Volatile Constituents of Blue-rose

Research Article

Received: 22 November 2012, Revised: 4 February 2013, Accepted: 4 February 2013 Published online in Wiley Online Library

(wileyonlinelibrary.com) DOI 10.1002/ffj.3153

Volatile constituents of blue-coloured hybridtea rose flowersAtsushi Joichi,a* Yasuko Nakamura,a Shinichiro Haze,a Takahiro Ishikawa,b

Hiroyuki Atoji,b Takashi Nishidab and Kazutoshi Sakuraib

ABSTRACT: The volatile constituents of ‘Blue Moon’ and ‘Blue Perfume’ rose flowers, which, on an olfactory basis, areclassified as a ‘blue type’ were analysed using AromascopeW technology (modified headspace technology) and solventextraction methods followed by gas chromatography–mass spectrometry analysis. One hundred and eighty componentswere identified in the headspace volatile components of ‘Blue Moon’ flower and 188 components were identified in solventextracts. Among them, geraniol, nerol, citronellol, 1,3-dimethoxy-5-methylbenzene and dihydro-b-ionol were identified asthe main odour components. On the other hand, in ‘Blue Perfume’, 165 components were identified in the headspacevolatile components and 150 components were identified in solvent extracts. Among them, geraniol, nerol, citronellol,neral, and geranial were identified as the major odour compounds. From both rose flowers, three components werenewly identified: 2-isopropyl-4-methylthiazole, (Z)-cyclododec-9-enolide (yuzu lactone), and methyl cis-(Z)-jasmonate.2-Isopropyl-4-methylthiazole and methyl cis-(Z)-jasmonate were identified in both of the headspace components andsolvent extracts of the two types of rose flower, and then yuzu lactone was identified only in solvent extracts as theone of the minor components. Several components identified in both flowers have asymmetric carbon atoms in their molecules,leading us to analyse their chirality. For the first time, the enantiomer ratios of linalool, (E)-nerolidol, theaspiranes anddihydro-b-ionol could be assigned by multi-dimensional gas chromatography–mass spectrometry. The results were asfollows in both rose flowers. The ratio of the (S)-enantiomer vs. the (R)-enantiomer of linalool was 8:92. Only the (S)-enantiomerwas detected for (E)-nerolidol and dihydro-b-ionol. The ratios of the (2R,5R)-enantiomer vs. the (2S,5S)-enantiomer intheaspirane A and the (2R,5S)-enantiomer vs. the (2S,5R)-enantiomer in theaspirane B were about 4:96. Copyright © 2013John Wiley & Sons, Ltd.

Keywords: hybrid tea rose; ‘Blue Moon’; ‘Blue Perfume’; volatile constituents; enantiomer

* Correspondence to: Atsushi Joichi, Shiseido Research Center (Shin-Yokohama),2-2-1, Hayabuchi, Tsuzuki-ku, Yokohama-shi, Kanagawa 224–8558, Japan.E-mail: [email protected]

a Shiseido Research Center (Shin-Yokohama), 2-2-1, Hayabuchi, Tsuzuki-ku,Yokohama-shi, Kanagawa, 224-8558, Japan

b Takasago International Corporation Corporate Research & DevelopmentDivision, 4-11, Nishiyawata 1-chome, Hiratsuka City, Kanagawa, 254-0073, Japan

IntroductionThe rose has been admired since ancient times because of thebeauty of its shape and colour, as well as its scent. Natural oilsobtained from Rosa x damascena, grown in Bulgaria and Turkey,and Rosa x centifolia, grown in southern France and Morocco,by using steam distillation or solvent extraction are used inperfumery and more than 350 components have beenreported.[1–6] Hybrid tea (HT) roses are mainly the result ofnatural and artificial hybridization between European andChinese roses over a long time.[7] As a result, the analysesrevealed many distinctive notes in the HT rose; in particular,the classified blue-coloured HT rose showed a characteristicodour reminiscent of a combination consisting of damask rosenotes, tea-like and lemon-like fresh notes. In this study, volatilecomponents of the ‘Blue Moon’ and ‘Blue Perfume’ cultivars,with characteristic rose notes represented by blue-coloured HTrose flowers, were analysed by using gas chromatography–massspectrometry (GC-MS), and the presence of major odourcompounds was revealed.

