ESSENTIAL OILS FROM SPICES GROWN IN ALBERTA.

ANISE OIL (PIMPINELLA ANISUM)

M. B. EMBONGT, D. HADZIyEV, and S. MOLNAR
Department of Food Science, University of Alberta, Edmonton, Aha. T6G 2N2 and
Alberta Horticultural Research Centre, Brooks, Alta. T}J 0J0.
Received I3 Sept. 1976, accepted 30 Dec. 1976.
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EnrnoNc, M. B., Hnozryev, D. aNo Mor_ren, S. l9'lj. Essential oils from

spices grown in Alberta. Anise oil (Pimpinella anisum). Can. J. plant Sci. 57:
681-688.

The annual plant anise (Pimpinella anisum L.), grown in Alberta over five
consecutive seasons, was rated as medium for its herb vigor and growth, and the
yield and quality of its essential oil was determined. The average yield of
steam-distilled oil, calculated on a dry weight basis, was 1.05-l .lvo for seed and
1.2-l.35vo for the whole crushed plant. The number of identified constituents, as
determined by using a Tl-Gl-chromatography method, was l6 for the plant, and | 2
for the seed oil, with corresponding percentages ofi4.7 and 84.6, respectively. The
onfy major oil constituent was trans-anethole which was 57 .4va of whole plant and
75.2Vo of seed oil. The other constituents of plant oil, present in amounts of I-5Vo
were crs-anethole, carvone, B-caryophyllene, dihydrocarvyl acetate, estragole and
limonene. compositional data obtained for world market oils, which are used for
For personal use only.

flavoring by domestic food industries, suggested that the Alberta anise oils are of
sufficient quality to substitute for the imported oils.

L'6tude porte sur I'analyse quantitative et qualitative de I'huile essentielle extraite

de I'anis vert (Pimpinella anisum L.) cultiv6 en Alberta pendant cinq campagnes
cons6cutives. calcul6 sur la matibre sdche, le rendement moyen d'huile obtenu par
distillation h la vapeur s'est situ6 entre 1.05 et l.lvo pour la graine et entre 1.2 et
l.35vo pour la plante entibre broy6e. A I'analyse combin6e paichromatographie sur
couche mince et en phase gazeuse, l6 compos6s ont 6t6 identifi6s pour l'huile de
plante entibre et l2 pour I'huile extraite de la graine, repr6sentant respectivement
74.7 et84.6vo de la composition totale des huiles. Le seul compos6 majeur de I'huile
6tait le trans-an1,thole qui constitue 57.4vo de I'huile de plante entibre et75.2vo de
I'huile de la graine. Les autres composanrs de I'huile de la plante, comptant pour
l-SVo du total, 6taient le cis-an6thole, le carvone, le B.caryoptryllane, le
dihydrocarvyl acltate, l'estragole et le limonbme. si I'on compare ces donn6es avec
la composition connue des huiles vendues.sur le march6 mondial, il semble que
l.'huile d'anis produite en Alberta soit d'assez bonne qualit6 pour remplacer
6ventuellement les huiles import6es.

