Final Project MANISH

MAKING OF PERFUME WITH
ESSENTIAL OILS
A Project, submitted to KBC North Maharashtra University, Jalgaon.
In Partial Fulfillment of the requirement for award of the degree of,
Submitted By:
PACHPANDE MANISH SANJAY

PATIL KUNDAN SUNIL
(Final Year B .Pharm)
University Seat No.
515859, 515860
Under Guidance of:
PROF. MR. RAHULSING. U. KHAIRNAR
KAI. YASHODABAI DAGADU SARAF COLLEGE OF

PHARMACY, SAKEGAON.
BUSAWAL DIST. JALGAON - 425201
2023-24
CERTIFICATE
This is to certify that Research Project entitled, “Making Of Perfumes
With Essential Oils ”By Pachpande Manish Sanjay, Patil
Kundan Sunil in the partialfulfillment of Degree Of Bachelor of

Pharmacy, has been carried out in KYDSCT’s College Of Pharmacy,
Sakegaon, under my guidance and to my satisfaction.
This Review Project is now ready for evaluation.
Prof. Mr. Rahulsing. U. Khairnar Dr. P. R. Patil

Guide Principal
ACKNOWLEDGMENT
NO research is ever they outcome of single individual's efforts. This

work is also no exception. Some of the personalities known and
unknown have contributed to and made it possible for me to present
inthis shape. I am heartily grateful to all of them.
I am highly thankful to Dr.Parag R. Patil, . Principal, KYDSCT college of
Pharmacy,Sakegaon (Bhusawal) who continuous help and providing
valuable suggestion for completing this project.
With a feeling of profound pleasure, I gratefully own my sincere thanks

to academic guide Prof. Mr. Rahulsing. U. Khairnar , KYDSCT
collegeof pharmacy, Sakegaon (Bhusawal) for providing all the facilities
and encouragement in completing my project work.
I am very thankful to staff members of KYDSCT college of pharmacy,
Sakegaon (Bhusawal), who are great support and very helpful to me
throughout my entire project.
Last, but not the least, I express my gratitude and apologize to anybody
who could not mention in this page.
PACHPANDE MANISH SANJAY
PATIL KUNDAN SUNIL

