FLAVOUR AND FRAGRANCE JOURNAL, VOL. 12, 423±432 (1997)

Volatile Leaf Oils of some South-western and Southern Australian Species of the Genus Eucalyptus (Series I). Part XVIII. A. Subgenus Monocalyptus. B. Subgenus Symphyomyrtus: (i) Section Guilfoyleanae; (ii) Section Bisectaria, Series Accedentes, Series Occidentales, Series Levispermae, Series Loxophlebae, Series Macrocarpae, Series Orbifoliae, Series Calycogonae; (iii) Section Dumaria, Series Incrassatae and Series Ovulares

C. M. Bignell,1 P. J. Dunlop1Ã and J. J. Brophy2 1Department of Chemistry, University of Adelaide, South Australia 5005, Australia 2Department of Organic Chemistry, University of New South Wales, Sydney, New South Wales 2052, Australia

The volatile leaf oils of Eucalyptus megacarpa F. Muell., E. calcicola Brooker, E. preissiana Schauer subsp. lobata Brooker & Slee, E. guilfoylei Maiden, E. accedens W. Fitzg., E. trivalvis Blakely, E. zopherophloia Brooker & Hopper, E. pruiniramis L. A. S. Johnson & K. D. Hill, E. redacta Brooker & Hopper ined., E. subangusta (Blakely) subsp. subangusta, E. subangusta (Blakely) Brooker & Hopper subsp. pusilla Brooker & Hopper, E. subangusta (Blakely) Brooker & Hopper subsp. cerina Brooker & Hopper, E. phaenophylla Brooker & Hopper subsp. interjacens Brooker & Hopper, E. loxophleba Benth. subsp. loxophleba, E. loxophleba Benth. subsp. lissophloia L. A. S. Johnson & K. D. Hill, E. loxophleba Benth. subsp. gratiae Brooker, E. semota C. J. Macph. & P. M. Grayling, E. glomerosa Brooker & Hopper, E. kingsmillii (Maiden) Maiden & Blakely subsp. alatissima Brooker & Hopper, E. leptopoda Benth. subsp. elevata K. D. Hill & L. A. S. Johnson, E. crucis Maiden subsp. lanceolata Brooker & Hopper, E. brevipes Brooker, E. captiosa Brooker & Hopper, E. brachycorys Blakely, isolated by vacuum distillation, were analysed by GC and by GC±MS. Many species contained a-pinene (0±27.4%), limonene (0±12.0%), a-phellandrene (0±16.1%), 1,8-cineole (1.9±78.2%), p-cymene (0.1±15.0%), b-caryophyllene (0±10.5%), aromadendrene (0±31.5%), bicyclogermacrene (0±42.3%) and torquatone (trace±6.7%) as principal leaf oil components. # 1997 John Wiley & Sons, Ltd.

Flavour Fragr. J. 12, 423±432 (1997) (No. of Figures: 0 No. of Tables: 2 No. of Refs: 16)

KEY WORDS: Eucalyptus megacarpa F. Muell.; Eucalyptus calcicola Brooker; Eucalyptus preissiana Schauer subsp. lobata Brooker & Slee; Eucalyptus guilfoylei Maiden; Eucalyptus accedens W. Fitzg.; Eucalyptus trivalvis Blakely; Eucalyptus zopherophloia Brooker & Hopper; Eucalyptus pruiniramis L. A. S. Johnson & K. D. Hill; Eucalyptus redacta Brooker & Hopper ined.; Eucalyptus subangusta (Blakely) subsp. subangusta; Eucalyptus subangusta (Blakely) Brooker & Hopper subsp. pusilla Brooker & Hopper; Eucalyptus subangusta (Blakely) Brooker & Hopper subsp. cerina Brooker & Hopper; Eucalyptus phaenophylla Brooker & Hopper subsp. interjacens Brooker & Hopper; Benth. subsp. loxophleba; Eucalyptus loxophleba Benth. subsp. lissophloia L. A. S. Johnson & K. D. Hill; Eucalyptus loxophleba Benth. subsp. gratiae Brooker; Eucalyptus semota C. J. Macph. & P. M. Grayling; Eucalyptus glomerosa Brooker & Hopper; Eucalyptus kingsmillii (Maiden) Maiden & Blakely subsp. alatissima Brooker & Hopper; Eucalyptus leptopoda Benth. subsp. elevata K. D. Hill & L. A. S. Johnson; Eucalyptus crucis Maiden subsp. lanceolata Brooker & Hopper; Eucalyptus brevipes Brooker; Eucalyptus captiosa Brooker & Hopper; Eucalyptus brachycorys Blakely; ; leaf essential oil composition; torqua- tone; mono- and sesquiterpenoids; GC±MS

à Correspondence to: P. J. Dunlop.

CCC 0882±5734/97/060423±10$17.50 Received 3 December 1996 # 1997 John Wiley & Sons, Ltd. Accepted 3 February 1997 424 C. M. BIGNELL ET AL.