The food industry is constantly seeking new
methods of improving or redesigning its
products for both gustatory and financial
reasons. This has created a lucrative market
for spices, and their oleoresins and essential
oils for use by producers of soft drinks,
rhesent address (M.B.E.): Universiti Kebangsaan
Malaysia, Kuala Lumpur, Malaysia.
Can. J. Plmt Sci. 57: 6tf -688 (July 1977)
681
baked goods, desserts, confections, meats,
and alcoholic beverages. Currently, all of
these flavoring products, including the
essential oils from the whole plant or seed
of anise, are imported into Canada.,
There are two botanically different plants
whose fruits yield essential oils of similar
color, flavor, and, hence, chemical com-
position. Star anise oit is obtained from
Illicium verum Hooker (family Mag-
 682 CANADIAN JOURNAL OF PLANT SCIENCE
noliaceae), and the other oil, which is true
anise oil and has a finer and more delicate
flavor. is distilled from anise fruit, Pim-
pinetla anisum L. (family Umbelliferae).
This true anise is an annual plant indigenous
to the Middle East, but is widely grown in
Europe, the United States, China, and
Chile.
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The yield of oil from commercial dry
anise seed of various origins ranges from
1.9 to 3. l%o, with an average of 2.29Vo by
weight (Fisher et al. 1945). The quality and
the yield ofoils distilled from central, first-,
and second-order umbels usually differs. A
high yield of fine quality oil is obtained
when the seeds in the central umbels are
fully ripe (Tsvetkov 1970).
The quality of anise oil depends mainly
on the amoun t of trans -anethole. The higher
the amount of this constituent, the more
delicate and sweet the odor and flavor of the
oil (Guenther 1950).
For personal use only.
A detailed study relating to anise oil
composition, quality, and yield for Canada
or for other countries does not exist. A few
years ago (1972) the Agriculture Canada
Research Branch at Morden, Manitoba, and
the Provincial Horticultural Research
Center at Brooks, Alberta, began investiga-
tions to determine whether herbs and spices
can be successfully grown under Canadian
soil and climatic conditions. This study
reports the chemically assessed quality of
the essential oil of true anise grown in
Alberta, and its comparison with similar
oils used by the food industry and obtained
on the world market.
MATERIALS AND METHODS
(a) Plant Material
The certified seeds were supplied by Di Giorgi
Co. Inc., Council Bluffs, Iowa. During l97l-
1975, the annual herbaceous plant growing
season was from l6 May to 30 September. At the
time of harvest, usually after 2O September, the
schizocarp fruits, with two fused carpels separat-
ing the two one-seeded mericarps, were col-
lected with the pedicels and carpophores. After
air-drying for I day, the fruits, which were
grayish green to grayish brown in color, were
separated. ground in a Wiley laboratory mill,
and immediately steam-distilled.
Histological examination of the pericarp
within the mericarp showed many small dorsal
oilsecretory canals, i.e., vittae, and two large
commissural vittae. The convex dorsal side had
five light-colored longitudinal ridges and
numerous short hairs. The seed endosperm and
the embryo contained small globules of oil as
well as the usual aleurone grains' Also, the cross
section of pedicels and carpophores revealed the
existence of two vittae.
The plants were cultivated at Brooks in
southern Alberta in a sandy loam with available
nutrients in kg/ha of N:27, P:69, and K:565, and
with a soil pH of 7.9. The sulfate content was 75
ppm, while the, organic matter and free lime
iontents were low. The average weather data for
a growing season were: precipitation 14.02 cm'
hours of sunshine I ,203 ; except for the middle of
May and the second half of September' the
period was frost free. Irrigation was applied four
times per season.
Steam distillation was conducted in the field.
The oil collected was separated from the water,
dried over anhydrous sodium sulfate, and then
stored at 4 C until analYzed.
(b) Chemical Analysis
The oil composition was determined by thin-
layer chromatography, by gas-liquid chroma-
tography, alone and coupled with mass spec-
trometry, and by infrared spectrophotometry.
TuIN-Lnven cHRoMATocRApnv (TLC). A slurry
of 25 g of MN-Kieselgel N (without binder) and
60 ml water was spread on five 2O x 2o-cm
plates at a thickness of 300 p. The plates were
dried at room temperature for I h and then
