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Effect of Drying on the Chemical Composition of the

Egyptian Cupressus macrocarpa (Hartw.ex Gordon)
Essential Oils and their Biological Characteristics
a

Ahmed H. El-Ghorab , Khaled F. El-Massry & Hamdy A. Shaaban

Flavor and Aroma Dept. National research Center , Egypt

Published online: 12 Mar 2013.

To cite this article: Ahmed H. El-Ghorab , Khaled F. El-Massry & Hamdy A. Shaaban (2007) Effect of Drying on the Chemical
Composition of the Egyptian Cupressus macrocarpa (Hartw.ex Gordon) Essential Oils and their Biological Characteristics,
Journal of Essential Oil Bearing Plants, 10:5, 399-411, DOI: 10.1080/0972060X.2007.10643573
To link to this article: http://dx.doi.org/10.1080/0972060X.2007.10643573

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Jeobp 10 (5) 2007 pp 399 - 411

399

ISSN 0972-060X

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Effect of Drying on the Chemical Composition of the Egyptian

Cupressus macrocarpa (Hartw.ex Gordon) Essential Oils and
their Biological Characteristics
Ahmed H. El-Ghorab, Khaled F. El-Massry* and Hamdy A. Shaaban
Flavor and Aroma Dept. National Research Center, Egypt
Received 25 February 2007; accepted in revised form 25 September 2007

Abstract: The essential oils of Cupressus macrocarpa (gold crest) grown on the north coast
of Egypt were subjected to analysis using gas chromatography-mass spectrometry (GC - MS). The
chemical compositions of essential oils of fresh and dry leaves were found to contain 43 components.
The main compounds in the volatile oils of fresh and dried leaves were neral (31 - 35%), hydroxy
citronellal (12 - 16%), geraniol (3 - 4%), piperitol (trans) (7 - 8 %), isobornyl isobutrate (0.7 - 6.61 %),
linalool (0.6 - 5.21 %), terpinyl acetate (0.10 - 3.27 %), myrcene (0.22 - 2.60 %), trans- ferrugiol (0.3 2.25%), abitol (0.4 - 2.18%) and eugenol dihydro (0.1 - 1.3%).
The volatile oils (fresh and dried leaves) and non- volatile extracts (EthOH and DCM) of c.
macrocarpa were further investigated for radical scavenging activities using 1.1-diphenylpicryl-2hydrazyl (DPPH) assay. Obviously, the volatile oils remarkably reduce DPPH radicals as well as
Eth.OH and DCM extracts, compared with BHA.
The antimicrobial efficacy of the c. macrocarpa volatile oils and non-volatile extracts when
examined by disc diffusion test showed that the DCM significantly inhibited the growth of all the
tested microorganisms. The antioxidant and antimicrobial activities could be due to the presence of
active compounds as neral, graniol, eugenol dihydro, carvacrol acetate and phenol (2,6-di methoxy).
The present study support the view that the characteristic volatile oil and non-volatile extracts
of the c. macrocarpa were highly bio available and could be suitable for using as antioxidant, antimicrobial and flavoring agent in the food industry.

Key words: Cupressus macrocarpa, antimicrobial activity, antioxidant activity, DPPH,

hydroxy citronellal and neral.
Introduction: Plant volatile oils have been well known since antiquity as possessing
biological activities. Chief amongst these are their antibacterial, antifungal and antioxidant
properties 1. Many authors in fact, have reported antimicrobial, antifungal, antioxidant and
radical-scavenging properties of spices and essential oils and, in some cases; a direct foodrelated application has been tested 2,3.
*Corresponding author (Khaled F. El-Massry)
E- mail: < kfarouk@yahoo.com >

