FLAVOUR AND FRAGRANCE JOURNAL, VOL.

1,33-35 (1985)

The Leaf Oil Terpene Composition of Eastern White Pine,

Pinus strobus L.
Ernst von Rudloff
Plant Biotechnology Institute, National Research Council of Canada, Saskatoon, Sask., Canada S7N 0 W9

Analysis by an on-line computerized gas-chromatrographic (g.c.)-mass spectral method of the steam-volatile leaf
oil of eastern white pine confirmed that a- and /3-pinene, and myrcene are the main monoterpenes, with lesser
amounts of camphene, car-3-ene, /3-phellandrene, limonene, terpinolene, a-terpineol, and bornyl acetate. In
addition, germacrene-D and caryophyllene were identified as the main sesquiterpenes; lesser amounts of
humulene and 6-cadinene were also found. Small or trace amounts of hexanal, hex-2-enal, tncyclene, o-cymene,
a-phellandrene, a-terpinene, p-cymene, y-terpinene, a-fenchol, pinocamphone, terpinen-4-01, a cadinol isomer,
manoyl oxide, and a kauranol isomer were identified. Germacrene-D tended to rearrange to other sesquiterpenes
on all but highly deactivated g.c. columns.

KEY WORDS Leaf oil Eastern White Pine Pinus strobus L. a- and b-pinene Myrcene Germacrene-D
Caryophyllene

INTRODUCIION decomposition of at least one major component.

Eastern white pine, Pinus sfrobus L.,*occurs over
a wide range in the northeastern United States
and southeastern Canada.'-3 Whereas some gene-
tic variability was referred to by Fowells,' no var-
ieties are known to occur within this range. Chis-
man and Lylos4 described a dwarf form, and a
variety of eastern white pine is found in Mexico
and Guatemala.'
Hanover' compared the oleoresin of eastern
white pine with that of western white pine, P.
monficolu Dougl., and reported on the mono-
terpene composition of the extracted needle oils.
The cortical and xylem monoterpenes of eastern
white pine were investigated by Thor et af.6 and
Wilkinson,' and Gerhold and Plant' analyzed the
monoterpene variation of the vapours of the
foliage of white pines and hybrids thereof. In our
chemosystematic studies of North American con-
ifers as based on leaf oil terpene analysis,' we
compared the steam-distilled leaf oil composi-
tions of pines (10 trees of each) of the subsection
haploxylon (white pines) and noted similarities in
the monoterpene patterns, but differences in
those of the sesquiterpenes. When we attempted
isolation and identification of the sesquiterpenes
of eastern white pine by the methods then in
use,9-" we encountered rearrangement and
*See also ref. 18. NRCC No. 24763.
We have now analyzed the leaf oil of another ten
eastern white pine trees from Ottawa, Ontario, by
more modern methods, including the on-line
computerized gas-chromatographic (g.c.)-mass
spectral (m.s.) method used in our more recent
chemosystematic s t ~ d i e s ' ~ , ' ~ .
EXPERIMENTAL
The foliage of ten 20-30 year-old eastern white
pine trees from Ottawa, Ontario, was sampled at
random from the crowns to give 200-300g of
leaves per tree. The foliage was collected in early
winter and transported in the dark and cold, and
stored at - 20 "Cuntil steam-distillation could be
carried out. The isolation of the steam-volatile oil,
as well as the g.c. and g.c.-m.s. methods for quan-
titative and qualitative analysis were described
previously. ''-I3 The instruments used were model
5750A Hewlett-Packard gas chromatographs
equipped with dual thermal conductivit detec-
tors and various packed columns, lo-' using
washed and deactivated Supelcoport (80-90
mesh) as solid support, and a model 4000 Fin-
nigan g.c.-m.s. unit, fitted with a 30 m high-
performance glass capillary column coated with
SP 2100 silicon polymer. The individual compo-
nents of the oil were identified by comparison of

0882-5734/85/010033-03$01.oo Received I9 March 1985

0 1985 by John Wiley & Sons, Ltd. Accepted 13 May 198.7
34 TERPENE COMPOSITION OF EASTERN WHITE PINE

Table 1. The ranges and means of the leaf oil terpene percentages
of ten eastern white pine trees from Ottawa, Ontario

