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Journal of Medicine and Pharmacy, Volume 9, No.3/2019
ANALYSIS OF COMMERCIAL CAJUPUT OILS EXTRACTED FROM
MELALEUCA CAJUPUTI POWELL BY GAS CHROMATOGRAPHY
Nguyen Thi Hoai, Hoang Xuan Huyen Trang
Hue University of Medicine and Pharmacy, Hue University
Abstract
Background: Cajuput oil is a traditional and famous product of Thua Thien Hue, it is considered as a
potential product, contributing to the economic development of Thua Thien Hue. Currently, there are many
different products of cajuput oil in the market. However, the quality of cajuput oils in the production facilities
has not been considered. The aim of the present study was to apply GC to the determination of the main
components of commercial cajuput oils from M. cajuputi species in the Thua Thien Hue province. Materials
and method: The chemical composition of 10 commercial cajuput oils extracted from leaves of Melaleuca
cajuputi Powell collected in the Thua Thien Hue province was determined by gas chromatography. Results and
conclusion: The major componentsofthe oils wereα-pinene, β-pinene, limonene, 1,8-cineoleand terpineol.
The result obtained demonstrate that 1,8-cineole is the main component,present in 10 samples with values
ranging from 15 to 59%.
Key words: Cajuput oil, Melaleuca cajuputi, 1,8-cineole, α-pinene, β-pinene, limonene, terpineol.
Corresponding author: Nguyen Thi Hoai, email: hoai77@gmail.com DOI: 10.34071/jmp.2019.3.1
Received: 8/11/2018, Revised: 5/6/2019; Accepted: 12/6/2019
1. INTRODUCTION
The Melaleuca L. genus belongs to the Myrtaceae
family, and is well-known for its use in the production
of medicinal essential oils. It is widely used in folk
medicine in Southeastern Asia to relieve aches and
pains, and repel insects [6]. The Melaleuca genus
contains hundreds of individual species, all with a
myriad of oil constituents and volatile compounds
present in leaves and other parts of the plants.
Leaves and stems of several Melaleuca species are
sources of essential oils with a strong aroma that
can be used for medicinal application.
Cajuput oil is extracted from leaves of Melaleuca
cajuputi by steam distillation and can be distilled
from leaves picked throughout the year, but the oil
content is lower in the rainy season than in the dry
season. The world production of cajuput oil in 1997
is estimated to about 600 tons, two major producing
countries being Indonesia (370 tons) and Vietnam
(100 tons) [3]. In Thua Thien Hue, Cajuput tree are
planted in many places such as Phu Loc, Phong
Dien, Huong Thuy… Cajuput oil is a traditional and
famous product of Thua Thien Hue, it is considered
as a potential product, contributing to the economic
development of Thua Thien Hue. Currently, there
are many different products of cajuput oil in the
market. However, the quality of cajuput oils in the
production facilities has not been considered.
Gas chromatography (GC) is a common analytical
technique in the qualitative and quantitative assays
of volatile organic compounds, and is especially
suitable for the determination of essential oil
components. The aim of the present study was
to apply GC to the determination of the main
components of commercial cajuput oils from M.
cajuputi species inthe Thua Thien Hue province.
This will hopefully contribute to the construction ofa
quality standard for commercial cajuput oil products
and aid in increasing their economical prospects.
2. MATERIALS AND METHODS
2.1. Chemicals
Heptane (99%), α-pinene (99%), β-pinene
(99%),limonene (97%), terpineol(isomer mixture,
95%) and α-cedrene (95%) were obtained from Sigma-
Aldrich (Stockholm, Sweden) and 1,8-cineole (99%)
was obtained from Alfa Aesar (Karlsruhe, Germany).
2.2. Cajuput oil samples
Commercial cajuput oil samples extracted from
Melaleuca cajuputi were randomly collected from
10 different manufacturers in Thua Thien Hue
province and coded from S1 to S10. S1 was collected
from Hue city, S2, S5, S6, and S8 from Phong Dien
town, S3, S4, S7,and S10 from Phu Loc town, and S9
from Huong Thuy town.
