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Volatile constituents of benzoin gums: Siam and Sumatra. Part 1

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DOI: 10.1002/ffj.1230

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328 X. FERNANDEZ
FLAVOUR ET AL.
AND FRAGRANCE JOURNAL
Flavour Fragr. J. 2003; 18: 328–333
Published online in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/ffj.1230

Volatile constituents of benzoin gums: Siam and

Sumatra. Part 1
Xavier Fernandez,1 Louisette Lizzani-Cuvelier,1* André-Michel Loiseau,1 Christine Périchet 2 and Claire
Delbecque2
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 February 2002
Revised 1 August 2002
Accepted 11 September 2002

ABSTRACT: The volatile extract composition of two different benzoin gums, Siam and Sumatra, were analysed
by GC–MS. Twenty components representing more than 99.1% of the oil from Siam and 29 components representing
more than 97.4% of the oil from Sumatra were analysed. The major components were benzyl benzoate (76.1–80.1%)
for the two oils and benzoic acid (12.5%), methyl benzoate (1.5%) and allyl benzoate (1.5%) for Siam, and styrene
(2.3%), cinnamic acid (3.5%) and benzyl cinnamate (3.3%) for Sumatra. Volatile compounds of oils and crushed
benzoins were also studied using solid-phase microextraction (SPME) employing carboxen/polydimethylsiloxane and
carbowax/divinylbenzene fibres. Copyright © 2003 John Wiley & Sons, Ltd.
KEY WORDS: benzoin gum Siam; benzoin gum Sumatra; balsam; GC–MS; benzyl benzoate; benzoic and cin-
namic acids; solid phase microextraction (SPME)

Introduction fixative properties. Its robust character is normally pre-

ferred to flavour tobacco products.2 Sumatra is also fre-
Benzoin gum is a balsam obtained from trees of the quently sold under four grades called A, B, C and D by
Family Styracaceae. It is produced mainly in Asia from some collectors and under only two or three grades by
places such as Indonesia, Sumatra, Java, Laos, Thailand others.
and Vietnam. Two types of benzoin gum exist in the Even if these resins are widely used in flavour and fra-
trade: benzoin gum Siam from Styrax tonkinensis Craib grance, there have been only a few studies on the chemi-
and benzoin gum Sumatra from Styrax benzoin Dryan- cal composition of Siam or Sumatra benzoin gums. In
der.1 Under normal conditions, the tree does not produce 1914, Reinitzed found that the major constituent of Siam
any resin; it is a pathological product that is only pro- is coniferyl benzoate 75–80%.3 Schroeder’s work shows
duced after deep incisions are made in the bark. that Siam is composed of coniferyl benzoate (65–75%),
Benzoin gum Siam is externally reddish yellow and p-coumaryl benzoate (10–15%), cinnamyl cinnamate
internally milky white. Its odour is pleasant, sweet- (0.5–6%), benzoic acid (12%), vanilline (0.3%) and
balsamic with a distinct note of vanilla. Siam is particu- siaresinolic acid (6%).4 Sumatra is reported to contain
larly useful in brown flavours, such as vanilla, chocolate, more cinnamic acid and cinnamates than Siam.5 One
nuts.2 This resin is graded according to the size of the study presented a comparative analysis of volatile com-
pieces. They occur in the trade in four different grades: pounds of Siam and Sumatra.2 In this study, 17 com-
1 (very large tears, without any foreign particles) to 4 pounds are described in the volatile fraction of Siam, the
(very small, powder-like). Sumatra comes in pieces that major products are benzyl alcohol (38.8%), benzoic acid
are reddish to greyish-brown in colour. Fine qualities (18.4%) and benzyl benzoate (39.3%). For Sumatra, 15
present a strong styrax-like odour, quite distinct from the compounds are presented, with benzyl alcohol (43.4%)
vanilla odour of the Siam variety. It is often used in soap and benzyl benzoate (50.7%) as the major constituents.
and detergent fragrances for this reason as well as its However, the experimental procedure to obtain the vola-
tile fractions is not described. One other study presented
volatile compounds in Sumatra but experimental proce-
dure and compounds ratio are not mentioned.6
* Correspondence to: L. Lizzani-Cuvelier, Laboratoire Arômes, Synthèses, In the present work we studied the chemical com-
Interactions, Faculté des Sciences de Nice Sophia-Antipolis, Parc Valrose,
06108 Nice cedex 2. position of the volatile extracts obtained from Siam and
E-mail: cuvelier@unice.fr Sumatra benzoin gums by direct GC injection of the oils.