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400

Recently, consumers and food manufacturers seek out products with all natural labels 4. Consequently, a lot of emphasis has been given to identification and incorporation of
novel natural antioxidants in food products. The area of natural antioxidants has developed
enormously in the past decade, mainly because of the increasing limitations on the use of
synthetic antioxidants and enhanced public awareness of health issues 5. The antioxidative
properties of common aromatic plants have been systematically investigated 6 and numerous
studies have been devoted to plant sources, not only for their possible utilization as antioxidative extracts, but also to look for new naturally efficient antioxidants. Several antioxidants
from plant leaves have been characterized and some of them have proved to be as good as
synthetic ones (BHA and BHT) 7.
Until recently, essential oils have been studied most from the view point of their flavor
and fragrance chemistry only for flavoring foods, drinks and other goods. Nowadays, however, essential oils and their components are gaining increasing interest because of their
relatively safe status, their wide acceptance by consumers, and their exploitation for potential multi-purpose functional use 8,9.
The literature outlines different approaches within this trend and both the biological
screening of new essential oils and the evaluation of new properties of already marketed oils
have been done. In both cases, different methodological approaches lead to scattered results, which are hardly comparable and often conflicting 10-13.
The cupressaceae family includes about 140 native species from moderate climate or
worm regions of both hemispheres. The genus Cupressus belongs to the Cupressoides
subfamily which contains about 12 native species from North America, Asia and Europe 14.
The genus Cupressus is thought to be represented by more than 20 species 15 of North
Africa, Asia, North America, and Mexico. Its geographic area is limited to the Northern
Hemisphere. This number is different to the Farjons classification, which lists 17 species 16.
Cupressus genus classification is not easy; thus, intergenetic and interspecific complexity is
important.
Several contributions have been reported on the chemical composition of volatile oils
from leaves of different Cupressus species as well as their biological activities 17,18. Sacchetti
et al. 19 found that the essential oil of Cupressus sempervirens showed antimicrobial activity against five food-spoilage yeasts. Antimicrobial activity have been also reported for essential oils of C. arizonica oil and C. torulosa 20,21.
To our knowledge, there is no information about the chemical composition, antioxidant
activity as well as the antimicrobial activity of leaf essential oil of Cupressus macrocarpa
(gold crest) grown in Egypt. Thus, the present study was planned to investigate the chemical
composition of the volatile oil of C. macrocarpa (Gold crest) grown in the north coast of
Egypt. Also, antimicrobial and antioxidant activities of volatile and non-volatile extracts of C.
macrocarpa were determined.
Materials and Methods
Materials: The C. macrocarpa plants were grown at the north coast of Egypt.
Butylated hydroxytoluene (BHA) was purchased from Sigma Chemical Co. (St. Louis, MO),
and dichloromethane (DCM) and ethanol (Et.OH) were from Fisher Scientific Co., Ltd.

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(Fair Lawn, NJ). 1,1-diphenyl picrylhydrazyl (DPPH) was purchased from TCI America
(Portland, OR, USA). All other solvents were from Aldrich (Germany).
Bacteria and fungi cultures were provided by the department of Microbiology, National research centre, Giza, Egypt.
Extraction of essential oil: Extraction of essential oil of C. macrocarpa was carried out by the steam distillation followed by liquid-liquid continuous extraction according to
the following method:
Fresh and dried leaves of C. macrocarpa (300 g each) was placed in a 3 L roundbottom flask with 1 L of deionized water. The solution was steam-distilled for 4 h. Each
distillate (900 mL) was extracted with 100 mL of DCM using a liquid-liquid continuous
extractor for 6 h. After that, each extract had been dried over anhydrous sodium sulfate and
the solvent was removed in a rotary flash evaporator (BUCHI 461 SWITZERLAND). The
distillation was stopped when the volume of extract was reduced to approximately 1 mL.
Then the solvent was further removed under a purified nitrogen stream until the volume was
reduced to 0.5 mL 22. Data represent the means + SD (The extraction method was carried
out in triplicate).
Extraction of non-volatiles: Dried and fresh leaves (200 g each) were extracted
with DCM (500 mL) and EtOH (500 mL) at room temperature for 24h. Each extract was
filtered and concentrated in a rotary evaporator under reduced pressure at approximately
500C and then stored in dark containers at -50C untill used.
Antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH): Radical scavenging activity of C. macrocarpa different extracts against stable DPPH radical was determined spectrophotometrically. When DPPH reacts with an antioxidant compound, which
can donate hydrogen, it is reduced. The changes in color (from deep violet to light yellow)
were measured at 517 nm on a UV/visible light spectrophotometer. Radical scavenging
activity of extracts was measured according to Miliauskas et al. 23 as described below.
Stock solutions of extracts (volatile oils, DCM and EtOH extracts) were prepared by
dissolving 0.1 g of each extract in 10 ml of methanol. The solution of DPPH in methanol
(6 x 10-5 M) was prepared daily, before UV/visible light spectrophotometer measurements.
Various concentrations of each extract (50, 100, 200 and 500 g/mL) were added to solutions (1 ml) of DPPH in methanol. The mixtures were shaken vigorously and left to stand at
room temperature for 30 min; the absorbances of the resulting solutions were measured
spectrophotometrically at 517 nm. In this assay, the percentage of DPPH reduction by
different extracts of C. macrocarpa was compared to that of BHA.
The experiment was carried out in triplicate. Radical scavenging activity was calculated by the following formula:
% Inhibition = [(AB - AA) / AB ] X 100
where: AB absorption of blank sample (t = 0 min);
AA absorption of tested extract solution (t = 30 min).

