CHAPTER 1

THE PROBLEM AND ITS BACKGROUND

1.1 Introduction

Cymbopogon citratus or commonly known as lemon grass is a tropical plant native to

tropical regions of Asian, American and Afican continents. Otherwise known as Squinant in

English, lemon grass is known by various other colloquial names throughout the world. Lemon

grass is a perennial monocotyledonous grass which can grow up to 6 feet in height and 4 feet in

width. It grows in clusters. It has long, slender, drooping bright green leaves that measures from

1.3-2.5 cm in width and 3 feet in length. Leaves are simple with entire margins. Flowers grow on

spikes. It has a lengthy inflorescence ranging from 30-60cm. The floral arrangement of this

scented grass gives it the name ‘Cymbopogon’. The members of Cymbopogon genus produce

volatile oils and thus are also known as aromatic grasses. A predominant feature of this grass

with strong lemon fragrance is due to high citral contents of its oil, it produces a diverse array of

bioactive compounds and exhibits a wide range of therapeutic activities. It has already found

applications in the cosmetic and perfumery industries due to its strong fragrance. It is also the

starting material for the manufacture of ionone’s, which produce Vitamin A. Lemon grass,

contains several bioactive compounds that impart medicinal value to it. Considerable evidence is

available for its medicinal properties that make it a potent herb for pharmacognostic applications.

This study tends to prove how does the components or properties of lemon grass provides things

that are used on making various drugs or medicines that helps to cure several diseases. The

therapeutic value of lemon grass and its essential oil may enable its use in herbal medicine in

future. This study would like to develop depth of understanding and which are distinct enough to

ensure that a certain kind of medicinal plant like lemon grass is used with the intention of

maintaining health to be administered for a specific condition. Although a lot of pharmacological

investigations have been carried out based on the ingredients present, but a lot more can still be

explored, exploited and utilized.

1.2. Conceptual/Theoretical Framework

Cymbopogon Citratus as a potent

herb for pharmacognostic
applications

Components of Cymbopogon Citratus

Attribute Therapeutic benefits Uses

How this thing does helps on curing

diseases

1.3. Statement of the problem

This study about Cymbopogon citratus as a potent herb for pharmacognostic applications

tends to reveal how does its component takes place on producing medicines for various diseases.

Due to this it deepens one’s understanding on the different ways in authentication of plants and

ensures reproducible quality of herbal plants specifically the lemon grass.

The study addressed the following specific questions:

1. Main problem:

A. What are the components of Cymbopogon Citratus?

2. Sub problem:

2.1 What are the benefits of its component?

2.2 How do its components cure diseases?

 2.3 What the effects of this plant in human body?

1.4. Significance of the study

This study focuses on the Cymbopogon citratus as a potent herb for pharmacognostic

applications. It also gives information to the readers for them to gain and have further knowledge

about medicinal plants like lemon grass. The study gives interest in some readers by giving them

enough uncommon facts when it comes to cymbopogon citratus. The result of this study will

contribute greatly to the benefit of the society. The result of this study would be a great help to

different kinds of individual, community, readers and future researchers.

1.5 Scope and Limitations

This study is delimited in knowing Cymbopogon citratus as a potent herb for

pharmacognostic applications. Thus, it will mainly focus on its components and uses specially its

pharmagostic applications. The purpose of this study is to determine how does the properties of

lemon grass contributes to the field of medicine. The population of the study will be the people

within all ages who wants to know about the lemon grass as a medicine. The study will be finish

on 2019.

