Method Development for Analysis of Essential Oils Authenticity using

Gas Chromatography-Mass Spectrometry (GC-MS)

Novi Nur Aidha

, Retno Yunilawati

and Irma Rumondang

Badan Penelitian dan Pengembangan Industri, Kementerian Perindustrian

Keywords: Essential Oil, Authenticity, GC-MS, Chemical Component.

Abstract: Essential oils widely used as fragrances and flavours in the food and cosmetics industry and also for the

medical and pharmaceutical fields for various effects. The demand increasing of essential oil caused the cases

of adulteration that affect the authenticity of essential oil. The authenticity is important in ensuring the quality

of essential oil. This study was aimed to analyse the authenticity of essential oil use Gas Chromatography-

Mass Spectrometry (GC-MS) by determining its chemical component. The experiment included repeatability,

accuracy, and limit of detection. GC performed on HP-5MS capillary column operated in 60⁰C-240⁰C

temperature programs. This method successfully applied to all types of essential oil with limit detection of

clove oil was 0.02 ppm, citronella oil was 0.033 ppm, patchouli oil was 0.005 ppm, and lemongrass was 0.016

ppm. All types of essential oil also have good repeatability and accuracy with these methods. This study will

facilitate the scientific community by enhancing the efficient method for essential oil.

1 INTRODUCTION

Essential oils are widely used as fragrances and flavor

in the food and cosmetics industry, and in the medical

and pharmaceutical fields for various effects

(Mohamed et al., 2018; Wany et al., 2013). The purity

of essential oils is very important in their use in

various fields. The demand increasing of essential oil

caused the cases of adulteration that affect the

authenticity of essential oil. The adulteration occurs

because of the prices for natural extracts higher than

those of synthetic materials. Adulteration also is

intended to gain volume or weight to get a higher

profit. Adulteration essential oils in various ways are

by mixing it using cheaper essential (Do et al., 2015),

add compound isolate or synthesis- dilution with inert

material (Ng et al., 2015; Ke et al., 2015; Schipilliti et

al., 2010) or add with other oil include nutmeg oil

contaminant with castor oil (Yunilawati et al., 2013),

lemongrass oil identified kerosene or coconut oil as

adulterants (Do et al., 2015) and sandalwood oil

diluted with cedarwood oil (Howes et al., 2004). The

adulteration can degrade the quality and can lead to

safety issues, health hazards, or noncompliance with

the natural grade.

The authenticity is important in ensuring the

quality of essential oil. Authenticity can be defined as

free from adulteration in the sense of absence of

foreign matter, but it also suggests free from

impurities. Control methods and standardization of

essential oils are required to check compliance with

the standards of quality. Many analytical techniques

to analysis the authenticity of essential oil including

isotope-ratio mass spectrometry (IRMS) (Schipilliti

et al., 2010), nuclear magnetic resonance

spectroscopy (NMR) (Cerceau et al., 2016), high-

performance thin-layer chromatography (HPTLC)

(Cerceau et al., 2016), high- performance liquid

chromatography (HPLC) (Gaonkar et al., 2016) and

gas chromatography (GC) (Esfahanizadeh et al.,

2018; Abualhasan et al., 2017; Beale et al., 2017;

Athar et al., 2013; Heuskin et al., 2009; Howes et al.,

2004; Shellie et al., 2002). GC is the analytical

technique for identification with controlled

conditions and can be directly coupled to a mass

spectrometer (MS) if information other than

fingerprint is needed. Each type of essential oil has

GC-MS qualitative fingerprint (Hu et al., 2006),

which compared with the literature. Therefore, GC-

MS has become a part of the routine testing for

essential oil and commonly used for detecting

adulteration of essential oil. Analysis using GC is

very profitable and efficient because easy, faster

separation, need short time, low cost, has sensitivity

and good detection limit for volatile compound

Nur Aidha, N., Yunilawati, R. and Rumondang, I.

Method Development for Analysis of Essential Oils Authenticity using Gas Chromatography-Mass Spectrometry (GC-MS).