activated at ll0 C for 1.5 h. After cooling to
room temperature in a desiccator, the plates
were ready to use. In qualitative assay, the
undiluted oil and its associated liquid standards
were spotted 1.5 cm from the plate edge, while
solid standards were dissolved at high concentra-
tion in ethyl acetate. The plates were developed
for 40 min with a solvent system of benzene:
ethyl acetate, 95:5 (v/v). Spots were visualized
by spraying with a solution of lVo vanillin in
conc. sulfuiic acid, followed by heating at 105 C
for l0 min. The Rf values and the colors of the
spots were used to determine the identity of the
compounds (Lawrence 1968; Stahl and Jork
1969; von Zacsko-Szitszand Sz6sz 1965).
 EMBONG ET
In preparative TLC work, a strip of oil was
spread on the plate. In addition to the usual
vanillin-sulfuric acid visualization, several
plates were sprayed with a solution oi,f O.5Eo
2,7-dichlorofluorescein in ethyl acetate saturated
with water, and the bands were viewed under
UV light. The positions of these component
bands were traced on transparent paper for
reference purposes. All the bands thuslocated
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were scraped from the plate, and the components
were extracted from the adsorbent by vigorous
shaking in ethyl acetate. The mixture was
centrifuged, then the supernatant was collected
and concentrated in a stream of nitroqen. Two or
three preparative plates *"r. n...rriry to obtain
sufficient quantities of the components for
furlher analysis.
Gns-rrquro cHRoMATocRApHv (GLC). A vol-
umeof I pl of the oil was injected into a Bendix
Model 2500 gas chromatograph (Bendix Instr.
Div., Ronceverte, W.Va.) fitted with two slass
cofumns.6 fr x l/8 inch i.d.. packed with l5To
ethylene glucol succinate (EGS) polymer on
For personal use only.
Chromosorb P, AW of particle size lOOl l2O
mesh. The operating conditions were nitrogen
flow rate, 60-ml/min, iniector temperature. 220
C, FID detector temperature, 240 i, and.oiu.n
temperature programmed from 50 to 195 C at
4"lmin, with the maximum temperature held for
15 min to get the final peaks. The chromato-
graph was operated in dual column mode.
Quantitation of the peaks was done by triangula-
tion with an accuracy equal to or better than
0.50k.
In qualitative work, each standard compound
used was dissolved in ethyl acetate and co-
injected with 0. I pl of the oil on both fresh and
aged columns. An aged column was one that had t
been used for more than 4 mo.
INrneneo specrnopHbroMErRy. Spectra of neat
oil and liquid standards were recorded on a
Perkin Elmer Model 2l IR-spectrophotometer.
The spectra for solid compounds were obtained
by using KBr pellets containing about lOVo of
the component.
Mess sppctneL ANALysls. Mass spectral record-
ings.
were done by injecting 0.3 pl of oil inro a
vanan Aerograph 1400 gas-liquid
graph fitted with
chromato_
a coiled stainless steel column,
6 ft. x 3l16 inch i.d. Conditions were as siven
above. This GL-chromatograph was coupled
with an MS-12 mass spectrometer (Assoc. Elec.
Ind., Manchester, U.K.).
AL.-ANISE OIL 683
Pure compounds used as references were:
anise acetone (99.OEo), and anisic aldehyde
(987o), supplied by Haarmann & Reimer Gmbh
(Hofzminden, Germany); estragole (4-allyl-
anisol, 98.lEo), /rdr?r-anethole (USP, m.p.
2l-23 C), anisyl alcohol (m.p. 23-25 C), and
p-anisic acid (99.9Vo, m.p. t83.0 C), obtained
from Aldrich Chem. Co., Milwaukee. Wisc.:
terpene hydrocarbons and alcohols were from K
& K Labs Inc., Plainview, N.Y., while other
chemicals were from Eastman Org. Chem.,
Rochester, N.Y., or Merck AG. Darmstadt.
Germany.
The commercial high quality anise seed oil
from Michigan was supplied by Kalsec Interna-
tional, Kalamazoo, Mich.
RESULTS
The yields of steam-distilled Alberta anise
oil were 1.05-1.l%o for the seed and
1.20-1.35Vo for the whole crushed plant,
calculated on a dry weight basis. The
composition of seed oil as found bv TLC is
illustrated in Fig. | . Thirteen spors were
obtained in Tl-chromatograms of anise oil
from Alberta as well as from Michigan. The
difference between the oils from these two
origins was shown by the size and color
intensity of these spots. Spots 13, 10, 9 and
6 of anise oil from Alberta were more
intense in color and larger in size than those
AN ISE OIL
A ti.on"n"
/
\-/ ^b'ffi",P.oo*
cugonol @"
C Q'o
Se onisc oldehydc
8t
1-lon iryl
Vo c clon e
B;
Se
p-oniric ocid
Ei
Fig. l Thin-layer chromatogram of anise seed
oil and some of its associated pure compounds.
 684 CANADIAN JOURNAL OF PLANT SCIENCE