1.6. Definition of terms

Cymbopogon citratus- lemon grass

Potent herb- strong or valuable medicinal plant

Pharmacognosy- study of medicines

Therapeutic benefits- response after treatment

Disease- an impairment of health or a condition of abnormal functioning

 CHAPTER 2
REVIEW OF RELATED LITERATURE

This chapter will provide the different facts, effects, and information about

cymbopogon citratus as a potent herb for pharmacognostic applications

(Sighn, 2010), Cymbopogon citratus is native to Sri Lanka and South India, and is now

widely cultivated in the tropical areas of America and Asia. The plant is used as a fragrance and

flavoring agent and in folk medicine as an antispasmodic, hypotensive, anticonvulsant, analgesic,

antiemetic, antitussive, antirheumatic, antiseptic and treatment for nervous and gastrointestinal

disorders and fevers. The plant is also used as an antibacterial, antidiarrheal and antioxidant, but

the mode of action for the different bioactivities has not been studied in detail. Cymbopogon

citratus contains various phytoconstituents such as flavonoids and phenolic compounds,

terpenoids and essential oils, which may be responsible for the different biological activities.

Hence, we can isolate some pure phytopharmaceuticals, which in turn can be used as lead

molecules for synthesizing the novel agents having good therapeutic activity. With regard to the

development of quality herbal medicine standardization of the extracts, phytopharmacology of

different extracts, isolation and characterization of active phytopharmaceuticals, elucidation of

the mechanism of action of the isolated compounds and clinical trials of the compounds are

much needed. In the changing global scenario, the interest toward plants with medicinal value is

increasing substantially in the primary healthcare system both in the developed and in the

developing countries. Therefore, the information will help scientists and researchers to screen the

compounds responsible for different bioactivities and to elucidate the molecular mechanism of

action.
 The chemical composition of the essential oil of Cymbopogon citratus varies according to

the geographical origin, the compounds as hydrocarbon terpenes, alcohols, ketones, esters and

mainly aldehydes have constantly been registered. The essential oil (0.2–0.5%, West Indian

lemon grass oil) consists of, mainly, citral. Citral is a mixture of two stereoisomeric monterpene

aldehydes; the trans isomer geranial (40–62%) dominates over the cis isomer neral (25–38%),.

Chemical structure of the major constituents of lemongrass essential oil.

Lemongrass consists of luteolin and its 6-C and 7-O–glycosides, isoorientin 2’-O-

rhamnoside and isolation of the flavonoids quercetin, kaempferol and apiginin from the aerial

parts. The phenolic compounds elimicin, catecol, chlorogenic acid, caffeic acid and

hydroquinone are also isolated from the plant.

Salvador(2008), Superficial mycoses of the skin are among the most common

dermatological infections, and causative organisms include dermatophytic, yeasts, and non-

dermatophytic filamentous fungi. The treatment is limited, for many reasons, and new drugs are

necessary. Numerous essential oils have been tested for both in vitro and in vivo antifungal

activity and some pose much potential as antifungal agents. By using disk diffusion assay, we

evaluated the antifungal activity of lemongrass oil and citral against yeasts of Candida species

(Candida albicans, C. glabrata, C. krusei, C. parapsilosis and C. tropicalis). This study showed

that lemongrass oil and citral have a potent in vitro activity against Candida spp.

Khayyat & Saddiq(2010), 6,7-Citral-epoxy derivative (a mixture of E and Z isomers with

respect to the C2 = C3 double bond) could be react with DNA base producing a major adduct.

The mixture of epoxides was condensed with 2 mol of cytosine to give the adduct through
condensation between aldehyde and amino groups. Antifungal and antibacterial studies were

carried out on citral and citral-epoxide. Studies on the antifungal

especially Penicillium italicum and Rhizopus stolonifer showed that citral and citral-epoxide

have good antibacterial action. Antimicrobial studies of P. italicum and R. stolonifer explained

also that citral and citral-epoxide have good antimicrobial activity. Citral epoxide shows high

activity against the growth of bacteria methicillin resistant Staphylococcus aureus (MRSA) and

fungi comparing by citral. The epoxide shows antibacterial activity more than the antibiotics

nalidixic acid (NA) and ampicillin (AP) and nitrofurantoin (NI). The results revealed that these

complexes are most effective against MRSA.

