J Am Oil Chem Soc (2008) 85:353–356
DOI 10.1007/s11746-008-1196-z
ORIGINAL PAPER
Santalum insulare Acetylenic Fatty Acid Seed Oils:
Comparison within the Santalum Genus
J.-F. Butaud Æ P. Raharivelomanana Æ
J.-P. Bianchini Æ E. M. Gaydou
Received: 7 April 2007 / Revised: 4 December 2007 / Accepted: 7 January 2008 / Published online: 24 January 2008
Ó AOCS 2008
Abstract The sandalwood kernels of Santalum insulare
(Santalaceae) collected in French Polynesia give seed oils
containing significant amounts of ximenynic acid, E-11octadecen-9-oic acid (64–86%). Fatty acid (FA) identifications were performed by gas chromatography/mass
spectrometry (GC/MS) of FA methyl esters. Among the
other main eight identified fatty acids, oleic acid was found
at a 7–28% level. The content in stearolic acid, octadec-9ynoic acid, was low (0.7–3.0%). An inverse relationship
was demonstrated between ximenynic acid and oleic acid
using 20 seed oils. Results obtained have been compared to
other previously published data on species belonging to the
Santalum genus, using multivariate statistical analysis. The
relative FA S. insulare composition, rich in ximenynic acid
is in the same order of those given for S. album or S.
obtusifolium. The other compared species (S. acuminatum,
S. lanceolatum, S. spicatum and S. murrayanum) are richer
in oleic acid (40–59%) with some little differences in
linolenic content.
J.-F. Butaud P. Raharivelomanana (&) J.-P. Bianchini
Laboratoire de Chimie des Substances Naturelles,
Université de la Polynésie Française, BP 6570,
98702 Faaa, Tahiti, French Polynesia
e-mail: phila.raharivelomanana@upf.pf; raharive@yahoo.com
E. M. Gaydou
Laboratoire de Phytochimie de Marseille, UMR CNRS 6171,
Université Paul Cézanne (Aix-Marseille III),
Faculté des Sciences et Techniques de Saint-Jérôme,
Avenue Escadrille Normandie Niémen,
13397 Marseille Cedex 20, France
Keywords Santalum insulare Santalaceae
Ximenynic acid Stearolic acid French Polynesia
Multivariate statistical analysis
Introduction
The Polynesian sandalwood, Santalum insulare Bertero ex
A. DC. (Santalaceae) is a small tree endemic to Eastern
Polynesia and has been used in fragrances, as medicine and
for religious purposes [1]. Studies on unusual acetylenic
fatty acids (FA) of Santalum seed oil genus began in the
1930s and most of them were identified by comparison with
those found in seed oils of the Ximenia genus (Olacaceae),
such as ximenynic acid, E-11-octadecen-9-ynoic acid, a
long chain acetylenic FA [2–4]. This rare ximenynic acid
previously named santalbic acid, was then identified and
reported in various genera of Santalaceae [4–9]. Proximate
and fatty acid composition changes in developing sandalwood (S. spicatum) seeds and the separation and
identification of ximenynic acid isomers in this seed oil as
their 4,4-dimethyloxazoline derivatives was achieved by
Liu et al. [10, 11]. Some works on triacylglycerols were also
conducted on S. album and S. spicatum [12–14].
In continuation of our study of the chemical variability
of S. insulare [1, 15], we focused on the FA profiles, since
an inverse relationship was demonstrated between the relative proportions of oleic and ximenynic acids in the case
of S. spicatum [16], S. acuminatum [4, 12, 17] or S. murrayanum [4, 16]. In this paper, as a part of our research
project on sandalwood, development and extension in the
Pacific islands, the FA composition of the S. insulare seed
oils are described for the first time. Results obtained from
twenty seed oils, have been compared, using multivariate
statistical analyses, with other Santalum species seed oils.
123
354
Table 1 Geographical origin
and physical characteristics
of S. insulare seeds
investigated
J Am Oil Chem Soc (2008) 85:353–356
Location
Number of seeds
Nuku Hiva (800–1000 m elevation)
15
Nuku Hiva (670 m elevation)
Tahuata (200–400 m elevation)
Sum or mean
Experimental Procedures
Ripe fruits of S. insulare were collected on Nuku Hiva
Island at two locations (670 and 800–1,000 m elevation)
and Tahuata Island (Marquesas archipelago) in 2002 and
2003. The dried seeds (Table 1) were ground and extracted
in a Soxhlet apparatus for 3 h with n-hexane. After
extraction, the solvent was removed by vacuum distillation
at 30 °C and FA methyl esters (FAME) were immediately
prepared by transesterification using 0.5 mol L-1 sodium
methoxide in anhydrous methanol at room temperature
overnight in a sealed tube under nitrogen [18].