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18: 328–333
 VOLATILE CONSTITUENTS OF SIAM AND SUMATRA BENZOIN GUMS 329

Volatile compounds of oils and crushed benzoin gums vial closed by a PTFE/silicone septum (Supelco, Cata-
were also studied by SPME. logue No. 27121-u). The containers were allowed to
stand overnight before any SPME sampling of the head-
space occurred. After exposure, the fibre was injected
Experimental into the gas chromatograph and left in the injection port
(equipped with a 0.75 mm i.d. inlet liner) for 4 min. The
Material injector temperature was set at 250 °C and operated in
the splitless mode for 4 min unless otherwise stated.
Benzoin gum Siam (from Styrax tonkinensis Craib) was After 4 min at 60 °C, the temperature was increased at a
collected in north Laos and graded as Benzoin Gum Siam rate of 2 °C min to 230 °C and then heated at 230 °C for
Grade 3, and was purchased in early 2001; Sumatra 45 min. Before sampling, each fibre was reconditioned
(from Styrax benzoin Dryander) was collected in Indone- for 15 min in the GC injection port at 230 °C.
sia and graded as Benzoin Gum Sumatra Grade B and
was purchased in early 2001.
HS–GC

Isolation of Volatile Extracts Benzoin gums were analysed by HS–GC to determine

the toluene and styrene titrations: a Hewlett-Packard
Siam and Sumatra benzoin gums (50 kg) were submitted 7694/6850 system equipped with a BP-1 column (30 m
to hydrodistillation in a 1000 l apparatus for 11 h using × 0.25 mm, 0.25 µm film thickness) was used. The
n-hexane (5 l) as a recovery solvent for the oil. Volatile headspace injector was static, with a working temperature
oils were obtained in 0.14% yield (52 g) for Siam and of 70 °C and a stabilization time of 30 min. GC oven
0.07% yield (27 g) for Sumatra. initial temperature was maintained at 35 °C for 10 min
then increased from 35 °C to 220 at a rate of 10 °C/min.
Injector and detector temperature were 250 °C; carrier
GC–MS gas, helium; split ratio, 1/10. The toluene and styrene
titrations were calculated by external calibration. Studies
The oil was analysed by GC–MS using a Hewlett- were realized with 2 g crushed benzoin gums. HS vials
Packard 5890/5970A system, with a HP1 column (50 m were allowed to stand overnight before any analysis.
× 0.20 mm, film thickness, 0.5 µm). GC oven initial tem-
perature was 60 °C and programmed to 220 °C at a rate
of 2 °C/min and then head at 220 °C for 45 min under Results and discussion
the following operation conditions: carrier gas, helium;
injector and detector temperatures, 250 °C; injected The components of the essential oils, the percentages of
volume, 0.4 µl (20% hexane solution); split ratio, 1:80. the constituents (by direct integration) and the retention
Retention indices were determined with C5 –C28 alkane indices are listed in Table 1. Chromatographic profiles of
standards as reference. The mass spectra were performed the volatile extracts revealed 20 identified constituents,
at 70 eV, mass range 35–400. Identification of the con- which represented 99.1% of the total GC–MS area for
stituents was based on comparison of the retention times Siam and 29 identified constituents which represented
with those of pure references and on computer matching 97.4% of the total GC–MS area for Sumatra. The analy-
against a commercial library (Wiley, MassFinder 2.1 sis revealed that Siam oil contained one aromatic car-
Library) and a home made mass spectra library built up boxylic acid (benzoic acid, 12.5%), eight esters (84.8%),
from pure substances and MS literature data.7–11 seven sesquiterpene hydrocarbons (0.4%). For the
Sumatra species, we observed two aromatic carboxylic
acid (benzoic and cinnamic acids, 5.2%), 10 esters
SPME Procedure (82.7%) and six sesquiterpene hydrocarbons (2.4%). The
major components are benzyl benzoate (76.1–80.1%) for
A manual SPME device and fibres were obtained from the two oils and benzoic acid (12.5%), methyl benzoate
Supelco (PA, USA). Two different fibres were used: (1.5%) and allyl benzoate (1.5%) for Siam, and styrene
carboxen/polydimethylsiloxane CAR/PDMS StableFlex (2.3%), cinnamic acid (3.5%) and benzyl cinnamate
(75 µm; Supelco, Catalogue No. 57334-u) and carbowax/ (3.3%) for Sumatra.
divinylbenzene CW/DVB (65 µm; Supelco, Catalogue Compounds present in oils were also studied by solid-
No. 57312). Before use, the fibres were conditioned as phase microextraction (SPME). This technique is based
recommended by the manufacturer. The fibre was ex- on the differential sorption of compounds to a fused-silica
posed to the headspace of crushed benzoin gum (10 g, 30 fibre coated with a polymeric organic liquid phase.12 Con-
min) or volatile extracts (3 g, 2 min) in a 40 ml amber siderable research has been done on the application of

