Evaluation of Volatiles from Ampelopsis brevipedunculata var ...

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Journal of Oleo ScienceCopyright ©2013 by Japan Oil Chemists’ SocietyJ. Oleo Sci. 62, (9) 645-655 (2013)

Evaluation of Volatiles from Ampelopsis brevipedunculata var. heterophylla UsingGC-Olfactometry, GC-MS and GC-pulsed Flame Photometric DetectorAtsuhiko Nakamura and Mitsuo Miyazawa＊

Department of Applied Chemistry, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka-shi, Osaka 577-8502, Japan

1 INTRODUCTION Ampelopsis brevipedunculata var. heterophylla is

widely distributed in Asia, where it is often used as a folk medicine. As an example, the drug efficacy of this plant has been reported for liver disease, specifically ethanol ex-tracts of fresh berries. The dried leaves and branches of A. brevipedunculata var. heterophylla have also been used as tea, whereas its fresh berries are usually immersed in alcohol in Japan.

Volatile sulfur compounds（VSCs）are trace components found in many plant aroma profiles of essential oils and are an important class of odor-active compounds, due to their extremely low odor threshold. Some of these volatiles de-pending on concentration can be responsible for off-flavors. In strawberry samples analyzed using a pulsed flame pho-tometric ditector（PFPD）, VSCs included methyl thiopropi-onate, ethyl thiobutanoate, methyl thiohexanoate, methyl（methylthio）acetate, ethyl（methylthio）acetate, methyl 2-

＊Correspondence to: Mitsuo Miyazawa, Department of Applied Chemistry, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka-shi, Osaka 577-8502, JapanE-mail: [email protected] March 12, 2013 (received for review January 21, 2013)Journal of Oleo Science ISSN 1345-8957 print / ISSN 1347-3352 onlinehttp://www.jstage.jst.go.jp/browse/jos/　　http://mc.manusriptcentral.com/jjocs

（methylthio）butyrate, methyl 3-（methylthio）propionate, ethyl 3-（methylthio）propionate, and methyl thiooctanoate1）. T h e r e f o r e , v o l a t i l e s u l f u r c o m p o u n d s i n A.brevipedunculata var. heterophylla were evaluated using a PFPD.

GC olfactometry（GC-O）is a technique that simultane-ously uses a physical detector and the human nose and is the most commonly used method for evaluation of odor-ants. In the aroma extraction dilution analysis method（AEDA）method, the sample is subsequently diluted until an odor is no longer perceived. The odor potency is ex-pressed as the flavor dilution（FD）factor. Thus, by means of sniffing serial dilutions of an essential oil, volatile com-pounds can be ranked according to odor potency2）. The significant contribution of each odorant to the characteris-tic flavor can be determined by the odor active value（OAV）. OAV is the ratio of concentration to the odor threshold of the compound, it is well-accepted that com-

Abstract: Ampelopsis brevipedunculata var. heterophylla is extensively cultivated in Asia, and the dried leaves and branches have a characteristic odor and have been used as a tea. To investigate the odorants contributing to the characteristic odor of A. brevipedunculata var. heterophylla, the aroma extraction dilution analysis method was performed through gas chromatography olfactometry. In addition, volatile sulfur compounds were evaluated using pulsed flame photometric detector. As a result, 86 compounds were identified in the oils of leaves and 78 in branches, accounting for 80.0% and 68.3%, respectively, of the compounds identified. The main compounds in the essential oil of leaves were palmitic acid (12.5%), phenylacetaldehyde (4.1%) and hexahydrofarnesyl acetone (3.9%). On the other hand, the essential oil of branches contained palmitic acid (12.7%), terpinen-4-ol (4.4%) and α-cadinol (3.7%). The total number of odor-active compounds identified in the leaf and branch oils was 39. The most odorous compounds of leaves and branches of A. brevipedunculata var. heterophylla were (E, Z )-2,6-nonadienal (melon, green odor), (E)-2-nonenal (grassy odor), phenylacetaldehyde (honey-like) and (E)-linalool oxide (woody odor).