Onawunmi & Ogunlana(2008), Lemon grass oil obtained by the steam distillation of the

fresh aerial parts of Cymbopogon citratus (DC.) Stapf was investigated for antibacterial activity.

The results obtained suggests that the oil possesses rapid bactericidal activity against Gram-

negative and Gram-positive bacteria, the latter being more susceptible. Other factors that could

affect the activity of the oil were also investigated. The dispersion of the oil in a dilution of

dimethyl sulphoxide which does not have an effect on the microorganisms studied provided a

convenient medium for the assay. It is being suggested that more information about the

antibacterial activity of essential oils could be obtained using such a methodology.

Olorunnisola, Asiyanbi, Hammed, Simsek(2014), The use of herbal preparations remained the

main approach of folk medicine to the treatment of ailments and debilitating diseases. Initial

intensive researches conducted on Lemongrass extracts (tea) may have showed conflicting

evidences, however the resurgence in claims of folk medicine practitioners necessitated further

inquiry into the efficacy of the tea. Lemongrass tea contains several biocompounds in its

decoction, infusion and essential oil extracts. Anti-oxidant, anti-inflammatory, anti-bacterial,

anti-obesity, antinociceptive, anxiolytic and antihypertensive evidences of lemongrass tea were

clearly elucidated to support initial pharmacological claims. Lemongrass tea was non-toxic, non-

mutagenic and receives wide acceptance among alternative medicine practitioners in several

developing countries. This review therefore presents previous research activities, technologies

and information surrounding bioactivities of lemongrass tea.

Tajidin(2012), The demand for lemongrass (Cymbopogon citratus) is for its high citral

content. Early or delayed harvesting of lemongrass affected essential oil and citral content. The

objective of the study was to determine the effects of three maturity stages at harvest of

lemongrass on essential oil, chemical composition and citral contents. The lemongrass plant was

planted using a randomized complete block design with four replications, at the University

Agriculture Park, Universiti Putra Malaysia. The plants were harvested at 5.5, 6.5, and 7.5

months after planting. After harvest, the essential oil, chemical composition and citral contents

were analysed using gas chromatography-mass spectrometry (GC-MS) analysis. There were

significant effects of maturity stages on essential oil and citral contents. Lemongrass harvested at

5.5 and 6.5 months after planting had significantly higher oil contents than those harvested at 7.5

months. A total of 65 compounds were detected from all the three stages of maturity. However,

only 13 compounds were present at each of the maturity stage. Among 13 compounds, only 7

compounds (β-myrcene, 3-undecyne, neral, geranial, nerol, geranyl acetate and juniper camphor)

had a concentration of greater than 1%. The citral content at 6.5 months after planting was higher

by 11.4% than at 5.5 months after planting. The citral content decreased by 5.4% when

lemongrass was harvested at 6.5 compared to at 7.5 months after planting. Citral content peaked

at 6.7 ± 0.3 months after planting. Thus, maturity stage at harvest influenced essential oil and
citral contents of lemongrass. Therefore, lemongrass should be harvested at the appropriate level

of maturity in order to achieve high quality essential oil and lower production cost.

Martinez-Castillo(2018), Pharmacological investigations. Summary of findings

Anti-amebic Effect

The essential oil in broth culture was active on Entamoeba histolytica. According to the study

Plants rich in flavonoids has compounds that can be use against infectious diseases cause by

protozoan parasites.

Antibacterial Activity

The chromatographic fraction of the essential oil in agar plate was active on Bacillus subtilis,

Escherichia coli, Staphylococus aureus and Salmonella paratyphi and Shigella flexneri. These

activities are shown in two of the three main components of the oil identified through

chromatographic and mass spectrometric methods. While the α-citral (geranial) and β-citral

(neral) components individually elicit an antibacterial action on gram-negative and gram-positive

organisms, the third component, myrcene, did not show any observable antibacterial activity on

its own.The extract was also active when the volatile oil extract was oxidized via the active

oxygen method.

