 438
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FIGURE CAPTION 439
440
Figu e 1. Expe imen al design 441
Figu e 2. Sco e plo o P incipal Componen Analysis (PCA) applied o quan i a i e 442
da a om esh and ozen u les (a wo di e en empe a u es: -20ºC, -80ºC and a 443
di e en imes: 1, 20, 40 and 60 days). 444
Figu e 3. Sco e plo o P incipal Componen Analysis (PCA) applied o quan i a i e 445
da a om only ozen u les 446
Figu e 4. A e age le els o 1-oc en-3-one ound in all u les analysed 447
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470
T u le 1
( ozen pe iod = 1 day)
T u le 2
( ozen pe iod = 20 days)
T u le 3
( ozen pe iod = 40 days)
T u le 4
( ozen pe iod = 60 days)
- 20 ºC
- 20 ºC
- 20 ºC
- 20 ºC
- 80 ºC
- 80 ºC
- 80 ºC
- 80 ºC
esh
esh
esh
esh
21
Compounds: 1=me hane hiol, 2=dime hylsul ide, 3=diace yl, 4=e hyl 3-me hylbu y a e, 5=dime hyldisul ide, 6=isoamyl alcohol, 7=1-oc en-3-one, 8=dime hyl isul ide, 9=1-
oc en-3-ol, 10=me hional, 11=3-e hyl-5-me hylphenol, 12=2-me hylisobo neol, 13=dime hylsul oxide, 14=p-c esol, 15=3-e hylphenol. C1, C20, C40, C60: F esh samples
(con ol), F1-20C: F ozen samples a -20ºC du ing 1 day, F1-80C: ozen samples a -80ºC du ing 1 day; F20-20C: ozen samples a -20ºC du ing 20 days, F20-80C: ozen
samples a -80ºC du ing 20 days, F40-20C: ozen samples a -20ºC du ing 40 days, F40-80C: ozen samples a -80ºC du ing 40 days, F60-20C: ozen samples a -20ºC
du ing 60 days and F60-80C: ozen samples a -80ºC du ing 60 days.
22
Compounds: 1=me hane hiol, 2=dime hylsul ide, 3=diace yl, 4=e hyl 3-me hylbu y a e, 5=dime hyldisul ide, 6=isoamyl alcohol, 7=1-oc en-3-one, 8=dime hyl isul ide, 9=1-
oc en-3-ol, 10=me hional, 11=3-e hyl-5-me hylphenol, 12=2-me hylisobo neol, 13=dime hylsul oxide, 14=p-c esol, 15=3-e hylphenol. C1, C20, C40, C60: F esh samples
(con ol), F1-20C: F ozen samples a -20ºC du ing 1 day, F1-80C: ozen samples a -80ºC du ing 1 day; F20-20C: ozen samples a -20ºC du ing 20 days, F20-80C: ozen
amples a -80ºC du ing 20 days, F40-20C: ozen samples a -20ºC du ing 40 days, F40-80C: ozen samples a -80ºC du ing 40 days, F60-20C: ozen samples a -20ºC
du ing 60 days and F60-80C: ozen samples a -80ºC du ing 60 days.
1
23
24
Table 1. Re en ion ime, a oma ic desc ip o and selec i e masses co esponding o
each o he i een compounds s udied.