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402

Chromatographic analysis (GC, GC-MS): A Hewlett Packard model 6890 gas

chromatograph equipped with a DB5 capillary column (30 m x 0.25 mm i.d. x 0.25 m df.),
FID detector was used. Analysis of volatile components was carried out under the following
conditions: the injector temperature was 2000C and the detector temperature was 2500C.
The column was programmed from 350C to 2200C at 30C / min. and held for 40 min. The
helium carrier gas flow rate was 29 cm/sec. Injections were in the splitless mode.
An HP model 6890 GC interfaced to an HP 5791A mass selective detector (GC-MS)
was used for mass spectral identification of the GC components at MS ionization voltage of
70 eV. A 30 m x 0.25 mm i.d. (df = 0.25m) DB-5 bonded-phase fused-silica capillary
column (J&W Scientific) was used for GC. The linear velocity of the helium carrier gas was
30 cm/s. The injector and the detector temperatures were 2500C. The oven temperature
was programmed from 35 to 2200C at 30C /min and held for 40 min.
Kovats indices were determined by co-injection of the sample with a solution containing homologous series of n-hydrocarbons (C8-C26) under the same conditions as described
above. The separated components were identified by matching with NIST mass-spectral
library data, and by comparison of Kovats indices with those of authentic components and
with published data 24. The quantitative determination was carried out based on peak area
integration.
Microbiological Activity
Test organisms: Pure cultures of the bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli) and fungi (Aspergillus niger, Aspergillus parasiticus
and Candida albicans) used for the testing antimicrobial activity were provided by the
department of Microbiology, National Research Center (NRC), Dokki, Giza, Egypt.
Inocula preparation: Mother cultures of each microorganism were set up 24 h before the assays in order to reach the stationary phase of growth. The tests were assessed by
inoculating Petri dishes from the mother cultures with proper sterile media, with the aim of
obtaining the microorganism concentration of 105 colony forming units (cfu/ml)
Antibacterial and antifungal activity testing: The microbial activity was tested
using disc-diffusion method. Whatman NO.1 filter paper discs, (5mm) in diameter, were
soaked with tested essential oil and non-volatile extracts (10 l), the discs were placed on
the surface of the cold solid medium in Petri dishes, inoculated with tested organisms. Sterile
trypticase soy agar (TSA) medium inoculated with the tested bacterial organisms is used
and potato dextrose agar (PDA) were inoculated with the tested bacteria and fungi, respectively. Incubation at 50C for one hour was carried out to permit good diffusion and then
transferred to an incubator at the temperatures and times as follows: for bacteria at 370Cfor
24 hours, for yeast at 270C for 48h. and for mould at 270C for 72h.25. After the incubation
period, the degree of growth inhibition was evaluated and compared with the growth inhibition results form the controls (gentamycin sulphate and nystatin for bacteria and fungi, respectively). Each assay was performed in triplicates on three independent experimental
runs.

