CHEMICAL COMPOSITION CHANGES DURING STORAGE OF LEMON BASIL ESSENTIAL OIL (Ocimum citriodorum sp.)

Dwiarso Rubiyanto*#, Chairil Anwar** dan Hardjono Sastrohamidjojo**

* Department of Chemistry, Faculty of Math and Natural Sciences,Islamic University of Indonesia, Jogjakarta, Indonesia

** Department of Chemistry, Faculty of Math and Natural Sciences,Gadjah Mada University, Jogjakarta, Indonesia

# e-mail : iwdarso@yahoo.com

ABSTRACT

Difference in post harvest treatment due to the pre distill process of some essential oil resources can lead to differences in chemical compositions included Daun Kemangi (Lemon basil (Ocimum citriodorum sp.)) essential oil. Specific compounds that taking the role of major constituents in lemon basil oil and control the special aromatic scents could be identified during storage time. Minor components in lemon basil essential oil took place as contributors that couldn’t be forgotten too. The unique characteristic and manner of each major and minor substituent in the presence of specific pre distillation process can lead to providing useful information to next step due to this essential oil utility. In this research, essential oil sample from lemon basil grown in 4 x 6 m2 garden was taken from Jogjakarta, Indonesia has been produced by steam distillation (SD) method. Five different storage time were conducted before distillation process running. Variation of storage time for pre distillation process including 0 day (fresh leaf), 1 day (stale condition), 7 days (very stale), 14 days (dry condition) and 30 days (very dry). The analysis of its main components has been performed using gas chromatography- mass spectrometry (GC-MS). Chemical composition vary due to increasing time although it could be seen that some compounds were remain chemically stable during storage. Some identical compounds could be figured out from each storage time period applied and some compounds could be potentially used as “storage-finger print-compound” of each time. There were approximately 71 accumulative compounds with vary concentration could be observed in lemon basil essential oil during storage. Citral which consist of citral A and citral B were the one of the primary compound with concentration of 63.17% - 79.08%.

Keywords: lemon basil essential oil, Kemangi, composition change, storage time, Ocimum citriodorum sp.

Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

Introduction

Extensive analytical and sensory evaluation is necessary to select the plant

cultivar/chemotype, the part of the plant growth and the stage of development for

harvest that gives the best yield of the particular oil character desired. The quality and

price of some oils are based on the percentage content of a single chemical component,

so separation and measurement of individual components is very important.

Indonesia was among of few countries that had very large variety of aromatic

plants and cultivars. Some of them had been known as an established essential oil

resources but many hadn’t been developed yet. There many other plants could be

potentially germinated. Its mega-biodiversity on types and varieties of them could bring

to prosperities and benefits to the nation and citizens.

The Ocimum genus belonging to the Lamiaceae family is characterized by a

great variability of both morphology and chemo-types, the ease of crosspollination leads

to a large number of subspecies, varieties and forms, which differ in essential oil

composition and morphological characters. (Omer, et.al., 2008) Basils (Ocimum spp.,

Lamiaceae) contain a wide range of essential oils rich in mono terpenoids and phenolic

compounds (Simon et. al., 1990; Phippen and Simon, 2000). The basil family includes

about 150 species from Asia, Africa, and South America. Here are a few examples :

Ocimum basilicum (Sweet basil); Ocimum basilicum var. citriodorum (Lemon basil);

Ocimum gratissimum (East Indian basil); Ocimum basilicum var. crispum (Curly basil);

Ocimum basilicum var. minimum (Greek basil); Ocimum sanctum (Holy or sacred

basil).

Basil has culinary and ornamental value as a herbs. Fresh or dried leaves can be

used in soups, stews, and sauces; meat, fish, and egg dishes; salads; and vegetables. The

leaves are used in flavored vinegars and teas. There was such opinion that these are

herbs that are best used in their fresh state since their flavors are easily lost soon after

picking from plants. It is best to pick these herbs when they are needed for use. They

also can be processed to produce flavored oils using blended with other vegetables oil.

