Isolation of Plant Essential Oil Using Steam

Distillation
Cyrille A. Amper
Chemistry Department, College of Humanities and Sciences,
De La Salle Medical and Health Sciences Institute, City of Dasmariňas, Cavite

18 FEBRUARY 2019

ABSTRACT

Essential oils are composed of pleasant-smelling organic compounds which are commonly used as perfume ingredients or
spices. Majority of these compounds belong to a group of secondary metabolites called terpenes. These can be extracted
from flowers, leaves, bark, roots, resin and peels of plants using steam distillation. In this experiment, simple distillation and
steam distillation were used to recover the organic solvent, Ethyl Acetate, and to isolate essential oils from a plant part,
respectively. In recovering ethyl acetate, the temperature of the vapor (65°C) and the initial volume (15.00mL) were
compared to the standard boiling point (77.1°C) and recovered volume (13.00mL) accumulating 86.67% percentage
recovery. On the other hand, in the isolation of the essential oil, the plant used was oregano leaf wherein the temperature of
the vapor (110.00°C) and mass of plant sample (16.00g) were compared to the standard boiling point of oregano leaf
(239.00°C) and recovered mass (0.17g) that resulted with a percentage yield of 1.0625% which is below the standard
percentage yield (1.20%). The isolated essential oil was tested with Baeyer’s solution which gained a positive result (brown).
At the end of the experiment it was concluded that the effectiveness of simple distillation was 86.67% which was assumed to
be caused by errors such as (1) impurities in the ethyl acetate or (2) the paper used. While in the extraction of essential oil
through steam distillation, due to the positive result in the Baeyer’s test, was successful in isolating the essential oils of
oregano.

Keywords: Essential oils, Terpenes, Distillation, Simple Distillation, Steam Distillation, Baeyer’s Reagent

Introduction: of enzymes. The oil is located in tiny compartments, either

Essential oils are volatile naturally occurring, complex
compounds characterized by a strong odor and are formed
by aromatic plants as secondary metabolites called
terpenes. They are liquid, volatile, rarely colored, lipid
soluble and soluble in organic solvents with a density
generally lower than that of water (Kalita, Bora, &
Sharma, 2013).
Essential oils are a natural, safe and cost-effective
therapy for a number of health concerns which has been
used for centuries in various cultures for medicinal and
health purposes (Axe, 2018). These are also commonly
used as perfume ingredients and spices since it is
composed of pleasant-smelling organic compounds that
can be found in aromatic plants (Vankar, 2004).
A typical essential oil is composed of hundreds of
constituents which is biosynthesized by the plant by means
within the plant tissue (wood, flower, seed, etc.) or in the
case of some leaves such as Salvia officinalis or Common
Sage, on the outside surface wherein gently rubbing the
leaf would cause the oil sacs to rupture (Tisserand, 2017).
The particles in essential oils come from distilling or
extracting the different parts of plants, including the
flowers, leaves, bark, roots, resin and peels. In fact, just
one drop of an essential oil can have powerful health
benefits. Typically created through the process of
distillation — which separates the oil and water-based
compounds of a plant by steaming — essential oils are
highly concentrated oils that have a strong aroma. In fact,
sometimes they are called volatile aromatic oils because of
their high concentration of the aromatic compounds. (Axe,
2018).
 2

Essential oils have a high boiling point yet it is Steam Distillation

considered volatile. This is due to the fact that essential
oils are mixtures of hundreds of components, thus some
components would vaporize at room temperature inducing
the scent of the oil and some would not due to its high
boiling point. The boiling point, therefore, is the maximum
or the highest temperature possible for the essential oil, yet
this high amount of heat would decompose the compound.
(Tisserand, 2017).
Distillation is a widely used method for separating the
component substances from a liquid mixture by selective
evaporation and condensation. To separate a mixture of
liquids, the liquid can be heated to force components,
which have different boiling points, into the gaseous state
(“Distillation”, 2019). In the experiment, simple
distillation and steam distillation will be used to recover
Steam distillation is the most common process of
extracting essential oils from plants. It is a method of
distilling compounds which are heat-sensitive. The
temperature of the steam is easier to control than the
surface of a heating element. This allows a high rate of
heat transfer without heating at a very high temperature
(“Distillation”, 2019). The advantage of steam distillation
is that the volatile components can be distilled at
temperatures lower than the boiling points of their
individual constituents and are easily separated from the
condensed water (“Basic Principles of Steam Distillation”,
1995). However, the yield of essential oils is only 0.1%-
25%, depending on the plant used (Robins, n.d.)
2 7

ethyl acetate and to isolate essential oil, respectively. 3

5
11
Simple Distillation (out)
8
Simple distillation is a procedure by which two liquids 11 4
with different boiling points can be separated. Simple (in) 6

distillation can be used effectively to separate liquids that

have at least fifty degrees difference in their boiling 1

points (Yoder, 2019).