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18: 328–333
330 X. FERNANDEZ ET AL.

Table 1. Chemical composition of the volatile extracts of Siam and Sumatra benzoin gums
Compoundsa RI Siam benzoin Sumatra benzoin

Styrene* 876 – 2.3

Benzaldehyde* 932 0.4 0.9
6-Methyl-5-hepten-2-one 962 – tr
Phenyl acetaldehyde 1011 – tr
1,8-Cineole* 1025 0.1 0.4
Acetophenone* 1037 – 1.8
Methyl benzoate* 1071 1.5 0.5
Benzoic acid* 1138 12.5 1.7
Ethyl benzoate* 1147 1.1 0.2
Allyl benzoate* 1232 1.5 0.9
(E)-Cinnamaldehyde* 1235 – 0.7
Propyl benzoate* 1247 0.2 0.1
4-Ethylguaiacol 1254 0.1 0.6
Isobutyl benzoate* 1306 tr –
Eugenol* 1331 1.1 0.8
Dihydroeugenol* 1344 tr tr
α-Copaene 1385 tr tr
β -Elemene 1394 tr –
Cinnamic acid* 1396 – 3.5
Isoamyl benzoate* 1415 0.1 –
trans-α-Bergamotene 1417 – 0.2
3-Methylbut-3-enyl benzoate* 1422 0.3 –
β-Caryophyllene* 1430 tr –
Ethyl cinnamate* 1436 – 0.1
Unknownb 1470 0.3 2.2
Valencene 1483 tr tr
Ledene 1493 tr tr
∆-Guaiene 1502 0.1 tr
Allyl cinnamate* 1524 – 0.5
(E)-Nerolidol 1550 – 0.4
Isobutyl cinammate* 1601 – 0.1
Benzyl benzoate* 1742 80.1 76.1
Cinnamyl benzoate* 2033 – 1.4
Benzyl cinnamate* 2048 – 3.3
(E)-Cinnamyl-(E)-Cinnamate* 2344 – 0.9
a
Compounds are listed in order of their elution from a HP1 column.
b
Unknown: 204(M+, 22.1), 189(100), 147(12.1), 134(11.3), 119(14.1), 107(16.3), 105(26.5), 93(14.9), 91(22.7), 79(12.1), 77(12.4), 55(9.8), 41(12.9).
RI, retention indices as determined on HP1 using the homologous series of n-alkanes; tr, trace (< 0.1%).
* Structure confirmed by standard compound injection.