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403

Determination of Minimum Inhibitory Concentration (MIC): The minimum inhibitory concentration (MIC) is defined as the minimum level of essential oil or extract
concentration that produces about 90% reduction in the growth (populations) of microbial
colonies. The MIC was determined by the microdilution agar plate method 26 according to
the following:
Ninety mL of tempered TSA were agitated vigorously with essential oil or extract to
achieve the following final oil or extract concentrations: 3.0, 2.8, 2.5, 2.2, 2.0, 1.5, 1.0, 0.6,
0.5, 0.25, 0.15, 0.10, 0.05 and 0.025 ml./100 mL. Approximately 15 mL of these mixtures
were transferred along with 1 mL of microorganism inoculum to agar plates. The plates
were incubated for 24h at 37 0C for bacteria, and 48h at 27 0C for yeast and for 72h at 27 0C
for mold. Afterwards, the number of colonies in each plate was determined. Each assay was
performed in triplicates on three independent experimental runs.
Results and Discussion
Effect of drying on the chemical composition of the essential oil: The yields of
the essential oils of C. macrocarpa were 0.5 + 0.07, 0.75 + 0.04 and 0.83 + 0.03% for
fresh leaves (FL), freeze dried leaves (FDL) and dried leaves (DL), respectively. The volatile oil of C. macrocarpa leaves (gold crest), known in Egypt as Lemon Sarw, was obtained
by hydrodistillation followed by solvent-solvent extraction. The volatile oils obtained were
subjected to analysis using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The volatile constituents of the different oils were identified by their
retention indices and mass spectrum in comparison with those of standard synthetic compounds. The results of the chemical analysis are presented in table 1.
About 43 compounds representing 99.7% of the essential oil were identified. GC and
GC-MS analysis revealed that the major constituents of the extracted oils (FL, FDL and DL)
were: neral (31- 35%), hydroxy citronellal (12- 16%), geraniol (3 - 4%), piperitol (trans) (7
- 8 %), isobornyl isobutrate (0.7 - 6.61 %), linalool (0.6 - 5.21 %), terpinyl acetate (0.10 3.27 %), stemon (0.21- 3.5 %), menthene (3-P) (1.93 - 3.27 %), menthene (1-P) (0.3 2.89%), vertocitral (cis) (0.19 - 2.70 %) myrcene (0.22 - 2.60 %), trans-ferrugiol (0.3 2.25%), abitol (0.4 - 2.18%), sabinene (0.2 - 1.93%), eugenol dihydro (0.1 - 1.3%) and allyl
hexanoate (0.18 -1.21%) as listed in table 1.
To best of our knowledge, there are no reports on the biological activities of our tested
plants, grown in the north coast of Egypt. However, there are some reports related to the
essential oil composition of C. macrocarpa grown in different parts of the world 18.
The effect of drying on the chemical composition of plants and vegetables has been
previously studied 27. These studies show that the drying method is an important factor for
the changes in the concentrations of volatile compounds.
Regarding the effect of drying by different methods on the cupressus macrocarpa
(gold crest) volatile oil compositions; it was found that the volatiles in all samples (FL, FDL
and DL) were similar regarding the qualitative pattern but displayed slightly quantitative
differences. In fresh, dried at room temperature and freeze dried C. macrocarpa gold crest
leaves oils, neral was the most abundant compound by margin 33.18, 35.78 and 31.34 %
(based on area percent) for all samples respectively (table 1).

404

In order to obtain a more systematic overview about the effect of drying method on
the chemical composition of the leaves oil, the analyzed compounds were grouped into compound classes and their yields (based on area percent) (Fig. 1).
The yields of the monoterpene hydrocarbon fraction were almost the same in the FL
extract (7.9 %) and the FDL extract (6.09 %) while the extracted oil dried at room temperature (DL), showed a significant increase (13.52 %).
Monoterpenes (M) are considerably higher in DL oil at room temperature in comparison with FDL and this mostly due to the higher concentration of p-cymenene (4.01% 0.33%), camphene (3.51% - 0.22%) and menthene (3-p) (3.27% - 2%), respectively. The
results revealed that the remarkable variation in the concentration of some compounds could
be ascribed to the effect drying method. The amount of sesquiterpenes (S) was low in all
extracts (1.48, 0.92 and 1.14% for fresh leaves, dried leaves at room temperature and
freeze dried), respectively. The amount of light oxygenated components (LOC) showed
non-significant variation in all the extracted oils (Table 1 and Fig. 1). The total yield of the
heavy oxygenated components (HOC) in the fresh (9.38%) and freeze dried oils (13.63 %)
is considerably higher in comparison with the extracted oil at room temperature (DL) (5.44%)
(Fig.1).
90
80
Fresh leaves oil
Dry leaves oil at room temperature
Freez dried leaves

70
60
50
40

Area %

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Khaled F. El-Massry et al. / Jeobp 10 (5) 2007 pp 399 - 411

30
20
10
0
Heavy oxygenated Light oxygenated
compounds
compounds

Sesquterpene

Monoterpene

Figure (1). Effect of drying on the main chemical classes in the essential oils of the
Cupressus macrocarpa leaves
Volatile aroma compounds are the most sensitive components in the process of food
drying. Huopalahti et al. 27 found that the reduction of the flavor extract after drying of dill
herb was significant (from 1.7 to 3.9 times in the case of freeze-drying, and from 6.7 to 11.2

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405

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times in the case of air-drying.