Index
Sr.No Content Page Number
1 Abstract 1
2 Introduction 2
3 Classification of scents 4
4 Analysis of essential oils 5
5 Isolation of oils from natural sources 5
6 Concretes/absolutes and essential oils 6
7 Natural versus artificial mix 6
8 Essence oils 7
9 Carrier oils and alcohol 8
10 fixatives 8
11 Molecular structure of aromatic compound 10
and their sensory properties
12 Olfactive properties and chemical identities of 11
floral compounds
13 Perfume blending 14
14 Some aspects of adulteration of natural 15
isolates
15 Materials and methods 17
16 Results 20
17 Conclusion 25
18 References 26
MAKING OF PERFUMES WITH ESSENTIAL OILS
Abstract
A study was undertaken to extract essential oils from flowers and leaves
of the Lavender plant (Lavanda officinalis) using water distillation
method. The extracted oils were further mixed with different proportions
of methanol, ethanol and formaldehyde to make perfumes.The study of a
perfume includes extraction of scented ingredients from botanicals,
behavior of chemical components, and careful blending of scents to
achieve the desired composition. Although there was no significant
difference in the amount of essential oils extracted from flowers and
leaves, fruits, roots, seeds, or bark of the lavender plant (P=0.08),
flowers gave higher yields of essential oils than leaves. Growing and
harvesting conditions are optimized for the production of the best
fragrances. In addition, there was no significant difference in the
percentage composition of the different compounds in the essential oils
extracts from flowers and leaves (P=0.46). Moreover, a mixture of
methanol and formaldehyde gave the highest quality of perfumes in the
final products compared to when methanol, ethanol or a mixture of
ethanol and formaldehyde is used. Lavender plant is a potential source of
essential oils for making perfumes. There is a need for testing the
essential oils of lavender plant from Egerton University for other uses
such as their antimicrobial properties.
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INTRODUCTION
A perfume is a substance that emits and diffuses a fragrant odor. It is a
very volatile liquid distilled from a plant part. The essential ‘plant
distillates’ (essential oils) interrelate with the human body by four
distinct modes of action – pharmacological, physiological, psychological
and spiritual. Our body uses the aromatic molecules (essential oils) in
two ways: (1) through our olfactory system which is connected to the
brain where our most primal feelings, urges and emotions reside, and (2)
by absorption of the low molecular weight compounds of essential oils
through skin. Lavender plant’s flowers and leaves produce essential oils
that can be put to many uses such as making of perfumes, flavoring of
foods, and treating of diseases (Bachhav and Shankar, 2013). Analgesics,
antimicrobial and anti-inflammatory effects of lavender essential oils
have been reported from many parts of the world (Fornari et al., 2012).
Previous studies have shown that essential oils from lavender are
composed of cotronellal (57%), citronellol (15.89%), citronellyl acetate
(15.33%) coupled with other organic compounds (Kaskoniene et al.,
2013). The fore mentioned compounds have indicated a high potential
for wide usage, both domestic and industrial which include but is not
limited to antimicrobial, antifungal and anti-candida properties
(Sekoussounon, 2012). In addition the essential oils have shown
expectorant and cough stimulating activity. The disinfecting properties
of essential oils from lavender plant has made them gain allot of
popularity in treating wounds, cleaning hospital apparatus and scrubbing
floors in hospitals (Vignesh et al., 2016). Besides, the essential oils
possess herbicidal, insecticidal, antihelmintic, anti-tumor and anti-leech
properties not to mention their inclusion in integrated pest management
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(Sambath et al., 2012). Beauty
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therapy is one of the fastest growing industries in the 21st century. This
has created high demand for perfumes (Mann et al., 2013). As a result,
millions of people have fallen prey of unscrupulous business people who
have taken advantage of the high demand to sell below standard beauty
materials (Jakhetia et al., 2011). The materials have proven to have dire
consequences for many of their users, creating a need for coming up with
quality, safe to use substances such as perfumes (Velavan et al., 2013).
The use of lavender essential oils in making of perfumes saw the wake of
the day back in ancient Chinese civilization where people would simply
rub flowers and leaves of the plant on their bodies. Since then, the use of
essential oils from lavender plant has gained popularity (Enzo, 2011).
There are various methods of extracting of essential oils from plant
samples. Expression is the oldest and the least complex. Plant materials
are manually pressed until all the oils are squeezed out (Coelho et al.,
2012). Following at a close range is another method, popularly known as
effleurage which involves spreading of plant materials on glass sheets
coated with grease. The glass sheets are placed between wooden frames
in tiers. The plant materials are removed by hand and changed until the
grease has absorbed their fragrance (Herrero et al., 2013). A similar
method to effleurage is maceration in which warmed fats are used to
soak up flower smell (Patinha et al., 2013). The grease and fats are in
turn dissolved in alcohol to obtain the essential oils. In an additional
method called solvent method, plant materials are placed in rotating
tanks and drums. Benzene or petroleum ether is poured over the plant
materials so as to extract the essential oils (Madziga et al., 2010). The
method that is currently witnessing allot of usage is steam distillation
(Kabiru et al., 2016). In this method, steam is passed through plant
materials to convert the essential oils to gas.
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Classification of Scents
Scents are classified as notes based on their olfactory character. A
perfume is a unique mixture of top, middle, and base notes designed to
give a particular harmony of scents. The following basic groups are listed
in order of decreasing vapor pressure, or volatility. a) The top notes are
those that are detected and fade first providing freshness to the blend.
They are responsible for the customer's first impression, and hence, in a
way, the selling note of a perfume. They are light scents, lasting 5-10
minutes, and extracted from plant material such as cardamom seed, basil,
bergamot, citronella, coriander, eucalyptus, ginger, grapefruit,
lemongrass, lemon, orange, lime, marigold, peppermint, sage, spearmint,
tangerine and tea tree. The more commonly used are those extracted
from citrus sources or grasses. b) The middle notes are those that last for
several hours and are the most prominent within the fragrance. They are
usually combinations of spicy, floral or fruit scents, extracted from
ambrete seed, black pepper, carrot seed, cassia, chamomile, cinnamon,
clove, fir, cypress, juniper, marjoram, pine, rose, rosemary, thyme and
yarrow. c) The base notes give a perfume the depth and last the longest.
They include the extract of plant material such as amyris, anise, angelica
root, clary sage, fennel, geranium, lavender, lavandin, balsam,
cedarwood, frankincense, jasmine, myrrh, patchouli, rosewood,
sandalwood, vetiver and ylang ylang. Musks extracted from musk deer,
musk rats and civet are scents derived from animal sources.
A perfume is a substance that emits and diffuses a fragrant odor. Scents
are classified as notes based on their olfactory character. A perfume is a
unique mixture of top, middle, and base notes designed to give a
particular harmony of scents.
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Analysis of Essential Oils

Essential oils should be subjected to both qualitative and quantitative
tests to know its purity. Oils are tested in four stages – the first stage is
sensory evaluation in which the smell, viscosity, color and clarity of the
oil are assessed. For example, rose oil which appears too mobile at low
temperature would be regarded as adulterated. This is a viscosity
assessment. The second stage is an odor/smell test, which helps to
determine if an oil is really what it is claimed to be, since certain
adulterants can be identified by this test. In the next stage, physical
parameters such as specific gravity, optical rotation and refractive index
are determined. If these test results are satisfactory, the oil is then
subjected to GC-MS evaluation.
Essential oils should be subjected to both qualitative and quantitative
tests to know its purity. Oils are tested in four stages.
Isolation of Oils from Natural Sources

The production method of the oil depends on the botanical source.
Several methods are employed in modern perfumery.Essential oils of
citrus plants may simply be expressed (squeezed) from the peels. The
majority of essential oils are obtained by steam distillation or hydro-
distillation. Three types of hydrodistillation are employed in the
essential oil industry: l Water distillation, l Water and steam distillation
and l Direct steam distillation. In order to isolate an essential oil from an
aromatic plant, the plant material is packed into a still, sufficient
quantity of water is added and the mass is brought to a boil, or live
steam is injected into the plant charge. Due to the action of hot water
and steam, the essential oil will be freed from the oil glands in the plant
tissue. Other
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essences are obtained by extraction methods. They are as follows: l

