70 X. FERNANDEZ ET AL.

Copyright © 2004 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 70–73

FLAVOUR AND FRAGRANCE JOURNALFlavour Fragr. J. 2005; 20: 70–73Published online 27 July 2004 in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/ffj.1370

Chemical composition of the essential oils from Turkishand Honduras Styrax

Xavier Fernandez,* Louisette Lizzani-Cuvelier,1 André-Michel Loiseau,1 Christine Perichet,2

Claire Delbecque2 and Jean-François Arnaudo2

1 Laboratoire Arômes, Synthèses, Interactions, Faculté des sciences de Nice Sophia-Antipolis, parc Valrose, 06108 Nice cedex 2,France

2 CHARABOT, Research Natural Products, 10, Av. Yves-Emmanuel Baudoin, 06130 Grasse, France

Received 14 April 2003; Revised 29 June 2003; Accepted 7 July 2003

ABSTRACT: The essential oil composition of two different styrax gums, Asian and American were analysed by GC and

GC-MS. Fifty-eight components representing more than 99.4% of the oil from Asian styrax gum and 57 components

representing more than 99% of the oil from American styrax gum were identified. The major components were styrene

(70.4 and 30.9% respectively), α-pinene (19.4 and 19.6%) and βββββ-caryophyllene (0.2 and 20.2%) for the two oils. These

results show that we could identify an Asian from an American styrax by relative amounts of styrene and βββββ-caryophyllene

in the essential oils. Copyright © 2004 John Wiley & Sons, Ltd.

KEY WORDS: Asian styrax; Liquidambar orientalis Mill.; American styrax; Liquidambar styraciflua, balsam; GC; GC-MS;

styrene; α- and β-pinene; β-caryophyllene; cinnamic esters

* Correspondence to: X. Fernandez, Laboratoire Arômes, Synthèses, Inter-

actions, Faculté des Sciences de Nice Sophia-Antipolis, Parc Valrose, 06108

Nice cedex 2, France.

E-mail: xavier.fernandez@unice.fr

Introduction

Styrax sometimes called storax, is a balsam obtained

from trees of the genus Liquidambar.1 Two types of

styrax exist in the trade: Asian styrax from L. orientalis

Mill., native to Asia Minor, and American styrax from

L. styraciflua, native to Mexico and Central America.

Styrax is a semi-solid, sticky material characterized

by a balsamic and styrene-like odour; it is often used in

floral-type perfumes.2 Essential oils can be obtained from

both types of styrax and are more often used by the

fragrance industry than the oleoresins.

Styrax presents various medicinal properties: it is used

for the treatment of skin diseases, as antiseptic, parasitic

and expectorant.3 Even if these resins are widely used in

flavour and fragrance, cosmetics and pharmacy, there are

only few studies on the chemical composition of styrax.

In 1877, Miller4 detected cinnamyl and 3-phenylpropyl

cinnamates in Asian Styrax. A more recent work showed

that the volatile fraction of American Styrax was composed

of cinnamyl cinnamate (38%), 3-phenylpropylcinnamate

(32.3%), cinnamic acid (4.8%), cinnamyl alcohol (4.1%),

3-phenylpropyl alcohol (3.1%), styrene and β -caryo-

phyllene (1.8%).5 Another study presented the volatile

compounds in Asian styrax identified in literature: 36

compounds were found. However, the experimental

procedure and compounds ratio were not mentioned.6

In the present work we studied and compared the

chemical composition of the essential oils obtained

from Asian and American styrax by GC and GC/MS

analysis.

Experimental

Material

American Styrax from Honduras (L. styraciflua) and

Asian Styrax (L. orientalis Mill.) from Turkey, certified

as natural, were supplied by Charabot S.A. (Grasse,

France). They were purchased in early 2001.

Isolation of essential oils

Styrax from Turkey and Honduras (200 g) were subjected

to hydrodistillation using a Clevenger-type apparatus

(4 h) to yield 0.6% oil from Asian gum and 0.8% from

American gum. The oil was dried over anhydrous sodium

sulphate and stored at low temperature (−12 °C) prior to

analysis.

Analytical GC

GC analysis was carried out using a Hewlett-Packard

5890 serie II gas chromatograph, under the follow-

ing operation conditions: vector gas, N2; injector and

detector temperatures, 250 °C; injected volume, 0.1 µl;

ESSENTIAL OILS FROM STYRAX 71

Copyright © 2004 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 70–73

Table 1. Chemical composition of Asian and American Styrax

Compoundsa RIb Asian (% ± SDc) American (% ± SDc)

