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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 hybrid tea rose owers


Atsushi 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 owers, which, on an olfactory basis, are classied as a blue type were analysed using AromascopeW technology (modied headspace technology) and solvent extraction methods followed by gas chromatographymass spectrometry analysis. One hundred and eighty components were identied in the headspace volatile components of Blue Moon ower and 188 components were identied in solvent extracts. Among them, geraniol, nerol, citronellol, 1,3-dimethoxy-5-methylbenzene and dihydro-b-ionol were identied as the main odour components. On the other hand, in Blue Perfume, 165 components were identied in the headspace volatile components and 150 components were identied in solvent extracts. Among them, geraniol, nerol, citronellol, neral, and geranial were identied as the major odour compounds. From both rose owers, three components were newly identied: 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 identied in both of the headspace components and solvent extracts of the two types of rose ower, and then yuzu lactone was identied only in solvent extracts as the one of the minor components. Several components identied in both owers have asymmetric carbon atoms in their molecules, leading us to analyse their chirality. For the rst time, the enantiomer ratios of linalool, (E)-nerolidol, theaspiranes and dihydro-b-ionol could be assigned by multi-dimensional gas chromatographymass spectrometry. The results were as follows in both rose owers. The ratio of the (S)-enantiomer vs. the (R)-enantiomer of linalool was 8:92. Only the (S)-enantiomer was detected for (E)-nerolidol and dihydro-b-ionol. The ratios of the (2R,5R)-enantiomer vs. the (2S,5S)-enantiomer in theaspirane A and the (2R,5S)-enantiomer vs. the (2S,5R)-enantiomer in theaspirane B were about 4:96. Copyright 2013 John Wiley & Sons, Ltd. Keywords: hybrid tea rose; Blue Moon; Blue Perfume; volatile constituents; enantiomer

Introduction
The rose has been admired since ancient times because of the beauty of its shape and colour, as well as its scent. Natural oils obtained 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 in perfumery and more than 350 components have been reported.[16] Hybrid tea (HT) roses are mainly the result of natural and articial hybridization between European and Chinese roses over a long time.[7] As a result, the analyses revealed many distinctive notes in the HT rose; in particular, the classied blue-coloured HT rose showed a characteristic odour reminiscent of a combination consisting of damask rose notes, tea-like and lemon-like fresh notes. In this study, volatile components of the Blue Moon and Blue Perfume cultivars, with characteristic rose notes represented by blue-coloured HT rose owers, were analysed by using gas chromatographymass spectrometry (GC-MS), and the presence of major odour compounds was revealed.

The selection criteria were essentially the elegance, sophisticated and strength of the typical blue rose character in their scent. Blue Moon had a rich rose-like, lemon-like, woody and phenolicspicy note, while Blue Perfume had a rich rose-like, lemon-like and fresh note with green fruity aspects.

Collection of the Rose Flower Scent


One to two owers which were just ready to open and had a good smell as judged by ve perfumers, were selected and enveloped in a TedlarW bag equipped with a glass column (i.d. 8 mm) packed with TenaxW TA (6080 mesh, 400 mg). The inlet of an air pump was connected to the TenaxW TA glass column in which the headspace gas was enriched by pumping at 400 ml/min. The owers enveloped in the TedlarW bag and the one undeveloped had no characteristic difference in appearance throughout the trapping. In order to study the seasonal changes of the volatiles emitted by the living owers, this procedure was repeated twice at the beginning of November in 2008 and the end of May in 2009 at Keisei Rose Garden.

Experimental
Selection of Typical Blue-type Rose Flowers
Among 700 stocks studied, we selected Blue Moon and Blue Perfume, two types of popular rose owers cultivated by Keisei Rose Nurseries Inc. (Keisei Rose Garden) located in Yachiyo city in Chiba prefecture.