Extraction with cold fat, l Extraction with hot fat (maceration), and l
Extraction with volatile solvents. Many scents that are difficult or costly
to obtain from natural sources are produced synthetically.
The majority of essential oils are obtained by steam distillation or hydro
distillation.
Concretes/ Absolutes and Essential Oils

Essential oils and absolutes are different. Distinguishing the essential oils
from concretes or absolutes (fragrances obtained by methods other than
steam distillation or pressure) is sometimes very difficult. The essential
oils are free-flowing, much like water, have no oily feel to them,
evaporate quickly and are a lot more expensive than the concretes,
absolutes and oil blends. Essential oils should be stored in tightly sealed,
dark colored glass bottles because light, air and excessive heat will
quickly degrade them. However, when obtaining essential oils in small
quantities, clear glass is permissible providing the bottles are kept in the
dark until ready for use. Because essential oils are highly concentrated
plant extracts, they have potent therapeutic value. Each drop is equal to at
least one ounce of plant matter.
Distinguishing the essential oils from concretes or absolutes (fragrances
obtained by methods other than steam distillation or pressure) is
sometimes very difficult.
Natural versus Artificial Mix

Many aromatic oils from plants contain a few major constituents, several
minor ones and a larger number of trace compounds (called elements in
perfumery parlance). It is virtually impossible to totally reconstruct such
a complex combination of components which would include all the trace
compounds. Most ‘nature identical’ oils are said to be 96% accurate,
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with
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the remaining 4% of trace compounds being non-duplicated. It is the

synergistics of the specific combination of hundreds of constituents
naturally present in each plant (including trace compounds) that give the
essential oils their valuable therapeutic/healing properties.
It is the synergistics of the specific combination of hundreds of
constituents naturally present in each plant (including trace compounds)
that give the essential oils their valuable therapeutic/healing properties.
Essence Oils
Essential oils not extracted via steam or pressure are referred to as
essence oils. Essence oils are derived in a variety of ways from flowers,
barks, leaves, roots or resins.
Concretes:
The aromatic plant matter is extracted by organic solvents such as
hydrocarbons. Concretes are more stable and concentrated than
unmodified essential oils. Resinoids: Oils from natural resins such as
frankincense, myrrh, amber, benzoin, etc. are extracted using
hydrocarbon solvents. Occasionally the ethanol soluble fraction of a
resinoid is called an absolute.
Absolutes:
They are obtained by taking the concretes through a second process of
ethanol extraction or liquid carbon dioxide. With the latter process, the
oils produced are of excellent aroma quality and entirely free of
unwanted solvent residues or nonvolatile matter.
The products obtained from all the above methods go through a final
process of solvent removal. These techniques are used worldwide for
higher yield or to obtain products that cannot be obtained by any other
process. They also produce a more true to natural fragnance. An example
of this is the case with jasmine concretes or absolutes which would lose
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their natural quality by hot water or steam distillation process.

Numerous extracts, each one having a distinctive character, are used in
combination to formulate a fragrance. Some overlap in tones or notes in
two or more of these scent extracts is possible. For example, rosewood
and ylang ylang have middle to base character and clary sage and fennel
have top to middle character.
Numerous extracts, each one having a distinctive character, are used in
combination to formulate a fragrance.
Carrier Oils and Alcohol

Many essential oils are much too strong and often expensive to use
without dilution. To moderate such attributes carrier oils are used to
dilute and carry the fragrance for use as a perfume. Carrier oils come
from nuts and seeds and are usually pressed out of commercially
produced crops. The best carrier oils are stable to oxidation and have
little or no fragrance of their own, and they include oils from sweet
almond, apricot, peach kernel, sesame seed, soybean, sunflower,
avocado, and jojoba. Some of these oils can be used as the only carrier
while others are only safe for skin use at 10-25% of the total amount.
Alcohol is added to perfumes to carry the fragrance by evaporation as
well as dilute the ingredients. Perfumes contain 25% fragrance and
about 75% alcohol and other diluents (carrier oils) by volume while
cologne is composed of more than 90% diluent.
Fixatives
Fixatives are scented components that act to hold the fragrance together
and regulate rate of evaporation for the fragrant components. The
fixatives often provide a base note and character to the mixture. One of
the most important steps in the manufacture of a fine perfume is the
addition of a fixative which enables the essential oils to retain their
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fragrance for a longer period of time. Fragrances remain fresh if they are
protected from light, heat and oxygen for about three years from the date
of manufacture. They are categorized based on the source into seven
different classes.
(see Box 1).
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Molecular Structure of Aromatic Compounds and