12
2 6
9
3 10
11 7
(out)

5
8 Figure 2: Steam Distillation Set-up
11
(in)
1. Heating Source
2. Iron Stand
4 3. Iron Clamp
10 9 4. 250 mL Round Bottom Flask
12 5. Safety tube
6. Compound to be distilled
7. Thermometer
8. Condenser
9. Ice Bath
10. Immersion Pump
11. Rubber Tubing
12. Distillate/Receiving Flask (250 mL Erlenmeyer Flask)
Figure 1: Simple Distillation Set-up
Reagents
1. Heating Source
2. Iron Stand
3. Iron Clamp Ethyl Acetate
4. 250 mL Round Bottom Flask
5. Still Head
6. Rubber Stopper Ethyl acetate is a colorless, with fruity odor liquid. Its
7. Thermometer melting and boiling point are -83.6 ºC and 77.06 ºC and its
8. Condenser
9. Ice Bath
density is 0.894 g mL-1. Ethyl acetate is not soluble in
10. Immersion Pump water, but it is soluble in most of the organic solvents such
11. Rubber Tubing as benzene, acetone, chloroform and toluene. (“Ethyl
12. Distillate/Receiving Flask (250 mL Erlenmeyer Flask)
acetate Formula”, 2019).

Oregano Leaf
Oregano or Origanum vulgare is a culinary and medicinal
herb from the mint, or Lamiaceae family. It has been used
in medicine and cooking for thousands of years. It adds
flavor, and it may have a number of health benefits. The
chemicals that give the herb its unique and pleasant smell
are thymol, pinene, limonene, carvacrol, ocimene, and
caryophyllene.
Figure 3: Terpenes of Origanum vulgare
Oregano has an average percentage yield of 1.2% essential
oil with a boiling point of 239 degree Celsius (The
Essential Oil Company, 2019).
Baeyer’s Reagent
Baeyer's reagent, named after the German organic
chemist Adolf von Baeyer, is used in organic chemistry as
a qualitative test for the presence of unsaturation, such as
double or triple bonds. It is an alkaline solution of
cold potassium permanganate, which is
powerful oxidant making this a redox reaction. Reaction
with double or triple bonds (-C=C- or -C≡C-) in an organic
material causes the color to fade from purplish-pink to
brown (“Baeyer's reagent”, 2017).
Objectives:
At the end of the experiment, the students should be able
to:
1. Recover an organic solvent efficiently using simple
distillation; and
2. Isolate essential oil from grinded plant parts.
Methodology:
Remember to wear lab gown and protective goggles at
all times, avoid inhaling the reagents especially ethyl
acetate, and to wash hands thoroughly with soap and water
before leaving and entering the laboratory.
Before proceeding to the experiment, bring at least 15
grams of one (1) of the following plant materials: star anise
fruit, ginger rhizome, cumin seed, cinnamon bark, basil
leaves, oregano leaves, rosemary leaves, thyme leaves,
3
chamomile flowers, or any material approved by the
instructor; prepare the following reagents: ethyl acetate,
diethyl ether, Baeyer’s reagent, and deionized water; and
borrow one (1) distillation set-up (which includes a heating
source, 2-3 iron stands, one still head, a condenser, and 1-2
rubber stoppers), one (1) thermometer, one (1) 250mL
distilling flask, one (1) stainless water bath, three (3) iron
clamps, one (1) mortar and pestle, one (1) aspirator, one
(1) 10mL graduated cylinder, one (1) 100mL graduated
cylinder, one (1) 10mL serological pipette, one (1) 125mL
erlenmeyer flask which will also serve as the receiver, one
(1) pasteur pipette, one (1) water circulator with two (2)
rubber tubings, and one (1) evaporating dish. After
completing the materials and reagents needed, proceed and
start Experiment A: Simple Distillation of an Organic
Solvent.
A. SIMPLE DISTILLATION OF AN ORGANIC
SOLVENT
Figure 4: Ethyl Acetate
a Assemble the simple distillation apparatus (See figure
1). Recheck if: the thermometer is attached, cold water is
flowing through the condenser, and that all connections are
tight. Afterwards, transfer 15 small pieces (the size of a 25-
cent coin) of crumpled used paper and 3 pieces of boiling
chips to the 250 mL round bottom flask then add 15 mL of
ethyl acetate using the serological pipette. Heat the mixture
gently wherein the distillate should be dropping at a rate of
1 drop per second in the receiver. Record the temperature
of the ethyl acetate vapor. Stop heating when all ethyl
acetate have been collected, and reserve the set-up for part
B retaining the paper and boiling chips inside the round
bottom flask. Lastly, measure the volume of the distillate
and calculate the percent recovery to determine the
efficiency of distillation.
B. STEAM DISTILLATION OF ESSENTIAL OIL
 4

