Volatile Compounds and the Changes in Their Concentration Levelstable

eadda f-lvrbe Lnadadohr 08.Ti off- This 2008). others and (Landaud beer off-flavor as sometimes is regarded compounds these of amount higher products, a food containing many Although beer of 2005). aromas the (Kondo to contribute beer compounds standard these does than compounds fur for preference consumer a and price taste. low light its only of because larity and beer ( ( content, 4 beer into standard malt categorized (100%), mainly beer on are types—all-malt Japan Based in content. beverages alcohol malt beer-like on based beer mnsi er(ih n tes20) PEi solvent-free and a extraction is simultaneous SPME for 2006). allows others that el- trace technique and of extraction (Pinho analysis mi- beer the phase in for used ements solid been as have such (SPME) croextraction techniques threshold. preparation low-odor a deter- sample have for they Several important because extremely profile, only are flavor present the they are mining beer, compounds in amounts flavor compounds trace stale flavor in most stale Although of for beer. amount conducted in the been (Vander- decreasing have unpleasant and studies sometimes Many detecting be 2006). can others and which haegen beer, of taste the reported. been nonmalt not in has volatiles stan- beer of of study those comprehensive from flavor a However, different the beer. be Therefore, dard must 2002). beer others nonmalt of and stability (Gijs and flavor respon- also stale is a pH for low and sible 2007), others and (Guido affects strongly stability beer flavor in polyphenol of amount The (Shimizu 2002). beer others of standard and in amount than total beer nonmalt and in lower pH is The polyphenol 2005). (Kondo dur- fermentation conditions yeast starvation ing nitrogen to attributable mainly is flavor B T Concentrations H. Malt Varying Beers Containing in Storage during Their Levels in Concentration Changes the and Compounds Volatile JFS ute erdcinwtotpriso sprohibited is permission without reproduction Further [email protected]). (E-mail: Tsuji with author to are inquiries Direct Authors 739- Japan. Higashihiroshima 0046, Kagamiyama, 10/8/2009. 3-7-1 Brewing, Accepted of Inst. 7/2/2009, Research Natl. Submitted 20090630 MS o:10.1111/j.1750-3841.2009.01428.x doi: C < 5) n oml er(%.Lwml erhsgie popu- gained has beer Low-malt (0%). beer nonmalt and 25%), gn fbe samjrqaiypolmbcuei changes it because problem quality major a is beer of Aging sul- and alcohol of amount higher a contains beer Low-malt 09Isiueo odTechnologists Food of Institute 2009 hssuy eoio a eago akrcniaergrls fteml content. malt the of in regardless analyzed candidate beer marker nonmalt good most a in be storage may during Nerolidol increase study. not this did beer. compounds standard these and all-malt of in concentrations those Additionally, from different were beer nonmalt 2-methoxy-4-vinylphenol—in and cinnamate, 7tree,1 loos 1ais uas rm opud,5croys n te opud.A nlssof analysis An compounds. other stir compounds— and flavor carbonyls, the stale 5 of using levels compounds, concentration aroma by the 6 that analyzed furans, suggested were beer 6 esters, aged acids, malt compounds—25 11 90 of alcohols, identified amounts 14 We terpenes, different method. 