Key words: Ampelopsis brevipedunculata var. heterophylla, GC-MS, GC-MS/O, aroma extraction dilution analysis (AEDA), pulsed flame photometric detector (PFPD)

A. Nakamura and M. Miyazawa

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pounds with high OAV contribute more to the aroma of food3）.

Nevertheless, to the best of our knowledge, no investiga-tion of the essential oil and odor compounds of A. brevipe-dunculata var. heterophylla have been reported to data. This study was performed to investigate the characteristic odor of the A. brevipedunculata var. heterophylla essen-tial oil using AEDA method, OAV and PFPD.

2 EXPERIMENTAL2.1 Plant material

The leaves and branches of Ampelopsis brevipeduncu-lata var. heterophylla were obtained from Fukushima Pre-fecture, Japan in July 2011.

2.2 Isolation of the essential oilDried leaves（100 g）and branches（100 g）of A. brevipe-

dunculata var. heterophylla were hydrodistilled for 2 hours in a Likens-Nickerson type apparatus. The oils were dried over anhydrous sodium sulfate. The yield of oil was 40 mg for leaves（0.004％）and 12 mg for branches（0.012％）. The oils were kept in sealed sample tubes and stored at －30℃ in a freezer prior to analysis.

2.3 Gas chromatography-mass spectrometry（GC-MS）Oil sample analysis was performed on an Agilent Tech-

nologies 6890 gas chromatograph coupled to an Agilent Technologies 5973 mass selective detector. GC conditions were: An HP-5MS phenylmethylpolysiloxane capillary column（30 m×0.25 mm i.d., film thickness 0.25 μm）and a DB-WAX column（15 m×0.25 mm i.d., film thickness 0.25 μm）; the column temperature was 40－260℃ at a rate of 4℃ /min, then a hold at 260℃ for 5 min. The carrier gas was He at a flow rate of 1.5 mL/min. The injector and de-tector temperatures were 270 and 280ºC, respectively. The ionization voltage was 70 eV and the split ratio was 1:10. A 1 μL aliquot of oil was injected for each sample.

2.4 GC-OGC-O was performed on the same coupled Agilent Tech-

nologies 6890/5973 system as above. GC was performed using the same capillary column as above, under the same run conditions, except the He carrier gas flow rate was 1.5 mL/min.

2.5 GC-PFPDVSC analysis was performed using the same coupled

Agilent Technologies 6890/5973 system as above equipped with a sulfur-specific PFPD（model 5380 PFPD, OI Analyti-cal Co., College Station, TX）. The GC conditions were the same as those used for GC-MS.

2.6 AEDAThe highest dilution（8 mg/mL）was assigned an FD-fac-

tor of 1. The volatile oil was stepwise diluted（1:1, v/v）by addition of diethyl ether, and aliquots of the dilutions（1 μL）were evaluated. The process stopped when no aroma was detected by assessors. The result was expressed as the FD factor, which is the ratio of the concentration of the odorant in the initial volatile oil to its concentration in the most diluted volatile oil in which the odor is still detectable by GC-O4, 5）. An odorant with a high FD factor can be con-sidered as an important contributor to the characteristic odor.

2.7 Identi�cation and quanti�cation of compoundsThe compounds were identified on the basis of compari-

son of their retention indices（RIs）and mass spectra from the Mass Finder 4 and NIST02 libraries, published data6）, our previous studies7－14） and（RIs）of standards or RIs re-ported in the literature. The RIs were calculated using a homologous series of n-alkanes C5-C27 on the HP-5MS column. The oil composition was quantitated by GC FID（with a flame ionization detector）by defining the total of the oil as 100％. Quantitative analysis of active aroma com-ponents of the oils were performed on the basis of calibra-tion curves for hexanal（2）, furfural（4）, （E）-2-hexanal（5）, 2-hexanal（6）, hexanol（8）, heptanal（9）, methional（11）, benzaldehyde（13）, dimethyl trisulfide（14）, 1-octen-3-ol（15）, 6-methylhept-5-en-2-one（16）, 2-pentylfuran（17）, α-terpinene（22）, hexanoic acid（23）, phenylacetaldehyde（28）, （Z）-linalool oxide（33）, 3,3,6-trimethyl-1,5-heptadi-en-4-ol（34）, （E）-linalool oxide（35）, linalool（36）, 3,5-octa-dien-2-one（39）, nonanal（40）, phenylethyl alcohol（43）, δ-camphor（46）, （E,Z）-2,6-nonadienal（49）, （E）-2-nonenal（51）, borneol（53）, terpinen-4-ol（54）, p-methylacetophe-none（56）, p-cymen-8-ol（57）, α-terpineol（58）, methyl sa-licylate（59）, myrtenol（60）, nerol（68）, p-vinylguaiacol（76）, γ-nonalactone（80）, （E）-β-damascenone（83）, α-ionone（87）, geranyl acetone（89）, β-ionone（92）, within the con-centration range 0.5－1000 μg/mL）. The weight- percent of each compound was calculated based on the response factors of the detector.