Experimental

Selection of Typical Blue-type Rose Flowers

Among 700 stocks studied, we selected ‘Blue Moon’ and ‘Blue Perfume’,two types of popular rose flowers cultivated by Keisei Rose NurseriesInc. (Keisei Rose Garden) located in Yachiyo city in Chiba prefecture.

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The selection criteria were essentially the elegance, sophisticated andstrength of the typical blue rose character in their scent. ‘Blue Moon’had a rich rose-like, lemon-like, woody and phenolic–spicy note, while‘Blue Perfume’ had a rich rose-like, lemon-like and fresh note with greenfruity aspects.

Collection of the Rose Flower Scent

One to two flowers which were just ready to open and had a good smellas judged by five perfumers, were selected and enveloped in a TedlarW

bag equipped with a glass column (i.d. 8 mm) packed with TenaxW TA(60–80 mesh, 400 mg). The inlet of an air pump was connected tothe TenaxW TA glass column in which the headspace gas was enrichedby pumping at 400 ml/min. The flowers enveloped in the TedlarW

bag and the one undeveloped had no characteristic difference inappearance throughout the trapping.

In order to study the seasonal changes of the volatiles emitted by theliving flowers, this procedure was repeated twice at the beginning ofNovember in 2008 and the end of May in 2009 at Keisei Rose Garden.

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Table 1. Components identified in hybrid tea rose flower (‘Blue Moon’ and ‘Blue Perfume’)

No. RT (min) Compound Gas chromatography area (%)‘Blue Moon’ ‘Blue Perfume’

Headspace Solvent extract Headspace Solvent extract

1 3.40 Acetaldehyde 0.05 ND 0.05 0.032 3.80 Butan-2-one 0.05 ND ND ND3 3.90 Ethanol 0.03 0.01 0.01 0.014 4.00 But-3-en-2-one 0.02 ND 0.02 ND5 4.06 Pentanal t 0.31 t 0.696 4.24 Decane 0.01 t 0.01 t7 4.39 2-Methylbut-3-en-2-ol 0.01 t 0.01 ND8 4.48 Butyl formate t ND ND ND9 4.75 Butyl acetate 0.01 t 0.01 t10 5.10 Isopentyl acetate t ND 0.01 ND11 5.10 Hexanal t 0.01 0.01 0.1112 5.12 Uecane 0.01 t t 0.0113 5.40 Butyl alcohol ND t ND ND14 5.40 2-Methylbutyl acetate t ND ND ND15 5.85 Myrcene 0.01 0.01 0.01 0.0716 6.08 Heptanal t ND t 0.2617 6.09 Pentyl acetate 0.01 t ND ND18 6.24 3-Methylbut-3-en-1-yl acetate 0.01 t t t19 6.30 2-Methylbutanol ND ND ND t20 6.31 Isopentyl alcohol ND 0.02 0.05 0.0121 6.31 Senecionaldehyde 0.16 t 0.21 0.0122 6.49 Dodecane 0.01 t 0.01 0.0123 6.49 Limonene 0.01 0.02 0.01 0.0124 6.66 (E)-Hex-2-en-1-al 0.01 0.09 0.01 ND25 7.00 cis-Ocimene t t ND ND26 7.01 6-Methylheptan-2-one 0.01 t ND ND27 7.06 Pentyl alcohol 0.01 0.02 0.01 0.0128 7.20 3-Methylbut-3-en-1-ol t 0.02 0.01 0.0129 7.23 Prenyl acetate 0.02 0.01 0.06 t30 7.30 trans-Herboxide t t ND ND31 7.32 trans-Ocimene t t t t32 7.65 Hexyl acetate 0.36 0.12 0.49 0.0734 7.75 p-Cymene t t 0.02 ND35 7.80 ‘(Z)-4,8-Dimethylnona-1,3,7-triene’ t t ND ND36 7.99 Octanal 0.01 t 0.02 t37 8.35 (E)-Hex-3-en-1-yl acetate 0.02 0.31 0.03 0.0138 8.40 Tridecane 0.02 t 0.01 0.0139 8.47 ‘(E)-4,8-Dimethylnona-1,3,7-triene’ 0.02 t ND ND40 8.56 (Z)-Hex-3-en-1-yl acetate 0.31 0.31 1.32 0.6341 8.66 6-Methylhept-6-en-2-one 0.02 t ND ND42 8.66 3-Methylpentanol 0.02 t 0.03 0.0143 8.92 (E)-Hex-2-en-1-yl acetate 0.13 0.38 0.49 0.1244 8.99 6-Methylhept-5-en-2-one 0.11 0.01 0.22 0.0245 9.11 Heptyl acetate t t ND t46 9.11 Anisole 0.05 0.03 0.02 0.0447 9.24 Hexanol 0.10 0.31 0.06 0.1048 9.45 2-Isopropyl-4-methylthiazole (1) t t t t49 9.48 cis-Rose oxide 0.04 0.02 0.08 t50 9.59 (E)-Hex-3-en-1-ol 0.01 0.01 0.03 0.0751 9.72 ‘3,3-Dimethylcyclohexanone’ 0.01 ND ND ND52 9.84 trans-Rose oxide 0.01 ND 0.03 t53 9.91 Heptyl acetate ND t t t54 10.00 (Z)-Hex-3-en-1-ol 0.11 0.04 0.03 t55 10.40 Benzyl methyl ether 0.02 ND 0.02 ND56 10.40 Nonanal 0.08 0.02 0.02 0.0257 10.57 Rose furan 0.07 ND 0.11 0.05