However, some authors have found the changes in aroma and concentration of volatile constituents during drying to be less considerable, others demonstrated the complexity of
the effect of drying on the equilibrium between aroma compounds in the essential oil storing
particles of the plant and head space vapours 28.
Cupressus macrocarpa (gold crest) leaves volatile oils has a characteristic flavor
due to the presence of many components (hydroxy citronellal, geraniol and neral) with highly
sensorial properties at a lower threshold. The flavor compounds of cupressus macrocarpa
(gold crest) consist mainly of light oxygenated compounds (77.42 - 79.81 %) for all extracted oils.
Antioxidant activity of essential oil: Natural extracts are in increasing demand
from the manufacturers of foods, cosmetics and pharmaceuticals. Thus the importance of
conducing studies on essential oils, lies not only in the chemical characterization but also in
the possibility of linking the chemical contents with particular functional properties.
In this regard, it is advisable to use methods for the assessment of biological activities
that not only highlight aromatic or preservative activities but also correlate with functional
properties potentially useful for pharmaceuticals, nutriceuticals and cosmetic applications.
Following this idea, we have used a convergent approach that has taken into account the use
of complementary methods to assess radical-scavenging and antioxidant properties (one of
them is DPPH assay) which are a very important for health claims in nutriceutical products11 .
The essential oil and various extracts were subjected to screening for their possible
antioxidant activity by DPPH free radical scavenging system.
The DPPH radical-scavenging activities of the C. macrocarpa (gold crest) (grown in
the north coast of Egypt) essential oils (FL, DL at room temperature and FDL) as well as its
non-volatile extracts (dichloromethane; DCM and ethanol ; EtOH extracts) and of synthetic
antioxidants (BHA) are shown in Fig. 2.
Fresh leaves (FL), dried at room temperature and freeze dried Cupressus essential
oils notably reduced the concentration of DPPH free radical, with an efficacy slightly lower
than that of BHA (87 + 0.32%). Their values, in fact, ranged from 68 0.53%, 75 + 0.31 and
75.7 + 0.28 % inhibition, respectively.
The free radical scavenging activity of ethanol extract (EtOH) was slightly higher in
activity than other extracts (Fig.2).
From the above results, it seems to be clear that the effect of drying method on the
antioxidant activity of the volatile oils of cuperssus macrocarpa (gold crest) significantly
not affected. This could be attributed to the presence of higher percentage of biologically
active components as light oxygenated compounds specially those with phenolic nucleus as
geraniol, neral, carvacrol acetate, phenol 2.6 dimethoxy, and eugenol dihydro. Otherwise, the
slightly higher activity of freez dried C. macrocarpa essential oil due to the higher concentration of neral (31.43%) and geraniol (3.33%) (Table1).
Our results are consistent with Choi et al. 29, who found that neral and geraniol (citral
isomers) are the most abundant compounds in Cuperssus citratus essential oil and have

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406

Fresh leaves (FL)

Dry leaves at room temperature (DL)

Inhibition %

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Freez dried leaves (FDL)

Essetial oil

Ethanol extract

Dichloro methane
extract

BHA

Figure 2. Inhibitory effect of the volatile oils of Cupressus macrocarpa (gold crest)
leaves samples measured by the DPPH assay
remarkable radical-scavenging efficacy. This result is in agreement with the poor performance given by other oils with similar patterns and by single monoterpenic hydrocarbons 12.
Overall results were better than those provided by the radical-scavenging activity and
some of the oils with high terpenic percentages were more effective, probably as a consequence of a higher specificity of the assay for lypophilic compounds.
Antioxidants minimize the oxidation of lipid components in cell membranes or inhibit
the volatile organic compounds and the conjugated diene hydroperoxides arising from linoleic acid oxidation that are known to be carcinogenic. Especially polar extracts exhibited
stronger activity than non-polar ones. Similar and moderate activities of non-polar extracts
can be attributed to the presence of several types of compounds belonging to different
classes, such as oleoresins in hexane extract 30, sterols and their derivatives, flavones and
flavonoids in dichloromethane extract (DCM) 31. Due to their complex nature, more detailed
studies employing bioassay-guided fractionation would be worthwhile to explore the nature
of active extracts prepared.