INTRODUCTION recorded by the integrator in 31 minutes. GC±MS was performed on a VG Quattro mass spectro- Continuing our investigation of indigenous meter operating at 70 eV ionization energy. Australian eucalypts,1 we have examined the leaf The GC column in this case was a DB-Wax oils of three species from Subgenus Monocalyptus, (60 m 0:32 mm). Compounds were identi®ed by one from Section Guilfoyleanae, four from Series their GC retention indices to known compounds Accedentes, one from Series Occidentales, four and by comparison of their mass spectra, either from Series Levispermae, four from Series Loxo- with known compounds or with published phlebae, three from Series Macrocarpae, one from spectra.8±10 Series the Orbifoliae, one from Series Calycogo- Only one of the species (E. brachycorys) was nae, one from Series Incrassatae and one from analysed with GC±MS. The oil components of the Series Ovulares. In this study we have followed the rest were identi®ed using normalized retention system of M. I. H. Brooker and D. A. Kleinig.2 times. For this purpose the column was calibrated Locations of all species, which are native to south- by assuming times for three markers, 1,8-cineole, western and southern Australia, are listed in octadecane (OD added to the ether) and torqua- Table 1. A more detailed description of the occur- tone. The raw retention times were ®rst normalized rence of some of these eucalypts has been given to 525 s for OD, and times before and after OD elsewhere.2 adjusted by assuming linearity and using 99s for To our knowledge, accounts of the leaf oils cineole and 997s for torquatone. Torquatone was of only E. megacarpa,3 E. accedens,3 E. loxophleba found to be present in almost all oil samples; when subsp. loxophleba,4;5 E. loxophleba subsp. lisso- this was not the case a sucient amount was added phloia,5 E. gratiae5 and E. semota5 have been to the oil solution to obtain that reference point. published previously. The normalized retention times of the column were identi®ed with oil components analysed previously by GC±MS for over 100 Eucalyptus species: some EXPERIMENTAL of these results have been published.1 In addition, Kova t's indices were obtained (see Table 2) by Samples of clean, mature leaves were picked from running a mixture of C10±C25 alkanes on the SGE- single trees and, after freezing with liquid nitrogen, BP20/GC column. were reduced to a ®ne powder using a stainless steel All GC analyses were performed in duplicate Waring Blender (Model no. SS110). This procedure and the retention times and percentage composi- was necessary to rupture the oil glands. The dry tions of each component averaged. Duplicate powder was then vacuum-distilled so that the leaf times were discarded if they di€ered from each oil condensed on to a gold-plated copper rod other by more than 1 second. Components which maintained at approximately 758C. Complete contributed less than 0.06% to the ®nal analyses details of this procedure have been published were not considered (an arbitrary but practical previously.6;7 All oils obtained were colourless to decision). pale yellow and lighter than water. Table 1 lists the oil yields (wt%, dry weight) for the 24 species studied. RESULTS AND DISCUSSION The GC analyses were performed with an HP5890 Series II unit operated in conjunction with Freshly isolated oils obtained by vacuum distil- an HP3396 Series II integrator. The `on-column' lation of powdered leaves from single trees were injection technique was used with an SGE BP20 analysed by GC and by GC±MS. The results capillary column of (25 m  0:33 mm i.d., and ®lm for the 24 species are listed in Table 2; only thickness 0:5 mm). The carrier gas was hydrogen those components with concentrations greater with an inlet pressure of 25 kPa: the ¯ow rate was than 0.05% are reported. The principal compo- 2:0 cm3=min. The oven was programmed to rise nents in the oils were the monoterpenes a-pinene from 808C to 2208C at 58=min, and the inlet (0±27.4%), b-pinene (0±7.2%), 1,8-cineole (1.9± temperature set to 838C and increased at the same 78.2%) and p-cymene (0.1±15.0%). Apart from rate as the column. Using these conditions and a 1,8-cineole, the main oxygenated monoterpenes 1:0 ml sample of 0.4% solution of oil in puri®ed dry detected were pinocarvone (0±5.2%), trans-pino- diethyl ether, essentially all the components were carveol (0±16.6%) and a-terpineol (0±5.8%).

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) VOLATILE LEAF OILS OF EUCALYPTUS SPECIES 425

Table 1. Oil yields from the Eucalyptus speciesa

Species and locality Oil yield (wt%, dry weight) Subgenus Monocalyptus E. megacarpa F. Muell.c tr. Dunsborough, E. calcicola Brooker 0.33 Currency Creek Arboretum, South Australia E. preissiana Schauer subsp. lobata Brooker & Slee 0.31 Currency Creek Arboretum, South Australia Subgenus Guilfoyleanae E. guilfoylei Maidenb tr. (S35858'48@/E116853'12@) Series Accedentes E. accedens W. Fitzg. 0.90 near Toodyay, Western Australia E. trivalis Blakely 0.96 Waite Arboretum, South Australia E. zopherophloia Brooker & Hopper 1.72 Currency Creek Arboretum, South Australia E. pruiniramis L. A. S. Johnson & K. D. Hill 3.07 Currency Creek Arboretum, South Australia Series Occidentales E. redacta Brooker & Hopper ined.b 0.45 (S34827'12@/E118852'33@) Series Levispermae E. subangusta (Blakely) subsp. subangusta 3.32 Currency Creek Arboretum, South Australia E. subangusta (Blakely) Brooker & Hopper subsp. pusilla Brooker & Hopper 1.26 Currency Creek Arboretum, South Australia E. subangusta (Blakely) Brooker & Hopper subsp. cerina Brooker & Hopper 3.12 Currency Creek Arboretum, South Australia E. phaenophylla Brooker & Hopper subsp. interjacens Brooker & Hopper 0.38 Currency Creek Arboretum, South Australia Series Loxophlebae E. loxophleba Benth. subsp. loxophleba 1.56 Waite Arboretum, South Australia E. loxophleba Benth. subsp. lissophloia L. A. S. Johnson & K. D. Hill 1.88 Kalgoorlie, Western Australia E. loxophleba Benth. subsp. gratiae Brooker 3.03 Currency Creek Arboretum, South Australia E. semota C. J. MacPh. & P. M. Graylingb 0.79 (S25837'12@/E119823'15@) Series Macrocarpae E. glomerosa Brooker & Hopperb 1.56 (S28852'56@/E130827'43@) E. leptopoda Benth. subsp. elevata K. D. Hill & L. A. S. Johnsonb 2.07 (S28829'20@/E128811'03@) E. kingsmillii (Maiden) Maiden & Blakely subsp. alatissima Brooker & Hopperb 0.66 (S28818'45@/E127814'15@) Orbifolia Group E. crucis Maiden subsp. lanceolata Brooker & Hopper 3.75 Currency Creek Arboretum, South Australia Series Calycogonae E. brevipes Brooker 0.69 Currency Creek Arboretum, South Australia Series Incrassatae E. captiosa Brooker & Hopperb 0.37 (S33846'13@/E118846'35@) Series Ovulares E. brachycorys Blakely 0.75 (S30806'49@/E115859'30@) a The specimens of these species were authenticated by Mr M. I. H. Brooker, Australian National Herbarium, or Dean Nicolle, Currency Creek Arboretum. b Botanical voucher specimens have been deposited at the South Australian Herbarium by Dean Nicolle, who collected the leaves of these species. c The leaves of E. megacarpa were supplied by Mr Don Grace of Dunsborough, Western Australia.