2.3. Gas chromatography (GC-FID)
The cajuput oils were analysed using a Hewlett-
Packard 6890 gas chromatography equipped with
a flame ionization detector. A 30 m × 0.32 mm i.d.
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Journal of Medicine and Pharmacy, Volume 9, No.3/2019
quartz capillary column with 0.25 µm film thickness
DB-WAX stationary phase was used. The carrier gas
was helium with a flow rate of 2.5 ml/min. Diluted
sample of 1.0 µL were injected in the split (1:15)
mode. After injection, the column temperature was
programmed to rise from 100 to 230 oC at a rate of
10oC/min and was finally kept at 230 oC for 5 min.
The injector and the detector were both kept at
250 °C. Detector signals were recorded using the
Orchrom software version 1.0.
2.4. Prepared the standard, internal standard
and samples
In order to avoid pipetting errors, all substances
and heptane solution aliquots mentioned in the
following were accurately weighed with a resolution
of 0.1 mg and all concentration values were corrected
according to the actual weights thus obtained.
Stock solutions of α-pinene, β-pinene, limonene,
1,8-cineole and terpineol were prepared by
dissolving about 100 mg of each substance in 10 mL
aliquots of heptane. Seven standard solutions in the
concentration range from 0 to 10 mg/mL were then
prepared by mixing aliquots of the stock solutions
and heptane. Sample solutions were prepared by
dissolving about 100 mg of the cajuput oils in 10 mL
aliquots of heptane. A stock solution of α-cedrene
(the internal standard) was prepared by dissolving
about 100 mg in 5 mL heptane. Finally, 2 mL aliquots
of all standard solutions and all oil sample solutions
were mixed with 0.2 mL aliquots of the internal
standard stock solution, resulting in an internal
standard concentration of about 2 mg/mL in each
mixture.
3. RESULTS AND DISCUSSION
3.1. Identification of sample components
Figure 1 shows the chromatograms obtained
from a standard solution and acajuput oil sample.
The sample components were identified from their
retention times. In order to make sure that there
were no light-weight components that were hidden
by the solvent peak, a pure oil sample was injected
without the heptane solvent at a higher split rate
(1:100) using a temperature program running
from 50 to 230 oC at 10 oC/min and with the final
temperature kept for 15 min. The results showed
that there were no significant peaks eluting at lower
or higher temperatures.
Figure 1. Chromatograms of (A) 5 mg/mL standard solution and (B) cajuput oil sample S4.
Peak identities are 1 =α-pinene, 2 = β-pinene, 3 = limonene, 4 = 1,8-cineole, 5 = α-cedrene (the internal
standard), and 6,7 and 8 = terpineol isomers.
3.2. Percentage of compounds in essential oil
samples
Table 1 shows the analysis results obtained from
the cajuput oil samples. All cajuput oil samples
have the same main component patterns.An
estimate of the percentages of the major sample
componentscan be made by calculating the sum of
all peak areas in a chromatogram and dividing each
individual peak area by this sum.α-pinene, β-pinene,
limonene, 1,8-cineole, terpineol have biological
activities such asantimicrobial [1], anti-inflammatory
[5], [2], antifungial [8]. These ingredients, especially
1,8-cineole, provide the pharmacological effects of
cajuput oil.
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Table 1. Weight percentage of the major components in the essential oil samples*)
Sample α-Pinene β-Pinene Limonene 1,8-Cineole Terpineol
S1 0.4 ± 0.1 0.7 ± 0.0 4.2 ± 0.1 59.3 ± 0.4 13.1 ± 0.1
S2 2.6 ± 0.1 1.1 ± 0.1 5.3 ± 0.1 42.9 ± 0.5 9.7 ± 0.1
S3 6.5 ± 0.1 3.6 ± 0.1 2.8 ± 0.0 20.0 ± 0.2 10.9 ± 0.2
S4 5.6 ± 0.1 4.2 ± 0.1 3.0 ± 0.1 22.7 ± 0.4 11.2 ± 0.2
S5 2.8 ± 0.3 1.4 ± 0.2 5.8 ± 0.1 43.4 ± 0.1 9.0 ± 0.1
S6 2.9 ± 0.1 1.5 ± 0.1 5.4 ± 0.0 45.7 ± 0.6 9.0 ± 0.1
S7 6.1 ± 0.1 5.2 ± 0.1 3.2 ± 0.1 15.2 ± 0.0 8.9 ± 0.1
S8 2.5 ± 0.1 1.4 ± 0.1 5.9 ± 0.1 48.8 ± 0.8 11.7 ± 0.1
S9 2.6 ± 0.1 2.1 ± 0.0 4.8 ± 0.1 42.7 ± 0.3 12.9 ± 0.1
S10 8.4 ± 0.1 9.6 ± 0.1 4.4 ± 0.1 31.2 ± 0.2 10.8 ± 0.2
*) Average ± the standard deviation.