17 with spectrometry brewed chromatography-mass extraction–gas beers sorptive in bar compounds Volatile ABSTRACT: e otiigasalaon fml spoue nJapan, on in taxes produced imposes government is Japanese malt the of because primarily amount small a containing eer UIAND SUJI ewrs er at BE tl lvr oaiecompounds volatile flavor, stale SBSE, malt, beer, Keywords: :Fo Chemistry Food C: .M A. IZUNO Introduction R > 7) o-atbeer low-malt 67%), irnlo CSn 0-29,9dcn1o CSn 13019-22-2), nr (CAS 9-decen-1-ol 106-22-9), nr (CAS 505-10- citronellol nr 2-undecanol (CAS 123-25-1), 3-(methylthio)-1-propanol 2), nr 1653-30-1), (CAS nr (CAS succinate acid (CAS 76649-16- diethyl 2-methyl-butyric nr 503-74-2), 116-53-0), (CAS nr nr (CAS 4-decenoate acid ethyl isovaleric 98-00-0), furfuryl 6), nr 150-84-5), (CAS nr de- 2035-99- (CAS alcohol nr ethyl acetate (CAS 3,7-dimethyl-6-octen-1-yl 112-12-9), n-caprylate isoamyl 6), nr 110-38-3), (CAS nr (CAS 5-methyl-2-furaldehyde 2-undecanone canoate 79-31-2), 620-02-0), nr nr (CAS (CAS acid 111- nr isobutyric (CAS 1-octanol 87-5), nonanoate 78-70-6), nr ethyl (CAS linalool 628-99-9), 123-29-5), nr nr (CAS (CAS 2-nonanol 104-76-7), 2-furaldehyde nr 2-ethyl-1-hexanol (CAS 64-19-7), 112-14-1), nr nr (CAS acetate (CAS octyl 98-01-1), nr acid 1-heptanol (CAS acetic 106-32-1), 53535-33-4), nr nr (CAS nonanal octanoate (CAS 821-55-6), ethyl nr 124-19-6), (CAS hep- nr 2-nonanone (CAS ethyl 112-06-1), ac- 142-92-7), nr heptyl 111-27-3), (CAS nr nr etate (CAS (CAS 1-hexanol 106-30-9), acetate nr (CAS hexyl tanoate 123-66-0), nr hexanoate ethyl (CAS 123-51-3), nr (CAS nr 3-methyl-1-butanol (CAS isobutyrate 2050-1-3), isoamyl 123-35-3), nr (CAS myrcene nr 123-92-2), (CAS 108-64- acetate isopentyl nr 78-83-1), nr (CAS (CAS isovalerate 2-methyl-1-propanol ethyl 5), 105-54-4), nr (CAS butyrate ethyl ntefbr slmtd trbrsrtv xrcin(BE,o the on (SBSE), extraction sorptive bar Stir limited. is coated fibers be can the that on material sorptive of amount the because sensitivity, limited but reproducibility good very has SPME concentration. Chemicals noted. also was gas storage during with beers these the SBSE in changes Additionally, using of pattern by (GC-MS). malts spectrometry of chromatography-mass amounts different containing beer. in 2003) others and (Ochiai carbonyls and acids fatty of analysis the to (Hor applied very been in present has are SBSE that amounts. compounds small detect to us allows hand, other sbtlaeae(hmclAsrcsSrie[A r110-19-0]), nr [CAS Service Abstracts (Chemical acetate Isobutyl nti td,w oprdvltl opstoso beers of compositions volatile compared we study, this In α kadohr 07,treod Ksiooadohr 2006), others and (Kishimoto terpenoids 2007), others and ak ´ freee(A r526-) -eao CSn 112-30-1), nr (CAS 1-decanol 502-61-4), nr (CAS -farnesene o.7,N.1 2010 1, Nr. 75, Vol. aeil n Methods and Materials β -damascenone, — ORA FFO SCIENCE FOOD OF JOURNAL γ nnlcoe ethyl -nonalactone, C 79

C: Food Chemistry (Continued). %) in total area ( Percent nd nd to 0.002 nd to 0.005 0.003 to 0.01 nd nd nd nd –nd nd to 0.027 nd to 0.035 nd to 0.041 nd to 0.157 0.013 to 0.05 0.01 to 0.039 0.018 to 0.035 nd to 0.01 0.007 to 0.014nd to 0.004 0.06 to 0.006 0.005 to 0.012 nd to 0.056 nd to 0.009 0.01 to 0.148 nd to 0.02nd to 0.03 0.001 to 0.104 0.004 to 0.003 0.017 to 0.012 0.014 to 0.014 0.01 to 0.012 0.016 to 0.021 nd to 0.039 nd to 0.05nd to 0.067nd to 0.226 nd to nd 0.02 nd to 0.052nd to 0.035 nd to 0.011 nd to 0.029 0.021 to 0.033nd to 0.071 nd to 0.025 nd to 0.023 0.025 to 0.06 0.003 to 0.082 0.012 to 0.023 0.011 to 0.056 0.008 to 0.017 nd to 0.047 nd to 0.033nd to 0.226 nd to 0.012nd to 0.013 nd to 0.133nd to 0.029 nd nd to 0.011 0.002 to 0.012 nd nd to 0.003 nd to 0.024 0.006 to 0.01 nd nd to to 0.007 0.34 0.007 to 0.008 0.204 to 0.69 0.087 to 0.199 0.056 to 0.103 0.079 to 0.211 0.094 to 0.39 0.106 to 0.285 0.063 to 0.0780.029 to 0.09 0.06 to 0.091 0.004 to 0.009 0.006 to 0.011 0.01 to 0.011 0.06 to 0.11 0.084 to 0.156 0.076 to 0.117 0.094 to 0.17 –nd to 0.022 0.023 to 0.039 0.033 to 0.047 0.022 to 0.04 11 to 25.533 16.567 to 18.867 18.1 to 22.733 15.167 to 23.767 0.065 to 0.123 0.036 to 0.087 0.051 to 0.0630.159 to 0.408 0.048 to 0.066 0.058 to 0.179 0.098 to 0.1440.018 to 0.061 0.095 to 0.144 0.006 to 0.031 nd nd 0.006 to 0.045 nd to 0.011 nd to 0.008 nd to 0.002 0.039 to 0.047 0.025 to 0.044 0.013 to 