3 Results and Discussion3.1 Constituents of essential oil from Ampelopsis brevi-

pedunculata var. heterophyllaThe essential oils obtained by hydrodistillation from the

leaves and branches of Ampelopsis brevipedunculata var. heterophylla had a yield of 0.040％ and 0.012％（w/w）, re-spectively. The leaf oil had a green, fresh and sweetish odor, while the branch oil had a green, woody and fresh odor. In leaf oil, 107 compounds were identified and in branch oil, 93 compounds, accounting for about 80.0％ and

Essential oil with characteristic odor in Ampelopsis

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No compoundsa) RI-5b) RI-Wc) peak area %d) Identification methodf)

Reference sourceg)leaf branch

1 pyridine 764 1413 0.1 － RI, MS Aldrich2 hexanal 798 1306 0.9 1.0 RI, MS Wako3 3-furanmethanol 819 1421 0.1 － RI, MS TCI4 furfural 829 1373 1.1 0.2 RI, MS Wako5 (E)-2-hexenal 848 － 2.2 0.1 RI, MS Wako6 2-hexenal 852 －－ 0.2 RI, MS Wako7 (E)-2-heptene 866 － 0.6 － RI, MS Wako8 hexanol 886 1405 － 0.5 RI, MS Wako9 heptanal 900 － 0.2 － RI, MS Wako

10 2-butoxyethanol 905 1342 0.3 0.2 RI, MS Wako11 methional 906 －－ tre) RI, MS Wako12 (E)-2-heptenal 954 1336 tr － RI, MS Wako13 benzaldehyde 958 1424 0.3 0.5 RI, MS Aldrich14 dimethyl trisulfide 968 －－ tr RI, MS Wako15 1-octen-3-ol 979 1396 0.1 0.2 RI, MS Wako16 6-methylhept-5-en-2-one 985 1337 0.3 － RI, MS Wako17 2-pentyl-furan 989 － 0.4 0.4 RI, MS Wako18 1-ethylcyclohexene 996 － 0.2 － RI, MS19 yomogi alcohol 1000 － 1.1 － RI, MS20 α-fenchene 1001 －－ 0.2 RI, MS Wako21 (E,E)-2,4-heptadienal 1010 1402 0.6 0.1 RI, MS Aldrich22 α-terpinene 1014 － 0.7 0.5 RI, MS Wako23 hexanoic acid 1015 1740 0.7 － RI, MS Aldrich24 o-cymene 1022 1298 0.1 0.7 RI, MS Aldrich25 limonene 1026 1305 0.1 － RI, MS Wako26 2-ethylhexanol 1030 1433 2.0 2.5 RI, MS Wako27 benzyl alcohol 1037 1758 0.3 0.4 RI, MS Aldrich28 phenylacetaldehyde 1044 1546 4.1 1.2 RI, MS Wako29 γ-terpinene 1056 －－ 0.9 RI, MS Wako30 (Z)-sabinenehydrate 1065 1482 － 0.5 RI, MS Aldrich31 heptanoic acid 1066 1843 tr － RI, MS Wako32 4-methyl-benzaldehyde 1067 － 0.1 － RI, MS Other33 (Z)-linalool oxide 1071 1408 0.9 0.9 RI, MS Wako34 3,3,6-trimethyl-1,5-heptadien-4-ol 1083 － 0.1 － RI, MS35 (E)-linalool oxide 1087 1679 0.9 2.1 RI, MS Wako36 linalool 1100 1488 2.1 0.7 RI, MS Wako37 3-mercaptohexanol 1123 － tr tr RI, MS38 (Z)-p-menth-2-en-1-ol 1097 －－ 0.8 RI, MS Other39 3,5-octadien-2-one 1102 1516 0.1 0.2 RI, MS Other40 nonanal 1103 1395 0.4 0.5 RI, MS Wako41 6-methylhepta-3,5-dien-2-one 1105 － 0.4 － RI, MS Other42 2,6-dimethylcyclohexanol 1107 1505 0.8 0.3 RI, MS Aldrich43 phenylethyl alcohol 1115 1790 0.3 0.4 RI, MS Wako44 unknown 1174 － 0.5 1.2 RI, MS45 (E)-p-menth-2-en-1-ol 1120 －－ 0.3 RI, MS Other46 δ-camphor 1140 1529 － 0.7 RI, MS Aldrich47 4-ketoisophotone 1142 － 0.2 － RI, MS Other48 camphene hydrate 1150 －－ 0.4 RI, MS Other49 (E,Z)-2,6-nonadienal 1152 1553 0.2 0.1 RI, MS Aldrich50 isoborneol 1156 1672 － 0.1 RI, MS Aldrich51 (E)-2-nonenal 1158 1524 0.3 0.3 RI, MS Wako