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Table 1. (Continued)



58 10.75 Tertadecane 0.01 t 0.02 ND59 10.93 (Z)-Hex-2-en-1-ol 0.01 0.01 0.01 0.1160 11.02 3-(4-Methylpent-3-enyl)furan 0.03 ND 0.10 ND61 11.53 Neryl methyl ether t t 0.09 ND62 11.54 Acetic acid 0.05 0.22 0.02 t63 11.63 trans-Linalool oxide (furanoid) t t 0.01 0.0164 11.73 (E)-Oct-2-en-1-ol t ND ND ND65 11.73 Heptanol 0.02 0.02 0.01 t66 11.95 ‘10-Methylene-2,6,6-trimethyl-1-

oxaspiro[4.5]decane’t t ND ND

67 12.06 6-Methylhept-5-en-2-ol 0.01 0.01 0.06 0.0168 12.30 Geranyl methyl ether ND ND 0.01 ND69 12.30 a-Cubebene 0.11 t 0.04 t70 12.39 ‘Nerol-1,5-oxide’ 0.02 t ND ND71 12.61 Citronellal 0.03 t 0.02 t72 12.75 a-Longipinene t t ND ND73 12.95 a-Ylangene ND ND 0.01 t74 13.25 a-Copaene 0.38 t 0.18 t75 13.25 Decanal 0.10 t 0.03 0.0176 13.43 Theaspirane A 0.05 0.11 0.01 0.0277 13.63 Pentadecane 0.04 t 0.06 0.1178 13.63 Formic acid 0.01 0.01 ND t79 13.73 Camphor 0.02 0.03 0.03 t80 13.73 Benzaldehyde 0.04 0.01 0.09 ND81 13.99 b-Bourbonene 0.64 t 0.34 ND82 14.00 Propionic acid ND t ND ND83 14.15 (E)-Non-2-en-1-al ND 0.03 ND ND84 14.27 a-Gurjunene t ND t t85 14.27 Pentadec-1-ene 0.03 t ND ND86 14.44 b-Cubebene 0.05 t 0.03 t87 14.44 Theaspirane B 0.02 0.18 0.01 0.1288 14.44 Linalool 0.55 0.03 0.55 0.1489 14.60 Octanol 0.01 t 0.02 1.3890 15.00 Methyl citronellate 0.01 t 0.01 ND91 15.16 Isopulegol ND 0.01 ND ND92 15.25 ‘(E,E)-Hexa-2,4-dien-1-al’ 0.01 t 0.01 ND93 15.56 ‘6-Methylhepta-3,5-dien-2-one’ 0.03 t 0.03 ND94 15.90 b-Elemene 0.12 t 0.05 0.0595 16.11 b-Caryophyllene 0.18 0.20 0.10 0.2096 16.11 ‘g-Valerolactone (pentan-1,4-olide)’ 0.02 t ND ND97 16.27 Prenyl tiglate ND t 0.02 ND98 16.40 Aromaderene 0.01 t 0.03 t99 16.58 Hexadecane 0.01 t 0.03 t100 16.65 Citronellyl formate 0.01 0.01 0.01 0.01101 16.95 Methyl neroate 0.02 0.01 0.02 t102 16.95 Phenylacetaldehyde 0.01 t ND ND103 17.16 (E)-Dec-2-en-1-al 0.01 0.01 ND ND104 17.16 Acetophenone 0.04 t 0.06 ND105 17.19 Menthol ND ND 0.04 ND106 17.66 Nonanol 0.02 ND 0.02 t107 17.84 Citronellyl acetate 0.20 0.04 0.29 0.56108 17.97 Neryl formate t ND 0.03 ND109 17.98 (E)-b-Farnesene 0.03 t ND ND110 18.05 4-Methoxystyrene ND t 0.02 t111 18.06 a-Humulene 0.03 t 0.02 ND