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) 426 C. M. BIGNELL ET AL.

Table 2. Compounds identi®ed and their percentage occurrence (40:05%) in the leaf oils of the Eucalyptus speciesa Slee & er Hill ta D. ook Br K. ta & subangus ined. loba Hopper & subsp. Johnson er Hopper subsp. S. ely) & ook er A. Br Muell. er ely Fitzg. . L. ook (Blak . F Schauer ook Maiden Br W ta Blak Br amis ophloia carpa cta eissiana ccedens eda trivalvis zopher pruinir r subangus mega calcicola guilfoylei a pr à E. E. E. E. E. KI E. E. E. E. E. 1 a-Pinene 1038 0.8 13.8 2.3 ± 3.2 10.1 9.0 23.1 4.8 27.4 2 a-Fenchene 1084 ± ± ± ± ± 0.08 ± ± ± ± 3 Camphene 1085 ± ± ± ± ± 0.06 ± ± ± ± 4 4-Methylpent-2-yl acetate 1109 ± ± 5 b-Pinene 1128 0.06 0.7 0.3 ± 0.06 0.2 0.2 0.4 0.07 0.8 6 Sabinene 1149 ± ± ± ± ± ± ± ± ± ± 7 4-Methylpentan-2-ol 1168 ± ± 8 Myrcene 1175 ± 0.4 0.5 ± ± 0.2 0.3 0.4 ± 0.3 9 a-Phellandrene 1186 0.1 11.0 0.1 ± ± 0.4 ± 0.2 0.1 0.3 10 Isobutyl isovalerate 1199 ± 0.1 0.1 ± ± ± ± ± ± ± 11 a-Terpenene 1205 0.07 0.2 ± ± ± ± ± ± ± ± 12 Limonene 1219 7.4 12.0 2.7 0.08 0.09 2.1 2.7 2.4 0.4 2.2 13 1,8-Cineole 1231 11.4 31.3 38.4 1.9 53.3 51.6 71.5 35.0 42.7 35.4 14 b-cis-Ocimene 1248 ± 0.4 ± ± ± 0.3 0.3 ± ± ± 15 g-Terpinene 1266 0.2 0.6 0.3 ± 0.06 0.2 0.4 0.1 ± 0.2 16 b-trans-Ocimene 1271 ± ± ± ± ± ± ± ± ± ± 17 p-Cymene 1292 2.2 6.9 0.08 0.1 0.8 1.3 0.2 0.2 0.8 0.9 18 Terpinolene 1300 10.2 0.7 0.06 0.06 ± 0.1 0.09 ± ± 0.1 19 Isoamyl isovalerate 1304 ± ± ± ± ± ± ± 0.2 ± 0.1 20 a-p-Dimethyl styrene 1457 0.6 ± ± ± ± ± ± ± ± ± 21 d-Elemene 1470 ± ± ± 0.3 ± ± ± ± ± ± 22 a-Cubebene 1480 0.1 ± 0.8 ± ± 0.2 ± 0.3 ± ± 23 cis-Linalol oxide 1485 0.2 ± ± 0.06 ± ± ± ± ± ± 24 Bicycloelemene 1495 0.1 ± 0.6 ± ± ± ± ± ± ± 25 a-Copaene 1505 0.1 ± 0.3 ± 0.1 ± ± 0.1 0.1 0.4 26 a-Campholenic aldehyde 1511 ± 0.06 ± ± 0.06 0.2 ± ± ± 0.08 27 a-Gurjunene 1545 0.2 0.08 2.1 ± ± 0.4 ± 2.1 0.2 0.2 28 Linalol 1564 3.5 0.2 ± ± ± ± ± ± ± ± 29 trans-p-Menth-2-en-1-ol 1578 0.3 0.1 ± ± ± ± ± ± ± 0.09 30 Camphor 1580 ± ± ± ± 0.06 ± ± 0.08 0.07 ± 31 Pinocarvone 1590 ± ± ± 0.1 3.6 0.5 ± 0.1 3.4 0.5 32 Fenchol 1598 ± ± ± 0.9 0.3 0.3 ± ± 0.2 ± 33 b-Elemene 1602 0.08 ± ± ± ± ± 0.06 ± ± 0.08 34 b-Gurjunene 1606 ± ± 0.3 ± 0.2 0.2 ± 0.7 0.2 ± 35 b-Caryophyllene 1611 0.6 1.0 0.2 0.09 0.7 0.3 0.06 ± 1.5 1.4 36 Terpinen-4-ol 1617 ± 1.3 2.7 ± ± ± ± ± ± 1.6