Antidiarrheal Activity

Cymbopogon citratus stalk decoction reduced the fecal output in a dose-dependent manner.
Antifilarial Activity

Fresh leaves were active on Setaria digitata.

Antifungal Activity

Lemon grass oil is active against such dermatophytes such as Trichophyton mentagrophytes, T.

rubrum, Epidermophyton floccosum and Microsporum gypseum, and is among the most active

agents against human dermatophytes. Other studies reported that lemon grass oil is active against

keratinophilic fungi, 32 ringworm fungi and food storage fungi.Lemongrass oil is also effective

as a herbicide and as an insecticide because of these naturally occurring antimicrobial effects.

Anti-inflammatory Activity

The hot water extract of the dried leaves administered intragastrically to rats was active when

compared with carrageenin-induced pedal edema.

Antimalarial Activity

The essential oils of Cymbopogon citratus were found to produce 86.6% suppression in the

growth of Plsmodium berghei when compared with chloroquine (taking inhibition by

chloroquine as 100%).

Antimutagenicity

The ethanolic extract of lemon grass extract exhibits an antimutagenic activity in various models

and retards the growth of fibrosarcoma cells transplanted in mice in association with the

prevention of lung metastasis. Theplant extract is known to show inhibition on the formation of

azoxymethane-induced DNA adducts and aberrant crypt foci in the rat colon. Inhibitory effects
of the plant extract on the early phase of hepatocarcinogenesis after initiation with

diethylnitrosamine were seen in 344 male Fischer rats.

Antimycobacterial Activity

The essential oil in agar plate was active on Mycobacterium smegaris.

Antinociceptive Effect

The essential oil of Cymbopogon citratus possesses a significant antinociceptive activity.

Comparing the results obtained with three different experimental models of nociception (hot-

plate, acetic acid-induced writhings and formalin test), we can speculate that the essential oil acts

both at the peripheral and at the central levels.

Antiprotozoan Activity

A dose-dependent antiprotozoan effect of the essential oil of Cymbopogon citratus could be

observed on two strains of Crithida deanei.

Ascaricidal Activity

The fresh leaf essential oil has an ascaricidal activity.

Free Radical Scavengers and Antioxidant Effects

Methanol, MeOH/water extracts, infusion and decoction of Cymbopogon citratus were shown to

have free radical scavenging effects by measuring the bleaching of the 1, 1-diphenyl-2-picryl-

hydrazyl (DPPH) radical, scavenging of the superoxide anion and inhibition of the enzyme

xanthine oxidase and lipid peroxidation in human erythrocytes.

Hypocholesterolemic Effect

The elevated cholesterol concentration was significantly lowered in the animals given the plant

extract. This reduction was found to be dose dependent. This result shows that the extract

possesses a hypocholestecolemic potential.

Hypoglycemic and Hypolipidemic Effects

A fresh leaf aqueous extract of Cymbopogon citratus administered in normal rats lowered the

fasting plasma glucose and total cholesterol, triglycerides, low-density lipoproteins and very

low-density lipoprotein dose dependently while raising the plasma high-density lipoprotein level

in the same dose-related fashion, but with no effect on the plasma triglyceride levels.

Larvicidal Activity

The fresh leaf essential oil has a larvicidal activity.

Neurobehavioral Effect

The essential oil was evaluated for sedative/hypnotic activity through pentobarbital sleeping

time, anxiolytic activity by elevated plus maze and light/dark box procedures and anticonvulsant

activity through seizures induced by pentylenetetrazole and maximal electroshock. The essential

oil was effective in increasing the sleeping time, the percentage of entries and time spent in the

open arms of the elevated plus maze as well as the time spent in the light compartment of the

light/dark box. In addition, the essential oil delayed clonic seizures induced by pentylenetetrazole

and blocked the tonic extensions induced by maximal electroshock, indicating the elevation of

the seizure threshold and/or blockage of the seizure spread.