Volatile constituents of blue-coloured hybrid tea rose flower

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112 18.19 Neral 1.01 1.30 0.73 1.18113 18.32 ‘Neral-2,3-oxide’ 0.10 t 0.12 0.01114 18.64 a-Terpineol ND 0.01 ND t115 18.70 g-Muurolene 0.03 t 0.02 0.03116 18.70 Methyl geranate 0.33 0.26 0.85 t117 18.70 ‘Geranial-2,3-oxide’ 0.05 0.03 0.10 0.10118 18.70 Borneol 0.05 0.03 0.02 0.01119 18.75 Anisyl alcohol 0.01 0.01 0.03 0.01120 19.00 Geranyl formate 0.05 0.02 0.02 t121 19.36 Germacrene D 5.90 0.11 2.13 0.07122 19.44 Benzyl acetate ND ND 0.02 0.51123 19.45 Piperitone 0.03 t ND ND124 19.59 Pentanoic acid ND 0.43 ND ND125 19.61 Neryl acetate ND ND 0.51 0.29126 19.67 b-Selinene 0.05 t ND ND127 19.67 Heptadecane 0.12 t 0.10 t128 19.67 Geranial 4.16 6.74 1.68 6.27129 19.74 b-Bisabolene 0.12 t 0.03 ND130 19.94 Naphthalene 0.02 t ND ND131 19.94 (Z)-Heptadec-8-ene 0.10 0.12 0.12 t132 20.00 Bicyclogermacrene 0.10 0.12 0.02 t133 20.16 ‘(E,E)-a-Farnesene’ t ND 0.02 t134 20.25 (E)-Undec-2-en-1-al 0.01 t ND ND135 20.25 p-Methyldimethylbenzyl alcohol 0.04 0.01 0.09 0.07136 20.50 Geranyl acetate 1.02 0.45 2.15 1.41137 20.50 d-Cadinene 0.12 t 0.01 0.10138 20.63 Methyl salicylate ND t ND ND139 20.76 Citronellol 7.94 3.41 5.83 0.98140 21.16 a-Cadinene ND ND t t141 21.19 g-Isogeraniol 0.22 t 0.36 0.03142 21.69 Nerol 9.83 3.82 19.23 10.34143 21.70 ‘(E,E)-Deca-2,4-dien-1-al’ ND 0.04 ND ND144 21.81 Isonerol 0.13 0.01 0.21 0.02145 21.88 Isogeraniol 0.16 0.01 0.39 0.02146 21.88 2-Phenyethyl acetate 0.05 0.04 0.05 0.02147 22.16 Octadecane t ND ND ND148 22.50 Dihydro-b-ionone 0.28 0.38 0.02 t149 22.60 Hexanoic acid ND 0.03 0.01 0.02150 22.61 Calamenene 0.03 t 0.02 t151 22.74 ‘1,3-Dimethoxy-5-methylbenzene (DMMB)’ 1.15 1.74 0.66 0.59152 23.15 Geraniol 44.51 38.94 47.91 34.35153 23.30 ‘cis-3,7-Dimethyloct-7-en-1-ol 3,6-oxide’ 0.11 0.15 0.21 ND154 23.32 Geranyl acetone t t 0.06 t155 23.42 ‘cis-3,7-Dimethyl-3,6-epoxyoct-7-en-1-yl acetate’ 0.03 t ND ND156 23.42 Benzyl alcohol 0.07 0.12 0.17 0.05157 23.86 ‘trans-3,7-Dimethyloct-7-en-1-ol 3,6-oxide’ 0.08 t 0.09 ND158 24.06 Epi-4-cubebol 0.05 0.04 ND ND159 24.35 2-Phenylethyl alcohol 0.09 1.17 0.11 0.13160 24.80 a-Corocalene 0.10 t ND ND161 25.07 Nonadecane 0.38 t 0.27 0.01162 25.19 6-Hydroxydihydrotheaspirane 0.01 t ND ND163 25.25 b-Ionone ND 0.04 ND ND164 25.44 Nonadec-1-ene 2.78 t 0.50 t165 25.44 Cubebol 0.10 t 0.02 t166 25.44 ‘3,7-Dimethylocta-1,5-diene-3,7-diol’ 0.05 t 0.03167 25.60 b-Ionol ND 0.03 ND ND