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) VOLATILE LEAF OILS OF EUCALYPTUS SPECIES 427 Hopper Hopper Hopper & & Hill & Hopper er er er & D. ook er ook ook K. Br Br Br Johnson & ook S. Br A. tissima pusilla cerina cens L. ala Johnson Hopper & S. & subsp. subsp. interja A. er er Hill subsp. L. ook D. ely ook yling subsp. a Br Hopper Hopper Br K. Blak Gr & & ta ta tiae & a er er M. gr loxophleba lissophloia . Hopper eleva P ook ook & Hopper lanceola & Br Br Hopper Maiden er & subsp. subsp. subsp. & subsp. ely er ely) ely) cph. ook er subsp. er Br Ma ook Blak (Blak (Blak Benth. Benth. Benth. ook ook s J. Br (Maiden) Benth. Br ta ta Br C. osa Maiden chycory a evipes subangus br br subangus semota glomer captiosa crucis loxophleba phaenophylla loxophleba loxophleba leptopoda kingsmillii E. E. E. E. E. E. E. E. E. E. E. E. E. E. 18.1 9.3 8.1 4.4 1.5 9.8 0.2 2.3 0.4 1.27 8.1 2.7 7.7 3.2 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0.1 ± ± ± 0.2 ± ± ± 0.09 ± ± 14.0 4.4 1.5 ± ± ± ± ± ± ± ± 0.6 0.4 0.4 0.1 ± 0.06 0.08 0.07 ± ± 0.3 0.6 0.06 7.2 ± 0.2 ± ± ± ± ± ± ± ± ± ± ± 0.2 ± 0.6 ± 0.1 ± ± ± ± ± ± ± ± 0.2 0.2 0.3 0.1 ± ± ± ± ± ± ± 0.6 ± ± 0.4 16.1 0.8 0.06 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 3.1 0.6 0.6 0.9 ± 2.4 ± ± ± ± 1.0 2.5 0.6 0.4 56.6 6.2 1.8 25.2 63.0 72.4 61.1 61.6 55.3 76.8 61.6 78.2 37.9 25.0 ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0.1 0.09 0.07 0.08 ± 0.1 ± ± ± ± ± 0.2 ± ± ± ± ± ± ± 0.07 ± ± ± ± ± ± ± ± 0.6 2.8 15.0 0.2 1.9 1.0 3.0 1.6 1.0 1.1 1.0 0.3 0.3 9.4 ± 0.09 ± 0.5 0.6 ± ± 0.06 0.4 ± 0.1 ± ± ± ± ± ± ± ± 0.6 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0.06 ± 0.6 0.08 ± ± ± ± ± ± ± ± 0.06 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0.2 ± ± ± ± ± ± ± ± ± ± 0.3 ± ± 0.2 ± ± ± 0.1 ± 0.08 0.2 ± 0.1 0.4 0.1 ± 0.1 ± ± ± ± ± ± ± ± ± ± ± ± 0.3 0.2 1.8 ± ± ± ± ± ± ± 0.06 ± ± ± ± ± ± ± ± ± ± ± ± ± 0.5 0.1 0.3 ± ± 0.06 ± ± ± ± ± ± ± ± ± ± 0.1 ± ± ± 0.3 ± ± ± ± ± ± ± ± ± ± 0.4 ± 0.09 0.3 3.1 0.4 0.3 5.1 5.2 3.3 1.6 ± 3.3 2.5 ± ± ± ± 0.5 ± 0.8 0.1 0.7 0.1 ± ± ± 1.5 ± ± ± ± ± 0.1 ± ± 0.06 ± ± ± 0.2 ± ± ± ± 1.1 ± ± ± 0.06 ± ± ± 0.1 ± ± 0.6 10.5 6.4 0.1 0.07 ± ± ± ± ± ± ± 0.4 0.2 0.5 ± 1.0 ± ± 0.9 0.2 ± ± 0.4 ± 1.0 ± 0.7 Table continues on page 428

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) 428 C. M. BIGNELL ET AL.