Table 1　Chemical compounds of the essential oil from Ampelopsis brevipedunculata var. heterophylla.


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52 pinocarvone 1160 1570 － 0.2 RI, MS Other53 borneol 1166 1604 0.2 1.9 RI, MS Wako54 terpinen-4-ol 1176 1522 0.3 4.4 RI, MS Wako55 octanoic acid 1180 1949 tr － RI, MS Aldrich56 p-methylacetophenone 1184 1793 0.1 0.4 RI, MS Wako57 p-cymen-8-ol 1186 1752 0.1 0.2 RI, MS Other58 α-terpineol 1190 1619 1.1 0.5 RI, MS Aldrich59 methyl salicylate 1193 1652 － 1.0 RI, MS Aldrich60 myrtenol 1195 1694 － 0.4 RI, MS Wako61 safranal 1198 － 0.2 － RI, MS Aldrich62 decanal 1203 1677 0.2 0.5 RI, MS TCI63 (E)-piperitol 1206 －－ 0.2 RI, MS64 β-cyclocitral 1219 1523 0.6 0.2 RI, MS Aldrich65 benzothiazole 1222 1811 0.2 0.3 RI, MS Wako66 cumin alcohol 1225 1979 － tr RI, MS Aldrich67 bornylene 1227 － 0.4 － RI, MS68 nerol 1255 1767 0.8 － RI, MS Wako69 3-isopropylbenzaldehyde 1238 －－ 0.2 RI, MS Wako70 unknown 1255 － 0.7 － RI, MS71 unknown 1277 － 1.4 － RI, MS72 nonanoic acid 1292 1638 1.0 － RI, MS Aldrich73 indole 1298 2355 0.2 － RI, MS Wako74 (Z)-cinerolone 1306 － 0.3 － RI, MS75 2,4-decadienal 1310 1722 tr － RI, MS Wako76 p-vinylguaiacol 1315 － 1.2 0.6 RI, MS Aldrich77 guaiacol 1330 1738 － tr RI, MS Wako78 dehydro-ar-ionene 1351 － 0.3 － RI, MS Other79 α-ionene 1354 － tr － RI, MS80 γ-nonalactone 1361 1903 － 0.4 RI, MS Other81 2-ethyl-3-hydroxyhexyl 2-methylpropanoate 1372 1775 0.2 － RI, MS82 decanoic acid 1380 2466 tr － RI, MS Wako83 (E)-β-damascenone 1382 1789 0.2 tr RI, MS Wako84 1,2-dihydro-1,4,6-trimethyl-naphthalene 1388 － 0.1 － RI, MS85 6,10-dimethyl-2-undecanone 1402 － 0.1 － RI, MS Aldrich86 unknown 1413 － 0.7 － RI, MS87 α-ionone 1426 1750 0.8 0.2 RI, MS Wako88 (Z)-β-farnesene 1441 1717 tr － RI, MS Wako89 geranyl acetone 1451 1853 1.5 0.8 RI, MS Wako90 unknown 1473 － 0.5 0.4 RI, MS91 curcumene 1480 1759 － 0.2 RI, MS Other92 β-ionone 1485 1760 2.3 0.9 RI, MS Wako93 α-muurolene 1498 1754 － 0.1 RI, MS Other94 unknown 1519 － 1.2 0.6 RI, MS95 δ-cadinene 1521 1756 － 0.3 RI, MS Other96 unknown 1526 － 1.4 0.6 RI, MS97 dihydroactinidiolide 1532 2257 2.2 0.2 RI, MS Other98 unknown 1546 －－ 2.1 RI, MS99 (Z)-nerolidol 1561 1997 － 0.1 RI, MS Wako100 dodecanoic acid 1568 2494 － 0.1 RI, MS Aldrich101 caryophyllene oxide 1583 1967 1.9 － RI, MS Wako102 unknown 1594 － 0.4 － RI, MS