* Correspondence to: Atsushi Joichi, Shiseido Research Center (Shin-Yokohama), 2-2-1, Hayabuchi, Tsuzuki-ku, Yokohama-shi, Kanagawa 2248558, Japan. E-mail: atsushi.jouichi@to.shiseido.co.jp
a Shiseido Research Center (Shin-Yokohama), 2-2-1, Hayabuchi, Tsuzuki-ku, Yokohama-shi, Kanagawa, 224-8558, Japan b Takasago International Corporation Corporate Research & Development Division, 4-11, Nishiyawata 1-chome, Hiratsuka City, Kanagawa, 254-0073, Japan

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A. Joichi et al. Table 1. Components identied in hybrid tea rose ower (Blue Moon and Blue Perfume) No. RT (min) Compound Gas chromatography area (%) Blue Moon Blue Perfume Headspace Solvent extract Headspace Solvent extract 0.05 0.05 0.03 0.02 t 0.01 0.01 t 0.01 t t 0.01 ND t 0.01 t 0.01 0.01 ND ND 0.16 0.01 0.01 0.01 t 0.01 0.01 t 0.02 t t 0.36 t t 0.01 0.02 0.02 0.02 0.31 0.02 0.02 0.13 0.11 t 0.05 0.10 t 0.04 0.01 0.01 0.01 ND 0.11 0.02 0.08 0.07 ND ND 0.01 ND 0.31 t t ND t ND 0.01 t t ND 0.01 ND t t ND 0.02 t t 0.02 0.09 t t 0.02 0.02 0.01 t t 0.12 t t t 0.31 t t 0.31 t t 0.38 0.01 t 0.03 0.31 t 0.02 0.01 ND ND t 0.04 ND 0.02 ND 0.05 ND 0.01 0.02 t 0.01 0.01 ND 0.01 0.01 0.01 t ND ND 0.01 t ND t ND 0.05 0.21 0.01 0.01 0.01 ND ND 0.01 0.01 0.06 ND t 0.49 0.02 ND 0.02 0.03 0.01 ND 1.32 ND 0.03 0.49 0.22 ND 0.02 0.06 t 0.08 0.03 ND 0.03 t 0.03 0.02 0.02 0.11 0.03 ND 0.01 ND 0.69 t ND ND t ND 0.11 0.01 ND ND 0.07 0.26 ND t t 0.01 0.01 0.01 0.01 ND ND ND 0.01 0.01 t ND t 0.07 ND ND t 0.01 0.01 ND 0.63 ND 0.01 0.12 0.02 t 0.04 0.10 t t 0.07 ND t t t ND 0.02 0.05

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57

3.40 3.80 3.90 4.00 4.06 4.24 4.39 4.48 4.75 5.10 5.10 5.12 5.40 5.40 5.85 6.08 6.09 6.24 6.30 6.31 6.31 6.49 6.49 6.66 7.00 7.01 7.06 7.20 7.23 7.30 7.32 7.65 7.75 7.80 7.99 8.35 8.40 8.47 8.56 8.66 8.66 8.92 8.99 9.11 9.11 9.24 9.45 9.48 9.59 9.72 9.84 9.91 10.00 10.40 10.40 10.57

Acetaldehyde Butan-2-one Ethanol But-3-en-2-one Pentanal Decane 2-Methylbut-3-en-2-ol Butyl formate Butyl acetate Isopentyl acetate Hexanal Uecane Butyl alcohol 2-Methylbutyl acetate Myrcene Heptanal Pentyl acetate 3-Methylbut-3-en-1-yl acetate 2-Methylbutanol Isopentyl alcohol Senecionaldehyde Dodecane Limonene (E)-Hex-2-en-1-al cis-Ocimene 6-Methylheptan-2-one Pentyl alcohol 3-Methylbut-3-en-1-ol Prenyl acetate trans-Herboxide trans-Ocimene Hexyl acetate p-Cymene (Z)-4,8-Dimethylnona-1,3,7-triene Octanal (E)-Hex-3-en-1-yl acetate Tridecane (E)-4,8-Dimethylnona-1,3,7-triene (Z)-Hex-3-en-1-yl acetate 6-Methylhept-6-en-2-one 3-Methylpentanol (E)-Hex-2-en-1-yl acetate 6-Methylhept-5-en-2-one Heptyl acetate Anisole Hexanol 2-Isopropyl-4-methylthiazole (1) cis-Rose oxide (E)-Hex-3-en-1-ol 3,3-Dimethylcyclohexanone trans-Rose oxide Heptyl acetate (Z)-Hex-3-en-1-ol Benzyl methyl ether Nonanal Rose furan