0.061 0.012 to 0.03 0.028 to 0.038 0.012 to 0.031 0.004 to 0.036 nd to 0.002 + 0.006 to 0.09 nd to 0.022 nd nd to 0.038 0.002 to 0.01 nd to 0.008 nd to 0.001 0.002 to 0.007 7.753 to 15.733 7.74 to 12.133 6.993 to 8.853 6.04 to 10.043 1.637 to 8.157 2.697 to 5.333 4.467 to 4.747 3.003 to 3.407 –0.01 to 0.0240.037 to 0.142 0.018 to 0.0410.004 to 0.006 nd to 0.074 0.022 to1.067 0.047 to 1.267 nd to 0.004 0.0050.014 to to 0.023 0.039 0.0140.018 to to 0.997 0.046 0.045 to 2.407 0.002 to 0.007 0.009 0.002 to to 0.115 0.035 1.084 to nd 3.307 to 0.043 0.002 to 0.005 0.004 to 0.041 1.079 to 3.953 0.025 to nd 0.287 to 0.037 0.004 to 0.021 0.152 to 0.258 nd to 0.026 0.106 to 0.061 0.088 to 0.4592.313 to 5.367 0.171 to 0.289 nd 3.933 to 6.073 0.228 to 0.353 5.123 to 7.803 nd to 0.176 3.91 to 7.287 0.074 to 0.1750.169 to 0.3310.005 to 0.031 0.184 to 0.265 0.224 to 0.3070.007 to 0.576 nd to 0.027 0.142 to 0.233 0.241 to–2.18 0.311 to 4.067 nd to 0.191 nd to 0.108 0.001 to 0.169 0.256 to 3.407 0.277 to 5.6170.032 to 0.162 nd nd to 0.01 3.523 to 0.033 5.42 to 0.0860.004 to 0.011 4.293 0.052 to to 5.637 0.089 0.006 to 0.009 nd to 0.329 0.011 to 0.0410.032 to 0.295 0.054 to 0.077 0.003 to0.002 0.009 to 0.057 nd to 0.119 0.018 to 0.267 nd to nd 0.006 to 0.032 0.112 to 0.23 nd to 0.01 0.171 to 0.307 nd 0.007 to 0.011 nd to 0.075 + + + –0.058 to 0.119 0.088 to 0.118 0.095 to 0.12 0.104 to 0.133 –0.014 to 0.088 0.046 to 0.121–0.011 to 0.057 0.107 to 0.15 0.033 to 0.109 0.208 to 0.257 0.088 to 0.127 0.1 to 0.159 + + + + + + –6.913 to 14.733 11.367 to 16.533 14.667 to 16.2 12.8 to 16.7 + b b Vol. 75, Nr. 1, 2010 b b — b a a a a a b a a a a a a a a a a a a a a a a a a a a a a a b a a a a a a a a a a a a a a a a a a a acid 2-methyl-propanoate -Damascenone nd to 0.068 nd to 0.085 0.069 to 0.072 0.028 to 0.09 -Farnesene nd to 0.121 nd to 0.088 0.056 to 0.081 0.023 to 0.066 -Calacorene nd to 0.01 nd nd nd to 0.002 -Farnesene -Ionene nd nd nd nd α β α α β JOURNAL OF FOOD SCIENCE 80 1846 Ethyl laurate 1680 Isovaleric acid 1552 Linalool 16991705 Ethyl 9-decenoate Methyl 3,7-dimethyl-2,6-octadienoic 0.065 to 0.739 0.213 to 0.701 0.319 to 1.77 0.231 to 0.982 1721 2-Undecanol 1459 1-Heptanol 1592 5-Methyl-2-furaldehyde 1681 2-Methyl-butyric acid 17981819 Ethyl phenylacetate 9-Decen-1-ol –nd nd to 0.005 0.002 to 0.007 0.002 to 0.005 1219 3-Methyl-1-butanol 1522 2-Nonanol 18741896 3-Hydroxy-2,4,4-trimethylpentyl Ethyl-3-phenylpropionate 1297 Methyl 2-methyl-heptanoate 1403 2-Nonanone 1143 Isopentyl acetate 1342 Ethyl heptanoate 1243 Ethyl hexanoate 1929 1963 (E)-2-Methyl-2-pentenoate 15491561 Furfuryl acetate 1-Octanol 1609 2-Undecanone 16631667 1-Nonanol1671 Isoamyl n-caprylate 1673 Furfuryl alcohol Ethyl 4-decenoate 1725 3-(Methylthio)-1-propanol nd to 0.0211768 Citronellol 1832 nd nd nd to 0.006 103611001100 Ethyl butyrate 1105 Ethyl isovalerate 2,7-Dimethyl-1,6-octadiene 2-Methyl-1-propanol 12071207 D-Limonene Isoamyl isobutyrate 1284 0.755 to 2.11 Hexyl acetate 1346 0.231 to 0.835 Methyl 4-methylene-hexanoate 1430 0.348 to 0.548 nd to 0.006 3-(4-Methyl-3-pentenyl)-furan 0.355 to 0.891 14841494 Octyl acetate nd1545 2-Ethyl-1-hexanol Ethyl nonanoate nd nd to 0.005 1778 3-Tridecanone 1443 Ethyl octanoate 1753 18491852 Geraniol Hexanoic acid nd to 0.023 nd to 0.021 0.004 to 0.014 nd to 0.01 1328 3-Methyl-2-buten-1-ol 1383 Heptyl acetate 1923 2-Phenylethanol 1169 Myrcene 1574 1579 Isobutyric acid 16211646 2-Decanol Ethyl decanoate 1674 16841686 Ethyl benzoate Diethyl succinate nd to 0.1051764 1-Decanol nd to 0.0381827 2-Phenylethyl acetate nd nd nd to 0.127 nd to 0.066 nd nd 1303 Ethyl 4(E)-hexenoate 1357 1-Hexanol 1407 Nonanal 146414641482 Acetic acid Isopentyl hexanoate 2-Furaldehyde nd to 0.003 0.073 to 0.116 nd to 0.007 0.035 to 0.106 0.002 0.041 to to 0.008 0.094 0.001 0.023 to to 0.006 0.085 C 1007 Isobutyl acetate Volatile compounds and the changes in their concentration levelsTable . 