Table 1　Continued.


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103 ledol 1602 2032 － 0.1 RI, MS Other104 6,7-epoxide humulene 1608 － 0.4 － RI, MS Other105 unknown 1637 2129 0.5 8.5 RI, MS106 τ-cadinol 1642 2167 0.7 － RI, MS Other107 α-copaene 1646 2081 0.2 － RI, MS Other108 unknown 1649 2079 tr 5.1 RI, MS109 τ-muurolol 1655 2068 1.2 1.9 RI, MS Other110 α-cadinol 1656 2066 － 3.7 RI, MS Other111 allo-aromadendrene 1658 1642 0.4 － RI, MS Aldrich112 ar-tumerone 1664 －－ 0.2 RI, MS Other113 cyclotetradecane 1674 －－ 0.8 RI, MS114 tetradecanol 1678 2096 － tr RI, MS Wako115 α-bisabolol 1682 1683 － 0.2 RI, MS Aldrich116 hexadecanal 1710 2121 0.2 0.8 RI, MS TCI117 tetradecanoic acid 1758 2677 tr － RI, MS Wako118 2,3,6-trimethyl-1,4-naphthoquinone 1760 － 0.2 － RI, MS119 unknown 1766 － 0.4 1.5 RI, MS120 hexahydrofarnesyl acetone 1842 － 3.9 1.1 RI, MS Other121 pentadecanoic acid 1860 2781 tr － RI, MS Wako122 diisobutyl phthalate 1865 2508 0.5 1.1 RI, MS Wako123 farnesylacetone 1915 － 2.3 0.6 RI, MS Wako124 methyl palmitate 1921 2223 0.2 0.1 RI, MS Wako125 16-hexadecanolactone 1927 2990 0.1 － RI, MS Aldrich126 isophytol 1943 2327 0.5 － RI, MS Wako127 butyl isobutyl phthalate 1959 － 0.7 － RI, MS Wako128 palmitic acid 1985 2896 12.5 12.7 RI, MS Wako129 methyl oleate 2086 2914 － tr RI, MS Wako130 methyl linoleate 2092 3212 tr － RI, MS Wako131 oleic acid 2102 3108 tr － RI, MS Wako132 phytol 2111 2624 7.1 0.9 RI, MS Wako133 octadecanoic acid 2172 3088 tr － RI, MS Wako134 linoleic acid 2214 3153 3.7 3.0 RI, MS Wako135 tricosane 2300 2300 － 0.7 RI, MS Wako136 tetracosane 2400 2400 0.1 0.2 RI, MS Wako137 pentacosane 2500 2500 0.1 1.2 RI, MS Wako138 hexacosane 2600 2600 2.8 － RI, MS Wako139 heptacosane 2700 2700 0.2 1.2 RI, MS TCI140 octacosane 2800 2800 － 0.1 RI, MS Wako141 squalene 2819 3056 0.1 0.5 RI, MS Wako142 nonacosane 2900 2900 0.4 3.7 RI, MS Wako

total 86.5 87.1a) Compounds are listed in order of their elution time from a HP-5MS column. Presence of compound is indicated by its GC/FID percentage. b) RI, retention indices determined on HP-5MS columns, using the homologous series of n-alkanes (C8-C27).