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Volatile constituents of blue-coloured hybrid tea rose ower Table 1. (Continued) No. RT (min) Compound Gas chromatography area (%) Blue Moon Blue Perfume Headspace Solvent extract Headspace Solvent extract 0.01 0.01 0.03 t 0.05 t t 0.02 t 0.01 ND 0.11 0.02 0.03 t ND 0.38 0.10 0.05 0.04 0.01 0.02 0.04 0.64 ND ND t 0.03 0.05 0.02 0.55 0.01 0.01 ND 0.01 0.03 0.12 0.18 0.02 ND 0.01 0.01 0.01 0.02 0.01 0.01 0.04 ND 0.02 0.20 t 0.03 ND 0.03 t 0.01 ND t 0.22 t ND 0.02 t 0.01 ND t t t t ND t t 0.11 t 0.01 0.03 0.01 t t 0.03 ND t t 0.18 0.03 t t 0.01 t t t 0.20 t t t t 0.01 0.01 t 0.01 t ND ND 0.04 ND t t t 0.02 0.01 0.10 0.09 0.02 0.01 ND 0.01 ND 0.06 0.01 0.04 ND 0.02 ND 0.01 0.18 0.03 0.01 0.06 ND 0.03 0.09 0.34 ND ND t ND 0.03 0.01 0.55 0.02 0.01 ND 0.01 0.03 0.05 0.10 ND 0.02 0.03 0.03 0.01 0.02 ND ND 0.06 0.04 0.02 0.29 0.03 ND 0.02 0.02 ND 0.11 ND ND t 0.01 ND t ND 0.01 ND t ND t ND t t 0.01 0.02 0.11 t t ND ND ND ND t ND t 0.12 0.14 1.38 ND ND ND ND 0.05 0.20 ND ND t t 0.01 t ND ND ND ND t 0.56 ND ND t ND

58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111

10.75 10.93 11.02 11.53 11.54 11.63 11.73 11.73 11.95 12.06 12.30 12.30 12.39 12.61 12.75 12.95 13.25 13.25 13.43 13.63 13.63 13.73 13.73 13.99 14.00 14.15 14.27 14.27 14.44 14.44 14.44 14.60 15.00 15.16 15.25 15.56 15.90 16.11 16.11 16.27 16.40 16.58 16.65 16.95 16.95 17.16 17.16 17.19 17.66 17.84 17.97 17.98 18.05 18.06

Tertadecane (Z)-Hex-2-en-1-ol 3-(4-Methylpent-3-enyl)furan Neryl methyl ether Acetic acid trans-Linalool oxide (furanoid) (E)-Oct-2-en-1-ol Heptanol 10-Methylene-2,6,6-trimethyl-1oxaspiro[4.5]decane 6-Methylhept-5-en-2-ol Geranyl methyl ether a-Cubebene Nerol-1,5-oxide Citronellal a-Longipinene a-Ylangene a-Copaene Decanal Theaspirane A Pentadecane Formic acid Camphor Benzaldehyde b-Bourbonene Propionic acid (E)-Non-2-en-1-al a-Gurjunene Pentadec-1-ene b-Cubebene Theaspirane B Linalool Octanol Methyl citronellate Isopulegol (E,E)-Hexa-2,4-dien-1-al 6-Methylhepta-3,5-dien-2-one b-Elemene b-Caryophyllene g-Valerolactone (pentan-1,4-olide) Prenyl tiglate Aromaderene Hexadecane Citronellyl formate Methyl neroate Phenylacetaldehyde (E)-Dec-2-en-1-al Acetophenone Menthol Nonanol Citronellyl acetate Neryl formate (E)-b-Farnesene 4-Methoxystyrene a-Humulene