DOI: 10.5220/0009956000410046

In Proceedings of the 2nd International Conference of Essential Oils (ICEO 2019), pages 41-46

ISBN: 978-989-758-456-5

(Vargas Jentzsch, 2019; Al-Rubaye et al., 2017;

Chauhan, 2014).

This study was aimed to analyse the authenticity

of essential oil use Gas Chromatography-Mass

Spectrometry (GC-MS) by determining its chemical

component. The essential oil used are patchouli,

citronella, clove, massoia, and lemongrass oil, and the

experiment included repeatability, accuracy, and

limit of detection. The first experiment is validating

the methods to ensure that it has reproducible and

reliable results, and the results can be used to assess

the quality, reliability, and consistency of the results

of the analysis.

2 MATERIALS AND METHODS

2.1 Materials

Essential oils were used in this experiment are

patchouli, citronella, clove, massoia, and lemongrass

were obtained from Indonesia. The standard of

essential oil was used are patchouli, citronella, and

clove from France. The solvent was used is methanol

(Merck).

2.2 Equipment

Gas chromatography with a mass spectrometer

detector (GCMS) Agilent 6890 series with capillary

column HP-5MS, 30 m x 0.25 mm id x 0.25 µm film

thickness. Helium gas was used as the carrier gas at a

constant pressure of 65 kPa. The essential oil was

injected with a volume of 1 µL in a split ratio of 1:25

and a solvent delay of 2 minutes. The increasing oven

temperature was programmed from 60-240°C with a

step of 3°C per minute until reaching 240°C.

2.3 Methods

2.3.1 Repeatability

1 µl of the essential oil (patchouli, citronella, clove,

massoia, and lemongrass) was diluted in 1 ml

methanol, then injected of 1 µl into GC-MS.

Repeatability is done by injecting essential oils 7 in

times.

2.3.2 Accuracy

1 µl of the essential oil was diluted in 1 ml methanol

(patchouli, citronella, clove oil from Indonesia as a

sample, and patchouli, citronella, clove oil from

France as a standard), then injected of 1 µl into GC-

MS. The chromatogram data were compared between

the essential oil from Indonesia and the standard.

2.3.3 Limit of Detection

The detection limit is done by injecting essential oils

with various concentrations. Variable concentrations

of clove, citronella, lemongrass and patchouli oil

were made are 0.1 ppm; 0.05 ppm; 0.033 ppm; 0.025

ppm; 0.02 ppm and 0.016 ppm by diluted 1 µl of the

essential oil into methanol (10 ml; 20 ml; 30 ml; 40

ml; 50 ml and 60 ml). Especially for patchouli oil,

various concentration was also made to 0.005 ppm.

The limit of detection was determined based on the

lowest concentration that can be detected by the

instrument. That concentration was observing the

height of the major component in essential oil.

3 RESULT AND DISCUSSION

3.1 Methods Developments for

Analysis of Essential Oil using

GC-MS

The development methods for the analysis of the

essential oil using GC-MS can be used to identify

chemical compounds in essential oils, regardless of

the type of essential oil, and also to analyze the

authenticity of essential oils. The essential oils were

used are patchouli, citronella, clove, lemongrass, and

massoia oil because these are the major of essential

oil produced in Indonesia (Ministry of Trade

Republic of Indonesia, 2011). In this study, the

method was created and optimized internally in the

previous experiment. The analyze using GC Agilent

6890 with HP-5MS column and the performance of

condition programs of GC-MS have been optimized

and verified as have done by Cardoso et al. (2018) and

Athar et al. (2013). The method validating included

repeatability, accuracy, and limit of detection of the

essential oil similar with the previous method were

reported by Cardoso et al. (2018); Esfahanizadeh et

al. (2018); Abualhasan et al. (2017); Chauhan (2014);

Athar et al. (2013).