c) RI, retention indices determined on DB-WAX columns, using the homologous series of n-alkanes (C8-C27).

d) Peak area (%) was related to total detected compounds by GC-MS. e) tr = trace (<0.1%). f) Identification methods: RI, retention indice; MS, mass spectrum. g) Reference materials were obtained from commercial source and our previous studies: Wako, Wako Pure Chemical Industries Ltd. (Osaka, Japan); TCI, Tokyo Kasei Kogyou Co. Ltd. (Tokyo Japan); Aldrich, Sigma-Aldrich, St. Louis, MO; Other, our previous studies 32, 80, 97, 120 from Miyazawa et al. (2008); 38, 45 from Kameoka et al. (1976); 39, 41, 93, 95, 110 from Sukhontha Sukhonthara et al. (2009); 47, 78 from Utsumi and Miyazawa (2011); 48 from Kameoka et al. (1994); 52, 57, 91, 112 from Kashima et al. (2011); 103, 104, 107, 109 from Miyazawa et al. (2007). h) Numbers 18, 19, 34, 37, 44, 63, 67, 74, 79, 81, 84, 113, 118 mass spectrum and RI agreed with literature data (David W, 1990; Basta A, 2007; Shunying Z, 2006; Lin J, 2002; Alonzo G, 2000; Topal U, 2008; Lalel Herianus JD, 2003; Gomez E, 1993; Formisano C, 2010; Liu J, 2006; Patharakorn T, 2010)

Table 1　Continued.


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68.3％ of the total oil content, respectively（Table 1）. The main compounds from essential oil of leaves were palmitic acid（12.5％）, phenylacetaldehyde（4.1％）and hexahydro-farnesyl acetone（3.9％）. The leaf oil consisted mainly of alcohols（23.2％）, followed by aldehydes（11.6％）, acids（16.9％）and hydrocarbons（6.6％）. The main compounds

from essential oil of branches ware palmitic acid（12.7％）, terpinen-4-ol and α-cadinol（3.7％）. The branches oil con-

sisted mainly of alcohols（25.8％）, followed by, aldehydes（5.9％）, acids（15.8％）and hydrocarbons（10.8％）.

3.2 Identi�cation of Ampelopsis sulfur volatilesSulfur volatiles have an important role in many plant

flavors. A PFPD chromatogram of sulfur volatiles from leaves and branches is shown in Fig. 1 and Fig. 2. In this study, four sulfur compounds were identified. Specifically,

Fig. 1　Comparison of sulfur and TIC chromatograms from Ampelopsis brevipedunculata var. heterophylla leaves.

Fig. 2　Comparison of sulfur and TIC chromatograms from Ampelopsis brevipedunculata var. heterophylla branches.


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Table 2　 Odor-active compounds of Ampelopsis brevipedunculata var. heterophylla.

No. compoundsa odorb FD-factorc, d

leaf branch2 hexanal green 32 44 furfural burnt 1 －5 (E)-2-hexenal green 32 16 2-hexanal green － 18 hexanol green － 19 heptanal fatty 2 －11 methional potato － 3213 benzaldehyde sweet 2 114 dimethyl trisulfide garlic, fishy － 415 1-octen-3-ol mushroom 2 116 6-methylhept-5-en-2-one green 2 －17 2-pentylfuran sweet 1 122 α-terpinene lemon-like 1 123 hexanoic acid sweaty 1 －28 phenylacetaldehyde honey-like 64 3233 (Z)-linalool oxide woody 8 3234 3,3,6-trimethyl-1,5-heptadien-4-ol burnt 1 －35 (E)-linalool oxide woody 4 6436 linalool floral 4 239 3,5-octadien-2-one dust 1 140 nonanal green 16 1643 phenylethyl alcohol floral 2 246 δ-camphor camphor, wood － 149 (E,Z)-2,6-nonadienal melon, green 128 3251 (E)-2-nonenal grassy 64 6453 borneol camphor, wood － 154 terpinen-4-ol green 4 3256 p-methylacetophenone sweet 1 157 p-cymen-8-ol citric, musty 1 158 α-terpineol floral 1 159 methyl salicylate mint － 1660 myrtenol mint － 1668 nerol floral 4 －76 p-vinylguaiacol spicy 1 180 γ-nonalactone floral － 183 (E)-β-damascenone sweet 32 887 α-ionone violet 4 189 geranyl acetone floral 4 492 β-ionone sweet, fruity 32 16