from Michigan. The opposite was true for

spots | , 4, 5 and 7. There were two major
spots, 12 and 13, for oil from Alberta, but
only one, number 12,for Michigan oil. The
spots of anise oil were tentatively identified
by running a chromatogram of anise oil and
its associated standards and comparing the
Rf values and colors. Additional clarifica-
Can. J. Plant Sci. Downloaded from www.nrcresearchpress.com by 36.70.101.130 on 12/30/19

Fig. 2. Gas-liquid chromatogram of anise

tion of spot identities was obtained by
isolation of individual bands after prepara-
tive TLC, followed by their analysis using
the combination of GLC and mass spec-
trometry. All these data are presented in
Table l.
From Fig. I and Table I it can be
concluded that the major oil constituents are
located in spots numbered from l0 to 13.
Spot 10 contained camphor and eugenol.
There were five constituents in spot I l, the
major one being peak 63 (see Fig. 2,
Gl-chromatogram). The other constituents
For personal use only.
wfiole plant oil on the aged column' For
separatiron conditions see Materials and
Mithods. The peaks of several trace constituents
are not presented. Peaks identified: o-pinene
(l), c-phellandrene (4), myrcene (5), limonene
(6), fenchone (14), camphor (18), linalool (2O),
B-caryophyllene (28), estragole (29)' dihy-
drocarvyl acetate (33), cis-anethole (34), cat-
vone (j5), trans-anethole (38), p-anisic acid
(44), anisyl alcohol (46), eugenol (51)' and
anisyl acetone (64).

were peaks 57, thujone, and 58, p-cresol'
Thujone was found more towards the upper
part of the spot. Spot 12 was the most
intense in size and color. and contained six
constituents, trans-anethole and estragole
being the major ones. Constituent 34,
cis-anethole, was located in the upper part
of the spot, and constituent 54 primarily in
the lower part. Spot number 13 was highly
heterogeneous, and contained all the ter-
pene hydrocarbons' In all, 34 constituents
were located in this spot, limonene'
B-caryophyllene and p-cymene being the
major ones.

Table I . Thin-layer chromatography data of anise whole plant or seed oils from Alberta

Tentative identity heparative TLC-GLC-MS

Spotno. Rfvalue ofthespotf data

I 0.02 p-anisic acid p-anisic acid

2 0.0s
J 0.12
4 0.14 Anisyl acetone Anisyl acetone + two minor constituents,
t6 & r7t
5 0. l8 Bomeol Borneol
6 o.2r Linalool Linalool
7 o.24
8 0.32 Carvone and p-anisaldehyde Carvone, p-anisaldehYde
9 0.35 Fenchone Fenchone
l0 0.40 Camphor, eugenol Camphor, eugenol
ll o.47 Thujone Thujone, constituent 63, 57, 58 &p-cresol
t2 0.61 frans-anethole, esfagole rans-anethole, estragole, anisyl alcohol,
constituents 54,50 & 34 (cis anethole)
13 o.72 Terpene hydrocarbons : limonene, Limonene, B-caryophylleneP-cymene +
myrcene, B-caryophyllene & 1 I minor and 20 trace constituents

P-cymene

tBased on Rf values ofpwe reference compounds.

(see Fig. 2).
*The numbers refer to the peak numbers asiigned on a Gl-chromatogram of the oil
 EMBONG ET AL.-ANISE OIL 685

The Gl-chromatogram on an aged col-

umn of anise whole plant oil from Alberta is AN r5E Ott
presented in Fig. 2. The major peak was 38
(trans-anethole), and the medium peaks
were 6
(limonene), 29 (estragole), and
unidentified peaks 32, 37, and 63. Myrcene
(5) and limonene (6) were not resolved, nor
were B-caryophyllene (28) and estragole
(29). Peak 7 was slightly separated from
Can. J. Plant Sci. Downloaded from www.nrcresearchpress.com by 36.70.101.130 on 12/30/19

limonene (6). Peaks 32, 33 (dihydrocarvyl

acetate), 34 (cis-anethole), 35 (carvone),
36, 37 , and 38 (trans-anethole) were
bunched closely together. The separation of
the constituents of seed oil from Michigan
on the aged column is illustrated in Fig. 3.
In this oil the medium peaks were
(limonene), and 44 (p-anisic acid).
63 and 64 were not found in Michiean oil.
The aged column gave peaks thit were
sharper in shape than those for the fresh
column.
The separation of the constituents on the
For personal use only.
Fig. 3. Gas-liquid chromatogram of anise seed
oil from Michigan on the aged column. For peak
identities see Fig. 2.
The identity of the peaks in the above
6 chromatograms was established by co-
Peaks chromatography using pure compounds as
references, and by analysis of mass spectral
data. Some of the latter data are presented
in Table 2.
The infrared spectra of anise seed and
whole plant oils from Alberta and Michigan

fresh column took a longer time to run than
on the aged column. Also, the sequence of
elution of the components was different
from that on the aged column. Myrcene was
separateb from limonene, and B-caryo-
phyllene from estragole. However, es-
tragole and dihydrocarvyl acetate were
eluted as one peak and were not well
separated from peak 37. Peak 7 was eluted
together with limonene. Cis-anethole, car-
vone, peak 36, and trans-anethole were
well separated on this column.
oil were typical for anise seed oil, and were
highly complex (Fig. a). The recordings
were dominated by the spectra of trans-
anethole, the major constituent, and other
benzene derivatives. The bands in these
spectra could be associated with those
typical of pure t ran s - anethole, cls - anethole,
eugenol, p-anisaldehyde, p-cymene,
thymol, anisyl alcohol, estragole, camphor,
linalool, and safrole. Due to their complex-
ity, the spectra could not be used as a means
of distinguishing the better quality oil.