Table 2. Continued Slee & er Hill ta D. ook Br K. ta & subangus ined. loba Hopper & subsp. Johnson er Hopper subsp. S. ely) & ook er A. Br Muell. er ely Fitzg. . L. ook (Blak . F Schauer ook Maiden Br W ta Blak Br amis ophloia carpa cta eissiana ccedens eda trivalvis zopher pruinir r subangus mega calcicola guilfoylei a pr à E. E. E. E. E. KI E. E. E. E. E. 37 Aromadendrene 1620 2.6 ± 5.3 0.4 7.3 5.3 2.5 14.3 6.0 ± 38 a-Bulnesene 1629 0.2 ± 0.7 ± 0.2 0.2 0.08 0.5 0.3 0.06 39 cis-p-Menth-2-en-1-ol 1645 0.3 0.2 0.06 ± ± ± ± ± ± 0.1 40 Myrtenal 1647 ± ± ± 0.9 0.06 ± ± ± ± ± 41 allo-Aromadendrene 1659 0.7 0.2 2.9 0.2 1.4 1.6 0.6 2.2 1.6 0.3 42 trans-Pinocarveol 1671 ± ± ± 0.4 15.8 2.5 0.5 0.4 10.5 0.6 43 Humulene 1684 ± ± ± 0.1 ± 0.06 ± ± 0.4 0.09 44 d-Terpineol 1687 0.1 0.1 0.3 ± 0.3 ± 0.2 0.1 ± ± 45 Cryptone 1690 0.1 ± ± ± ± ± ± ± ± 0.09 46 cis-Piperitol 1692 ± ± ± ± ± ± ± ± ± ± 47 Neral 1702 3.2 ± ± ± ± ± 0.1 0.5 ± 0.6 48 Viridi¯orene 1704 0.2 0.06 9.9 29.6 0.07 0.8 0.1 0.5 0.7 0.2 49 a-Terpineol 1711 5.8 4.3 ± 0.9 0.2 0.8 1.2 0.9 0.3 0.5 50 Borneol 1717 ± ± 0.2 1.4 0.3 0.4 ± ± 0.2 ± 51 Verbenone 1725 ± ± 0.09 ± ± ± ± ± ± ± 52 b-Selinene 1731 0.2 ± 0.5 ± 0.4 0.1 0.1 0.4 0.2 0.3 53 a-Selinene 1735 0.6 ± 0.5 ± ± ± ± 0.3 0.07 0.2 54 A Muurolene 1742 ± ± ± 0.1 0.6 0.2 0.06 0.07 0.3 ± 55 Piperitone 1744 1.0 0.4 1.1 0.9 0.3 ± 2.3 0.4 ± ± 56 Bicyclogermacrene 1747 ± ± ± ± ± 3.0 ± ± 6.2 8.3 57 Carvone 1750 ± ± 0.08 ± 0.1 ± ± ± 0.1 ± 58 trans-Piperitol 1761 0.7 0.2 ± 0.4 ± 0.1 0.2 0.06 ± 0.1 59 d-Cadinene 1774 0.3 ± 0.8 0.1 0.2 0.6 0.1 0.5 0.2 0.2 60 g-Cadinene 1779 0.2 ± 0.1 0.07 ± ± ± ± 0.09 0.06 61 Myrtenol 1807 ± ± ± 5.8 0.3 0.06 ± 0.2 0.2 0.08 62 Cadina-1,4-diene 1810 ± ± ± ± ± ± 0.07 ± ± ± 63 trans-p-Mentha-1(7),8-dien-2-ol 1811 ± ± ± ± 0.8 0.4 0.1 ± 0.7 0.1 64 Nerol ± ± ± ± ± ± ± ± ± ± 65 Calamenene 1847 ± ± ± 0.3 0.09 0.5 ± ± ± ± 66 trans-p-Mentha-1,8-dien-6-ol 1850 0.2 0.06 ± ± 0.3 0.1 ± ± 0.3 0.09 67 Geraniol 1859 0.4 1.3 0.2 ± ± ± 0.1 0.08 ± ± 68 p-Cymen-8-ol 1865 5.4 ± 0.07 0.1 0.1 0.06 0.09 0.3 0.09 0.2 69 cis-p-Mentha-1,8-dien-6-ol 1881 ± ± ± 0.2 ± ± ± ± ± ± 70 cis-p-Mentha-1(7),8-dien-2-ol 1903 0.2 ± ± 0.1 0.8 0.4 ± ± 0.7 ± 71 Calacorene 1932 ± 0.07 ± ± ± 0.07 ± 0.08 ± 0.1 72 Palustrol 1938 ± 0.5 0.4 ± 0.09 0.2 0.07 0.1 0.2 ±