a) Compounds are listed in order of their elution time from a HP-5MS column.b) Odor quality perceived through the sniffing pot.c) The sample concentration (10 mg/ml) was assigned FD-factor 1.d) FD factor on HP-5MS column.


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methional, dimethyl trisulfide, 3-mercaptohexanol and benzothiazole, among which benzothiazole was present in concentrations high enough to be seen in the MS total ion current chromatogram. The other small sulfide peaks remain unidentified.

3.3 GC-O, AEDA and OAVA total of 39 odor-active compounds were identified in

both the leaf and branch oils（Table 2）. In leaves from A.brevipedunculata var. heterophylla, 30 odor active compounds were identified. A comparison of the gas chro-matogram and the corresponding FD chromatogram of the odor-contributing compounds is shown in Fig. 3. The strongest aroma compound, which had an FD factor of 128, was（E,Z）-2,6-nonadienal（peak 49; melon, green odor）. Other strong aroma compound（E）-2-nonenal（51; grassy

Fig. 4　 Gas chromatogram and aromagram (FD factor) of volatile oil from Ampelopsis brevipedunculata var. het-erophylla branches: 11, methional; 28, phenylacetaldehyde; 33, (Z)-linalool oxide, 35, (E)-linalool oxide; 49, (E,Z)-2,6-nonadienal; 51, (E)-2-nonenal; 54, terpinen-4-ol.

Fig. 3　 Gas chromatogram and aromagram (FD factor) of volatile oil from Ampelopsis brevipedunculata var. hetero-phylla of leaf parts : 2, hexanal; 5, (E)-2-hexenal; 28, phenylacetaldehyde; 49, (E,Z)-2,6-nonadienal; 51, (E)-2-nonenal; 83, (E)-β-damascenone; 92, β-ionone.


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Table 3　 Odor activity values of odor-active compounds in Ampelopsis brevipedunculata var. heterophylla.

No. compoundsconcentration (ppb) odor thresholda OAVb

leaf branch (ppb) leaf branch2 hexanal 3520 1173 5 704 2354 furfural 4512 － 776 6 －5 (E)-2-hexenal 8648 179 17 509 116 2-hexanal － 199 17 － 128 hexanol － 622 250 － 29 heptanal 968 － 3 323 －11 methional － 64 0.2 － 32013 benzaldehyde 1212 635 350 3 214 dimethyl trisulfide － 16 0.1 － 16015 1-octen-3-ol 212 234 1 212 23416 6-methylhept-5-en-2-one 1276 532 68 19 817 2-pentylfuran 1460 532 6 243 8922 α-terpinene 2732 612 85 32 723 hexanoic acid 2688 － 1840 1 －28 phenylacetaldehyde 16212 1474 4 4053 36933 (Z)-linalool oxide 540 1111 320 2 334 3,3,6-trimethyl-1,5-heptadien-4-ol 3408 － ndc －－35 (E)-linalool oxide 540 2483 nd －－36 linalool 3276 257 1.5 2184 17139 3,5-octadien-2-one 492 869 6 82 14540 nonanal 8460 646 1 8460 64643 phenylethyl alcohol 1640 436 620 3 146 δ-camphor － 864 460 － 249 (E,Z)-2,6-nonadienal 1276 96 0.02 63800 480051 (E)-2-nonenal 956 354 0.08 11950 442553 borneol － 2276 140 － 1654 terpinen-4-ol 1024 5203 6400 <1 156 p-methylacetophenone 1280 479 21 61 2357 p-cymen-8-ol 412 257 nd －－58 α-terpineol 384 631 330 1 259 methyl salicylate － 1210 40 － 3060 myrtenol － 520 nd －－68 nerol 4388 － 680 6 －76 p-vinylguaiacol 3264 709 3 1088 23680 γ-nonalactone 816 202 76 11 383 (E)-β-damascenone 4772 58 0.01 477200 580087 α-ionone － 476 27 － 1889 geranyl acetone 3104 910 150 21 692 β-ionone 6164 1092 0.1 61640 10920

a) Odor threshold measured in water solution and represented μg of compound / kg of water (ppb).b) OAV = Odor activity value (concentration divided by odor threshold).c) nd = not determined.