Cultural methods which cause changes in basil essential oil content and composition

thus have the potential to also affect the aroma and taste of the fresh product. (Johnson,

et. al., 1998)

Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

Basil essential oils production still limited because most people used in fresh or

blended conditions. As a part of Ocimum species, compared to other like basils leaf,

lemon basil essential oil was rarely found in market because most people used this fresh

herb as part of culinary to complete of food processing and as an ornamental herb. But,

this pungent-strong lemon odor and taste-leaves can produced to such an essential oil

having great chemical component and physicochemical properties that can be

converted to another chemical compounds.

One of Ocimum citriodorum (Lemon basil) variety was known as Kemangi in

Indonesia had been long time used in culinary. It’s cheap and easily founded makes

people plant them in their yard or garden because its colors and smells that natural and

fresh. It is believed that Kemangi has physiology activity in traditional healing methods

like reducing muscle tension, stomachache, headache, and giving pleasant-relaxing

scents. Using Kemangi as source of essential oil production was very potential way to

get new ingredient for some purpose.

The essential oil of Ocimum basilicum was screened for its composition and

some central nervous system activities (viz., sedative, hypnotic, anticonvulsant, local

anesthetic). When tested in mice, OB essential oil had no effect on motor activity up to

a dose of 1.2 mLkg-1 at 90 min post administration. (Ismail, 2006) People knew that

some essential oils could act as a specific chemical that responded by other species like

insects or mammals as pheromone, deterrent, attractant, and having some biologically

action or effects as antifungal, antimicrobial, and etc. Many researches aimed to find the

effective way on pest control especially using deterrent or antifeedant effect against

insects (Aerts and Mordue, 1997; Ahmad, 1993; Carrol, 1994; Cowles, et. al., 1990;

Govindacharl, 1996; Rubiyanto, 2008). So, composition of the essential oil of Daun

Kemangi should be evaluated and controlled to find out whether differences in pre

distill treatment could impact those chemical composition. It has been considered that

usually people using Daun Kemangi in freshly state. In a point of view that concerning

lemon basil oil as source of chemical substances, it is important to examine if there were

any changes of chemical composition of different time of storage in a post harvest

treatment comparing them in the fresh state.

Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

Materials and methods

Fresh samples of lemon basil (O. citriodorum) or daun Kemangi were collected from 4

x 6 m2 garden taken from specific location in Jogjakarta Indonesia which was planted in

rainy season in October 2008. This plant was harvested each time which particular

storage time would be done. The samples (leaf and stem) were cut and left to air dry in

0 day (fresh leaf), 1 day (stale condition), 7 days (very stale), 14 days (dry condition)

and 30 days (very dry). Plant materials were then subjected to distillation using semi

macro steam distillation (SD) apparatus that consist of stainless steel boiler tank (20 L

of capacity) as steam production, connected to sample Kemangi leaves container (4 to 5

kg dry sample maximum capacity). Vapor produced was passed out through water

condenser and outlet to oil collector. The oily layers obtained were separated and

purified using sodium sulphate anhydrous. Analysis of components has been carried out

using GCMS-QP2010S SHIMADZU (EI, Rtx-5MS, 30 m x 0.25 mm ID, He).

Results and discussion

Steam distillation methods were very common in means of obtaining esential

oils. Essential oils producer must take a best effort to collect essential oils as much as

they can. The key to get optimum yield of essential oil collection is to maximize water

steam penetrated to samples and discovered suitable state or condition of the samples

like fresh, stale, dry or frozen .

Even there are hundreds of Ocimum species, but mostly only 2 – 3 types that

can be found in daily life in Jogjakarta province as well as used in cullinary and dishes

or just an ornamental herbs. (Rubiyanto, 2006)

Figure 1. Left to right : Ocimum citriodorum (lemon basil), Ocimum basilicum “Canum sims” (purple basil) and Ocimum basilicum “Lime” (sweet basil)

Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

In this research, all harvesting was done after lemon basil plants reached enough age for

cropping, means between 4 – 6 months. Each plant approximately needs 2 – 3 months

after seeding to reach strong rod and leaf form completely blossomed. Leaf harvesting

should be set continuously to prevent flowering because it could lead to retard leaf

developing that affected oil content.

Post harvest treatment including mechanically (stripping, cutting, abrasive),

thermally (drying, sunning, freezing), or aging for periods would affect the oil contents.