Infrared spectra (IR) of FAME were recorded on a Jasco
FT-IR 460 Plus spectrophotometer.
A Hewlett Packard 5890 Series gas chromatograph
equipped with a flame ionisation detector (FID) and a fused
silica capillary column (25 m long, 0.25 mm i.d.) coated
with Carbowax (CW20M, 0.2 lm phase thickness) was
used for analyses. Temperatures were 220 °C for column
and 260 °C for inlet and detector ovens. Available FAME
were used for quantitative external standards. Identifications of fatty acids were carried out using mass
spectrometry of their FAME.
Gas chromatography–mass spectrometry was performed
using the same GC as described above coupled to a mass
selective detector (MSD) HP 5970 B Series. Helium was
Fig. 1 Mass spectra of the
ximenynic methyl ester
obtained by GC/MS of the
S. insulare FAME seed oil
123
1
Length (mm)
Width (mm)
Weight (g)
25.9
25.1
6.73
19
18
2.73
4
20.2
19.2
3.28
20
24.5
23.6
5.8
used as the carrier gas, ion source 220 °C, ionising voltage
70 eV. Results were compared to the NBS75K data bank
and to published results for ximenynic acid [11].
Principal component analysis was performed using the
dataset composed of the eight samples (mean of results
published on each Santalum species and mean of this work
on S. insulare) and seven variables (palmitic, palmitoleic,
stearic, oleic, linoleic, linolenic, and ximenynic acids)
transformed into centered and reduced variables (standardized PCA). Data were processed with XLSTAT
program version 7.5 (Addinsoft, France).
Results and Discussion
The geographical origin and the physical characteristics of
S. insulare seeds collected on two islands of French
Polynesia are given in Table 1. Among the ten FAMEs,
nine were identified by GC/MS. The mass spectra of the
main peak gave a molecular ion (M+ m/z 292) in agreement
with the molecular formula of ximenynic acid methyl ester
C19H32O2. Furthermore the occurrence of a significant
fragment (50%) m/z 150 [CH2=C=CH-CH=CH–(CH2)5–
CH3]+ and a basic peak at m/z 79 [CH2=C=CH–CH=CH–
CH2]+ confirmed unambiguously the 11-octadecen-9-ynoic
acid methyl ester (Fig. 1). An IR absorption of the oil at
J Am Oil Chem Soc (2008) 85:353–356
355
Table 2 Range of fatty acid composition of S. insulare kernels
investigated from Marquesas Islands
Meana
SD
Min.
Max.
Palmitic
1.0
0.2
0.8
1.4
Palmitoleic
0.6
0.1
0.4
0.8
Stearic
1.0
0.3
0.5
1.7
18.1
5.4
7.1
27.8
Linoleic
0.5
0.1
0.3
0.8
Linolenic
1.0
0.2
0.6
1.3
Stearolic
Arachidic
1.4
0.4
0.5
0.1
0.7
0.3
3.0
0.5
1.0
0.2
0.7
1.7
74.5
5.9
64.1
86.1
Fatty acid
Oleic
unknown
Ximenynic
a
Mean of 20 samples
955–956 cm-1 indicated the presence of the trans double
bond in the enyne system. Other small fragments were
in agreement with results previously published [11]. The
cis-ximenynic isomer previously characterized in minute
amount in S. spicatum [11] was not detected in these
S. insulare seed oils. Another acetylenic FA was identified
as stearolic acid (octadec-9-ynoic acid). One peak in small
amounts (0.7–1.7%) remained unidentified. Among the
nine FA identified (Table 2), two of them, ximenynic and
oleic acids represented about 95% of the relative percentage of these oils. The relative proportion of ximenynic acid
presents a large variation within the 20 S. insulare samples
from 64.1 up to 86.1% with an average of 74.5%. In the
case of oleic acid, the average was 18.1% with a minimum
of 7.1 and a maximum of 27.8%. A high negative correlation (0.99) between these two acids was observed and the
sum of these two acids was remarkably stable around 92–
94%. Such inverse correlation was previously observed in
the case of S. spicatum [16] and no significant change was
observed within elevation or between the two islands
investigated.