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odor）, phenylacetaldehyde（28; honey-like odor）, hexanal（2; green odor）, （E）-2-hexenal（8; green odor）, （E）

-β-damascenone（83; sweet odor）and β-ionone（92; sweet, fruity odor）had FD factors of 64 or 32. In contrast, furfural, heptanal, benzaldehyde, 1-octen-3-ol, 6-methylhept-5-en-2-one, 2-pentylfuran, α-terpinene, hexanoic acid, （Z）-lin-alool oxide, 3,3,6-trimethyl-1,5-heptadien-4-ol, （E）-linalool oxide, linalool, 3,5-octadien-2-one, nonanal, phenylethyl alcohol, terpinen-4-ol, p-methylacetophenone, p-cymen-8-ol, α-terpineol, nerol, p-vinylguaiacol, α-ionone and geranyl acetone had FD factors between 1 and 16. Conse-quently, the main aroma of the leaf oil was green, sweet odor. On the other hand, 33 odor-active compounds were identified in branches from A. brevipedunculata var. het-erophylla. A comparison of the gas chromatogram and the corresponding FD chromatogram of the odor-contributing compounds is shown in Fig. 4. There were two strong aroma compounds with an FD factor of 64, （E）-linalool oxide（35; wood odor）and（E）-2-nonenal（51; grassy odor）. The other strong aroma compound methional（11; potato odor）, phenylacetaldehyde（28; sweet odor）, （Z）-linalool oxide（33; wood odor）, （E,Z）-2,6-nonadienal（51; melon, green odor）and terpinen-4-ol（54; green odor）. In contrast, hexanal, （E）-2-hexenal, 2-hexanal, hexanol, benzaldehyde, dimethyl tr isul f ide, 1-octen-3-ol , 2-pentyl furan, α-terpinene, linalool, 3,5-octadien-2-one, nonanal, phenyl-ethyl alcohol, δ-camphor, borneol, p-methylacetophenone, p-cymen-8-ol, α-terpineol, methyl salicylate, myrtenol, p-vinylguaiacol, γ-nonalactone, （E）-β-damascenone, α-ionone and β-ionone had FD factors between 1 and 16. Consequently, the main aroma of branch oil was a woody and green odor.

In order to determine the relative contribution of each compound to the aroma of A. brevipedunculata var. het-erophylla, the OAV was employed, taking into account the concentration and odor threshold of each compound. The results are shown in（Table 3）. Due to the unavailability of odor threshold data in the literature, the OAV of 3,3,6-tri-methyl-1,5-heptadien-4-ol, （E）-linalool oxide, and p-cy-men-8-ol could not be determined. In the leaf oil, （E）-β-damascenone had the highest OAV（477,200）, followed by（E,Z）-2,6-nonadienal（63,800）, β-ionone（61,640）, （E）-2-nonenal（11,950）, nonanal（8,460）, phenylacetaldehyde（4,053）and p-vinylguaiacol（1,088）. In particular, （E,Z）

-2,6-nonadienal, （E）-2-nonenal, （E）-β-damascenone and phenylacetaldehyde had high FD-factors, and thus, were considered the main aroma compounds of leaf oil. In the branch oil, β-ionone had the highest OAV（10,920）, fol-lowed by（E）-β-damascenone（5,800）, （E,Z）-2,6-nonadi-enal（4,800）and（E）-2-nonenal（4,425）. Whereas the main aroma compounds of branch oil had a wood odor, （Z）-lin-alool oxide and（E）-linalool oxide had low OAV or could not be determined.

AcknowlegmentThis work was supported by Grant-in-Aid from the Japan

Society for the Promotion of Science（No. 24658055）

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