1 . --- Compounds . identified in fresh beers containing different amounts ofRI malt. Compound NM LM HM AM 1670 3,7-Dimethyl-6-octen-1-yl acetate C: Food Chemistry += ICmon ML MAM HM b LM a RI NM Compound 1-Dodecanol 1973 RI . Continued. --- . 1 . Table levels concentration their in changes the and compounds Volatile cd(A r120-) -ehx--iypeo CSn 7786- nr (CAS 2-methoxy-4-vinylphenol nonanoic 61-0), 103-36-6), 112-05-0), nr nr (CAS (CAS acid cinnamate acid octanoic ethyl 40716-66-3), 124-07-2), nr nr (CAS (CAS trans-nerolidol 118-71-8), 104-61-0), nr nr (CAS maltol 112-53-8), nr (CAS 1-dodecanol 60-12-8), nr (CAS Peaks beer. ethyl-3-phenylpropionate 2-phenylethanol 2021-28-5), GC-MS. all-malt 142-62-1), nr (CAS (B) with nr in (CAS SBSE 106- or acid by nr beer hexanoic (CAS obtained nonmalt laurate 33-2), beer ethyl (A) 103-45-7), in all-malt nr high (B) (CAS acetate are and 2-phenylethyl concentrations beer whose nonmalt compounds represent (A) arrows of by chromatograms indicated ion --- Total 1 Figure 42Lui cdn o005n o018n o009007t 0.307 to 0.017 0.019 to nd 0.148 to nd 0.095 to nd nd 0.016 to nd 5-Hydroxymethyl-2-furaldehyde acid Lauric 0.032 to nd 2520 2492 0.033 to nd acid Octanoic alcohol Caryophyllenyl tetradecanoate Ethyl 2069 2058 2054 08Nerolidol 2048 2039 32Farnesol acid 9-Decenoic 2362 2342 2-Methoxy-4-vinylphenol 2206 cinnamate Ethyl 2140 15Nnni acid Nonanoic 1,5,5,8-Tetramethyl-3,7- 2175 2165 2436 2222 94Maltol 1974 20Dcni acid Decanoic 3,7,11-Trimethyl-6,10- 2280 2273 2236 NM nd – dnie ntebsso ohms pcrldt n eeeta compounds. beer. referential in and time data 1st spectral the mass for both identified of Compound basis the on Identified = = = A B = oe ocnrto nnnatbe hni l-atbe,a ofimdb NV ( ANOVA by confirmed as beer, all-malt in than beer nonmalt in concentration lower ihrcnetaini oml erta nalml er scnre yAOA( ANOVA by confirmed as beer, all-malt in than beer nonmalt in concentration higher oasrtninidxo PINWXcolumn. HP-INNOWAX on index retention Kovats Intensity (% in max height) Intensity (% in max height) detected. not oml 0) M o-at( low-malt LM: (0%), nonmalt 12.5 12.5 α 25 25 bsbll(A r7612-) eaocai CSnr (CAS acid decanoic 72691-24-8), nr (CAS -bisabolol γ δ α α -Dodecalactone -Bisabolol -Cadinol -Nonalactone cycloundecadien-1-ol dodecadien-1-ol 1 10 10 a 2 3 4 a a 5 a , b a 6 a a a a 7 a a a , b 8 < 5) M ihml ( high-malt HM: 25%),

9 10 a 11 12 a

3 20 20 γ 14 nnlcoe(CAS -nonalactone 15 > 16 + + + 014t .9 .7 o056022t .402 o0.353 to 0.21 0.34 to 0.222 0.368 to 0.165 0.516 to 0.071 0.157 to 0.122 0.199 to –0.104 0.181 to 0.031 0.034 to –0.006 .0 o001n o005008t .1 .0 o0.008 to 0.008 0.015 to 0.008 0.308 5.367 to to 0.099 1.537 0.021 to nd 0.025 0.615 to to nd 0.088 4.09 to 1.39 0.242 to 0.054 0.051 0.039 to to 3.627 4.783 0.006 nd to to 2.713 1.092 0.219 to 0.021 1.763 to 3.41 1.293 to 0.035 2.32 to nd 2.977 to 2.037 0.035 to 0.006 2.597 to –1.35 .7 o018007t .6 .2 o00800 o0.143 to 0.03 0.068 to 0.024 0.061 to 0.037 0.198 to 0.073 7) n M l-at(100%). all-malt AM: and 67%), .3 o007n o008n o008n o0.016 to nd 0.018 to nd 0.028 to nd 0.067 to 0.034 .3 o036018t .7 .3 o026002t 0.096 to 0.042 0.236 to 0.034 0.273 to 0.128 0.316 to 0.232 .8 o014005t .5 .1 o009007t 0.059 to 0.017 0.029 to 0.016 0.058 to 0.015 0.124 to 0.084 n o00500 o00 .2 o006003t 0.062 to 0.023 0.066 to 0.027 0.04 to 0.01 0.015 to –nd dt .0 .3 o015016t .7 .2 o0.179 to 0.122 0.016 to 0.005 0.174 to 0.116 0.016 to 0.013 0.165 to 0.032 0.038 to 0.004 0.308 to nd 0.008 to nd dt .7 dt .9 .7 o01009t 0.13 to 0.019 0.1 to 0.077 0.193 to nd 0.071 to nd dt .4002t .2 .1 o003009t 0.024 to 0.009 0.023 to 0.014 0.021 to 0.012 0.04 to nd 0.154 to 0.078 0.101 to 0.076 0.122 to 0.02 0.09 to nd