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168 26.06 Dihydro-b-ionol 1.91 5.19 t 0.04169 26.37 Caryophyllene oxide 0.04 ND ND ND170 26.70 Phenol 0.21 t 0.09 t171 27.10 Methyl eugenol ND ND 0.30 t172 27.37 ‘Geraniol-6,7-epoxide acetate’ ND ND 0.02 0.05173 27.52 Eicosane 0.05 0.02 0.01 t174 27.60 (Z)-Cyclododec-9-enolide (yuzu lactone) (2) ND t ND t175 27.67 ‘Geranial-6,7-oxide’ 0.03 t t 0.05176 27.84 Eicos-1-ene 0.10 0.02 ND ND177 27.94 ‘3,7-Dimethyl-6,7-epoxyoctanol �1)’ 0.02 t 0.04 t178 27.98 (E)-Nerolidol ND 0.03 ND 0.01179 28.16 ‘3,7-Dimethyl-6,7-epoxyoctanol �2)’ 0.03 t 0.02 t180 28.29 ‘2,7-Germacradien-1-ol’ ND ND 0.04 t181 28.29 Octanoic acid ND 0.01 0.03 ND182 28.32 Cubenol 0.05 0.11 ND ND183 28.40 ‘Germacra-2,7-dien-1-ol’ 0.03 ND ND ND184 28.41 Cubenol ND ND 0.02 0.07185 28.67 Epi-cubenol 0.03 0.03 0.03 t186 28.74 ‘Nerol-6,7-oxide’ 0.08 0.04 0.21 0.01187 28.74 ‘Nerol-2,3-oxide’ 0.10 0.04 0.18 0.05188 28.88 Elemol ND ND 0.04 0.07189 29.29 ‘Geraniol-2,3-oxide’ 0.21 0.59 0.28 0.18190 29.93 ‘Caryophylla-2(12),6-dien-5-yl acetate’ 0.06 ND ND ND191 30.06 Heneicosane 0.14 t 0.14 t192 30.12 ‘Geraniol-6,7-oxide’ 0.18 0.31 0.19 0.06193 30.23 Spathulenol 0.02 t ND ND194 30.34 Heneicos-1-ene 0.19 t ND ND195 30.46 ‘1,3,5-Trimethoxybenzene (TMB)’ 0.18 0.31 0.13 0.71196 30.72 Mint sulfide t ND 0.03 ND197 30.78 Eugenol ND 0.03 ND ND198 31.10 t-Cadinol 0.08 0.31 0.01 t199 31.19 Nonanoic acid ND 0.03 ND ND200 31.49 t-Muurolol 0.10 0.04 0.04 0.09201 31.61 Torreyol 0.04 t 0.02 t202 31.65 ‘(2Z,6Z)-Farnesol’ ND ND ND 0.02203 32.29 a-Eudesmol t t 0.02 0.11204 32.30 b-Eudesmol t t 0.03 0.14205 32.48 Methyl hexadecanoate ND 0.02 ND ND206 32.55 a-Cadinol 0.08 t 0.03 0.25207 32.60 ‘(2Z,6E)-Farnesal’ 0.14 t 0.24 t208 32.79 Citronellic acid ND 0.01 0.01 0.01209 33.40 (E)-Dihydrofarnesol ND t ND ND210 33.42 Ethyl hexadecanoate ND 0.04 ND t211 33.75 Neric acid ND 0.03 ND ND212 33.85 Tricosane t ND t ND213 33.98 ‘(2E,6E)-Farnesal’ 0.44 0.09 0.02 t214 34.51 ‘(2Z,6E)-Farnesol’ ND 0.02 0.02 0.04215 34.57 Tricosane 0.03 t t t216 34.87 Geranic acid 0.10 5.39 t 6.21217 35.61 ‘(2E,6E)-Farnesol’ 0.11 0.57 0.29 1.44218 36.03 Methyl cis-(Z)-jasmonate (3) 0.01 0.02 0.01 0.01219 36.95 Indole t t ND t220 38.50 Dodecanoic acid ND 0.11 0.02 0.11221 40.54 Methyl linolenate ND 0.11 ND t222 41.04 Octadecanol ND ND ND t223 41.54 Ethyl linolenate ND 0.53 ND ND