Terpene % Range Mean %

Tricyclene 0.1- 0.6 0.3

a-Pinene 21.2-57.8 33.1
Camphene 1.7- 4.6 2.5
8-Pinene 15.8-35.5 26.3
Myrcene 5.8-16.2 13.3
Car-3-ene 0.4- 3.0 1.1
/3-Phellandrene 1.7- 5.3 2.7
Limonene 1.2- 2.4 1.6
Terpinolene 0.5- 1.1 0.7
a-Fenchol trace- 0.3 0.1
Terinen-4-01 trace- 0.3 0.2
a-Terpineol 0.2- 2.7 1.7
Bornyl acetate 0.2- 2.1 1.4
8-Caryophyllene 1.9- 5.4 3.8
Humulene 0.2- 1.1 0.7
Germacrene-D 5.0-19.6 9.1
6-Cadinene trace- 0.7 0.3
Cadinol isomer trace- 0.9 0.4
Manoyl oxide trace- 0.4 0.2
Kauranol isomer 0.1- 0.6 0.3
Other’ 0.2- 0.4 0.2

% Yield of Oil? 0.1- 0.4 0.2

~~ ~ ~~~~~~~ ~~

*Trace components (0-0.1 %): o-cymene. a-phellandrene,

a-terpinene, p-cymene, y-terpinene, unidentified (mot. wt. 152),
pinocamphone, sabinol (?), umbellulone (?), 8-cadinene or
muurolene. The non-terpenoid artifacts hexanal (trace-0%) and
hex-2-enal (trace-0.3%) are not included in the percentage compo-
sition calculations.
t Based on fresh weight of leaves

retention times (tR) and m.s. of reference com-
pounds and by computerized comparison with the
Finnigan m.s. 1ibra1-y.’~The major terpenes iden-
tified are listed in Table 1 according to the t on
the SP 2100 glass capillary column. The identity
of germacrene-D,15*’6 which has a m.s. similar to
some cadinene-muurolene isomers, was con-
firmed by comparison of the f R and m.s. of the
main sesquiterpene hydrocarbon of the oil of
young Douglas-fir leaves, which Kepner et a[.”
have shown to be germacrene-D. The quantita-
tive composition (relative percentages) was
obtained by summation of the m.s. ion counts for
each g.c. peak. This agreed well with the means
obtained on the four packed columns, using digi-
tal peak integration with a Hewlett-Packard
model 3354 Lab Data system.
RESULTS AND DISCUSSION
Table 1 lists the identified terpenes, the mean
percentages, as well as the percentage ranges as
recorded for the ten eastern white pine trees. The
monoterpenes, including their relative amounts,
are in good agreement with previous findings,’,’
except that the tree-to-tree variability was higher
in the trees sampled in this study. The fact that
Hanover’ obtained his data with only 5 g leaf
samples indicates that within-tree variation is
small. Whereas a- and /I-pinene appear to be the
main leaf oil constituents for the pines of the
haploxylon, significant amounts of myrcene,
P-caryophyllene, and germacrene-D are charac-
teristic for the eastern white pine.
Of the few minor or trace components that
could not be positively identified (see Table l),
the cadinol isomer had a m.s. similar to that of
torreyol, and the diterpene alcohol to kauranol.
The unidentified monoterpene, mol. wt. 152, and
pinocamphone may be autoxidation products,
since they were absent in freshly distilled samples;
in such samples o-and p-cymene were present
only in trace amounts. The non-terpenoid artefact
hex-2-enal found in most conifer leaf oils as a
result of cutting the leaves’ was present in less
 ERNST VON RUDLOFF
than 0.3% amounts, which is low for a pine
species. Hexanal was found in trace amounts.
Germacrene-D is known to rearrange to
cadinen-muurolene isomers under acidic condi-
tions and to /?-bourbonene in the presence of
light.” Hence the rearrangements to more com-
plex sesquiterpene mixtures that we encountered
with older g.c. columns using washed Chromop
sorb G as solid support’ are not unexpected.
Work with washed and neutralized, as well as
deactivated solid supports or capillary columns is
essential for a reliable analysis. The possibility for
similar chemical changes during steam-distillation
exists as well if the macerated plant material is not
neutralized prior to distillation.
Acknowledgements-The supply of the pine foliage by Dr. L.
W. Argus, and the excellent assistance in the g.c.-m.s. analyses
by L. R. Hogge and D. Olson are gratefully acknowledged.
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