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) VOLATILE LEAF OILS OF EUCALYPTUS SPECIES 429 Hopper Hopper Hopper & & Hill & Hopper er er er & D. ook er ook ook K. Br Br Br Johnson & ook S. Br A. tissima pusilla cerina cens L. ala Johnson Hopper & S. & subsp. subsp. interja A. er er Hill subsp. L. ook D. ely ook yling subsp. a Br Hopper Hopper Br K. Blak Gr & & ta ta tiae & a er er M. gr loxophleba lissophloia . Hopper eleva P ook ook & Hopper lanceola & Br Br Hopper Maiden er & subsp. subsp. subsp. & subsp. ely er ely) ely) cph. ook er subsp. er Br Ma ook Blak (Blak (Blak Benth. Benth. Benth. ook ook s J. Br (Maiden) Benth. Br ta ta Br C. osa Maiden chycory a evipes subangus br br subangus semota glomer captiosa crucis loxophleba phaenophylla loxophleba loxophleba leptopoda kingsmillii E. E. E. E. E. E. E. E. E. E. E. E. E. E. ± 1.3 ± 31.5 0.7 ± ± 2.6 4.9 ± 1.9 ± 4.2 0.1 ± ± ± 1.0 ± ± ± 0.08 0.1 ± 0.08 ± ± ± 0.08 ± ± 0.07 ± ± 0.8 ± ± ± 0.1 0.07 ± 0.1 ± ± ± ± 0.07 ± 0.08 0.3 0.1 0.4 0.09 ± ± 3.2 0.2 1.3 1.6 3.8 0.3 ± 0.1 0.6 0.5 ± 0.6 0.2 0.8 0.2 0.3 ± ± 1.1 10.7 1.5 ± 10.5 12.5 6.0 1.5 ± 16.6 5.5 0.2 1.0 ± 0.2 ± ± 0.1 0.2 0.4 ± ± 0.3 0.2 ± ± ± ± ± 0.3 0.2 0.4 ± ± 0.4 0.5 ± ± ± 0.09 ± 0.4 ± 0.2 ± ± 0.1 ± 0.2 ± ± ± 7.1 ± ± ± ± ± ± ± ± ± ± 0.1 ± ± ± ± ± 0.2 0.7 ± ± 0.07 ± ± ± ± ± 0.1 ± ± 1.0 0.1 0.4 ± 0.06 0.1 ± ± ± ± ± 0.2 ± 1.8 ± ± ± 0.06 2.7 0.1 ± ± ± 2.3 3.5 1.5 0.3 ± ± 0.1 ± 0.5 ± ± 0.1 0.7 0.1 ± ± 0.9 0.2 ± ± 0.06 0.08 ± ± 0.9 0.3 0.06 0.4 0.09 ± ± 1.9 0.2 ± 0.08 0.7 ± 0.07 ± 0.1 0.2 ± 0.2 ± 0.5 ± 0.1 ± 0.1 0.4 ± ± 0.07 ± ± ± ± ± 0.4 ± 0.2 ± ± 0.1 0.5 0.2 ± 0.4 0.3 0.1 ± ± ± 0.2 ± ± ± 1.1 0.09 0.3 2.6 0.2 0.08 0.06 0.3 0.3 0.5 0.2 1.9 42.3 7.2 ± ± ± ± ± ± ± ± ± ± ± 0.06 ± ± ± 0.1 0.06 0.3 0.3 0.2 0.2 0.08 ± 0.1 0.5 ± ± 0.2 ± ± 0.2 ± ± ± ± ± 0.1 ± 0.1 0.1 0.1 ± 0.4 ± 0.07 0.09 ± ± ± 0.1 0.06 0.1 0.2 ± 0.4 ± ± ± ± 0.06 ± ± ± ± ± ± ± ± ± ± ± 0.2 ± ± ± 0.3 0.4 0.1 ± ± 2.5 ± ± ± 0.3 ± ± 0.1 ± ± ± ± ± ± ± 0.1 ± ± ± 0.8 0.4 ± 0.6 ± 0.5 0.2 0.07 0.9 0.4 ± ± ± ± ± ± 0.07 ± ± ± ± ± ± ± ± 0.1 ± ± ± 0.06 ± ± ± ± ± 0.5 ± 0.1 ± ± ± 0.1 0.1 0.06 0.3 0.1 0.2 0.3 ± ± 0.7 ± ± 0.2 ± ± ± ± ± ± ± ± 0.8 ± ± ± ± ± 0.2 0.09 ± 0.7 0.4 0.1 0.4 0.1 ± 0.2 0.9 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0.09 0.9 0.08 ± 0.06 0.09 0.3 0.2 ± 1.0 ± ± ± 0.2 ± 0.06 ± ± ± ± ± ± ± ± ± ± ± ± 0.1 ± ± ± 0.1 ± 0.07 ± ± ± 0.08 Table continues on page 430

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) 430 C. M. BIGNELL ET AL.

Table 2. Continued Slee & er Hill ta D. ook Br K. ta & subangus ined. loba Hopper & subsp. Johnson er Hopper subsp. S. ely) & ook er A. Br Muell. er ely Fitzg. . L. ook (Blak . F Schauer ook Maiden Br W ta Blak Br amis ophloia carpa cta eissiana ccedens eda trivalvis zopher pruinir r subangus mega calcicola guilfoylei a pr à E. E. E. E. E. KI E. E. E. E. E. 73 Caryophyllene oxide 1994 2.7 0.9 ± 0.4 0.3 ± ± ± 0.2 0.1 74 b-Phenylethyl propionate 2001 0.3 ± ± 0.4 ± ± 0.06 ± ± ± 75 C15H26O 2008 0.2 0.06 0.2 0.1 0.1 0.1 ± 0.2 0.2 ± 76 C15H26O 2011 ± ± ± 5.0 ± ± ± ± ± ± 77 C15H26O 2023 0.3 ± 1.0 0.4 0.4 0.6 0.2 1.0 0.5 0.1 78 C15H26O 2040 0.2 ± 0.4 3.2 0.08 0.2 0.07 0.2 0.2 ± 79 C15H26O 2050 2.1 0.1 ± 1.2 ± ± ± ± ± ± 80 C15H26O 2070 0.5 0.07 1.4 0.3 0.1 0.3 0.08 0.2 0.5 ± 81 C15H26O 2076 0.2 ± 0.06 0.2 ± ± ± 0.09 ± ± 82 Globulol 2085 0.9 0.2 2.7 1.1 1.5 2.8 1.0 4.3 2.6 0.3 83 Elemol 2089 ± ± ± ± ± ± ± ± ± ± 84 Viridi¯orol 2093 0.5 0.2 1.5 0.6 0.4 0.8 0.4 1.3 1.3 0.3 85 Guaiol 2104 0.3 ± 0.2 ± ± 0.06 ± 0.5 ± ± 86 C15H26O 2110 0.3 0.06 0.8 0.3 0.2 0.4 0.2 0.7 0.5 0.06 87 C15H26O 2120 0.8 0.1 2.8 0.4 0.1 0.6 0.3 0.8 0.6 0.07 88 Spathulenol 2136 1.5 0.6 0.8 3.5 1.2 1.6 0.3 0.3 3.0 0.3 89 C15H24O 2153 0.09 ± 0.06 0.3 ± ± ± 0.06 0.08 ± 90 g-Eudesmol 2177 1.5 0.2 1.2 0.7 ± ± 0.1 0.09 0.06 0.7 91 d-Cadinol 2187 0.8 0.09 ± 0.5 ± ± 0.1 ± ± 0.8 92 C15H26O 2197 0.8 0.1 ± 0.4 ± ± 0.07 ± ± 0.5 93 C15H26O 2223 0.09 0.5 ± 0.09 ± ± ± ± ± ± 94 C15H26O 2229 0.2 0.07 ± 0.3 ± 0.08 ± ± 0.1 ± 95 a-Eudesmol 2234 4.5 0.7 3.1 1.0 0.08 0.2 0.4 0.09 0.2 3.3 96 C15H26O 2240 ± ± ± ± ± 0.2 ± ± ± ± 97 b-Eudesmol 2244 5.6 0.6 3.5 2.5 1.0 0.1 0.8 0.1 1.3 5.3 98 C15H26O 2263 ± ± ± ± ± ± ± ± ± ± 99 C15H26O 2269 0.2 ± 0.06 0.8 ± 0.1 ± 0.09 0.1 0.07 100 Farnesyl acetate 2271 ± ± ± 0.1 ± ± ± ± ± ± 101 Jacksonone 2280 ± ± ± 0.2 ± ± ± ± 0.06 ± 102 Tasmanone 2303 0.4 ± ± 4.6 ± ± ± ± ± ± 103 Isobicyclogermacral 2325 ± ± ± 0.8 ± ± ± ± 0.6 0.2 104 (E,E)-Farnesol 2366 0.4 0.07 ± 0.7 ± ± ± ± ± 0.07 105 Lateriticone 2388 0.9 ± ± ± ± ± ± ± ± ± 106 Torquatone 2424 1.7 0.1 0.4 6.7 0.06 0.2 0.06 tr. 0.3 tr.