 Ranade & Thiagarajan(2015), Lemon grass has been traditionally used to remediate a

plethora of medical conditions. This is due to the broad spectrum of secondary metabolites that it

produces. It has been used to treat fever, cough, elephantiasis flu, leprosy, malaria and digestive

problems among many other illnesses. The use of lemon grass in Ayurveda is still relevant today

due to its therapeutic value. Conventional medicine has a lot of adverse effects.

Therefore plant-based medicine has become a popular alternative for synthetic medicine. Thus,

this herbaceous

plant may find many applications in the pharmaceutical industry. Indonesian Scientists have

investigated and confirmed the ability of β-citronellol-the major component of “Sereh Wangi”

(colloquial name for Lemon grass in Indonesia) oil, to bring about a reduction in weight of rats

fed with a high fat diet. Inhalation of vapors of β-citronellolenhances the sympathetic nerve

activity of the rats that leads to the increased activity in the adipose tissue resulting in weight

loss. The findings of this study were significant due to the fact that β-citronellol caused a

reduction in body mass without affecting the concentration and activity of the liver enzymes.25

Weight loss is only one of the few pros of citronella grass. Maintenance of oral health is an

important aspect of daily routine. Gingivitis and periodontitis are oral health conditions caused

due to dental plaque. Many reports confirm that these dental caries are risk factors for ischemic

stroke and cardiovascular disease. The antagonistic activity of lemon grass against the planktonic

and biofilm forms of Candida dubliniensis, a common oral pathogen has been reported.

Citronella grass may be used in formulating herbal drugs for oral healthcare. Medical conditions

like hyperlipidemia, hypercholesteremia and hyperglycemia lead to metabolic disorders like

obesity and diabetes mellitus. It has been reported that lemon grass is bestowed with

hypolipidemic, hypocholesteremic and hypoglycemic properties. Consumption of plant extracts

have shown to bring about a reduction in plasma cholesterol and very low density lipids, both of

which are highly correlated with heart disease. Hypoglycemic condition in rats was achieved

after 42 days of administration of 500mg/kg/day lemon grass extract. The mechanism of action is

howevernot clear. In addition to these benefits, several reports have confirmed the anti-

inflammatory, anticonvulsant and anxiolytic effects of lemon grass extracts. Additionally, the

antagonistic activity of lemon grasstowards different pathogenic bacteria, protozoa and fungihas

also been reported. Leishmaniasis is severe disease that affects the global human population

annually on a large scale. It has been reported that the promastigotes of Leishmania infantum

undergo programmed cell death upon exposure to citral, a major constituent of lemon grass oil.

Also, reports suggest the anti-proliferative effect of citronella oil on the anexic amastigotes of

Leishmaniasp. Lemon grass, may be foreseen as an anti-protozoan drug of the future. Theability

of Cymbopogon species to inhibit the growth and aflatoxin production in Aspergillus sp. and in

Penicillium citrinum, thereby reducing the deterioration of melon seeds, has been reported. It is

also seen that the combination of silver nanoparticles and the oil have synergistic inhibitory

action on growth of pathogens like - Escherichia, Staphylococcus, Moraxella, Enterococcus and

Candida, Citronella oil, also exhibits potent antifungal activity against Candida sp. and

Aspergillus niger by showing inhibitory zones in the range of 35 to 90mm. Also, the oil shows

similar effects to 50 mg/kg synthetic oral drug diclofenac when administered in mice suffering

from carragennan induced edema. In vivo anti-inflammatory action of the oil is also evident

when it is topically applied on croton oil induced edematous mice. Lemon grass can be used in

preparations of topical skin creams and in the manufacture of plant based oral drugs. Research on

anti-microbial and anti-inflammatory activities of lemon grass, after its GC-MS analysis, has
revealed its major constituents as limonene, nerol,gerianal, geraniol and myrcene. Hence it is

inferred that these compounds may be responsible for its microbicidal and anti-inflammatory

effects. There is a lot of data that attests the anti-inflammatory effects of monoterpene aldehydes.