can be concluded that the ethyl acetate used was

contaminated with impurities. The decrease in volume
agrees with the conclusion for the accumulated low vapor
temperature. Another possible error might be the 15 small
pieces of crumpled paper added to the distilling flask since
the paper used was a yellow pad paper wherein ink is
present that might have reacted with the ethyl acetate.

Table 2: Steam Distillation of Essential Oil

Figure 5: Grinded Oregano Leaf
Name of plant and part used Oregano- leaf
Temperature of the vapor 110.00 degree Celsius
Assemble the steam distillation set-up (See figure 2).
Standard Boiling Point of 239.00 degree Celsius
Grind the plant part into a fine powder using mortar and Essential Oil
pestle then transfer approximately 15 grams to the round Mass of evaporating dish 47.48 grams
bottom flask. Record its exact mass. Add deionized water Mass of evaporating dish 49.35 grams
until the flask of the compound to be distilled is 60-70% and essential oil
filled. Recheck if: the thermometer is attached above the Mass of essential oil 0.17 grams
flask of the grinded plant part, cold water is flowing Mass if plant sample 16.00 grams
through the condenser, and that all connections are tight. % essential oil 1.0625%
Afterwards, heat the mixture gently ensuring that the rate Standard % yield 1.20%
Color of essential oil with Brown
drop of the distillate is 1 drop per second. Record the
Baeyer’s reagent
temperature of the essential oil vapor. When the distillate
stops dropping into the receiver, remove the heat and add
5mL diethyl ether shaking the flask twice. Let the distillate In the experiment, 16.00 grams of grinded Oregano-leaf
sit until a layer is formed. Carefully draw the upper layer underwent steam distillation. Through this, the boiling
using a Pasteur pipette taking care not to disturb the water point of the extracted essential oil of Oregano leaf was
layer and transfer into pre-weighed evaporating dish. After reduced from 239.00 degree Celsius to 110.00 degree
extraction is complete, place the dish under the fume hood Celsius. The mass however, was reduced to a great amount
and allow the ether to evaporate. Obtain the mass of the wherein only 1.0625% was accumulated which is below
evaporating dish with essential oil and calculate the and somehow close to the average percentage yield.
percentage of essential oil present in the plant material.
Lastly, add 1 drop of essential oil and 3 drops of Baeyer’s Table 3. Table of Reaction
reagent in a clean test tube, shake thoroughly, and take
note of the color change. Reactant Product
Mno4- Mn2+
Pink Brown
Results and Discussions:

Table 1: Simple Distillation of an Organic Solvent

Temperature of the vapor
Standard Boiling Point of
Ethyl Acetate
Original volume of ethyl
acetate
Volume of recovered ethyl
acetate
% Recovered Ethyl Acetate
An unsaturation test was performed through the addition of
Baeyer’s reagent wherein the isolated essential oil turned
65.00 degree Celsius
77.10 degree Celsius into a brown color-
15.00 mL
13.00 mL
86.67%

After performing simple distillation on ethyl acetate, it

was observed that the temperature of the vapor, 65 degree
Celsius, was lower compared to the standard boiling point
of ethyl acetate which was 77 degree Celsius. On the other
hand, the original volume of ethyl acetate, 15.00 mL, was
Figure 6: Baeyer’s test of the Essential Oil
reduced to 13.00 mL. The percentage of recovered ethyl
acetate is calculated to be 86.67%. With the data given it