(Gang, et.al., 2001) After storage for certain period then the lemon basil samples were

subjected to steam distillation for 3 - 4 hours. The essential oil yield and its density then

measured in each process. Data showed that lemon basil essential oil could get the

optimum yield and density (compared to basil essential oil in common with density :

0.89 – 0.90 g/mL) in 3 – 7 days of storage. Detail results as follows :

0 1 3 7 14 300.0000

0.2000

0.4000

0.6000

0.8000

1.0000

0.6818

0.5125

0.91360.8989

0.83800.8688

0.16 0.12

1.00

0.570.52

0.35

Density, g/mLYield, %

Storage days

Figure 2. Correlation between essential oil yield and density in each storage time

Two classes of compounds, terpenoids and phenylpropenes, make up the bulk of

essential oils and contribute to the particular properties of many spices and herbs. The

analysis of chemical composition of lemon basil essential oil was run on GC-MS using

Rtx-5-MS as the stationary phase and electron impact (EI) ionization.

Some compounds showed small traces (below 1%) as a characteristic in storage

periods (Table 1) which meant that there was substituent showed up in first periode of

storage ( 0 – 7 days) but the other hand, there was showed up in middle (7-14 days) or

end period (14 – 30 days). These changes could be caused by inter conversion among

substituent that be induced by oxidation or microorganism activities. Aging by leaving

lemon basil samples to dry would bring in decreasing physical form of leaf surface that

could be indicated with pale color, growing darkness and lack of humidity. Flexibility

Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

and freshness would be replaced by rough and crisp texture. This process could be

continued by changing in chemical composition of essential oil materials. Day to day

changing also showed us that some compounds remained stable and some higher

quantities of constituent could be potential for scientific reason due to explore new

sources from plants material as well as an individual compound or as an aggregate.

Table 1. Chemical composition of between periods of storage

Period, days 0 - 7 0-14 1-14 1-30 14 - 30

Compounds

Cis-geraniolBeta-

pinenoxidePhelandral Beta- Ocimene

Trans-geraniol2,6-dimethyl

heptanalAlpha-

gurjunene

Trans -3,5-dimethyl-1,6-

octadiene

Alpha.-copaene

2,6-dimethyl-1,6-octadiene

Beta-sesqui phellandrene

Neric acidAlpha-

humulene2,3-dimethyl-3-buten-2-ol

Germacrene D

Geranyl acetate

Beta-bisabolene

Alpha-cadinol

Chromatograms showed some characteristic profiles and amount of substituent

influenced by time of storage that could be associated to lemon basil essential oil

composition. Identification of compounds were obtained from mass spectra (MS)

analysis. Totals compounds in accumulation of difference storage time should be 71

compounds. Thus, those chemicals would contribute the overall characteristic of lemon

basil essential oil storage. From each chromatogram of storage condition would be

summarized that there were identical compounds found in all days of storage (Figure 4)

which most of them present in higher concentration (above 1%) and some compounds

could be called as “storage-finger print-compound” meant only specific compounds

should be discovered in particular storage time (Figure 5) although they revealed in

small quantities (below 1%). It showed 12 compounds that identically in figure 4 which

are : (a) 6-methyl-5-hepten-2-one, (b) Linalool, (c) 2,2-Dimethyl-3,4-octadienal, (d)

Trans-4,5-epoxi- caran, (e) Alpha terpineol, (f) Cis-citral, (g) Trans-citral, (h) Trans-

caryophyllene, (i) Alpha.-bergamotene, (j) (E)-beta-Farnesene, (k) Cis-alpha-

bisabolene, (l) (-)-Caryophyllene oxide

Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

72

12

2

61

33

9 0 - 7 days0 - 14 days0 - 30 hari days1 - 14 days1 - 30 days14 - 30 daysaccumulation of specific dayspread

Figure 3. Amounts of chemical compounds in each period of storage

Figure 4. Chromatographic profiles of identical components in all storage time; A:0 day, B: 1 day, C: 7 days, D: 14 days and E: 30 days.

Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

Figure 5. Specific compounds that found in individual storage time

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Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor

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Presented in the International Seminar on Essential Oil 2009, (ISEO 2009) 26th-28th Oct 2009, IPB International Convention Center, Bogor