their Sensory Properties
Many benzene derivatives possess aromatic, spicy and specific odors.
These aromatic compounds include mono- and poly substituted benzene
derivatives, such as benzaldehyde, anethole and anisaldehyde, eugenol
and vanillin, safrole and heliotropin. The following substituents are
found in these aromatic compounds: aldehyde (-CHO), hydroxy (-OH),
alkoxy (RO-), allyl (- CH2 -CH=CH2 ), propenyl (-CH=CHCH3 ) and
dioxymethylene (-O-CH2 -O-).
The sensory qualities of the allyl- and propenyl-substituted benzene
derivatives are very characteristic, and they are character-impact
compounds for certain natural isolates. The allyl substituted benzene
derivatives can be oxidized to the corresponding aldehydes, which also
possess characteristic odor qualities.
Various studies on the relation between the sensory properties and
structural features of aromatic compounds have been carried out.
Threshold values and odor qualities of a series of alicyclic and aromatic
aldehydes have been investigated. It has been found that aldehydes
bearing terminal phenyl or cycloalkyl substituents exhibit threshold
minima at eight carbon atoms. A similar trend is observed for straight
chain aliphatic aldehydes. Straight alkyl chains resulted in lower
threshold values than the cyclic structures.
Various studies on the relation between the sensory properties and
structural features of aromatic compounds have been carried out.
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Table 1.
Olfactive Properties and Chemical Identities of Floral

Compounds
The chemical identities of molecules show odor-structure relationship
clearly. Chemical compounds with a fresh, floral odorcharacter are found
in tertiary monoterpene alcohols and their lower esters. The compounds
have branched chain with methyl groups on the 2-, 3-, 6- or 7-position.
The presence of one or two double bonds increases their freshness and
naturalness.
Quantitatively the most important chemical compound with a fresh floral
odorcharacter is linalool.
Linalool, Tetrahydrolinalool and their Acetates
Quantitatively the most important chemical compound with a fresh
floral odor-character is linalool, 3,7-dimethyl-1,6-octadien- 3-ol, which
is a tertiary monoterpene alcohol. Linalool consists of two enantiomers,
which are (R)-(-)-linalool or licareol and (S)- (+)-linalool or coriandrol.
Licareol occurs in many flowers, such as bitter orange flower and
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lavender flower and has a fine fresh floral odor-character. Coriandrol

occurs in coriander fruits and has a more herbal, floral odor-note. The
acetate of (R)-(-)- linalool has an even more fresh floral, slightly
fruity, odor and occurs in many plant materials, such as bergamot peel
and leaves of bitter orange. Linalool and its acetate are used in
alcoholic and cosmetic perfumery, where it shows a good performance
and
stability. Tetrahydrolinalool, 3,7-dimethyloctan-3-ol, is used in functional
perfumery, because of its stability in more aggressive media.
Tetrahydrolinalool and its acetate have also floral olfactive properties,
though with a slightly fatty and earthy undertone.
Myrcenol, Ocimenol and their Acetates
Myrcenol, 2-methyl-6-methylene-7-octen-2-ol, and its acetate have
natural, fresh, floral odors, even more delicate than those of linalool and
its acetate.
Ocimenol, 2,6-dimethyl-5,7-octadien-2-ol, and its acetate have
remarkably fresh floral olfactive qualities and are preferred to myrcenol
and its acetate.
Linalool and its acetate are used in alcoholic and cosmetic perfumery,
where it shows a good performance and stability.
Dihydromyrcenol and its Acetate
Dihydromyrcenol, 2,6-dimethyl-7-octen-2-ol, is olfactively somewhat
reminiscent of linalool. This compound and its acetate are often used as
substitutes for linalool and its acetate, because they are stable in
aggressive media, where linalool is oxidised, e.g. in detergent compounds
containing perborate and tetraacetylethylene diamine (TAED).
The perfume blending does not follow the physical or chemical rules or
properties of the materials.
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Tetrahydro-myrcenol and its Acetate

Tetrahydro-myrcenol, 2,6-dimethyloctan-2-ol, and its acetate are
sometimes used instead of tetrahydro-linalool and its acetate, because
they have a less earthy note and are also stable in all media. The
compounds have fresh floral, slightly fatty, odors.
Perfume Blending
The perfume blending does not follow the physical or chemical rules or
properties of the materials. The procedure for the development of a
perfume can be illustrated by the example of rose perfume. Rose
perfume formula of Maurer is made by mixing the following
components: Geraniol – 30 parts, Citronellol – 25 parts, Phenylethyl
alcohol – 25 parts, Linalool – 5 parts.
The four compounds occur naturally in rose flowers. Geraniol lends a
petal-like, slightly herbaceous, tea-leaf-note to the base. Citronellol
lends fresh, rosy floralness, while phenylethyl alcohol gives a deep
sweetness, and linalool imparts a certain woody floralness. These four
materials can make a perfect rose oil. If added in moderate quantities,
these compounds cannot be detected in rose oil by mere routine analysis.
In such cases, the analyst will have to rely upon careful olfactory tests
which require much training and a thorough chemical examination of
the suspected oil. This then needs to be analyzed on GC-MS for traces of
adulterants.
Gas chromatography alone is not a definitive tool for determining the
purity of essential oils. This can be smartly manipulated by an
adulterator by choosing an adulterant which has similar chemical
constituents. The retention time (RT) in the gas chromatogram and the
optical rotation (OR) of a compound can be made to match by adding
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adulterants with similar values for RT and OR. It is in fact very difficult
to differentiate
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the pure oil and the impure oil by merely these techniques. GC-MS is a
better diagnostic tool as the mass fragmentation pattern will be
different for
different compounds even when they have the same RT and OR.
The analyst will have to rely upon careful olfactory tests which require
much training and a thorough chemical examination of the suspected
oil.The adulteration of natural isolates in fact covers a wide range of
actions, such as: standardizing, reinforcement, liquidizing, reconstitution,
commercializing.
Some Aspects of the Adulteration of Natural Isolates