Table 2. Percentages of quantified components in 10 cajuput oil samples from Vietnam.
% min % max % average
α-pinene 0.4 ± 0.1 8.4 ± 0.1 4.1
β-pinene 0.7 ± 0.0 9.6 ± 0.1 3.1
Limonene 2.8 ± 0.0 5.9 ± 0.1 4.5
1,8-cineole 15.2 ± 0.0 59.3 ± 0.4 37.2
Terpineol 8.9 ± 0.1 13.1 ± 0.1 10.7
Table 2 summarizes the percentages of the
quantified components in the Vietnamese essential
oil samples. It can be seen that 1,8-cineole is the major
constituent of the cajuput oils, with concentrations
varying from 15% to 59%. The quality of cajuput oils
is decided from the concentration of 1,8-cineole. The
best cajuput oils have concentrations of 1,8-cineole
over 40% according to Vietnamese phamacopoeia
IV [4] and Local technical regulation Hue Cajeput oil
[7]. Based on these standards, only 6/10 samples
meet the standards. The remaining 4 samples (S3,
S4, S7 and S10) had a lower 1,8-cineole content
than the standards, especially the S7 with a very
low 1.8-cineole content of only 15%. In addition
to the 1.8-cineole, the study also quantified other
components such as α-pinene, β-pinene, limonene
and terpineol. Those are also considered as the high-
content ingredients, affecting the quality and aroma
of cajuput oil. The content of these ingredients is
also unstable in the oil samples.
From the results it can be seen that 10 cajuput
oils were collected randomly from different
locations in Thua Thien Hue province have the
same main components. However, the content
of components in the samples is not stable.
Some samples have very low content of main
components, which greatly affects the quality of
the oil. Environmental factors such as temperature,
relative humidity, soil type, total duration of sunlight
exposition and wind regime can greatly affect the
proportion of 1,8-cineole present in cajuput oils.
In addition, differences in production processes
between manufacturers of commercial cajuput oils
is a cause for variation sin the 1,8-cineole content.
The production processes includes collecting raw
materials, distilling and preserving cajuput oils
in different facilities. Each production facility has
its own procedures that lead to differences in the
quality of cajuput oil. Some samples have very low
content of components, especially 1,8-cineole, which
may be due to the crude extraction technique, the
extraction conditions are not guaranteed, resulting
in not yet extracting the components from material.
Therefore, it is necessary to have researches from
the process of planting, harvesting, extracting and
preserving for the best quality of cajuput oil. This
will ensure the quality of oil is stable, creating
prestige in the market, helping to develop products
and bringing high economic efficiency. This will help
cajuput oil become a strategic product of Thua Thien
Hue province. Our results will hopefully contribute
to make a premise to control the main components
(especially 1,8-cineole) in commercial cajuput oils
from the Thua Thien Hue province to ensure their
efficiency in use for medical treatment.
4. CONCLUSIONS
Quantitative analysis in the yield of essential oils
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Journal of Medicine and Pharmacy, Volume 9, No.3/2019
has widely ecological and economic importance.
In this study we investigated the content and
principal component of cajuput oil from Melaleuca
cajuputi, grown in Thua Thien Hue province.
Our study provides evidence that 1,8-cineole is
main component of commercial cajuput oil from
Melaleuca cajuputi species. It’s also identified the
percentage of five terpenoids including α-pinene,
β-pinene, limonene, 1,8-cineole and terpineol in
each cajuput oil sample.
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