this technique for volatile flavour analysis.13–15 Several other sesquiterpene hydrocarbons. On the other hand,
fibres are available in accordance with compounds to SPME analyses are not able to identify low volatile
studied and Bicchi’s works show that the most effective compounds such as benzyl cinnamate, cinnamyl benzoate
fibres for head-space-SPME are those consisting of two or cinnamyl cinnamate. The ratios obtained are not sig-
components, a liquid (PDMS) for the less polar compo- nificant and are dependent only on compound affinities
nents and a solid (DVB or CAR) polymer coating for the with the fibre.
more polar compounds.16 Compounds found here in benzoin volatile extracts
In this work, we have selected two fibres: carboxen/ from Laos and Indonesia are in good agreement with
polydimethylsiloxane (CAR/PDMS) and carbowax/divinyl- those found in the previously published work for aro-
benzene (CW/DVB). CAR/PDMS fibre is recommended matic esters and acids.2 However, we do not observe the
for low molecular weight compounds17 and has been presence of benzyl alcohol, nevertheless described as
described as the best fibre to characterize volatile com- very abundant in Moyler’s works.2 This observation can
pounds such as menthol.18 The mixed coating CW/ be explained by the procedure used to obtained the vola-
DVB was tried, as it was considered suitable for the tile fractions. We also observe that volatile fractions
analysis of polar semivolatiles.19 Compounds present obtained do not contain vanillin, which has nevertheless
in oil with these two fibres are listed in Table 2. Due been described as present at trace level among the vola-
to fibre selectivity and the low volatility of benzyl tile compounds of Siam and Sumatra benzoin gums.2,6
benzoate, SPME studies lead to the identification of To check the presence of benzyl alcohol and vanillin
new compounds in oil such as ethanol, toluene, α- and in benzoin gums, we performed SPME analyses on
β-pinene, linalool oxide, terpinen-4-ol, α-ylangene and crushed Siam and Sumatra benzoin gums with CAR/

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18: 328–333
 VOLATILE CONSTITUENTS OF SIAM AND SUMATRA BENZOIN GUMS 331

Table 2. SPME studies of Siam and Sumatra benzoin volatile extracts

Compoundsa Siam benzoin Sumatra benzoin

Fibre Ab Fibre Bc Fibre A Fibre B

Ethanol 4.5 4.2 2.7 3.1
Toluene 13.5 1.2 tr –
Styrene 2.5 0.5 61.3 51.0
Benzaldehyde 13.6 28.2 13.2 16.5
α-Pinene 19.1 – – –
6-Methyl-5-hepten-2-one – 0.3 0.7 0.9
β -Pinene 1.4 0.4 – –
Trimethylbenzene 0.3 0.3 – –
Phenyl acetaldehyde – – 0.1 0.3
α-Phellandrene 0.2 – – –
Isoterpinolene – 0.1 – –
p-Cymene 2.2 1.0 0.2 0.2
1,8-Cineole 15.0 5.8 6.1 4.3
Benzylformate – 0.1 – –
Acetophenone 0.2 tr 6.2 11.2
γ -Terpinene 0.1 0.2 – –
Linalool oxide 1.8 1.0 0.1 tr
Methyl benzoate 16.0 33.7 3.5 3.6
Linalool – 0.3 – –
Benzoic acid 1.3 0.9 0.2 tr
Ethyl benzoate 1.4 5.3 0.6 0.7
Terpinen-4-ol tr 0.1 0.1 0.1
Allyl benzoate 0.7 5.7 0.4 1.1
(E)-Cinnamaldehyde – – – 0.3
Propyl benzoate – 1.0 0.1 0.2
4-Ethylguaiacol – 0.2 0.1 0.4
Isobutyl benzoate – 0.1 – tr
Eugenol tr 0.6 tr 0.2
∆-Elemene 0.9 0.5 0.3 0.2
α -Ylangene 1.0 0.1 tr tr
α -Copaene 1.6 0.9 0.4 0.3
β -Elemene 0.7 0.5 0.2 0.1
trans-α -Bergamotene – – 0.3 0.3
β -Caryophyllene 0.6 0.4 0.1 0.2
Unknownd 1.1 0.7 2.5 2.7
Germacrene D – 0.3 – –
Valencene 0.1 tr tr tr
α -Muurolene tr 0.3 – –
Ledene 0.2 0.3 tr 0.1
∆-Guaiene – 0.1 – –
∆-Cadinene tr 0.1 tr 0.1
Benzyl benzoate – 1.6 – 0.3
a
Compounds are listed in order of their elution from a HP1 column.
b
Carboxen/polydimethylsiloxane fibre (75 µm).
c
Carbowax/divinylbenzene fibre (65 µm).
d
See Table 1.
tr, trace (< 0.1%).