17 18 dn dn o0.001 to nd nd nd nd 19 20 21 eeprhsdfo oy hmclIdsr o,Ld (Tokyo, Ltd. Co., Industry C21-25) Chemical Japan). and Tokyo C17-20, from and purchased C12-16, 4602-84-0), were (C7-11, nr mixtures (CAS farnesol standard Industry, 623-17-6), paraffin nr Chemical (CAS fur- acetate Pure 556-82-1), furyl nr Wako (CAS 3-Methyl-2-buten-1-ol Japan). from (Tokyo, nr Ltd. purchased (CAS were 5-hydroxymethyl-2-furaldehyde and 67-47-0) 713-95-1), 14436-32-9), nr nr (CAS (CAS acid 9-decenoic 334-48-5), 22 30 30 23 P P < < 24 0.01). 0.01). o.7,N.1 2010 1, Nr. 75, Vol. 25 26 Percent 27 ( )i oa area total in %) — 40 40 ORA FFO SCIENCE FOOD OF JOURNAL 16... Ethyl phenylacetate 10... Ethyl nonanoate 26... 21... 1-Dodecanol 15... 3-Tridecanone 14... Citronellol 13... Isovarelic acid 11... Isobutyricacid 9... 2-Ethyl-1-hexanol 6... 3-Methyl-2-buten-1-ol 3... 3-Methyl-1-butanol 2... Isoamyl isobutyrate 1... 2-Methyl-1-propanol 22... Caryophyllenyl alcohol 20... (E)-2-Methyl-2-pentenoate 2-methyl-propanoate 18... 3-Hydroxy-2,4,4-trimethylpentyl 27... Farnesol 25... 2-Methoxy-4-vinylphenol 24... Ethyl cinnamate 23... Octanoicacid 19... Ethyl-3-phenylpropionate 17... Hexanoic acid 12... Isoamyl n-caprylate 8... Ethyl octanoate 7... Heptylacetate 5... Hexyl acetate 4... Ethyl hexanoate α -Cadinol Time (min) Time (min) δ -dodecalactone C 81

C: Food Chemistry = C/min C/min, ◦ ◦ C. (A) ◦ C(for1min)to ◦ -nonalactone, -damascenone, Figure 3 --- Changes in concentrations of stale ﬂavors in beer samples during storage at 30 β (B) γ (C) 2-methoxy-4- vinylphenol, and (D) ethyl cinnamate. Catarateof12 ◦ 0.25 mm; film thickness NM1 NM2 NM3 NM4 LM1 LM2 LM3 LM4 LM5 HM1 HM2 HM3 AM1 AM2 AM3 NM1 NM2 NM3 NM4 LM1 LM2 LM3 LM4 LM5 HM1 HM2 HM3 AM1 AM2 AM3 C. Then, the temperature of ◦ = 150 C/min. The desorbed compounds − ◦ 4 4 150 to 230 C (for 1 min) at a rate of 15 ◦ ∗ − ∗ 30 m; i.d. = ∗ 2 2 -Nonalactone Time (weeks) Time (weeks) γ Ethyl cinnamate C (for 4 min) to 100 ∗ ◦ 0.5 mm. (Gerstel, Mulheim an der Ruhr, Germany). Ten m), and helium was used as the carrier gas. The velocity of C(for4min)atarateof60 μ ◦ × SBSE was performed by using a “Twister” with dimensions of A thermal desorption system (TDS2, Gerstel) was used for des- A gas chromatograph (Agilent Technologies, Santa Clara, Calif., ∗ were cryofocused in the CIS 4 at SBSE Analysis of compounds in beer 10 orption of compounds from theintroduced Twister. The into desorption a tube thermodesorption was thermally unit. desorbed Then, by the heating stir bar TDS2 was from 20 milliliters of each beer sample wereing poured the into Twister a and 10 sealed mL with a vial siliconewas by operated septum. us- at Then, 750 the rpm Twister for 1 h for extraction at room temperature. and maintained for 4 min.into The a trapped GC column compounds in were the injected splitless mode. U.S.A.; N6890) and5973N) a were used mass for separation spectrometer andgraph detection. The (Agilent was gas equipped chromato- Technologies; with an HP-INNOWAXlent capillary Technologies; column length (Agi- the carrier gas was 1 mL/min. Thelows: temperature 30 program was as fol- 230 the CIS 4 was increased from 0.25 01 01 0 0

0.4 0.3 0.2 0.1

0.12 0.08 0.04

% in total area total in % area total in % PC1 (56%) AM3 AM+HM NM1 NM2 NM3 NM4 NM1 NM2 NM3 NM4 LM1 LM2 LM3 LM4 LM5 HM1 HM2 HM3 AM1 AM2 AM3 LM1 LM2 LM3 LM4 LM5 HM1 HM2 HM3 AM1 AM2 AM3 NM2 HM3 HM2 AM2 4 4 LM5 ∗ ∗ Vol. 75, Nr. 1, 2010 HM1 — AM1 LM4 LM2 LM3 ∗ ∗ 2 2 PC2 (27%) LM1 LM -Damascenone Time (weeks) Time (weeks) β ∗ ∗ 2-Methoxy-4-vinylphenol NM4 ∗ ∗ NM1 NM3 01 01 0 0 JOURNAL OF FOOD SCIENCE 0.2 0.6 0.4

0.20 0.15 0.10 0.05 0.25