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A. Joichi et al. Table 1. (Continued) No. RT (min) Compound Gas chromatography area (%) Blue Moon Blue Perfume Headspace Solvent extract Headspace Solvent extract 1.01 0.10 ND 0.03 0.33 0.05 0.05 0.01 0.05 5.90 ND 0.03 ND ND 0.05 0.12 4.16 0.12 0.02 0.10 0.10 t 0.01 0.04 1.02 0.12 ND 7.94 ND 0.22 9.83 ND 0.13 0.16 0.05 t 0.28 ND 0.03 1.15 44.51 0.11 t 0.03 0.07 0.08 0.05 0.09 0.10 0.38 0.01 ND 2.78 0.10 0.05 ND 1.30 t 0.01 t 0.26 0.03 0.03 0.01 0.02 0.11 ND t 0.43 ND t t 6.74 t t 0.12 0.12 ND t 0.01 0.45 t t 3.41 ND t 3.82 0.04 0.01 0.01 0.04 ND 0.38 0.03 t 1.74 38.94 0.15 t t 0.12 t 0.04 1.17 t t t 0.04 t t t 0.03 0.73 0.12 ND 0.02 0.85 0.10 0.02 0.03 0.02 2.13 0.02 ND ND 0.51 ND 0.10 1.68 0.03 ND 0.12 0.02 0.02 ND 0.09 2.15 0.01 ND 5.83 t 0.36 19.23 ND 0.21 0.39 0.05 ND 0.02 0.01 0.02 0.66 47.91 0.21 0.06 ND 0.17 0.09 ND 0.11 ND 0.27 ND ND 0.50 0.02 0.03 ND 1.18 0.01 t 0.03 t 0.10 0.01 0.01 t 0.07 0.51 ND ND 0.29 ND t 6.27 ND ND t t t ND 0.07 1.41 0.10 ND 0.98 t 0.03 10.34 ND 0.02 0.02 0.02 ND t 0.02 t 0.59 34.35 ND t ND 0.05 ND ND 0.13 ND 0.01 ND ND t t ND

112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167

18.19 18.32 18.64 18.70 18.70 18.70 18.70 18.75 19.00 19.36 19.44 19.45 19.59 19.61 19.67 19.67 19.67 19.74 19.94 19.94 20.00 20.16 20.25 20.25 20.50 20.50 20.63 20.76 21.16 21.19 21.69 21.70 21.81 21.88 21.88 22.16 22.50 22.60 22.61 22.74 23.15 23.30 23.32 23.42 23.42 23.86 24.06 24.35 24.80 25.07 25.19 25.25 25.44 25.44 25.44 25.60

Neral Neral-2,3-oxide a-Terpineol g-Muurolene Methyl geranate Geranial-2,3-oxide Borneol Anisyl alcohol Geranyl formate Germacrene D Benzyl acetate Piperitone Pentanoic acid Neryl acetate b-Selinene Heptadecane Geranial b-Bisabolene Naphthalene (Z)-Heptadec-8-ene Bicyclogermacrene (E,E)-a-Farnesene (E)-Undec-2-en-1-al p-Methyldimethylbenzyl alcohol Geranyl acetate d-Cadinene Methyl salicylate Citronellol a-Cadinene g-Isogeraniol Nerol (E,E)-Deca-2,4-dien-1-al Isonerol Isogeraniol 2-Phenyethyl acetate Octadecane Dihydro-b-ionone Hexanoic acid Calamenene 1,3-Dimethoxy-5-methylbenzene (DMMB) Geraniol cis-3,7-Dimethyloct-7-en-1-ol 3,6-oxide Geranyl acetone cis-3,7-Dimethyl-3,6-epoxyoct-7-en-1-yl acetate Benzyl alcohol trans-3,7-Dimethyloct-7-en-1-ol 3,6-oxide Epi-4-cubebol 2-Phenylethyl alcohol a-Corocalene Nonadecane 6-Hydroxydihydrotheaspirane b-Ionone Nonadec-1-ene Cubebol 3,7-Dimethylocta-1,5-diene-3,7-diol b-Ionol