m/z*
e en ion ime (min)
a oma ic desc ip o
Rele an odo an s (by GC-O)
Dime hylsul ide
62
4.15
u le, sul u
Diace yl
43
10.14
bu e , c eam
E hyl 3-me hylbu y a e
88
13.76
ui , anise
Dime hyldisul ide
94
14.08
u le, sul u
Isoamyl alcohol
70
19.49
cheese
3-E hyl-5-me hylphenol
121
30.96
phenolic/lea he
p-C esol
107
40.56
phenolic/lea he
3-E hylphenol
122
41.98
phenolic/lea he
Mush oom- like no es
1-Oc en-3-one
70
22.93
mush oom
1-Oc en-3-ol
57
27.99
mush oom, ea h, mould
Mouldy/ea hy- like no es
2-Me hylisobo neol
95
31.48
mould, ea h
Boiled po a oes- like no es
Me hional
104
28.73
boiled po a oes
O he sul u ic compounds
Me hane hiol
47
3.70
cooked cabbage, ege al
Dime hyl isul ide
126
24.43
o en ood
Dime hylsul oxide
78
32.34
ga lic-like no e
*m/z: selec i e masses chosen o iden i ying each compound
25
Table 2. Senso y desc ip i e analysis o T. melanospo um (black u le) ozen a -20
ºC and a -80 ºC
Pa ame e
Day
F esh
F ozen*
a -20 ºC
a -80 ºC
A oma in ensi y
1
20
40
60
7.7 ± 1.3a
6.8 ± 1.7a,A
7.2 ± 1.5a,A
6.6 ± 0.9a,A
6.4 ± 1.9a,A
7.0 ± 0.7a,A
6.9 ± 0.7a,A
6.3 ± 0.8a,A
6.6 ± 1.1a,A
Cha ac e is ic esh
u le a oma
1
20
40
60
7.8 ± 1.3a
5.9 ± 1.8b,A
5.5 ± 0.7b,A
6.2 ± 1.1ab,A
4.8 ± 1.4b,A
5.6 ± 1.4b,A
6.0 ± 1.0b,A
5.2 ± 0.9b,A
5.1 ± 1.3b,A
Sul u
1
20
40
60
1.7± 0.8a
2.1 ± 0.5a,A
2.3 ± 0.6a,A
2.4 ± 0.6a,A
1.6 ± 0.8a,A
2.6 ± 0.7a,A
2.3 ± 0.4a,A
2.2 ± 0.5a,A
2.0 ± 0.9a,A
Mush oom
1
20
40
60
2.4 ± 1.1a
1.8 ± 0.4a,A
3.7 ± 1.0b,B
2.6 ± 1.1a,AB
2.0 ± 0.7a,A
1.8 ± 0.6a,A
2.7 ± 0.9a,A
2.4 ± 0.7a,A
2.2 ± 0.9a,A
Mould
1
20
40
60
1.0 ± 0.0a
1.0 ± 0.0a,A
4.0 ± 0.7b,B
1.0 ± 0.0a,A
1.0 ± 0.0a,A
2.7 ± 1.1b,A
1.0 ± 0.0ac,B
3.8 ± 0.4b,C
1.0 ± 0.0a,B
Animal
1
20
40
60
1.9 ± 0.9a
1.9 ± 0.5a,A
2.4 ± 1.0a,A
2.0 ± 1.0a,A
1.8 ± 1.0a,A
2.1 ± 0.7a,A
2.2 ± 0.9a,A
1.9 ± 1.2a,A
1.8 ± 1.2a,A
Boiled Po a oes
1
20
40
60
1.4 ± 0.5a
2.0 ± 1.1a,A
2.2 ± 0.8ab,A
2.4 ± 0.9a,AB
3.5 ± 0.7b,B
1.7 ± 0.7a,A
2.9 ± 0.5b,B
2.4 ± 0.5a,AB
2.8 ± 0.6b,AB
Bu e
1
20
40
60
1.5 ± 0.5a
1.2 ± 0.3a,A
1.3 ± 0.4a,A
1.4 ± 0.4a,A
1.4 ± 0.8a,A
1.4 ± 0.7a,A
1.4 ± 0.4a,A
1.4 ± 0.5a,A
1.4 ± 0.8a,A
Cheese
1
20
40
60
1.3 ± 0.7a
1.3 ± 0.4a,A
1.3 ± 0.4a,A
1.4 ± 0.6a,A
1.0 ± 0.0a,A
1.1. ±0.2a,A
1.6 ± 0.6a,A
1.3 ± 0.5a,A
1.0 ± 0.0a,A
*Values in he same line ollowed by di e en lowe case le e s show signi ican di e ences be ween
ba ches o he same day (p<0.05). In his case ozen samples we e always compa ed wi h he ini ial
alue o he esh sample (day 0). Values in he same column ollowed by di e en capi al le e s a e
signi ican ly di e en (p<0.05).