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407

Antimicrobial activities: As can be seen in Table 2, the volatile oil of C. macrocarpa

(gold crest) grown in the north coast of Egypt and its different non-volatile extracts showed
considerable antimicrobial activities. The DCM extract of dry (at room temperature), DCM
extract of fresh and EtOH extract of dry cupressus macrocarpa showed greater inhibition
level against tested microorganisms than essential oil of dry, fresh and EtOH extract of fresh
plant. The antimicrobial activity of the cupressus sp. oil could be due to the presence of
active compounds with phenolic, alcoholic and aldehydic nature such as neral (31.43 - 35.78%),
geraniol (3.11- 4.01%) and hydroxyl citronellal (12.59 - 16.59%). Our results are consistent
with those of Sacchetti et al.19, who reported that yeasts and fungi are markedly inhibited by
essential oils extracted from different cupressus sp. rich in active compounds as aldehyde,
ketones, phenolics etc. The MIC values for Cupressus macrocarpa non-volatile extracts
and essential oil were presented in Table (3). DCM extract of dry cupressus macrocarpa
(at room temperature) showed the least MIC value (0.55 - 0.7 ml/100ml), while the essential
oil from fresh plant showed the higher MIC value (0.75 - 0.85 ml/100 ml). Sacchetti et al., 19
studied MIC values for essential oil of other species as Cupressus sempervirens and found
that concentration between 0.6 and 0.8 ml/100ml were microbiocidal.
Obviously, the present investigation reinforcement that the volatile oils and non-volatile extracts of the Egyptian Cupressus macrocarpa (gold crest) were highly bioavailable.
Volatile oil have a characteristic flavor due to the presence of many components with strong
sensory properties at a low threshold, such as neral, geraniol and hydroxy citronellal. So, it
could be suitable for using as multifunctional flavoring compound with antioxidant and antimicrobial efficacies in the food and pharmaceutical industries after its safety appraisal which
is under investigation.

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Table (1). Effect of drying on the chemical composition of essential oil of
Cupressus macrocarpa (gold crest) leaves

No. KI

1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17

887
888
965
951
954
979
991
996
1025
1079
1085
1091
1093
1097
1099
1127
1131

Area
%
FL
0.14
0.39
0.17
0.32
0.21
1.93
1.93
0.28
2.89
0.19
1.02
0.32
0.52
0.39
5.21
0.24
0.14

Area
%
DL FDL

Compounds

Method of Identification

0.14
0.10
0.26
2.29
3.51
0.41
3.27
0.22
1.96
0.38
1.21
0.79
4.01
0.33
0.23
0.21
0.12

Santene
Pentanal (5-hydroxy)
Benzaldehyde
Octen-3-ol (-1-)
Camphene
Sabinene
Menthene (3-P)
Myrcene
Menthene (1-P)
Vertocitral (cis)
Allyl hexanoate
Camphenilone
p-Cymenene
-Pinene oxide
Linalool
Stemone
Ethyl hexanoic acid

(M)
(LOC)
(LOC)
(LOC)
(M)
(M)
(M)
(M)
(M)
(LOC)
(LOC)
(LOC)
(M)
(LOC)
(LOC)
(LOC)
(LOC)

0.40
0.15
0.16
0.15
0.22
0.22
2.00
2.60
0.32
2.70
0.18
1.68
0.33
0.85
0.64
3.54
0.17

KI & MS
KI & MS
KI & MS
KI & MS
KI & MS & AU
KI & MS
KI & MS
KI & MS & AU
KI & MS
KI & MS
KI & MS
KI & MS
KI & MS
KI & MS
KI & MS & AU
KI & MS
MS

Khaled F. El-Massry et al. / Jeobp 10 (5) 2007 pp 399 - 411

410

table 1. (continued)

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No. KI

Area
%
FL

Area
%
DL FDL

18
19
20
21
22
23
24

1209 7.89 7.90 8.32

1245 33.18 35.78 31.43
1255 3.11 4.01 3.33
1284 13.55 16.59 12.59
1297 0.16 0.36 0.50
1325 0.74 0.57 0.62
1346 1.06 2.60 0.53