Adulteration generally means the act of lowering the standard or
character of a product by the addition of one or more inferior ingredients.
Because there is a lot of confusion about the adulteration of natural
isolates, it seems worthwhile to discuss ‘how’ and ‘why’ this is
performed. Natural isolates may consist of essential oils, oleoresins,
gums,
absolutes or other natural extractives that contain several hundreds of
chemical compounds, which can be organoleptic substances (so-called
character-impact compounds), essential compounds, balance
compounds and artefacts. The adulteration of natural isolates in fact
covers a wide range of actions, such as: standardizing, reinforcement,
liquidizing, reconstitution, commercializing, which are discussed
below.
Standardizing: Natural isolates are mixtures of chemical compounds.
The concentration of the organoleptic characteristic compounds in these
mixtures can vary or deviate from standardized ranges, due to climatic
and/or ecological reasons. One can standardize the content of
characteristic substances by adding the products, isolated from another
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natural source or produced synthetically. Examples are,
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1. citral (eg. Litsea cubeba oil or synthetic) to standardize lemon oil;

2. 1,8-cineole (eg. Eucalyptus globulus oil) to standardize rosemary oil;
3. camphor (eg. camphor oil) to standardize labiate oils.
Reinforcement: Reinforcement is more or less an extension of
standardizing. When the quality of the natural isolates can be
improved, there is always the temptation of adding an excess amount
of the characteristic compound to improve the quality and to make the
end product of more ‘olfactive value for money’.
Liquidizing: Some natural isolates are solids or semi-solids as, for
instance, gums, oleoresins and certain absolutes. To liquidize these
materials many solvents are used, e.g., benzyl benzoate, propylene
glycol, triethyl citrate, isopropyl myristate, dialkyl phthalates,
isononylphenol,etc It is quite impossible to reconstitute the complete
natural product,because natural isolates consist of several hundreds of
chemical compounds, of which many would be unknown.
Reconstitution: There are natural isolates, such as rose oil, jasmine oil
or orange flower oil, which are too expensive for application in more
economic (cheaper) functional perfumery as, for instance, in perfumes
for soaps, detergents or other household products. Therefore, these
naturals are reconstituted, that is, the natural isolate is rebuilt
(compounded) with a mixture of natural or so-called nature-identical
chemical compounds, after a more or less thorough analysis of the
identities and quantities of the main constituents. It is clear that it is
quite impossible to reconstitute the complete natural product, because
natural isolates consist of several hundreds of chemical compounds, of
which many would be unknown.
Commercializing: By commercializing of a product is meant lowering
its quality in order to make it more profitable. It will be clear that the
‘reconstituted oils’ are also examples of ‘commercial oils’.Although
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commercializing of a product appears to be a fraudulent act, it ought not

necessarily be so. A buyer who cannot afford to pay the cost price of the
natural may be willing to buy a commercialized product. Today,
however, essential oils are often adulterated in a very clever manner and
the analyst may face considerable difficulty in finding the adulterants.
The adulterator has at his disposal a number of natural isolates of lower
priced essential oils. If added in moderate quantities, these compounds
cannot be detected in oils easily by simple analytical tools. It is very
difficult to find
out the authenticity of the pure oil by olfactic examination. A crude but
simple test for adulteration involves heating the essential oil with iodine,
is as follows: To 5 parts of the oil heated to 60°C and 1 part of powdered
iodine is gradually added, without allowing any further rise in
temperature. The mixture, on cooling, will deposit a mass of crystals if
the oil is adulterated. However, this is a very crude test which helps in
only knowing whether the oil is adulterated or not. Further, the
adulteration occurs mostly during the supply chain in India. At every
stage of the supply, adulteration may occur where the addition of an
adulterant can increase the profit from the oil by tenfold. It is therefore
important to realize and to devise an effective method of quality control
for ascertaining the purity of any oil, and to ensure that adulteration is
not affected along the whole chain from the producer to the retailer.
Even a price ceiling is required to be made by the government to let the
farmers get satisfactory financial returns for their product
The adulterator has at his disposal a number of natural isolates of
lower priced essential oils. If added in moderate quantities, these
compounds cannot be detected in oils easily by simple analytical
tools.
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MATERIALS AND METHODS
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Study area
Egerton University Main Campus is located 180 kilometers North West
of Nairobi and about 30 Kilometers from Nakuru on a 144 – hectare
piece of land donated by Lord Maurice Egerton of Tatton (1874 – 1958).
The coordinates of the university are: 0°22'11.0"S, 35°55'58.0"E
(Latitude:
-0.369734; Longitude: 35.932779). The University lies at an altitude of
approximately 2,250 meters above sea level (GoK, 2014).
Collection of plant samples
Lavender (Lavandula angustifolia) flowers and leaves were collect from
Egerton University main campus, Njoro, Kenya. The flowers and leaves
were separately washed with running tap water and finally with distilled
water to sterilize them.
Extraction of essential oils
A sample of 400g of fresh lavender flowers and leaves was separately
loaded into 2-Litre round bottom flask containing 1.5 liters of water
and placed on a heating mantle having a power rating of 450 watt and
timed. The samples were boiled with water which helps to release the
oil held within the matrix of the flowers and leaves. The volatile oils
evaporated along with the water into the condenser connected to the
flask at 100oC and atmospheric pressure (Figure 1). The condensed
steam and oils were collected in a separating funnel where the essential
oil and water were separated. The water was drained off gently and the
oils were separately collected in a 10ml measuring cylinder and
measured. The measurement was taken at an interval of 20minutes for
180minutes. The cumulative volume of the oil was measured and
recorded. The traces of water in the essential oils were removed by
adding 1 gram of magnesium sulfate in the oil as a drying agent after
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which the yield obtained was calculated using the formula given below
(Seid et al., 2014)
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𝐚𝐦𝐨𝐮𝐧𝐭 𝐨𝐟 𝐞𝐬𝐬𝐞𝐧𝐭𝐢𝐚𝐥 𝐨𝐢𝐥 𝐨𝐛𝐭𝐚𝐢𝐧𝐞𝐝 (𝐠)