Table 2. Mass spectral data of some major constituents of anise seed oil from Alberta

GLC peak Molecular Base Identity of

no. ion (M+) peak mief the compound

38 148 148 147 tt7 r33 7'7 12r rO5 9r trans-anethole

34 148 148 147 133 ll7 105 l2t 9t ll5 cis-anethole
29 148 148 r47 r2r 1t7 t33 7'7 105 9l Esfragole
46 138 138 LOg t37 121 7'7 94 r07 79 Anisyl alcohol
44 t52 t52 r35 77 10'1 92 63 t36 p-anisic acid
64 178 161 t76 133 1'15 162 ll8 44 77 Anisyl acetone
5l 1@ 164 149 57 7',1 148 103 136 Eugenol
6 136 68 93 67 94 136 79 t2r rW Limonene
35 IJU 82 64 M rO8 135 54 93 r07 Carvone
28 204 93 69 133 9l 4r '79 8r l2r B-caryophyllene

lln decreasins order of relative abundance

 686 CANADIAN JOURNAL OF PLANT SCIENCE
1{@ 126
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wavEtfNclH (MrcRN)
FREOUEMY
2@ r€@ r@ t/m t&
2Ji3Lrdibn1213r.15
WAVELNG]N
Fig. 4. Infrared spectra of anise seed oil from (a) Alberta, and (b) Michigan.
The percentage compositions of anise
For personal use only.
whole plant and seed oils from Alberta and
the high quality seed oil from Michigan are
given in Table 3. The number of identified
constituents was 16 for the whole plant and
12 for the seed oil from Alberta, and 15 for
the Michigan oil. the corresponding percen-
tages of these groups of known constituents
being 7 4.7 , 84.6, and 90.5 . The number of
unidentified constituents for Alberta whole
plant and seed oils, and Michigan seed oil
were 23, ll and 19, respectively, with
corresponding percentages of 25.3, 15.5
and 9.7 . The least number of trace con-
stituents was 15 in whole plant oil and l0 in
seed oil from Alberta, and 7 in Michigan
oil.
Also listed in Table 3 are the percentages
of total monoterpene hydrocarbons, al-
cohols, and ketones. The sesquiterpene
hydrocarbons were present only in small
amounts. In contrast to the small amount of
these terpene constituents, anise oil con-
tained high amounts of benzene-related
constituents. The most important and only
major oil constituent was trans-anethole,
which amounted to 57.4Vo of the whole
plant and 75.2Vo of the seed oil from
nm
I@I)
(dIqN)
Alberta, and 72.2Vo of the Michigan
seed
oil.
DISCUSSION
The quality of anise oil depends rpainly on
the amount of trans-anethole. Arctander
(1969) stated that trans-anethole is the
compound which imparts a very sweet
herbaceous warm odor and a sweet taste to
the oil. The amount of this compound in
essential oils of high quality anise seed has
been reported to be high, and to range
between 80 and 907o (Guenther 1950). The
Egyptian anise oil was reported by El-Deeb
et al. (1962) to contain 88.6Vo anethole.
Tsvetkov (1970) found an anethole content
of 87.3Vo in Bulgarian anise seed oil
obtained from the central umbels.
In this study, it
was found that the
trans-anethole content of Alberta anise oil
from the whole crushed plant was an
average of lSVo lower than the content in
seed oil. When the oils from seed are
compared, the Alberta oil quality is higher
than that from Michigan. This is also the
case with respect to the low cls-anethole
content found in oils from Alberta, l.lVo
vs. 2.5Vo from Michigan.
 EMBONG ET AL.-ANISE OIL 687

Estragole, an isomer of anethole, has a results obtained in this study showed that
sweet herbaceous anise-fennel odor, much anisyl acetone was not present in Michigan
more herbaceous than anethole, but without anise oil, while Alberta seed oil contained
the sweet flavor of anethole. Tsvetkov close to l%o , and the whole plant oil, O.3Vo.
(1970) reported that the estragole content of When the combined amounts of all these
the central, first-, and second-order umbels constituents are taken into consideration. it
was 8.6, 7.5, and 7.'l%o, respectively. In is obvious that anise seed oil from Alberta is
this study it was found that Alberta oils of higher quality than the imported oil from
contained a higher amount of estragole,
Can. J. Plant Sci. Downloaded from www.nrcresearchpress.com by 36.70.101.130 on 12/30/19