Total percentages 88.7 93.0 95.0 82.4 97.8 94.5 97.4 96.8 96.3 96.2

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) VOLATILE LEAF OILS OF EUCALYPTUS SPECIES 431 Hopper Hopper Hopper & & Hill & Hopper er er er & D. ook er ook ook K. Br Br Br Johnson & ook S. Br A. tissima pusilla cerina cens L. ala Johnson Hopper & S. & subsp. subsp. interja A. er er Hill subsp. L. ook D. ely ook yling subsp. a Br Hopper Hopper Br K. Blak Gr & & ta ta tiae & a er er M. gr loxophleba lissophloia . Hopper eleva P ook ook & Hopper lanceola & Br Br Hopper Maiden er & subsp. subsp. subsp. & subsp. ely er ely) ely) cph. ook er subsp. er Br Ma ook Blak (Blak (Blak Benth. Benth. Benth. ook ook s J. Br (Maiden) Benth. Br ta ta Br C. osa Maiden chycory a evipes subangus br br subangus semota glomer captiosa crucis loxophleba phaenophylla loxophleba loxophleba leptopoda kingsmillii E. E. E. E. E. E. E. E. E. E. E. E. E. E. 0.2 ± 4.2 ± 0.2 ± 0.3 ± ± ± ± ± 0.2 ± ± ± 0.6 ± ± ± ± ± 0.1 ± 0.1 ± ± ± ± ± ± 0.1 ± ± 0.06 0.1 0.2 ± 0.06 ± 0.2 0.2 ± ± ± 0.07 ± ± ± 0.08 ± ± ± ± ± ± ± ± 0.06 0.9 ± ± ± 0.4 1.0 0.1 0.1 ± 0.4 0.07 ± 0.1 0.7 0.1 ± ± ± 0.1 0.07 ± 0.07 ± 0.1 0.1 ± ± ± ± ± ± 0.2 ± ± ± ± ± ± 0.1 ± 0.08 0.4 0.1 0.07 ± 2.0 0.2 0.4 ± 0.07 ± 0.3 0.06 ± ± 0.07 0.06 ± ± 0.1 ± ± ± ± ± ± ± 0.1 0.3 0.8 2.1 0.6 0.2 0.8 2.0 4.9 0.5 0.7 0.2 1.9 0.3 ± ± ± ± ± ± ± ± ± ± 0.07 ± ± ± 0.2 1.0 0.5 0.2 0.07 0.2 0.7 0.5 0.3 0.3 0.09 0.8 0.1 ± ± ± 0.2 0.06 ± ± 0.07 0.08 0.07 0.09 ± 0.08 0.1 ± ± 0.3 0.3 0.1 ± 0.3 0.2 0.4 0.09 0.1 ± 0.4 1.5 ± 0.07 0.2 0.3 ± ± ± 0.3 0.2 0.2 0.2 0.06 0.4 ± 0.8 2.1 29.6 0.2 1.2 0.1 0.6 0.4 0.1 0.1 0.5 0.2 1.1 2.8 ± ± 0.1 ± ± ± 0.06 ± ± ± ± ± ± 0.1 0.4 ± 0.08 ± ± ± 0.2 ± ± ± 0.06 0.4 0.09 0.06 0.6 ± 0.1 ± ± ± 0.2 ± ± ± ± 0.1 0.5 ± 0.4 ± 0.2 ± ± ± ± ± ± ± 0.3 0.1 0.2 ± ± ± ± ± ± ± 0.2 ± ± ± ± ± ± ± ± ± 0.6 ± 0.1 ± ± 0.1 0.09 0.1 ± ± 0.07 0.3 2.7 ± 0.8 ± 0.1 0.1 0.6 0.2 0.1 0.1 1.4 1.2 0.4 0.6 ± ± ± ± ± ± 0.5 ± ± ± ± ± ± ± 5.1 ± 1.3 0.07 0.4 0.3 3.1 0.9 0.1 0.4 8.6 ± 6.6 1.2 ± ± 0.3 ± ± ± ± ± ± ± ± ± ± ± ± ± 0.3 ± 0.1 ± 1.1 0.09 0.2 0.07 0.06 ± 0.1 ± ± ± 0.3 ± ± ± 0.3 ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 0.08 ± ± ± ± ± ± 0.2 ± 0.2 0.08 ± 0.06 0.1 0.3 0.2 0.1 0.2 0.2 ± ± 0.4 ± ± ± ± ± ± 0.2 0.3 0.07 0.4 0.4 0.06 ± 0.1 ± ± ± ± 0.08 ± ± 0.08 ± 0.08 ± ± ± 0.9 ± ± ± ± ± ± ± ± ± ± ± tr. tr. 0.4 tr. 0.2 tr. 0.3 0.4 0.4 0.4 0.1 0.08 0.7 0.2