𝐲𝐢𝐞𝐥𝐝 𝐨𝐟 𝐞𝐬𝐬𝐞𝐧𝐭𝐢𝐚𝐥 𝐨𝐢𝐥 (%) = 𝐚𝐦𝐨𝐮𝐧𝐭 𝐨𝐟 𝐫𝐚𝐰 𝐦𝐚𝐭𝐞𝐫𝐢𝐚𝐥 𝐮𝐬𝐞𝐝 (𝐠) x 100
Determination of essential oil constituents

In order to identify the chemical constituents of the essential oils and
therefore determine their quality, the extracted essential oil samples
were analyzed using Gas Chromatography Mass Spectrometry (GC-MS)
Agilent 6890 gas chromatography instrument coupled to an Agilent
5973 mass spectrometer and an Agilent Chem. The following operating
parameters were used for the essential oil sample: capillary GC column
HP-5MS 5% phenylmethyl siloxane\(30 x 0.25mm i.d. x 0.25 mm film
thickness), a carrier gas Helium (flow rate 1.2mL min-1) and a split-less
injection mode. Injector temperature was 250°C; Oven temperature was
set initially at 50°C and then raised to 250°C at 10°C min-1 rate till the
end of the analysis. The eluted analyses detected using (5973 network)
mass selective detector and Electron Impact ionization (EID) was
carried out at 70 eV.
Making of perfumes
The essential oils from flowers and leaves were separately blended
together according to a formula given by Andreaet al. (2013). The oils
were mixed with varying proportions of methanol, ethanol and
formaldehyde to develop the scent. The perfume was aged for 4 months,
after which the quality was determined as high or low
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Figure 1. Essential oil distiller based on standards described by Chouitah

et al. (2011).
Data analysis
Data was presented using frequency tables. The relationship between
time and yield of essential oils was determined using Pearson correlation.
The differences in yield of essential oils and the percentage of
compounds
in the flowers and leaves were calculated using t-test. All the data
analysis procedures were carried out using Statistical Package for Social
Sciences (SPSS) version 17.0 software.
RESULTS
Extraction of Essential Oils
The percentage yield of essential oils varied from 4.5 after the sampled
were heated for 180 minutes to 0.5 after heating for 20 minutes in
flowers (Table 1). On the other hand, the percentage yield in leaves
ranged from
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3.5 after the samples were heated for 180 minutes to 0.1 after heating for
20 minutes. The weights of the plant samples, volume of distilled water
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and the heating temperature were maintained constant at 400g, 1.5 L,

100oC respectively. There was a relationship between heating time and
yield of essential oils from flowers (r=1) and leaves (r=0.99). Conversely,
there was no significant difference in the amount of essential oils
produced by flowers and leaves (P=0.08)
Constituents of essential oils
The percentage composition of myrcene in flowers was 4.02 and in
leaves 4.30, β-lin1oo1 (0.69, 0.70), nerol (20.00, 24.99), citral (16.09,
15.06),
limonene oxide (0.20, 0.91), cineole (10.29, 11.02), berbenol (2.10, 1.56)
and oleic acid (25.76, 24.86) (Table 2). There was no significant
difference in the percentage composition of the different compounds in
the essential oil extract from flowers and leaves (P=0.46).
Making of perfumes
The addition of a mixture of methanol and formaldehyde to the scented
products gave perfume of the highest quality (Table 3). In addition,
addition of methanol yielded a product of high quality. However, a
mixture of ethanol and formaldehyde gave low quality product just like
the addition of ethanol alone.
DISCUSSION
The induction period for extraction of essential oils was between 0-20
minutes. Induction period, is the time required to rupture the cells of the
plant material in order to release the oil and transport it to the condenser
(Girotra et al., 2013). In the present study, the induction period was
higher in the leaf as compared to the flower samples. This may be
attributed to differences in particle strengths, suggesting that flower
samples have higher particle strength than leaf samples (Solomon et al.,
2013). This necessitated the longer induction time in flower samples.
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The
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induction period observed in this study agrees with results obtained by