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Volatile constituents of blue-coloured hybrid tea rose ower Table 1. (Continued) No. RT (min) Compound Gas chromatography area (%) Blue Moon Blue Perfume Headspace Solvent extract Headspace Solvent extract 1.91 0.04 0.21 ND ND 0.05 ND 0.03 0.10 0.02 ND 0.03 ND ND 0.05 0.03 ND 0.03 0.08 0.10 ND 0.21 0.06 0.14 0.18 0.02 0.19 0.18 t ND 0.08 ND 0.10 0.04 ND t t ND 0.08 0.14 ND ND ND ND t 0.44 ND 0.03 0.10 0.11 0.01 t ND ND ND ND 5.19 ND t ND ND 0.02 t t 0.02 t 0.03 t ND 0.01 0.11 ND ND 0.03 0.04 0.04 ND 0.59 ND t 0.31 t t 0.31 ND 0.03 0.31 0.03 0.04 t ND t t 0.02 t t 0.01 t 0.04 0.03 ND 0.09 0.02 t 5.39 0.57 0.02 t 0.11 0.11 ND 0.53 t ND 0.09 0.30 0.02 0.01 ND t ND 0.04 ND 0.02 0.04 0.03 ND ND 0.02 0.03 0.21 0.18 0.04 0.28 ND 0.14 0.19 ND ND 0.13 0.03 ND 0.01 ND 0.04 0.02 ND 0.02 0.03 ND 0.03 0.24 0.01 ND ND ND t 0.02 0.02 t t 0.29 0.01 ND 0.02 ND ND ND 0.04 ND t t 0.05 t t 0.05 ND t 0.01 t t ND ND ND 0.07 t 0.01 0.05 0.07 0.18 ND t 0.06 ND ND 0.71 ND ND t ND 0.09 t 0.02 0.11 0.14 ND 0.25 t 0.01 ND t ND ND t 0.04 t 6.21 1.44 0.01 t 0.11 t t ND

168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223

26.06 26.37 26.70 27.10 27.37 27.52 27.60 27.67 27.84 27.94 27.98 28.16 28.29 28.29 28.32 28.40 28.41 28.67 28.74 28.74 28.88 29.29 29.93 30.06 30.12 30.23 30.34 30.46 30.72 30.78 31.10 31.19 31.49 31.61 31.65 32.29 32.30 32.48 32.55 32.60 32.79 33.40 33.42 33.75 33.85 33.98 34.51 34.57 34.87 35.61 36.03 36.95 38.50 40.54 41.04 41.54

Dihydro-b-ionol Caryophyllene oxide Phenol Methyl eugenol Geraniol-6,7-epoxide acetate Eicosane (Z)-Cyclododec-9-enolide (yuzu lactone) (2) Geranial-6,7-oxide Eicos-1-ene 3,7-Dimethyl-6,7-epoxyoctanol 1) (E)-Nerolidol 3,7-Dimethyl-6,7-epoxyoctanol 2) 2,7-Germacradien-1-ol Octanoic acid Cubenol Germacra-2,7-dien-1-ol Cubenol Epi-cubenol Nerol-6,7-oxide Nerol-2,3-oxide Elemol Geraniol-2,3-oxide Caryophylla-2(12),6-dien-5-yl acetate Heneicosane Geraniol-6,7-oxide Spathulenol Heneicos-1-ene 1,3,5-Trimethoxybenzene (TMB) Mint sulde Eugenol t-Cadinol Nonanoic acid t-Muurolol Torreyol (2Z,6Z)-Farnesol a-Eudesmol b-Eudesmol Methyl hexadecanoate a-Cadinol (2Z,6E)-Farnesal Citronellic acid (E)-Dihydrofarnesol Ethyl hexadecanoate Neric acid Tricosane (2E,6E)-Farnesal (2Z,6E)-Farnesol Tricosane Geranic acid (2E,6E)-Farnesol Methyl cis-(Z)-jasmonate (3) Indole Dodecanoic acid Methyl linolenate Octadecanol Ethyl linolenate

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A. Joichi et al. Table 1. (Continued) No. RT (min) Compound Gas chromatography area (%) Blue Moon Blue Perfume Headspace Solvent extract Headspace Solvent extract ND ND ND ND ND ND ND ND 1.31 ND 0.41 0.31 0.04 0.03 0.11 0.02 ND ND 0.11 t 5.92 t 1.09 ND

224 225 226 227 228 229

44.31 44.55 54.41 55.93 70.98 79.77

Tetradecanoic acid Pentadecanoic acid Hexadecanoic acid Hexadec-9-enoic acid Octadecanoic acid Linoleic acid

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

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

Solvent Extraction
Flowers of both Blue Moon and Blue Perfume were extracted with pentane for 24 h just after picking. The extract was dried over anhydrous sodium sulfate, ltered, and then concentrated to give the concentrate under reduced pressure. A sample of 1.08 g of concentrate was obtained from 650 g of Blue Moon owers and 4.44 g of concentrate from 1610 g of Blue Perfume owers. After removing the higher fatty acids and ower waxes by using column chromatography utilizing a glass column packed with silica gel 60 (70230 mesh), 267 mg of volatile concentrate was obtained from Blue Moon, and 1.57 g of volatile concentrate was obtained from Blue Perfume.

