Escobedo 1

Verification of Essential Oils Distilled from Mentha x pipereta

Abstract

Steam distillation was used to extract the essential oils from a wild sample of Mentha x

pipereta. The oils were then analyzed via Gas-Chromatography-Mass Spectroscopy. When

compared with expected values, the overall percentage yield of the specimen, as well as most of

the compounds found, were discovered to be within normal limits. Some anomalous compounds

and percentages were also discovered. These anomalies were found to warrant further

investigation and experimentation.

Introduction

Distillation is an ancient scientific technique that has been used for centuries to separate

the different components of a mixture. This separation is achieved through the exploitation of the

components different boiling points. In its simplest form, this is achieved by heating a mixture

of multiple components to a degree that would cause one or more of those components to escape

the mixture as a vapor. That vapor is then condensed and collected. This provides two different

substances; that which was removed from the original mixture and that which was left behind.

Originally used to produce alcoholic beverages, obtain oils for perfumes, and extract medicinal

products from plants, distillation was refined into an industrial process in the 19th century to

provide both products and raw materials of a higher purity (Vogelpohl 2015). Distillation

continues to be an invaluable industrial process that is still utilized across countless fields

including the production of fuels and medicine.

Distillation is utilized in this experiment as a method to extract the key components or

essential oils of a Mentha x pipereta specimen for scientific verification. The particular
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distillation process used for this experiment is steam distillation and is achieved with the use of a

round-bottomed flask, Liebig condenser, and receptacle as illustrated in Figure 1. According to

the United States National Library of Medicine, common results for an experiment such as this

yield an oil consisting of 29-48% menthol, 20-31% menthone, and 3-10% menthyl acetate.

Mentha x pipereta itself, typically yields approximately 0.1-1.0% essential oils.

Figure 1 Distillation Apparatus

Methods and Materials

Materials utilized for this experiment include a large sample of Mentha x pipereta, a

mortar and pestle, a separatory funnel, and a distillation apparatus. Tert-butyl methyl ether

(C5H12O) or MTBE and sodium sulfate (Na2SO4) were used during the extraction process. A

Shimadzu GCMS-QP2010 SE, equipped with autosampler and SH-Rxi-5SilMS column (30m x

0.24mm ID x 0.25µm df) was used to obtain a Gas-Chromatography-Mass Spectroscopy

(GC/MS). Specifications for the machine are found in Table 1.

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Methods for this experiment were adapted from Miller (2018). To begin the experiment,

a 9.905g sample of ground M. pipereta leaves were added to a round-bottomed flask which was

then filled to the halfway point with distilled water and allowed to soak for approximately ten

minutes. The sample was then distilled via steam method. Organic materials from the distillate

were extracted twice using 5mL of ether (Figure2) and then dried using sodium sulfate (Figure

3). Two milliliters (2mL) of the organic substance were set aside for GC/MS analysis and the

remainder was allowed to airdry for 48 hours. After the drying period, a mass was obtained for

the sample.

Oven program Isothermal, 100°C

Injector temperature 280°C
Ion source temperature 220°C
Flow rate 0.80mL/min
Split ratio 1.4
Solvent delay 2 minutes
Table 1 GC/MS Specifications

Figure 2 Extraction with Ether

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Figure 3 Drying with Sodium Sulfate

Results

Distillation of the sample yielded 0.012g or 0.12% of organic components after drying.

These components consisted of menthol, menthone, menthyl acetate, decanol, eugenol, and

pulegone as illustrated by the GC-MC results in Figure 4. Structures for all components can be

found in Table 2.
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Figure 4 GC-MS Results

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Menthol

Menthone

Menthyl Acetate

Eugenol

Pulegone

Decanol

Table 2 Compounds Extracted from Distillate

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Discussion

Analysis of the results of the distillation process confirmed the presence of the expected

compounds menthol, menthone, menthyl acetate, and pulegone. The percentage of menthol and

menthyl acetate, as well as the overall amount of essential oils yielded by the sample of M.

pipereta were found to be within expected parameters (as cited by United States Library of

Medicine). Menthone was found to be below expected amounts, while pulegone was slightly

higher than the expected range. According to the U.S. Library of Medicine, menthone usually

constitutes 20-31% of M. pipereta and pulegone accounts for 1-11%. Two other compounds,

eugenol and decanol, were also identified. Eugenol is an allylbenzene most often found in clove

oil and many other aromatic plant extracts (National Center for Biotechnology Information).

Decanol is a ten-carbon fatty alcohol naturally found in coconut oil, and is often used as a flavor

additive (National Center for Biotechnology Information).

The most likely reasons for compound percentage variance from norm is human error.

Since the sample was obtained from the wild, there is a high possibility that it would have had

contaminates on it. Not enough care was taken to clean the sample of foreign contaminates.

Additionally, the round-bottomed flask was not heated to a high enough temperature during the

distillation process. This likely resulted in too small of a sample to provide very accurate results.

There is also the possibility that the distillation apparatus was not properly cleaned of

contaminates.

Due to the results of this experiment yielding data that was out of expected ranges, the

next step in the process is to distill and analyze multiple samples of Mentha x pipereta from the

same source. Trial results similar to the original may point to the sample being a Mentha hybrid

rather than a pure strain. It may be necessary to consult a taxonomist for sample evaluation.
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References

Miller, C. 2018. CHEM 2423: Isolation of Essential Oils by Distillation. Accessed October 16,

2018.

National Center for Biotechnology Information. PubChem Compound Database; CID=3314,

https://pubchem.ncbi.nlm.nih.gov/compound/3314 (accessed Nov. 13, 2018). CID=8174,

https://pubchem.ncbi.nlm.nih.gov/compound/8174 (accessed Nov. 14, 2018).

United States National Library of Medicine. TOXNET Toxicology Data Network. Accessed

November 13, 2018. Retrieved from https://toxnet.nlm.nih.gov/cgi-

bin/sis/search2/r?dbs+hsdb:@term+@rn+@rel+8006-90-4

Vogelpohl, Alfons. 2015. Distillation: The Theory. Berlin/Boston: De Gruyter, Inc.. Accessed

October 23, 2018. ProQuest Ebook Central.