-which indicates that there is a presence of unsaturation
and double or triple bonds therefore the collected oil has
the presence of terpenes indicating that the isolation was
successful.
Conclusions:
Essential oils have been used for hundreds of years due to
its fragrance, flavor, and medicinal contributions. These
oils are found in tiny compartments within plant tissues
which is best extracted through a process called
distillation. Distillation is one of the methods of purifying
liquid organic compounds. In this process, the liquid
compound is converted to its vapor state at its boiling point
and condensed into the liquid state. Two types of
distillation were used in the experiment which has specific
differences. The first type used was Simple Distillation
which is used for immiscible liquids having a large
temperature difference (minimum difference: 50°C). This
type of distillation was used to recover the ethyl actetate.
The second type of distillation used was Steam
Distillation, which was used for purifying some high
boiling substances that decompose at their boiling point.
The advantage of using steam distillation is that the
essential oil can be isolated at a temperature below 100°C
therefore prevents decomposition. The result gained in the
simple distillation was 86.67% which did not follow the
assumption of a 100% recovery. This concludes that errors
were present such as (1) impurities or (2) the paper used.
While the result for the isolation of essential oil through
steam distillation was positive due to the positive result in
the Baeyer’s test (brown) which means it was unsaturated
and contains a double or triple bond, therefore a terpene.
It is recommended to not add paper but instead, place
atleast three boiling chips in the boiling flask. Before the
experiment, always check if all connections are tight and
not loose. Also, check if the thermometer is in place, and
the condenser must contain ice cold water and should
never have bubbles of air inside.
References:
Kalita, B., Bora, S., & Sharma, A. (2013). International
Journal of Research and Development in Pharmacy
and Life Sciences. PLANT ESSENTIAL OILS AS
MOSQUITO REPELLENT-A REVIEW,3(1), 741-
747. doi:10.3897/bdj.2.e1061.figure2f
Axe, J. (2018, July 26). 101 Essential Oil Uses and
Benefits. Retrieved February 15, 2019, from
https://draxe.com/essential-oil-uses-benefits/
5
Tisserand, R. (2017). About Essential Oils - Introduction
to Essential Oils - Tisserand. Retrieved February 17,
2019, from https://roberttisserand.com/essential-oils/
Vankar, P. S. (2004, April). Essential Oils and Fragrances
from Natural Sources. Retrieved February 17, 2019,
from
https://www.ias.ac.in/article/fulltext/reso/009/04/0030
-0041
Food and Agriculture Organization of the United Nations
(1995). Basic Principles of Steam Distillation.
Retrieved August 18, 2005, from
http://www.fao.org/docrep/V5350e/V5350e13.htm.
Distillation. (2019). Retrieved February 18, 2019, from
https://byjus.com/chemistry/distillation/
Yoder, C. (2019). Wired Chemist. Retrieved February 18,
2019, from
http://www.wiredchemist.com/chemistry/instructional/
laboratory-tutorials/distillation
Baeyer's reagent. (2017, May). Retrieved February 18,
2019, from
https://ipfs.io/ipfs/QmXoypizjW3WknFiJnKLwHCnL
72vedxjQkDDP1mXWo6uco/wiki/Baeyers_reagent.ht
ml
Ethyl acetate Formula. (2019). Retrieved February 18,
2019, from
http://www.softschools.com/formulas/chemistry/ethyl
_acetate_formula/416/
Examine.com. (2018, June 14). Origanum vulgare -
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Retrieved from
https://examine.com/supplements/origanum-vulgare/
Robins, W. (n.d.). Essential Oil Yields | AromaWeb.
Retrieved from
https://www.aromaweb.com/articles/essential-oil-
yields.asp
The Essential Oil Company. (2019). Percent Yield Guide
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https://www.essentialoil.com/pages/percentage-yield
Sample Calculations:

% recovered Ethyl Acetate = (volume of recovered ethyl acetate / original volume of ethyl acetate) x 100

= (13 mL/15mL) x 100

= 86.67%

Mass of Essential Oil = Mass of evaporating dish and Essential Oil - Mass of evaporating dish

% recovered Essential Oil = (mass of essential oil / mass of plant sample) x 100

= (0.17 g / 16.00 g) x 100

= 1.0625%