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224 44.31 Tetradecanoic acid ND ND 0.04 0.11225 44.55 Pentadecanoic acid ND ND 0.03 t226 54.41 Hexadecanoic acid ND 1.31 0.11 5.92227 55.93 Hexadec-9-enoic acid ND ND 0.02 t228 70.98 Octadecanoic acid ND 0.41 ND 1.09229 79.77 Linoleic acid ND 0.31 ND ND

The gas chromatography peak area percentage was determined by the AromacopeW method (dynamic headspace GC and solventextraction.t, trace, i.e. <0.01%; ND, not detected.Identification was by mass spectrometry. The retention times (RTs) agreed with those of authentic samples.

A. Joichi et al.

The volatile components adsorbed on the TenaxW TA underwentsolvent extraction followed by a desorption process. The volatilecomponents adsorbed on the TenaxW TA were eluted with diethyl ether.The resulting extract was concentrated at 40�C under nitrogen atatmospheric pressure and analysed by GC-MS.

Solvent Extraction

Flowers of both ‘Blue Moon’ and ‘Blue Perfume’ were extracted withpentane for 24 h just after picking. The extract was dried over anhydroussodium sulfate, filtered, and then concentrated to give the concentrateunder reduced pressure. A sample of 1.08 g of concentrate was obtainedfrom 650 g of ‘Blue Moon’ flowers and 4.44 g of concentrate from 1610 gof ‘Blue Perfume’ flowers. After removing the higher fatty acids and flowerwaxes by using column chromatography utilizing a glass column packedwith silica gel 60 (70–230 mesh), 267 mg of volatile concentrate wasobtained from ‘BlueMoon’, and 1.57 g of volatile concentrate was obtainedfrom ‘Blue Perfume’.

Gas Chromatography

GC analyses were carried out on a Agilent HP6890 gas chromatograph(Agilent Technologies, Santa Clara CA) equipped with a flame ionizationdetector and a 50 m� 0.25 mm� 0.15 mm BC-WAX (produced by GLSciences, Tokyo, Japan) coated capillary column. The oven temperaturewas programmed linearly from 70�C to 215�C at a rate of 4�C/min.

Gas Chromatography–Mass Spectrometry

GC-MS analyses were carried out using a GCMS-QP2010 (Shimadzu Co.,Kyoto, Japan) with an ionization energy of 27 eV and equipped withBC-WAX column (50 m� 0.25 mm i.d., film thickness of 0.15 mm; GLSciences). The injector and the ion source were set at 250�C and200�C, respectively. The spectrometer was used in scan mode. Thecarrier gas was helium with a constant pressure set at 100 kPa. Theoven temperature program was set from 70�C to 218�C at 4�C/min.Identification of the compounds was performed by comparing andmatching the retention times and mass spectra with original setsof standard data.

Multi-dimensional Gas Chromatography andMass Spectrometry

Multi-dimensional gas chromatography (MDGC) analyses were carriedout on a Shimadzu QP2010 equipped on the first gas chromatographwith a BC-WAX column (50 m� 0.25 mm� 0.25 mm; GL Sciences), and

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on the second gas chromatograph with b-DEX 225, b-DEX 325 (25 m0.25 mm� 0.25 mm; Agilent Technologies), or b-DEX sm (Restek,Bellefonte, PA, USA) combined with a MS detector. Chiral GC analyseswere performed using a MDGC/GCMS-2010 (Shimadzu). With thissystem, a heart-cut of the relevant fractions could be made as well as atransfer from the achiral column to the chiral one. The injector and theflame ionization detector temperatures were both 250�C. The carriergas was helium with a constant pressure set at 180 kPa. The oventemperature program was set from 70�C to 230�C at 5�C/min.

For citronellol analysis, the second gas chromatograph was equippedwith a b-DEX 225 column (30 m� 0.25 mm i.d., film thickness of 0.25 mm;Supelco, Bellefonte, PA, USA) and the oven temperature program was setfrom 70�C to 180�C at 1�C/min.

For citronellyl acetate, the second gas chromatograph was equipped witha b-DEX se column (30 m� 0.25 mm i.d., film thickness of 0.25 mm; Restek)and the oven temperature program was set from 70�C to 180�C at 1�C/min.