PDMS and CW/ DVB fibres. Several sorption times were tities of these compounds in benzoin gums, we performed
tested (10, 15, 30 and 45 min) and the best results were static HS–GC titrations of these compounds. We found a
obtained with a time of 30 min. Compounds present in quantity of 10 ppm toluene in Siam benzoin and trace
the headspace of Siam and Sumatra crushed benzoin level (< 1 ppm) in Sumatra benzoin. We found a quan-
gums are listed in Table 3. SPME analyses to crushed tity of 800 ppm styrene in Sumatra benzoin and a trace
Siam and Sumatra benzoin gum were able to identify level in Siam benzoin. These results show that these large
new volatile constituents in analogy of essential oil such quantities observed for toluene in Siam benzoin and
as benzyl alcohol, vanillin, acetic and propanoic acid, styrene in Sumatra benzoin was due to the high affinity
ethylbenzene and phenol. of these compounds with the CAR/PDMS fibre in oper-
Studies of head space with CAR/PDMS fibre gum ating conditions. The CAR/PDMS fibre represents a very
lead to the identification of a large quantity of toluene useful fibre for the detection of non polar hydrocarbon
(> 65%) in Siam benzoin and styrene (> 90%) in Sumatra compounds, but it is not recommended for qualitative
benzoin, in all our analyses. To determine the real quan- study of benzoin gums.

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18: 328–333
332 X. FERNANDEZ ET AL.

Table 3. SPME studies of crushed Siam and Sumatra benzoin gums

Compoundsa Siam benzoin Sumatra benzoin

Fibre Ab Fibre Bc Fibre A Fibre B

Ethanol 0.2 1.5 0.4 0.7
Acetic acid 4.1 1.1 0.8 0.9
3-Buten-2-ol 0.6 – – –
Propanoic acid 0.4 0.1 – –
Toluene 66.4 1.2 0.3 –
Ethylbenzene 0.2 – – –
Styrene 5.0 – 90.4 70.1
Benzaldehyde 4.4 5.9 1.3 3.2
α-Pinene 4.5 – 0.2 –
Phenol – – 0.7 2.3
6-Methyl-5-hepten-2-one – – – tr
Phenyl acetaldehyde – – – 0.1
Isoterpinolene – 0.1 – –
Benzyl alcohol 2.4 11.2 – 0.4
p-Cymene 0.4 0.3 0.3 0.3
1,8-Cineole 0.6 – 0.3 0.4
Acetophenone 0.4 – 1.5 7.6
Benzylformate 0.2 0.8 – –
Linalool oxide tr 0.3 – tr
Methyl benzoate 2.0 8.7 0.3 tr
Benzoic acid 4.8 43.4 0.6 3.9
Ethyl benzoate 1.4 12.1 – 1.3
Allyl benzoate – tr – tr
(E)-Cinnamaldehyde – – – 1.0
Propyl benzoate – 0.5 – 0.2
4-Ethylguaiacol – 0.5 – 0.1
Isobutyl benzoate – 0.7 – tr
Eugenol – 1.6 – 0.2
∆-Elemene 0.5 – – –
Vanillin 0.1 3.2 – 0.4
α -Copaene 0.1 – – –
β -Caryophyllene 0.3 – – –
Unknownd 1.7 0.7 2.3 7.5
Germacrene D – 0.1 – –
Benzyl benzoate – 4.1 – 0.3

a
Compounds are listed in order of their elution from a HP1 column.
b
Carboxen/polydimethylsiloxane fibre (75 µm).
c
Carbowax/divinylbenzene fibre (65 µm).
d
See Table 1.
tr, trace (< 0.1%).