Ethyl benzene 846/1130 0.1 0.5

Styrened 878/1270 70.4 ± 0.5 30.9 ± 0.3

α-Thujene 923/1039 tr 0.1

Benzaldehyded 930/1462 tr 0.6

α-Pinened 934/1019 19.0 ± 0.3 19.6 ± 0.3

Camphene 944/1071 0.7 0.6

Thuja-2,4(10)-diene 947/1116 0.1 1.5 ± 0.1

β-Pinened 972/1102 4.3 ± 0.1 4.1 ± 0.1

Myrcened 982/1208 0.4 tr

α-Phellandrene 996/1193 0.1 0.1

(E)-β-Methylstyrened 1008/1360 0.2 0.8p-Cymene 1011/1249 0.1 0.2

Limonened 1020/1182 1.2 ± 0.1 0.5 ± 0.1

Acetophenoned 1039/1583 0.2 3.2 ± 0.1

γ-Terpinened 1047/1191 0.1 0.1

Dehydro p-cymene 1071/1397 tr tr

Terpinolened 1077/1265 0.1 0.1

allo Ocimene 1094/1330 — tr

Isoborneol 1104/1649 tr —

3-Phenylpropanald 1122/1723 0.1 0.2

Pinocarveol 1128/1599 tr —

p-Ethylphenold 1137/2094 0.2 1.7 ± 0.1

Pinocamphone 1144/1504 tr —

Ethyl benzoated 1146/1620 — tr

Borneol 1148/1648 tr trTerpinene-4-old 1160/1528 0.1 tr

α-Terpineold 1170/1640 0.2 —

Estragold 1176/1756 tr 0.3

Myrtenold 1182/1723 tr 0.1

Verbenone 1185/1666 tr tr

Phenylpropylic alcohold 1197/1993 0.2 1.4

Allyl phenol (chavicol) 1229/2282 0.1 0.2

Cinnamaldehyded 1235/1953 tr tr

Methyl phenylpropionate 1246/1775 — tr

Unknown (1) 1254/1829 — 0.1

cis-Anethol 1257/1822 — tr

Cinnamic alcohold 1268/2195 0.3 0.9 ± 0.1

3-Phenylpropyl acetate 1347/1872 tr 0.1

α-Cubebene 1356/1425 tr 2.1α-Longipinene 1364/1427 0.1 —

α-Ylangene 1370/1441 tr 0.4

α-Copaene 1375/1452 tr 0.9

α-Cedrene 1389/1608 tr tr

β-Cedrene 1390/1565 tr 0.3 ± 0.1

Unknown (2) 1405/1640 — 0.1

Cinnamyl acetate 1406/2098 tr tr

cis-Caryophyllene 1409/1573 tr tr

Longifolene 1416/1520 0.1 0.9 ± 0.2

β-Caryophyllened 1422/1569 0.2 20.2 ± 0.2

Ethyl cinnamated 1436/2053 tr tr

Calarene 1450/1652 tr 0.7

Unknown (3) 1458/1630 — 0.1

α-Humulene 1465/1612 0.1 1.1 ± 0.2Germacrene D 1472/1593 tr 0.3 ± 0.1

Unknown (4) 1480/1633 tr 0.2

γ-Muurolene 1483/1652 tr tr

α-Muurolene 1492/1669 tr 0.9

Calamenene 1510/1790 tr 0.3

∆-Cadinene 1513/1701 0.1 1.6 ± 0.1

Valencene 1516/1644 tr 0.1

Unknown (5) 1523/1720 tr 0.3

α-Calacorene 1531/1850 tr 0.1

Caryophyllene oxided 1570/1936 tr 0.4

Unknown (6) 1628/1823 tr 0.3

∆-Cadinol 1639/2128 tr 0.2

Benzyl cinnamated 2042/- tr 0.13-Phenylpropyl cinnamate 2257/- 0.2 0.4

Cinnamyl cinnamated 2343/- 0.4 0.2

a Compounds are listed in order of their elution from a HP1 column.b RI, retention indices as determined on HP1 and HP20M column using the homologous series of n-alkanes.c SD, standard deviation.

tr, trace (<0.1%).d Structure confirmed by standard compound injection.

72 X. FERNANDEZ ET AL.

Copyright © 2004 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 70–73

Table 2. Mass spectrums of unknown compounds

Unknown RIa Mass spectrumb

compound

1 1254 152(M+, 3.9), 118(81.8), 117(100), 109(16.9), 92(12.2), 91(62.1), 79(14.9), 77(17.7), 65(11.7), 40(15.3)

2 1405 204(M+, 4.5), 161(11.7), 120(19.4), 119(100), 105(22.4), 93(85.3), 91(33.7), 79(12.6), 77(20.4),

69(31.5), 56(12), 41(22.6)

3 1458 204(M+, 23.7), 162(13.2), 161(100), 133(25.1), 120(15.5), 119(47.3), 107(16.4), 105(65.9), 93(30.2),

92(14.9), 91(40.5), 81(31.7), 79(24.7), 77(16.8), 69(23.3), 55(14.2), 41(21.3)

4 1480 222(M+, 47.9), 179(18.3), 164(31.4), 137(100), 126(34.6), 123(16.8), 121(40.2), 111(18.3), 109(46.3),

105(39.7), 95(34.8), 93(31.8), 81(34.4), 79(18.8), 71(18.4), 67(18.8), 55(20.2), 43(58.5)