For linalool, (E)-nerolidol, theaspiranes and cis-rose oxide, the secondgas chromatograph was equipped with a b-DEX 325 column (30 m0.25 mm i.d., film thickness of 0.25 mm; Supelco) and the oven tempera-ture program was set from 70�C to 180�C at 1�C/min.

For trans-rose oxide, the second gas chromatograph was equippedwith a Chiralsil DEX CB column (30 m� 0.25 mm i.d., film thickness of0.25 mm; Agilent) and the oven temperature program was set from70�C to 180�C at 1�C/min.

Results and Discussion

Major Volatile Components in ‘Blue Moon’ and ‘Blue Perfume’

The volatile components of two blue-coloured HT roses arelisted in Table 1. In the headspace analyses, 156 componentswere identified in the scent of ‘Blue Moon’ and 136 in the scentof ‘Blue Perfume’. Then, in the solvent extracts of the two roseflowers, 201 components were identified in ‘Blue Moon’ and 170components were identified in ‘Blue Perfume’. Monoterpenealcohols and their aldehydes, such as geraniol, nerol, citronelloland geranial, were identified as the major components (Figure 1).These monoterpene derivatives greatly contributed to the rich roseand the fresh lemon-like note. 1,3-Dimethoxy-5-methylbenzene anddihydro-b-ionol were identified as the major components in thescent of ‘Blue Moon’ flower. 1,3-Dimethoxy-5-methylbenzene,with a phenolic–spicy note, and dihydro-b-ionol, with a woodynote, were identified during the evaluation of this rose flower.

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Blue Moon

Blue Perfume

0 10 20 30 40

0 10 20 30 40

2

BH

TB

HT

solvent

1

1. germacrene D, 2. geranial, 3. geranyl acetate, 4. citronellol, 5. nerol, 6. 1,3-dimethoxy-5-methyl benzene, 7. geraniol, 8. nonadecane, 9. dihydro- -ionol

12 3

4

4

5

5

6

7

7

8

8

9

Figure 1. Gas chromatogram of the hybrid tea roses ‘Blue Moon’ and ‘Blue Perfume’. BHT (dibutyl hydroxy toluene)


On the other hand, the amounts of these two components werequite low in the scent of ‘Blue Perfume’.

Table 2. The enantiomeric ratio of the chiral components

Component and enantiomer Amount (%)Citronellol

3R —3S >99

Citronellyl acetate3R —3S >99

Linalool3R 923S 8

Dihydro-b-ionolR —S >99

(E)-Nerolidol3R —3S >99

cis-Rose oxide‘2S, 4R’ 99‘2R, 4S’ 1

trans-Rose oxide

Minor Key Volatile Components

The high contribution components for the odour of blue-type roseflower were studied by using gas chromatography–olfactometryand a new component was detected which has a faint pungent,vegetable and tropical fruit-like character. This componentwas determined as 2-isopropyl-4-methylthiazole (1) using GCand GC-MS. The GC retention time and the mass spectrum ofcompound 1 was the same as that of an authentic samplepurchased from Junsei Kagaku Co., Ltd (Tokyo, Japan). In fact,compound 1, which is found as one of the volatile componentsin tomato[8] and durian,[9] was identified in rose flower for thefirst time. It was confirmed that the addition of a small amountof compound 1 to duplicated rose-type oil and rose-typefragrance (‘Blue Moon’ type) showed good performance forthe original rose fragrance (data not shown).

(Z)-Cyclododec-9-enolide [(Z)-yuzu lactone] (2), which wasidentified in the solvent extracts of ‘Blue Moon’ and ‘BluePerfume’, was identified in the rose flowers for the first time.Compound 2, whichwas amacrocyclic lactonewith a camphorousnote, was found in yuzu (popular Japanese citrus),[10] Japanesemugwort,[11] golden orange (another Japanese citrus),[12] andlily of the valley.[13]

Then, methyl cis-(Z)-jasmonate (3), which is found in orchidflower[14] and lemon peel oil,[15] was also identified in ‘BlueMoon’ and ‘Blue Perfume’ oils collected by the headspace andsolvent extraction methods. This is the first time that compound 3has been found in rose flower. The addition of a small amount ofcompound 3 to a rose-type fragrance greatly enhanced thenatural odour of blue-coloured HT rose flower. This performancewas similar to that of compound 1.