Michigan. This conclusion suggests that the

4.5%io in the whole plant, and 4.OVo in the Alberta oil. if available in commercial
seed, than the l.UVo in Michiean seed oil. amounts, could easily replace the imported
Hence, this component adds tlo the quality oils which are currently used by the
of Alberta oil. domestic food industry. However, the
Anisyl acetone gives the oil an intensive average yield of anise seed oil from Alberta
sweet floral and slightly fruity, cherry- is l.l%o, which should be considered as
preserve-like odor of good tenacity. The low. Therefore, any feasibility study on

Table 3. The composition in percent ofanise seed oil

Alberta
Retention
time Whole Cornmercial
Constituent (rrazs-anethole :
For personal use only.

1.00)f plant Seed oil

,razs-anethole 1.00 57.4 75.2 72.2
cri-anethole 0.91 1.8 l.l 2,5
Anisyl alcohol t.23 0.2 0.4 0.9
Anisyl acetone 2.13 0.3 no
p-anisic acid r.20 0.2 0.4 4.3
Camphor 0.69 0.3 0.1 o.2
Carvone 0.93 2.1 0.3 2.1
B-caryophyllene 0.84 1.2 0.8 o.2
Dihydrocarvyl-acetate 0_91 1.4 o.9 o.4
Estragole 0.84 4.5 4.0 1.0
Eugenol 1 .33 0.1 o.2 0.6
Fenchone 0.58 0.0 0.0 0.5
Limonene 0.34 4.0 0.0 4.9
Linalool o.71 0.3 0.2 o.2
Myrcene 0.33 o.2 0.0
a-phellandrene 0.3 1 0.4 0.0 0.3
o-pinene 0.19 0.1 0.0 0. I
No. of identified constituents l6 l2 15
Conesponding 7o 74.71 84.56 90.48
No. of constituents unidentified 23 ll t9
Corresponding 7o 25-29 15.50 9.72
Minimum no. of constituents present
in traces 15 l0 7
Monoterpene hydrocarbons 6.24 0.30 6.39
Monoterpene alcohols 0.35 0. l8 0.18
Monoterpene ketones 2.43 0.50 2.-t5
Sesquiterpene hydrocarbons 1.22 0.84 o.23
Benzene-related constituents 64.59 82.16 8r.57
The results are an average offour growing seasons (1971-1974), and did not vary significantly between the growing
seasons.
tGLC separation data as on Fig. 2.
 688 CANADIAN JOURNAL OF PLANT SCIENCE

commercial production of oil must addi-
tionally involve the economical and agro-
logical aspects of production specifi-
- and per
cally, the yield of oil per plant
hectare. Nevertheless, the aim of this study
was achieved. since it has shown that a
good quality anise oil can be obtained even
with the short season and rather cool
Can. J. Plant Sci. Downloaded from www.nrcresearchpress.com by 36.70.101.130 on 12/30/19
raphy. J. Pharm. Sci. United Arab Republic 3:
63-78.
FISHER, L., TORNOW, P. A. and PROPER'
B . L. 1945 . The content and physical properties
of certain volatile oils. Bull. Natl. Formulary
Comm. 13: 6-10. (Chem. Abstr. 1945, 39:
t962).
GUENTHER, E. 1950. The essential oils. Vol'
IV. D. Van Nostrand Co., Inc', New York,

climatic conditions in Alberta. N.Y.pp. 563-570.

LAWRENCE, B. M. 1968. Thin laYer
ACKNOWLEDGMENTS chromatography. Part I. A review of the use of
This work was supported by a grant from the thin layer chromatography in essential oil
Alberta Agricultural Research Trust. The assis- analysis. Perfum. Essent. Oil Rec' 59:421-432.
tance of L. Steele in chemical analysis, and of T. STAHL, E. and JORK, H. 1969. TerPene
Leishman in field distillation of the oil is derivatives. essential oils, balsams and resins.
gratefully acknowledged. M. B. Embong was a Pages 206-238 in E. Stahl, ed. Thin layer
recipient of a Canadian Commonwealth Fellow- chromatography. 2nd ed. Springer Yerlag Inc.'
ship. New York, N.Y.
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