Conclusions under current study, using different fibres (PDMS, CW/

DVB and polyacrylate).
In conclusion, our GC–MS studies of Siam and Sumatra The direct and SPME analyses of Siam and Sumatra
volatile extracts led to the identification of 20 and 29 benzoin volatile extracts show the presence of ses-
compounds, representing 99.3% and 97.2% of the total quiterpene hydrocarbons at trace level. The fractionation
area. The major components were benzyl benzoate (76.1– of these oils, GC and GC–MS studies of the different
80.1%) for the two oils and benzoic acid (12.5%), methyl fractions obtained, will be the subject of a further
benzoate (1.5%) and allyl benzoate (1.5%) for Siam, publication.
styrene (2.3%), cinnamic acid (3.5%) and benzyl
cinnamate (3.3%) for Sumatra.
The headspace analyses by SPME of oils and crushed References
benzoin gums were able to identify new compounds, in
particular benzyl alcohol and vanillin, which were iden- 1. Coppen JJW. Perfum. Flavor., 1999; 24: 11.
tified in crushed Siam and Sumatra but not in the oils. 2. Moyler DA. Rev. Ital. EPPOS, 1998; 351.
This technique seems particularly adequate to the studies 3. Reinitzer F. Arch. Pharmacol., 1914; 252: 341.
4. Schroeder HA. Phytochemistry, 1968; 7: 57.
of balsams and SPME analyses of volatile constituents of 5. Pastorova I, de Koster CG, Boon JJ. Phytochem. Anal., 1997; 8:
different qualities of Siam and Sumatra benzoin gums are 63.

Copyright © 2003 John Wiley & Sons, Ltd. Flavour Fragr. J. 2003; 18: 328–333
 VOLATILE CONSTITUENTS OF SIAM AND SUMATRA BENZOIN GUMS 333

6. Boelens HM, de Rijke D, Haring HG. Perfum. Flavor., 1982; 6: 11. ESC 2000. The Complete Database of Essential Oils. Boelens
7. Aroma Chemical Information Service: The Netherlands, 1999.
7. Davies NW. J. Chromatogr., 1990; 503: 1. 12. Scheppers Wercinski SA. Solid Phase Microextraction, a Practi-
8. (a) Joulain D, König WA, The Atlas of Spectra Data of cal Guide. Marcel Dekker: New York, 1999.
Sesquiterpens Hydrocarbons. E.B.-Verlag: Hamburg, 1998. (b) 13. Yang X, Peppard T. J. Agric. Food Chem., 1994; 42: 1925.
Joulain D, König WA, Hochmuth DH. Terpenoids and Related 14. Steffen A, Pawliszyn J. J. Agric. Food Chem., 1996; 44: 2187.
Constituents of Essential Oils. Library of MassFinder 2.1, 15. Coleman WM, Lawrence BM. Flavour Fragr. J., 1997; 12: 1
2001. 16. Bicchi C, Drigo S, Rubiolo P. J. Chromatogr. A, 2000; 892: 469.
9. McLafferty FW, Stauffer DB. The Wiley/NBS Registry of Mass 17. Brunton NP, Cronin DA, Monaham FJ. Flavour Fragr. J., 2001;
Spectral Data. Wiley: New York, 1989. 16: 294.
10. Adams RP. Identification of Essential Oil Components by Gas 18. Coleman III WM, Lawson S. J. Chromatogr. Sci., 1998; 36: 401.
Chromatography/Mass Spectroscopy. Allured: Carol Stream, IL, 19. Pawliszyn J. Solid-phase Microextraction—Theory and Practice.
1995. Wiley-VCH: New York, 1997.

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