97.3 97.1 89.5 98.0 94.4 96.2 84.5 94.6 92.5 95.8 96.2 94.8 94.7 83.9

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997) 432 C. M. BIGNELL ET AL.

The principal sesquiterpenes encountered in REFERENCES these species were aromadendrene (0±31.5%), allo-aromadendrene (0±3.8%), bicyclogermacrene 1. C. M. Bignell, P. J. Dunlop and J. J. Brophy, (Series I) (0±42.3%) and the related alcohols, globulol Part XVII, Flavour Fragr. J. 12, 269 (1997), and earlier Parts cited therein. (0.1±4.9%) and spathulenol (0.1±29.6%). The 2. M. I. H. Brooker and D. A. Kleinig, A Field Guide to the aromatic ketone torquatone was detected (trace± Eucalypts, Vol. 2, Inkata Press, Melbourne (1990). 6.7%) in all of the 24 species. 3. R. T. Baker and H. G. Smith, A Research on the Eucalypts, Fifteen of the species reported in this paper Especially in Regard to Their Essential Oils, 2nd edn, belong to series we have studied previously; Government Printer, Sydney (1920). 11 11 4. J. J. Brophy, in D. J. Boland, J. J. Brophy and A. P. N. E. megacarpa, E. calcicola, E. pressiana subsp. House (eds), Eucalyptus Leaf Oils; Use, Chemistry, Distilla- lobata,11 E. redacta,12 E. subangusta subsp. sub- tion and Marketing, Inkata Press, Melbourne (1991). angusta,13 E. subangusta subsp. pusilla,13 E. sub- 5. P. M. Grayling and M. I. H. Brooker, Aust. J. Bot., 44, 1 angusta subsp. cerina,13 E. phaenophylla subsp. (1996). 13 14 6. R. B. Inman, P. J. Dunlop and J. F. Jackson, in Modern interjacens, E. glomerosa, E. leptopoda subsp. Methods of Analysis, New Series, Vol. 12, pp. 201±19, 14 14 elevata, E. kingsmillii subsp. alatissima, E. crucis ed. H. F. Linskens and J. F. Jackson, Springer-Verlag, subsp. lanceolata,15 E. brevipes,1 E. captiosa16 Berlin (1991). and E. brachycorys.1 Chemometric analysis of the 7. C. P. Milner, R. D. Trengove, C. M. Bignell and P. J. chromatographic data of all the oils of each series Dunlop, in Modern Methods of Plant Analysis, New Series, Vol. 19, pp. 141±58, ed. H. F. Linskens and J. F. Jackson, indicates in each case excellent agreement with the Springer-Verlag, Berlin (1997). botanic classi®cations which are based on morpho- 8. S. R. Heller and G. W. A. Milne, EPA/NIH Mass Spectral metric measurements. Data Base, U.S. Government Printing Oce, Washington Our oil analyses for E. loxophleba subsp. D.C. (1978, 1980, 1983). 9. E. Stenhagen, S. Abrahamsson and F. W. McLa€erty, lissophloia agreed to some extent with an analysis Registry of Mass Spectral Data, Wiley, New York (1974). 4 reported previously for E. loxophleba (sub-species 10. A. A. Swigar and R. M. Silverstein, Monoterpenes, Aldrich, not de®ned) but not with others;3;5 in general many Milwaukee, WI (1981). more components were identi®ed. 11. C. M. Bignell, P. J. Dunlop, J. J. Brophy and C. J. R. Fookes, (Series I) Part XIV, Flavour Fragr. J., 12, 177 (1997). 12. C. M. Bignell, P. J. Dunlop, J. J. Brophy and J. F. Jackson, Acknowledgements Ð The authors thank Mr Ian Brooker, (Series I) Part XI, Flavour Fragr. J., 11, 107 (1996). Australian National Herbarium and Dean Nicolle, Currency 13. C. M. Bignell, P. J. Dunlop and J. J. Brophy, (Series I) Part Creek Arboretum, South Australia, for identifying the species XV, Flavour Fragr. J., 12, 185 (1997). and helpful discussions. We are grateful to Dr Jennifer Gardner, 14. C. M. Bignell, P. H. Dunlop, J. J. Brophy and J. F. Jackson, Curator of the Waite Arboretum, for her interest in this (Series I) Part III, Flavour Fragr. J., 9, 309 (1994). study. This work was supported in part by a grant from the 15. C. M. Bignell, P. J. Dunlop, J. J. Brophy and J. F. Jackson, Australian Council for International Agricultural Research (Series I) Part IX, Flavour Fragr. J., 11, 95 (1996). (ACIAR) to JJB. 16. C. M. Bignell, P. J. Dunlop, J. J. Brophy and J. F. Jackson, (Series I) Part I, Flavour Frag. J., 9, 113 (1994).

# 1997 John Wiley & Sons, Ltd. Flavour and Fragrance Journal, Vol. 12, 423±432 (1997)