C for 1, 2, and 4 wk. After being stored, they were maintained at area total in % NM area total in % ◦ Fifteen commercial beers—4 nonmalt beers, 5 low-malt beers, Aged samples of beers were prepared by storing fresh beers at C until further analysis. CD AB ◦ 82 C Figure 2 Plots --- axes. of beer samples onto PCA component Volatile compounds and the changes in their concentration levelsBeer . samples . . 2 3 standard beers, and 3 all-malt beers—wereThese analyzed beers in are this produced by study. major breweries inin Japan and 350 supplied mL cans. 30 C: Food Chemistry 5min)atarateof15 nlsswr efre oeiiaetecnaiainpeaks. contamination blank the and eliminate times, to 3 performed performed were were analyses analyses All eV. 70 at mode pact ( ratio charge tnadbes n l-atbeswr nlzdb using by analyzed beers—were all-malt 3 and beers, standard 3 with performed variance were JMP6(SASInst.Inc.,Cary,N.C.,U.S.A.). analyses and of component analysis repli- area principal one-way by and a (ANOVA) peak results using these mean Comparisons analyses. from The cate calculated (NIST). were deviations AMDIS standard using by matogram 1978). (Halang others literature and the in reported procedure the with accordance compounds, in reference as calcu- (C10-C25) were samples paraffin indices using retention by those lated GC with The indices compounds. retention standard GC the their of and library NIST the in those dniiaino oaiecompounds volatile of Identification analysis quantitative and Identification . 150 . to . 100 levels concentration their in changes the and compounds Volatile h assetoee a e odtc oswt mass-to- a with ions detect to set was spectrometer mass The ite omrilbes4nnatbes o-atbeers, low-malt 5 beers, nonmalt beers—4 commercial Fifteen chro- ion total the from determined were percentages area Peak with spectra their comparing by identified were Compounds ◦ fr5mn tart f5 of rate a at min) 5 (for C C AB

% in total area m % in total area 0.01 0.02 0.03 / 0.1 0.2 0.3 z f3 o40adwsoeae nteeeto im- electron the in operated was and 400 to 35 of ) 0 0 eut n Discussion and Results 01 12 01 ◦ C/min. Diethyl succinate Time (weeks) Time (weeks) Nerolidol 2 ◦ /i,ad10t 240 to 150 and C/min, 4 4 ◦ AM3 AM2 AM1 HM3 HM2 HM1 LM5 LM4 LM3 LM2 LM1 AM3 AM2 AM1 HM3 HM2 HM1 LM5 LM4 LM3 LM2 LM1 NM4 NM3 NM2 NM1 NM4 NM3 NM2 NM1 C(for e u nweg,ti ste1ttm ht3tieaoewas 3-tridecanone that time 1st As the beer. is nonmalt this in knowledge, in only shown our detected is per was beer 3-Tridecanone all-malt or 1. nonmalt Figure fresh the ( of test chromatogram ANOVA one-way the ing mut fmalt of amounts different containing beers among Comparison estvt fti ehdfclttstedtcino uhtrace such of detection the The facilitates components. method delta-dodecalactone. this of and sensitivity alpha-bisabolol, (E)-2-methyl- 2-pentenoate, 3-tridecanone, acetate, 3-(4-methyl-3-pentenyl)-furan, 4(E)-hexenoate, 3,7-dimethyl-6-octen-1-yl 4-methylene-hexanoate, ethyl 2-methyl-1-heptanoate, methyl time beer—methyl 1st the for in detected were compounds Nine method. including by this detected using flavor flavors be beer could tangy 2-phenylethanol imparting and isovalerate, and compounds isobutyrate, fruity, other Many peppery, beer. imparts to linalool, myrcene, and example, for prenol, hop; The by flavor originated hexanoate. important commonly also ethyl compounds, is Terpene such and terpene. is compounds, octanoate, group major flavor ethyl secondary acetate, important isopentyl including primary ester, as The an 1). carbonyls, (Table is 5 identified group compounds, compounds—were aroma terpenes, other 6 6 17 furans, and 6 esters, acids, 11 compounds—25 alcohols, Ninety 14 GC-MS. with SBSE

% in total area us- samples among compositions volatile the compared We 0.01 0.20 0.30 0.35 0 01 o.7,N.1 2010 1, Nr. 75, Vol. lsdrn trg at 30 storage during samples beer in stituents con- in of concentrations --- Increase 4 Figure C nerolidol. (C) (B) nate, ◦ .()dehlsucci- diethyl (A) C. Time (weeks) α -Ionene α 2 inn,and -ionene, — ORA FFO SCIENCE FOOD OF JOURNAL P < .1 Tbe1.Tettlion total The 1). (Table 0.01) 4 AM3 AM2 AM1 HM3 HM2 HM1 LM5 LM4 LM3 LM2 LM1 NM4 NM3 NM2 NM1 C 83