other researchers (Beena and Humaira, 2015). In both the flower and leaf
samples, the yield of essential oils increased with time from 20 minutes
reaching a maximum after 180 minutes. This confirms Ficks’s second
law of diffusion about the final equilibrium achieved by the solute
concentrations in plant matrix and in the solvents after a certain time
(Pereira and Meireles, 2010). Fick indicates that extraction beyond final
equilibrium solute concentration leads to no significant improvement in
oil yield when prolonging the extraction time. These results concur with
a study carried out elsewhere by Verma et al., (2010)
Table 1. Yield of Essential Oil extracted from flowers and leaves of the lavender
plant
Plant Weight (g) Distilled H2O Heating time Temperature Yiel

part (L) (Min) (oC) d
(%)
Flower 400 1.5 20 100 0.5

400 1.5 40 100 1.0
400 1.5 60 100 1.5
400 1.5 80 100 2.0
400 1.5 100 100 2.5
400 1.5 120 100 3.0
400 1.5 140 100 3.5
400 1.5 160 100 4.0
400 1.5 180 100 4.5
leaves 400 1.5 20 100 0.1
400 1.5 40 100 0.3
400 1.5 60 100 0.5
400 1.5 80 100 1.0
400 1.5 100 100 1.5
400 1.5 120 100 2.0
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400 1.5 140 100 2.5

400 1.5 160 100 3.0
400 1.5 180 100 3.5
Table 2. GC-MS analysis of eucalyptus leave oil extracted using different methods
compounds 𝐜𝐨𝐦𝐩𝐨𝐬𝐢𝐭𝐢𝐨𝐧 (%)

𝐅𝐥𝐨𝐰𝐞𝐫𝐬 𝐋𝐞𝐚𝐯𝐞𝐬
Myrcene 4.02 4.30

β lin 1OO1 0.69 0.70
Nerol 20.00 24.99
Citral 16.09 15.06
Limonene oxide 0.20 0.91
Cineole 10.29 11.02
Berbenol 2.10 1.56
Oleic acid 25.76 24.86
Table 3. Quality of perfume from flowers and leaves of lavender plant after addition
of other ingredients
Ingredient Quality
Methanol High
Ethanol Low
Methanol + Highest
formaldehyde Ethanol + Low
formaldehyde
The percentage composition analysis of the essential oils carried out in

the present study was meant to determine the quality of the products
(Table 2). The results concurred with a previous study carried out in
Nigeria by Kabiru et al., (2016). This could be attributed to the sources
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of the materials under study being similar ecologically (Bencheqroun et

al.,
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2012). However, Gracelin et al., (2013) obtained different results from

the ones obtained in this study. The previous study targeted plant
materials collected from a semi-arid area while usthe current study was
carried out in an area that receives an adequate amount of rainfall. It is
also evident that the values obtained for the various components were
almost the same in both flower and leaves extracts. This disagrees with
the result obtained by Baba-Moussa et al., (2012) in Benin.
Thevariations could have emanated from differences in time of
harvesting and methods of processing the samples (Gupta et al., 2015).
A study carried out elsewhere obtained similar results on quality of
perfumes, after addition of other ingredientsas the current study (Bonou
et al., 2016) (Table 3). This could be attributed to the standard polarity
of methanol, ethanol and formaldehyde (Mariott, 2010). However, when
carrying a similar study, Kabiru et al., (2016) obtained contrasting
results with the ones obtained in this study. The possible reason could be
that the earlier study compared flower and leaves from different plants.
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CONCLUSION
This study shows that lavender plants from Egerton University are a
potential sources of essential oils.Essential oils are the natural volatile
compounds having loveable odor. The essential oils are isolated mostly
from the hydrodistillation method which is more suitable for this process
and easy to carry. Whole parts of the plants are used for the extraction of
plants. Steam distillation method is expensive than the hydrodistillation,
so it is less preferred. Essential oils have good medicinal applications and
used in the treatment of different diseases including the infectious
diseases, depression, anxiety, act as the antifungal, antimicrobial,
anticancer, and wound healing; they are also used in cosmetics and
perfume industries. In the field of heath, essential oils are used more
frequently and are mostly applied to the external body parts to relieve
the pain. In the field of fragrance, essential oils are used in the perfume
industry and due to attractive odor, the essential oils are used mostly in
this industry. It is used worldwide and due to their better usage, the world
essential oil market is growing rapidly and getting more importance day
by day.
The oils can be exploited economically for making of perfumes which
are sources of income. In addition, the perfumes can alleviate the
problem of side effects that comes after use of poorly made perfumes that
have penetrated our market. There is need for moving this study a notch
higher by producing the perfumes in large scale.
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REFERENCES
1) Andrea, C., Massimo, E. and Andrea, O. (2013). Supercritical Fluid
Extraction of Plant Flavors and Fragrances. Molecules; 18: 7194-
7238
2) Baba-Moussa, F., Adjanohoun, A.and Adeoti, K. (2012).
Antimicrobial properties and phytochemical profiling of essential oils
Extracted from traditonally used medicinal plants in Benin. Int. J.
Natural Product Sci.; 2 (4): 1-11.
3) Bachhav, R. and Shankar. (2013). Antinocicepive and
antiinflammatory activity of alkaloid fraction of Trichopus
zeylanicus Gaerntn. Inter. J. Pharm. Pharmaceu. Sci., 4(2): 1-5
4) Beena, J. and Humaira, A. (2015). Liver Protective Activity of
Indoneesiella echioides Against Carbon Tetrachloride (CCl4)- Induced
Hepatotoxicity in Rats. Pharmacol., 2: 416-429.
5) Bencheqroun, H., Ghanmi, K. M. and Satrani, B. (2012).
Antimicrobial activity of essential oils of Artemisia mesatlantica,
endemic plant to Morocco. Bul. Royal Society of Liège Science, 81:4-
21.
6) Bonou, J., Ahouandjinou, H., Baba-Moussa, F., Adéoti, Z.,
Dougnon, V., Metongnon, I., Gbenou, D., Toukourou, F. and Baba-
Moussa, L. (2016). Assessment of the antimicrobial activity of
essential oils from some Beninese medicinal plants: Influence of
different tweens. Issues in Biological Sciences and Pharmaceutical
Research; 4(6): 43-49.
7) Chouitah, O., Meddah, B. and Aoues, A. (2011). Chemical
composition and Antimicrobial Activities of the Essential Oil
from Glycyrrhiza glabra Leaves, Jeobp 14 (3):l284- 288.
8) Coelho, J. P., Cristino, A.F., Matos, P. G., Rauter, A. P., Nobre, B. P.,
Mendes, R. L., Barroso, J. G., Mainar, A., Urieta, J. S., Fareleira, J. M.,
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Sovova, H. and Palavra, A.F. (2012). Extraction of Volatile Oil from
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Aromatic Plants with Supercritical Carbon Dioxide: Experiments and