Gas Chromatography
GC analyses were carried out on a Agilent HP6890 gas chromatograph (Agilent Technologies, Santa Clara CA) equipped with a ame ionization detector and a 50 m 0.25 mm 0.15 mm BC-WAX (produced by GL Sciences, Tokyo, Japan) coated capillary column. The oven temperature was programmed linearly from 70 C to 215 C at a rate of 4 C/min.

on the second gas chromatograph with b-DEX 225, b-DEX 325 (25 m 0.25 mm 0.25 mm; Agilent Technologies), or b-DEX sm (Restek, Bellefonte, PA, USA) combined with a MS detector. Chiral GC analyses were performed using a MDGC/GCMS-2010 (Shimadzu). With this system, a heart-cut of the relevant fractions could be made as well as a transfer from the achiral column to the chiral one. The injector and the ame ionization detector temperatures were both 250 C. The carrier gas was helium with a constant pressure set at 180 kPa. The oven temperature program was set from 70 C to 230 C at 5 C/min. For citronellol analysis, the second gas chromatograph was equipped with a b-DEX 225 column (30 m 0.25 mm i.d., lm thickness of 0.25 mm; Supelco, Bellefonte, PA, USA) and the oven temperature program was set from 70 C to 180 C at 1 C/min. For citronellyl acetate, the second gas chromatograph was equipped with a b-DEX se column (30 m 0.25 mm i.d., lm 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 second gas chromatograph was equipped with a b-DEX 325 column (30 m 0.25 mm i.d., lm thickness of 0.25 mm; Supelco) and the oven temperature program was set from 70 C to 180 C at 1 C/min. For trans-rose oxide, the second gas chromatograph was equipped with a Chiralsil DEX CB column (30 m 0.25 mm i.d., lm thickness of 0.25 mm; Agilent) and the oven temperature program was set from 70 C to 180 C at 1 C/min.

Gas ChromatographyMass Spectrometry


GC-MS analyses were carried out using a GCMS-QP2010 (Shimadzu Co., Kyoto, Japan) with an ionization energy of 27 eV and equipped with BC-WAX column (50 m 0.25 mm i.d., lm thickness of 0.15 mm; GL Sciences). The injector and the ion source were set at 250 C and 200 C, respectively. The spectrometer was used in scan mode. The carrier gas was helium with a constant pressure set at 100 kPa. The oven temperature program was set from 70 C to 218 C at 4 C/min. Identication of the compounds was performed by comparing and matching the retention times and mass spectra with original sets of standard data.

Results and Discussion


Major Volatile Components in Blue Moon and Blue Perfume The volatile components of two blue-coloured HT roses are listed in Table 1. In the headspace analyses, 156 components were identied in the scent of Blue Moon and 136 in the scent of Blue Perfume. Then, in the solvent extracts of the two rose owers, 201 components were identied in Blue Moon and 170 components were identied in Blue Perfume. Monoterpene alcohols and their aldehydes, such as geraniol, nerol, citronellol and geranial, were identied as the major components (Figure 1). These monoterpene derivatives greatly contributed to the rich rose and the fresh lemon-like note. 1,3-Dimethoxy-5-methylbenzene and dihydro-b-ionol were identied as the major components in the scent of Blue Moon ower. 1,3-Dimethoxy-5-methylbenzene, with a phenolicspicy note, and dihydro-b-ionol, with a woody note, were identied during the evaluation of this rose ower.