5 1523 204(26.3), 161(52.4), 121(19.1), 120(19.6), 119(100), 105(67), 93(13.9), 92(14.8), 91(22.3), 77(10.1),

55(9.8), 41(12.1)

6 1625 204(48.3), 189(26.1), 161(100), 147(24.3), 133(31.6), 121(40.1), 119(46.7), 109(25.9), 107(29.5),

105(88), 95(32.1), 94(16.7), 93(38.2), 91(52.1), 81(38.1), 79(35.7), 77(25.1), 43(42.9), 41(29.7)

a Retention indices on HP1 column.b Only the most abundant ions are given.

splitless; HP1 column (J&W Scientific), polydimethyl-

siloxane (50 m × 0.20 mm i.d., film thickness 0.5 µm,

pressure 165 kPa); temperature programme 60–250 °C at

2 °C/min and 250 °C for 60 min. Retention indices were

determined with C5–C26 alkane standards as reference.

Relative amounts of individual components are based on

peak areas obtained without FID response factor correct-

ion. Three replicates were performed for each sample.

The average of these three values and the standard devi-

ation were determined for each component identified.

GC-MS

GC-MS analysis was accomplished by using a Hewlett-

Packard 5890/5970A system, with a HP1 (J&W Scien-

tific) column (50 m × 0.20 mm; film thickness, 0.5 µm)

and HP-20M (Carbowax 20M, J&W Scientific) column

(50 m × 0.20 mm; film thickness, 0.5 µm). Temperature

was programmed as above; the gas vector was He.

Retention indices were determined with C5–C26 alkane

standards as reference. The mass spectra were performed

at 70 eV of the mass range of 35–400. Identification of

the constituents was based on comparison of the reten-

tion times with those of pure references and on com-

puter matching against a commercial library (Wiley,

MassFinder 2.1 Library) and a home-made mass spectra

library built up from pure substances and MS literature

data.7–13

HS-GC

Styrax gums were analysed by HS-GC to determinate the

styrene titrations: a Hewlett-Packard 7694/6850 system

equipped with HP1 (J&W Scientific) column (30 m ×0.25 mm × 0.25 µm) was used. The Head-Space injector

was static, with a working temperature of 70 °C and a

stabilization time of 30 min. GC oven initial temperature

was maintained at 35 °C for 10 min then increased from

35 to 220 °C by a 10 °C/min rate. Injector and detector

temperature were 250 °C, the carrier gas was helium

and the split ratio was 1/10. The styrene titrations were

calculated by external calibration. Studies were carried

out with 2 g styrax.

Results and discussion

The components of essential oils, the percentage of each

constituent (by direct integration of FID responses) and

the retention indices are listed in Table 1 according to

their elution order on the HP-1 column. Chromatographic

profiles of the essential oils reveal 58 identified constitu-

ents, representing 99.4% of the total GC area for Asian

styrax, and 57 identified constituents, representing 99%

of the total GC area for American styrax. The analysis

revealed that Asian styrax oil contained six esters (0.6%),

11 monoterpene hydrocarbons (26.1%) and 17 sesqui-

terpene hydrocarbons (0.6%). For the American species,

we observed eight esters (0.8%), 12 monoterpene hydro-

carbons (26.4%) and 16 sesquiterpene hydrocarbons

(29.9%). The major components were styrene (70.4 and

30.9% respectively), α-pinene (19 and 19.6%) and β-

caryophyllene (0.2 and 20.2%) for the two oils.

Chemical composition of the two oils was similar,

however significant differences in the relative amounts

of major components were observed between the two

species. Styrene is the major compound in both oils,

but is more abundant in Asian styrax (70.4%) than

in American styrax (30.9%). On the other hand, β-

caryophyllene is more abundant in American styrax

(20.2%) than in Asian styrax (0.2%). These results show

that we could identify an Asian from an American styrax

by the larger amount of styrene and smaller amount of

β-caryophyllene.

However to determine the real importance of styrene in

these two resins, we performed static HS-GC titrations of

these compounds. We found 5500 ppm (0.55%) styrene

in Asian styrax and 5100 ppm (0.51%) in American

ESSENTIAL OILS FROM STYRAX 73

Copyright © 2004 John Wiley & Sons, Ltd. Flavour Fragr. J. 2005; 20: 70–73

styrax. These two values show that, even if the concen-

tration of styrene in the two essential oils is different,

they are similar in the resins.

New studies on different lots of the same styrax

(American or Asian) showed similar chemical composi-

tions but presented variations in major compound concen-

tration such as styrene, α-pinene and β-caryophyllene.

The method presented to identify an Asian from an

American styrax must be checked by a study of the vari-

ations as a function of time, method of harvesting and

origin of the resins. This study will be the subject of a

future publication.

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