Since some results have been done on the FA composition of various Santalum species seed oils, we used
multivariate statistical analyses for S. insulare classification with the other species. Table 3 give the FA content of
S. album [5, 12, 14], S. acuminatum [4, 12, 17], S. lanceolatum [12], S. murrayanum [4, 12], S. obtusifolium [9],
and S. spicatum, [4, 5, 12, 13, 16]. The data set used was
composed of 8 samples (the mean of results published
on each Santalum species and means of this work on
S. insulare) and 7 variables (palmitic, palmitoleic, stearic,
oleic, linoleic, linolenic, and ximenynic acids). A graphic
representation of the projection of variables and samples
onto the two first principal components is given in Fig. 2,
using principal component analysis. Axis 1, which represents 48.0% of the total information, is positively loaded
with linoleic acid (0.91), linolenic acid (0.77) and oleic
acid (0.84) and negatively loaded with ximenynic acid
(-0.72) and palmitoleic acid (-0.16). Differentiation of
S. spicatum and S. murrayanum from the other species is
done on this axis. On axis 2 (36.0% of the total information), which is positively loaded with palmitoleic acid
(0.91) and oleic acid (0.44) and negatively with ximenynic
acid (-0.67), the two species containing lower amounts of
ximenynic acid namely S. lanceolatum and S. acuminatum
are well differentiated from the three species rich in ximenynic acid, S. album, S. obtusifolium and S. insulare.
Differentiation of S. murrayanum and S. spicatum from
S. lanceolatum and S. acuminatum may be explained by a
higher content of linolenic acid.
Acetylenic acids such as ximenynic acid are known to
interfere with fatty acid metabolism in a variety of tissues
[19, 20]. Therefore the seed oil of S. insulare may be a
good source of ximenynic acid for cyclo-oxygenase and
lipoxygenase enzyme studies [21].
Table 3 Comparative composition of oil and main fatty acids of various Santalum species kernel oils
Species
album
acuminatum
Ref
[11] [13] [4] [11]
Palmitic
0.8
0.8
2–2.9
Palmitoleic 0.5
0.6
0.3–2.7
Stearic
1
0.4
1.1–2.3
Oleic
12.3 18
Linoleic
Linolenic
0.8
b
c
[3] [17] [11]
0.3–1.4
0.5
0–2.5
[11]
[3]
[8]
1.3
0.3
0.4
0.6–1.3
0.1
0.7
0.6
1.7
2.7
2.1
1.2
1.8–3.3
1.9
1.9
1.0
50.6–58.7 49.1 54.4
1.8
14.3
3.2
[4] This workb
1
c
3.9–5.7
[16] [11] [3]
2.3
54.8
0.6
[12]
insulare
2.4
2
2.4
a
3
43.8–57.7 50 49.9 26
0.7
Ximenynic 82.8 79
a
lanceolatum murrayanum obtusifolium spicatum
1.3
1.4
0.7
0.9–1.7
1.0
2.4
2.3
3.2
2.4–3.8
3.5
75 32.2–46.2 40 39.8 45.5
35.5
45
71.5
3.5
0.6
1.0
0.5
1.0
27.9–37.3 40.3 33.4 36.3 34 74.5
Other minor FA identified in this species: dihydrosterculic acid 0.1, gondoic 0.3, eicosadienoic 2.5, eicosatrienoic 0.2, eicosapentaenoic 4.3
Mean of 20 samples
Presence confirmed
123
356
J Am Oil Chem Soc (2008) 85:353–356
F2
S. lanceola tum
(36.0 %)
S. a cu mi na tum
18:0
16:1n-9
S. spic at u m
16:0
0.5
18:1n-9
- 1.5
- 0.5
S . i n su la r e
0.0
18:2n-6
Ximen
F1
(48.0 %)
18:3n-3
S. m u rra ya num
S. album
S . ob t u si f ol i u m
- 1.5
Fig. 2 Two dimensional plot of the FAME profiles of Santalum
genus kernel oils investigated by PCA. The amounts of variance
relative to each axis and the FAME providing contribution are given
Acknowledgments We wish to thank the Rural Development Service (SDR) of French Polynesia for its logistical support and for its
participation in this study. We thank the Ministère de l’Ecologie et du
Développement Durable (France) for a grant for this research project
included in the ‘‘Ecosystèmes Tropicaux’’ program.