C: Food Chemistry ˇ Cejka P. 2007. Determination of some beer ´ aM, References Conclusions -nonalactone, ethyl cinnamate, and 2- 0.05). γ < ık J, Jurkov ´ P ˇ Cul -value test ( -nonalactone were lower for nonmalt beers. Because these t -damascenone, γ ´ ak T, Kellner V, β n this study, we identified 90 compoundsious in amounts beer containing of var- malt by using SBSE with GC-MS. Compared to lease and 4-vinylguaiacoltions formation for feruloyl during esterase activity brewing52(3):602–8. in Saccharomyces and cerevisiae. J fermentation: Agric Food indica- Chem damascenone and dimethyl trisulfide productionChem through 50:5612–6. beer aging. J Agric Food of malt quality parametersFood and Chem 55:728–33. beer flavor stability: multivariate analysis. Jof Agric retention indices in temperature-programmed gas-liquidChem chromatography. Anal 13:1829–32. flavours by stir113(2):154–8. bar sorptive extraction and solvent backcompounds extraction. in J unhopped InstJ Brew beer Agric Food Chem and 54:8855–61. beers hopped withJ different Brew Soc hop Jpn 100:787–95. varieties. metabolism of methionine andMicrobiol other Biotech sulfur 77:1191–205. compounds in fermented food. Appl stale-flavor carbonyl compounds in beer byderivatization stir bar and sorptive extraction thermal withJ in-situ desorption-gas Chromatogr A chromatography-mass 986:101–10. spectrometry. traction in combinationanalysis with of beer gas volatile fraction. chromatography J Chromatogr with A 1121:145–53. massof spectrometry carbonyl compounds for by yeast flavor122–9. stability of Happoshu. J Am Soc Brew Chem aging. Food Chem 95:357–81. Delvaux FR. 2003a. Flavor-active esters:96:110–8. adding fruitiness to beer. J Biosci Bioengr IS, Winderickx J, Theveleinalcohol JM, acetyltransferase Delvaux genes ATF1, FR. Lg-ATF1, 2003b.a and broad Expression ATF2 range control levels of the of volatile formation esters. the of Appl yeast Environ Microbiol 69:5228–37. cannot be used as markersmalt to beer. Additionally, indicate nerolidol the is storagecan a period be good of used marker non- candidate for(Figure that indicating 4C). The the difference between storage concentrations period of nerolidolfore of be- and after any aging type was significant of inwith beer all groups of beers supported those of all-malt beer, concentrations ofhigher 14 alcohols, compounds, terpene including alcohol, andhigher, short whereas chain concentrations fatty of acids, 13 were esters, compounds, acetate including esters, ethyl and middlenonmalt beer. chain Three fatty groups corresponding acids, to weremalt nonmalt beers, lower and beers, standard in low- or all-malt beers werecomponent formed analysis. by principal methoxy-4-vinylphenol can be detectedwith SBSE-GC-MS. at The the results of samebeers a revealed time study that on the using the concentrations of agingfor these property compounds of except compounds impart stalestorage flavors, in nonmalt it beer seemsstandard are beer. that considerably different changes from during those in I Coghe S, Benoot K, Delvaux F, Vanderhaegen B, Delvaux FR. 2004. Ferulic acid re- Gijs L, Chevance F, Jerkovic V, Collin S.Guido LF, Curto 2002. AF, Boivin P, How Benismail N, low Goncalves CR, Barros pH AA. 2007. Correlation can intensify beta- Halang WA, Langlais R, Kugler E. 1978. Cubic spline interpolation for the calculation Hor Kishimoto T, Wanikawa A, Kono K, Shibata K. 2006. Comparison of theKondo H. odor-active 2005. Brewing yeast fermentation and performance in beer andLandaud Happoshu. S, Helinck S, Bonnarme P. 2008. Formation of volatile sulfur compounds and Ochiai N, Sasamoto K, Daishima S, Heiden AC, Hoffman A. 