Modeling. Molecules; 17: 10550– 10573.
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Products with Activity against Oral Bacteria: Potential Application in the
Prevention and Treatment of Oral Diseases. Hindawi Publishing
Corporation journal; 5: 15-25.
10) Fornari, T., Vicente, G. Vazquez, E. Garcia-Risco, M. R. and
Reglero, G. (2012). Isolation of essential oil from different plants
and herbs by supercritical fluid extraction. J. Chromatogr.; 12: 4–
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and Quantitative Analysis of Phytochemicals In Five Pteris Species Int.
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fluid technology: a promising approach in pharmaceutical research.
Pharm. Dev. Technol.; 18: 22–38
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(2013). Compressed fluids for the extraction of bioactive compounds.
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Final Project MANISH

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MAKING OF PERFUME WITH

PACHPANDE MANISH SANJAY

Under Guidance of:

PROF. MR. RAHULSING. U. KHAIRNAR

KAI. YASHODABAI DAGADU SARAF COLLEGE OF

Kundan Sunil in the partialfulfillment of Degree Of Bachelor of

This Review Project is now ready for evaluation.

Prof. Mr. Rahulsing. U. Khairnar Dr. P. R. Patil

NO research is ever they outcome of single individual's efforts. This

With a feeling of profound pleasure, I gratefully own my sincere thanks

PACHPANDE MANISH SANJAY

PATIL KUNDAN SUNIL

(Sambath et al., 2012). Beauty

Analysis of Essential Oils

Isolation of Oils from Natural Sources

essences are obtained by extraction methods. They are as follows: l

Concretes/ Absolutes and Essential Oils

Natural versus Artificial Mix

the remaining 4% of trace compounds being non-duplicated. It is the

their natural quality by hot water or steam distillation process.

Carrier Oils and Alcohol

Molecular Structure of Aromatic Compounds and

Olfactive Properties and Chemical Identities of Floral

lavender flower and has a fine fresh floral odor-character. Coriandrol

Tetrahydro-myrcenol and its Acetate

Some Aspects of the Adulteration of Natural Isolates

natural source or produced synthetically. Examples are,

1. citral (eg. Litsea cubeba oil or synthetic) to standardize lemon oil;

commercializing of a product appears to be a fraudulent act, it ought not

MATERIALS AND METHODS

𝐚𝐦𝐨𝐮𝐧𝐭 𝐨𝐟 𝐞𝐬𝐬𝐞𝐧𝐭𝐢𝐚𝐥 𝐨𝐢𝐥 𝐨𝐛𝐭𝐚𝐢𝐧𝐞𝐝 (𝐠)

Determination of essential oil constituents

Figure 1. Essential oil distiller based on standards described by Chouitah

and the heating temperature were maintained constant at 400g, 1.5 L,

induction period observed in this study agrees with results obtained by

Plant Weight (g) Distilled H2O Heating time Temperature Yiel

Flower 400 1.5 20 100 0.5

400 1.5 140 100 2.5

compounds 𝐜𝐨𝐦𝐩𝐨𝐬𝐢𝐭𝐢𝐨𝐧 (%)

Myrcene 4.02 4.30

The percentage composition analysis of the essential oils carried out in

of the materials under study being similar ecologically (Bencheqroun et

2012). However, Gracelin et al., (2013) obtained different results from

Sovova, H. and Palavra, A.F. (2012). Extraction of Volatile Oil from

Aromatic Plants with Supercritical Carbon Dioxide: Experiments and

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