Bioactive compounds such as citral, neral and geranial have been found to inhibit the production

of IL- 1β, an inflammatory cytokine released by macrophages, upon exposure to pathogens

thereby reducing inflammation. Furthermore, the bioactive compounds of lemon grasshave also

shown potent antagonistic activity against viruses. Anti-viral effects of lemon grass against

onenveloped murine virus and have been reported. Murine novovirus is a surrogate virus of

human novovirus, which is responsible for non-bacterial gastroenteritis epidemic worldwide. The

lemon grass oil and citral used in the study reduced viral infectivity by coating the viral capsid

and thus preventing it from binding to the host cell. Lemon grass and citral can be used to

sanitize food and surfaces to prevent viral infections. The bioactive compounds in citronella

grass have been investigated for their anti-cancerous properties also. An emulsion of citral and

lemongrass oil exhibited its anticancerous properties on cervical cell lines (HeLa and ME-180)

by reducing cell proliferation and by initiating apoptosis. Also, a change in the mitochondrial

membranepotential and increase in ROS production was observed the cancerous cells upon

exposure to the emulsion. It has also been reported that two lipopolysaccharides containing

(1→4) linked b-d-Xylofuranose moiety, extracted from lemon grass, brought about apoptosis

inSiha and LNCap reproductive cancer cell lines via theintrinsic pathway. Hence it is envisaged

that the constituents of lemon grass may be used to form potentanti-cancer drugs in future.
 CHAPTER 3
RESEARCH METHODS

The contents of this chapter shows the components of the plant, materials used for
experimentation and methods that has been conducted to finish this study.

COMPONENTS OF CYMBOPOGON CITRATUS

1.1. Nutritional component of Cymbopogon citratus

Nutritional Component Quantity

1. Carbohydrate 55%

2. Crude fat 5.10%

3. Crude protein 4.56%

4. Crude fiber 9.28%

5. Energy 350.55 al/100g

1.2 Mineral Content of Cymbopogon citratus

Mineral Quantity(mg/100g)

1. Sodium 54.8

2. Potassium 59.5

3. Calcium 39.5

4. Magnesium 70

5. Iron 0.024

6. Manganese 0.952

7. Zinc 121

8. Phosphorus 89.3

9. Phytate 11860

1.3: Bioactive components of Lemon grass oil

Chemical Constituent
1. Citral
2. Burneol
3. α-terpineol
4. ß-Myrcene
5. ß-O-Cimene
6. Allo-o-cimene
7. α-Pinene oxide
8. Myrcenol
9. t-Muurolol
10. Linalool
11. 1-Octyn-3-ol trans-
12. Chrysanthemal
13 3-Undecyne 3-carvomenthenone
14. Citronellal
15. Neral
16. trans-(-)-Carveol
17. Geranial
18. Nerol
19. Citronellol
20. Methyl-n-nonyl-ketone
21. Dextro-carvone
22. Geranic-acid
23. α-Bergamotene
24. Isolongifolene-4-5-9-10-dehydro Levo-ß-elemene
25. γ-Muurolene
26. α-Gurjunene
27. α-Muurolene
28. a-Amorphene ß-Sesquiphellandrene
29. α-Farnesene
30. α-Elemol
31. d-Cadinene
32. Germacrene-D
33. Valencene
34. Viridiflorol
35. α-Selinene
36. Humulene
37. α-Guaiene
38. t-Cadinol
39. ß-Eudesmol
40. (E,E)-Farnesal pimelyl dihydrazide
41. Di-n-octylphthalate
42. Geranyl-acetate

MATERIALS AND METHODS

Extraction of Lemon Grass Oil

Essential oils are extracted from flowers, herbs, trees and various other plant materials. These

oils contain a mixture of chemical compounds. Terpenes associated with aldehydes, alcohols and

ketones form the major chemical component of such essential oils. Also finds an application in

the pharmaceutical industry. The extraction of this essential oil is classified as clean technology.