Multi-dimensional Gas Chromatography and Mass Spectrometry


Multi-dimensional gas chromatography (MDGC) analyses were carried out on a Shimadzu QP2010 equipped on the rst gas chromatograph with a BC-WAX column (50 m 0.25 mm 0.25 mm; GL Sciences), and

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Volatile constituents of blue-coloured hybrid tea rose ower


7

Blue Moon

solvent
4 5

BHT

6 9 8

10

20
7

30

40

Blue Perfume

12 3
0 10 20

BHT

30

40

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

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 were quite low in the scent of Blue Perfume.

Minor Key Volatile Components The high contribution components for the odour of blue-type rose ower were studied by using gas chromatographyolfactometry and a new component was detected which has a faint pungent, vegetable and tropical fruit-like character. This component was determined as 2-isopropyl-4-methylthiazole (1) using GC and GC-MS. The GC retention time and the mass spectrum of compound 1 was the same as that of an authentic sample purchased from Junsei Kagaku Co., Ltd (Tokyo, Japan). In fact, compound 1, which is found as one of the volatile components in tomato[8] and durian,[9] was identied in rose ower for the rst time. It was conrmed that the addition of a small amount of compound 1 to duplicated rose-type oil and rose-type fragrance (Blue Moon type) showed good performance for the original rose fragrance (data not shown). (Z)-Cyclododec-9-enolide [(Z)-yuzu lactone] (2), which was identied in the solvent extracts of Blue Moon and Blue Perfume, was identied in the rose owers for the rst time. Compound 2, which was a macrocyclic lactone with a camphorous note, was found in yuzu (popular Japanese citrus),[10] Japanese mugwort,[11] golden orange (another Japanese citrus),[12] and lily of the valley.[13] Then, methyl cis-(Z)-jasmonate (3), which is found in orchid ower[14] and lemon peel oil,[15] was also identied in Blue Moon and Blue Perfume oils collected by the headspace and solvent extraction methods. This is the rst time that compound 3 has been found in rose ower. The addition of a small amount of compound 3 to a rose-type fragrance greatly enhanced the natural odour of blue-coloured HT rose ower. This performance was similar to that of compound 1.

equipped with a chiral column as the secondary column. The following results were found for both rose owers. The ratio of the (S)-enantiomer vs. the (R)-enantiomer of linalool was 8:92, although the existing ratio of linalool in rose oil was reported to be almost 50:50.[16] At this time, we do not have 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 citronellyl acetate, only the (S)-enantiomer was detected. For trans-rose oxide, only the (2R,4R)-enantiomer was detected. The results of the enantiomeric ratio of citronellol, citronellyl acetate, Table 2. The enantiomeric ratio of the chiral components Component and enantiomer Citronellol 3R 3S Citronellyl acetate 3R 3S Linalool 3R 3S Dihydro-b-ionol R S (E)-Nerolidol 3R 3S cis-Rose oxide 2S, 4R 2R, 4S trans-Rose oxide 2R, 4R 2S, 4S Theaspirane A 2R, 5R 2S, 5S Theaspirane B 2S, 5R 2R, 5S Amount (%) >99 >99 92 8 >99 >99 99 1 >99 4 96 97 3

Analysis of the Enantiomers of Blue-type Hybrid Tea Roses Several components identied in the two types of rose ower had asymmetric carbon atoms in their molecules, leading us to analyse their chirality. For the rst time, the enantiomeric ratio of linalool, (E)-nerolidol, theaspiranes, dihydro-b-ionol, citronellol and citronellyl acetate could be assigned by MDGC-MS

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A. Joichi et al. cis-rose oxide, and trans-rose oxide were suggested for proceeding 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)-enantiomer vs. the (2S)-enantiomer in theaspirane A and theaspirane B was about 4:96.
3. I. Watanabe, T. Yanai, S. Tamogami. In Proceedings of the 7th International Congress 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, and Aromatics, 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.

Conclusion
The characteristic volatile components of rose ower (blue type) were revealed by a combination of GC-MS and gas chromatographyolfactometry analyses. The enantiomeric distribution of the important odour-active components was determined using MDGC-MS. These analytical results would give very important information for creating fragrances close to the natural aromas.

References
1. G. Ohloff, Perfumer Flavorist 1978, 3, 11. 2. C. Mann, T. Smith. In Proceedings of the 7th International Congress of Essential Oils, Kyoto, Japan, 1977, p. 458 (abstract).

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Flavour Fragr. J. 2013