2003. Determination of Pinho O, Ferreira IM, Santos LH. 2006. Method optimization by solid-phase microex- Shimizu C, Ohno M, Araki S, Furusho S, Watari J, Takashio M. 2002. Effect of reduction Vanderhaegen B, Neven H, Verachtert H, Derdelinckx G. 2006. TheVerstrepen chemistry of KJ, beer Derdelinckx G, Dufour JP, Winderickx J, Thevelein JM,Verstrepen Pretorius KJ, IS, Van Laere SD, Vanderhaegen BM, Derdelinckx G, Dufour JP, Pretorius - γ -nonalactone -damascenone γ β -Damascenone, β -nonalactone is lower in γ 12%). Vol. 75, Nr. 1, 2010 — = -damascenone and β -ionene were detected only in the α -nonalactone, and ethyl cinnamate were not γ 20%) and nerolidol (CV JOURNAL OF FOOD SCIENCE = -damascenone, β The concentrations of 2-nonanol, 2-decanol, 2-undecanol, The concentration of 7 compounds was found to increase during Principal component analysis was performed to determine the Diethyl succinate and 84 detected in NM3, and 2-methoxy-4-vinylphenolin was NM2 not detected during storage (Figure 3). Therefore, these compounds aged samples, and their(Figure concentrations 4A increased and during 4B). It storage diethyl has succinate been increases during reported storage that (Vanderhaegen the anders oth- concentration 2006). of C Changes during storage Principal component analysis Volatile compounds and the changes in their concentration levelsdetected . in . beer. Concentrations . of 15 compounds inwere nonmalt beer higher than thoseisobutyric acid in and isovaleric all-malt acid beer. maynonmalt affect High the beer sensory concentrations because profile of of ofcentrations their of low-odor 13 threshold. Further, compoundsthose con- in in nonmalt all-malt beerThe beer. were flavor Most lower that than ofvinylphenol, which originated is these delivered from from malt compounds (Coghe ferulic and were others 2004). acid esters. was2-nonanone, and 2-methoxy-4- 2-undecanone inAM1—are 4 significantly samples—NM1, higher NM4, than LM3, thoseCompounds in contributing the to other hop 11 bitterness samples. 2-alkanones, would which could be be degraded further reduced to to(Vanderhaegen 2-alkanols and by others yeast 2006). Hence, theseon results may the depend type of hops present in them. storage (Figure 3 and 4). These compoundsation had (CVs) coefficients of of less vari- than 10%, with the exception of similarities in volatile compounds between1st the and samples. 2nd Only principal the componentsvariance. accounted On the for basis 83% of of variablescomponent, correlated the with 3 total the 1st groups principal correspondingbeers, to and standard nonmalt or beers, all-malt low-malt beersceptions were (Figure formed, 2). with The only 1st 2for principal 56% ex- component, of which the accounts total variance,acetate is and positively 2-phenylethyl acetate, correlated but with negatively isopentyl correlated withmethyl-1-butanol 3- and 2-phenylethanol. Alcoholof acetyltransferase yeast(Verstrepen produces and 3-methyl-1-butanol otherswhich from accounts 2003b). for 27% isopentyl The of the total 2ndethyl acetate variance, octanoate is and principal well ethyl correlated decanoate. with component, (CV nonalactone, 2-methoxy-4-vinylphenol, and ethylthe cinnamate major compounds are contributing to theand others aged 2002). flavor The concentration of of beer these compounds (Gijs during increased storage in all-malt and standardin beers, but nonmalt barely increased beerchanges (Figure in 3). the sensory These profiledifferent of from results all-malt nonmalt and strongly standard beer beer. during Additionally, suggest thetration concen- storage that are of these compoundsnonmalt except beer before for aging, the difference may increaseage. during Because stor- the formation of depends on temperature, nonmalt beer mayto be temperature. relatively resistant C: Food Chemistry