References
1. Butaud JF, Raharivelomanana P, Bianchini JP, Baron V (2003) A
new chemotype of sandalwood (Santalum insulare Bertero ex A
DC) from Marquesas Islands. J Essent Oil Res 15:323–238
2. Madhuranath MK, Manjunath BL (1938) Chemical examination
of the oil from the seeds of Santalum album (Linn). J Indian
Chem Soc 15:389–392
3. Smith CR (1970) Occurrence of unusual fatty acids in plants. In:
Progress in the chemistry of fats and other lipids, vol XI part I.
Pergamon Press, New York, pp 139–177
4. Hatt HA, Schoenfeld R (1956) Some seed fats of the Santalaceae
family. J Sci Food Agric 7:130–133
5. Hopkins CY, Chisholm MJ, Cody WJ (1969) Fatty acid components of some Santalaceae seed oils. Phytochemistry 8:161–
165
6. Spitzer V, Marx F, Maia JGS, Pfeilsticker K (1991) Curupita
tefeensis II Occurrence of acetylenic fatty acids. Fat Sci Technol
93:169
123
7. Spitzer V, De Bordignon LSA, Schenkel EP, Marx F (1994)
Identification of nine Acetylenic fatty acids, 9-hydroxystearic
acid and 9,10-epoxystearic acid in the seed oil of Jodina rhombifolia Hook and Arn (Santalaceae). J Am Oil Chem Soc
71:1343–1348
8. Sundarrao K, Jones GP, Rivett DE, Tucker DJ (1992) Occurrence
of santalbic acid in Exocarpus sparteus and Exocarpus aphyllus
seed oils. Oléagineux 47:91–92
9. Vickery JR, Whitfield FB, Ford GL, Kennet BH (1984) Ximenynic Acid in Santalum obtusifolium seed oil. J Am Oil Chem
Soc 61:890–891
10. Liu Y, Longmore RB, Kailis SG (1997) Proximate and fatty acid
composition changes in developing sandalwood (Santalum spicatum) seeds. J Sci Food Agric 75:27–30
11. Liu YD, Longmore RB, Fox JED (1996) Separation and identification of ximenynic acid isomers in the seed oil of Santalum
spicatum RBr as their 4,4-dimethyloxazoline derivatives. J Am
Oil Chem Soc 73:1729–1731
12. Liu YD, Longmore RB, Boddy MR, Fox JED (1997) Separation
and identification of triximenynin from Santalum spicatum R. Br.
J Am Oil Chem Soc 74:1269–1272
13. Rivett DE, Jones GP, Tucker DJ, Sedgley M (1984) The chemical
composition of kernels of Santalum species In: Jones GP (ed)
Proceedings of a colloquium on the food potential of seeds from
Australian native plants. Deakin University Victoria, Australia,
pp 75–92
14. Lie Ken Jie MSF, Pasha MK, Ahmad F (1996) Ultrasond-assisted
synthesis of santalbic acid and a study of Triacylglycerol species
in Santalum album (Linn) seed oil. Lipids 31:1083–1089
15. Butaud JF, Raharivelomanana P, Bianchini JP, Faure R, Gaydou
EM (2006) Leaf C-glycosylflavones from Santalum insulare
(Santalaceae). Biochem Syst Ecol 34:433–435
16. Liu Y, Longmore RB, Fox JED, Kailis SG (1995) A comparison
of kernel compositions of sandalwood (Santalum spicatum) seeds
from different Western Australian locations. Mulga Res Cent J
12:15–21
17. Jones GP, Birkett A, Sanigorski A, Sinclair AJ, Hooper HT,
Watson T, Rieger V (1994) Effect of feeding quandong (Santalum acuminatum) oil to rats on tissue lipids, hepatic cytochrome
P-450 and tissue histology. Food Chem Toxicol 32:521–525
18. Gaydou EM, Rasoarahona J, Bianchini JP (1983) A micromethod for the estimation of oil content and fatty acid composition with particular reference to cyclopropenoic acids. J Sci
Food Agric 34:1130–1136
19. Croft KD, Beilin LJ, Ford GL (1987) Differential inhibition of
thromboxane B2 and leucotriene 84 biosynthesis by two naturally
occurring acetylenic fatty acids. Biochem Biophys Acta 921:621–
624
20. Nugteren DH, Christ-Hazelhof E (1987) Naturally occurring
conjugated octadecatrienoic acids are strong inhibitors of prostaglandin biosynthesis. Prostaglandins 33:403–417
21. Liu YD, Longmore RB (1996) Effect of feeding sandalwood seed
oil on growth and SGOT activity in mice. Proc Nutrition Soc
Aust 20:182
Jean-François Butaud