‘2R, 4R’ >99‘2S, 4S’ —

Theaspirane A‘2R, 5R’ 4‘2S, 5S’ 96

Theaspirane B‘2S, 5R’ 97‘2R, 5S’ 3

Analysis of the Enantiomers of Blue-type Hybrid Tea Roses

Several components identified in the two types of rose flowerhad asymmetric carbon atoms in their molecules, leading us toanalyse their chirality. For the first time, the enantiomeric ratioof linalool, (E)-nerolidol, theaspiranes, dihydro-b-ionol, citronelloland citronellyl acetate could be assigned by MDGC-MS

Flavour Fragr. J. 2013 Copyright © 2013 John

equipped with a chiral column as the secondary column. Thefollowing results were found for both rose flowers.The ratio of the (S)-enantiomer vs. the (R)-enantiomer of

linalool was 8:92, although the existing ratio of linalool in roseoil was reported to be almost 50:50.[16] At this time, we do nothave any explanation for this discrepancy.For cis-rose oxide, the ratio of the (2S,4R)-enantiomer vs. the

(2R,4S)-enantiomer was 99:1. For citronellol and citronellylacetate, only the (S)-enantiomer was detected. For trans-roseoxide, only the (2R,4R)-enantiomer was detected. The resultsof the enantiomeric ratio of citronellol, citronellyl acetate,

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A. Joichi et al.

cis-rose oxide, and trans-rose oxide were suggested for pro-ceeding research.[16]

As shown in Table 2, only the (S)-enantiomer was detected for(E)-nerolidol, and dihydro-b-ionol. The ratio of the (2R)-enantiomervs. the (2S)-enantiomer in theaspirane A and theaspirane Bwas about 4:96.

ConclusionThe characteristic volatile components of rose flower (bluetype) were revealed by a combination of GC-MS and gaschromatography–olfactometry analyses. The enantiomericdistribution of the important odour-active components wasdetermined using MDGC-MS. These analytical results wouldgive very important information for creating fragrances closeto the natural aromas.

References1. G. Ohloff, Perfumer Flavorist 1978, 3, 11.2. C. Mann, T. Smith. In Proceedings of the 7th International Congress of Essen-

tial Oils, Kyoto, Japan, 1977, p. 458 (abstract).

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3. I. Watanabe, T. Yanai, S. Tamogami. In Proceedings of the 7th InternationalCongress of Essential Oils, Kyoto, Japan, 1977, p. 461 (abstract).

4. F. Buccellato, Perfumer Flavorist 1980, 5, 29.5. J. Garnero, Parfums Cosmetiques Aromes, 1982, 47, 31.6. A. Omata, K. Yomogida, S. Nakamura, T. Ota, T. Toyoda, A. Amano, S.

Muraki, Flavour Fragr. J. 1991, 6, 149.7. A. Joichi, K. Yomogida, K. Awano, Y. Ueda, Flavour Fragr. J. 2005

20, 152.8. C.-T. Ho, N. Ichimura., Lebensm-Wiss Technol. 1982, 15, 340.9. H. Weenen, W. E. Koolhaas, A. Apriyantono, J. Agric. Food Chem.

1990, 44, 3291.10. Y. Matsuura, G. Hata, S. Abe, I. Sakai, A. Abe, Technical Data of the 8th

International Congress of Essential Oils, Cannes, Grasse, France., 1980,p. 497.

11. K. Umano, Y. Hagi, K. Nakamura, A. Shoji, T. Shibamoto, J. Agric. Food.Chem. 2000, 48, 3463.

12. K. Katayama, H. Iwabuchi, Food and Food Ingredients J. Jpn. 2002,202, 55.

13. T. Watanabe, M. Omoto, H. Iwabuchi, In Proceedings of the 52nd

Symposium on the Chemistry of Terpenes, Essential Oils, andAromatics, Izumino, Itakura-machi, Ora-gun, Gunma, Japan, 2008,p. 337 (abstract).

14. A. Omata, S. Nakamura, K. Yomogida, K. Moriai, Y. Ichikawa,I. Watanabe, Agric. Biol. Chem. 1990, 54, 1029.

15. R. Nishida, T. E. Acree, J. Agric. Food. Chem. 1984, 32, 1001.16. P. Kreis, A. Mosandl, Flavour Fragr. J. 1992, 7, 199.

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Volatile Constituents of Blue-rose