3.2 Repeatability

The repeatability experiments were established in

order to evaluate the methods' trueness and precision,

respectively. The repeatability was determined by the

analytical procedure under normal conditions using

seven (7) repetition on the same day (intraday

precision). The precision of the development method

ICEO 2019 - 2nd International Conference of Essential Oil Indonesia

using GC-MS has established by comparing each

chromatogram of the essential oil, and it was

considered the peak profile of this compound. Figure

1. showed that all of the essential oil has good

repeatability. Compared with an earlier study which

is reported repeatability using GC-MS with triplicate

(Esfahanizadeh et al., 2018; Athar et al. 2013) and six

replicate (Cardoso et al., 2018).

(a)

(b)

(c )

(d)

(e )

Figure 1: Repeatability of clove oil (a); patchouli oil (b); citronella oil (c); lemongrass (d) and massoia oil (e).

Method Development for Analysis of Essential Oils Authenticity using Gas Chromatography-Mass Spectrometry (GC-MS)

3.2 Accuracy

Accuracy was determined by comparing the

chromatogram between essential oil from Indonesia

with essential oil from France as standard (clove,

patchouli, and citronella oil). Figure 2 showed that the

essentials oil has good accuracy. The methods

showed the spectra of Indonesian essential oil

matched with the standard. The spectra have similar

major components in each essential oil, although

there is a difference in the high area of the spectra

between Indonesian essential oils and standard. The

variation of components and its concentration of

essential oil depend on the type of regions. The main

constituent in clove oil is eugenol; in patchouli oil is

patchouli alcohol; and in citronella oil are citronellal,

citronellal, and geraniol.

The accuracy of this method is comparable with

previous research (Esfahanizadeh et al., 2018), the

accuracy of eugenol from clove oil compared with

standard eugenol (Sigma) (Athar et al., 2013),

patchouli from China (Hu et al., 2006) and citronella

oil compared with essential oil standard Sigma (Wany

et al., 2014). The methods of those research validated

the accuracy of one type of essential oil, but in this

research can be used to validate the accuracy of all

types of essential oil. If one of the peaks of the active

ingredient was absent or there was absence an active

component or impurities from other essential oils or

foreign matter, it can be ensured that the essential oils

are not authentic.

(a)

(b)

(c )

Figure 2: Accuracy of clove oil (a); patchouli oil (b);

citronella oil (c).

3.3 Limit of Detection (LOD)

The limit of detection was showed the lowest

concentration of essential oil could be detected using

GC-MS. This method can be used for 4 (four) types

of essential oil (clove, patchouli, citronella, and

lemongrass oil) with the different LOD showed in

Table 1. The value of LOD depended on the type of

GC and detector, sensitivity on separations, and the

experimental conditions (program) used. Athar et al.

(2013) reported the LOD of eugenol from clove oil

was 3µl/L used the same column and carrier gas but

different in experimental conditions. Jumepaeng et

al. (2014) obtained LOD of lemongrass values was in

the range from 0.3 to 0.6 µg/mL using GC FID. The

limit detection of citronella close with clove oil.

Patchouli oil has the lowest limit detection than other

essential oil.

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ICEO 2019 - 2nd International Conference of Essential Oil Indonesia

Table 1: Limit detection of essential oil.

Essential oil Limit of detection (ppm)

Clove oil 0.02

Patchouli 0.005

Citronella oil 0.033

Lemongrass oil 0.016

4 CONCLUSIONS

The validation method can be applied to all types of

essential oil for analysis of authenticity. The

repeatability and accuracy for the essential oil are

good with limit detection of clove, citronella,

patchouli, and lemongrass oil, which were 0,02 ppm;

0.033 ppm; 0.005 ppm and 0.016 ppm. This study will

facilitate the scientific community by enhancing the

efficient method for essential oil. Through this study

can maintain Indonesia's reputation in the trade sector

of essential oil, especially clove, patchouli, citronella,

lemongrass, and massoia oils, and to facilitate

identification of their purity and quality.

ACKNOWLEDGEMENTS

This research was supported by Center for Chemical

and Packaging (CCP). We also thank to Technical

Assistant EU-Indonesia Trade Support Programme

(TSP) for the training to support the research.

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ICEO 2019 - 2nd International Conference of Essential Oil Indonesia