25
26
27
28
29
30
31
32
33

1347
1351
1364
1368
1369
1374
1377
1428
1462

0.99
1.48
1.30
0.27
0.92
1.08
0.22
6.61
0.29

0.49
0.10
1.01
2.97
0.11
0.45
0.53
0.21
0.11

3.97
4.34
0.15
0.39
0.23
0.31
0.21
0.72
0.23

34 1473
35 1478

0.14
0.15

3.35
0.17

0.17
0.26

36
37
38
39
40
41
42
43

0.16
0.19
0.16
3.08
0.16
0.96
2.25
2.18

0.14
0.11
0.14
0.12
0.21
0.60
0.37
0.40

0.31
0.44
5.58
0.17
0.21
0.18
4.93
1.85

1479
2084
2128
2210
2287
2308
2328
2394

Compounds

Method of Identification

Piperitol (trans)
(LOC)
KI & MS
Neral
(LOC)
KI & MS
Geraniol
(LOC)
KI & MS
Hydroxy Citronellal
(LOC)
KI & MS
Vanilin (ortho)
(LOC)
KI & MS
Limonene aldehyde
(LOC)
KI & MS
Acetophenone
(LOC)
MS
(4-methoxy)
Phenol (2,6- dimethoxy) (LOC)
MS
Terpinyl acetate (a-)
(LOC)
KI & MS
Eugenol dihydro
(LOC)
KI & MS
Furfuryl hexanoate
(LOC)
KI & MS
Cyclosativene
(S)
KI & MS
Carvacrol acetate
(LOC)
KI & MS
-Copaene
(S)
KI & MS
Isobornyl isobutrate
(LOC)
KI & MS
4-Methyl-1,2,3,4-tetra- (HOC)
MS
hydrodibenzofuran-1-carboxylic Acid
Isobornyl n-butyrate
(HOC)
MS
Ionone (6-methyl(S)
KI & MS
gama-E-)
Menthyl lactate
(HOC)
MS
Abietadiene
(S)
KI & MS
Nezukol
(HOC)
KI & MS
Phyllocxladanol
(HOC)
KI & MS
Abietal (4-epi)
(HOC)
KI & MS
Isopimarol
(HOC)
KI & MS
trans-Ferruginol
(HOC)
KI & MS
Abietol
(HOC)
KI & MS

& AU
& AU
& AU
& AU

& AU

MS: tentatively identification by comparison with data obtained from NIST mass spectra
library.
AU: confirmed by comparison with mass spetrum of authentic compounds.
KI: confirmed by comparison with kovat index, FL (fresh leaves).
DL: (dry leaves at room temperature), FDL (freez dried leaves).
LOC: light oxygenated compounds, HOC: heavy oxygenated compounds.
S: sesquterpene, M: monoterpene.

19
22
23
20
25
23
35
*

Staphylococcus
aureus

20
25
26
22
28
26
40
*

Pseudomonas
aeruginosa

12
15
16
13
18
17
29
*

22
23
23
22
26
23
*
38

Escherichia Aspergillus
coli
niger
Zone of inhibition (mm)
16
18
19
18
22
20
*
29

Aspergillus
parasiticus

15
20
21
19
23
22
*
30

Candida
albicans

Staphylococcus
aureus

0.7
0.75
0.65
0.7
0.6
0.65

Extract assayed

DL oil
FL oil
EthOH DL
EthOH FL
DCM extract DL
DCM extract FL

0.65
0.75
0.6
0.7
0.55
0.6

Pseudomonas
aeruginosa

0.75
0.8
0.75
0.8
0.65
0.7

0.7
0.75
0.65
0.75
0.6
0.65

0.75
0.8
0.7
0.75
0.7
0.7

Escherichia Aspergillus Aspergillus

coli
niger
parasiticus
(MIC) values (ml/100 ml)

0.75
0.85
0.7
0.8
0.7
0.7

Candida
albicans

Table (3): Values of minimum inhibitory concentration (MIC) of volatile and non-volatile extracts of
Cupressus macrocarpa leaves

* not assayed.

FL oil
DL oil
Ethanolic extract DL
Ethanolic extract FL
DCM extract DL
DCM extract FL
Gentamycin sulphate (40mg/ml)
Nystatin (50-IU)

Extract assayed

Table (2): Antimicrobial activities of volatile and non-volatile extracts of Cupressus macrocarpa

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