Lemon grass contains 1-2% of essential oil on a dry weight basis. Lemon grass oil is also known

as citronella oil. Steam and hydro distillation are the conventional methods of its extraction.

These procedures are however time consuming. An innovative Microwave-Assisted

Hydrodistillation (MAHD) not only reduces the extraction time but also retains the quality of oil.

The benefits of microwave radiation aided oil extraction technique over hydrodistillation have

also been reported. Pressurized liquid extraction using nitrogen gas, is a novel technique and was

found to yield better quality of oil in comparison to Soxhlet extraction and hydrodistillation

methods. Supercritical extraction of citronella oil with CO2 under high pressure has also been

investigated.

A. Plant Sample
Lemongrass (Cymbopogon Citratus) leaves were collected. The plant sample was freshly cut,

10cm from the root, in the morning of the day they were collected. Lemongrass (Cymbopogon

Citratus), the percentage essential oil yield for the partially dried leaves was found to be higher

than that of the fresh leaves. Thus, once collected, the plant material was dried at room

temperature for maximum 4 days, then kept in a sealed plastic bag at ambient temperature and

protected from the light. Extraction yield increase by decreasing the particle size due to the

higher amount of oil released as the leave cells are destroyed by milling. In order to improve the

collection efficiency, the plant material was soaked in its distilled water for 30min before the

extraction performed.
B. Reagents
N-hexane was used as analytical grade reagent purchased from Research Lab.

C. Solvent Extraction Method

150g of the dry sample of lemongrass were weighed from the sliced lemongrass sample and

placed in a 1 lit clean flat bottom flask. 500ml of N- hexane solvent were poured into the flask.

The flask and content were allowed to stand for 36 hrs; this was done to extract all the oil content

in the lemongrass and for complete extraction. After which the extract was decanted into another

1 lit beaker. 200ml of Ethanol were added to extract the essential oil since essential oil is soluble

in Ethanol. The mixture was then transferred to 500ml separating funnel and separated by a

process called liquid/liquid separation process. The content of the separating funnel was and

allowed to come to equilibrium, which separated into two layers (depending on their different

density). The lower Ethanol extract and the upper Hexane layer were collected into two separate

250ml beaker and were placed in a water bath at 78oC. This was done to remove the Ethanol

leaving only the natural essential oil. The yield of oil was determined by weighing the extract on

an electronic weighing balance. The difference between the final weight of the beaker with

extract and the initial weight of the empty beaker gave the weight of essential oil.

D. Steam Distillation Method

150g of fresh lemongrass sample were placed into a 1 lit round bottom flask containing 250ml of

distilled water. The flask was fitted with a rubber stopper connected to condenser and heated.

Water at 0oC flowed counter currently through the condenser to condense the ensuring steam.

When the water reached 100oC it started boiling ripping off the essential oil from the

lemongrass. When the lemongrass got heated up, the essential oil that was extracted from the leaf

mixed with the water vapor. Both passed through the condenser and the vapor was condensed

into liquid. With the use of ice block, cooling was made possible and volatilization of the
essential oil was avoided. The condensate was directly collected using a 500ml beaker and then

poured into a separating funnel. This formed two layers of oil and water. The tap of the

separating funnel was opened to let out the water while the oil was immediately collected into a

100ml stoppered bottle. The bottle was closed tightly to prevent vaporization of the essential oil.

The oil was collected and the volume of oil obtained was weighed.

E. Formulation of Perfume

10ml of lemongrass essential oil extract were measured and placed in a 120ml beaker containing

5ml of Methanol. 5ml of the Fixatives were added to the mixture (to improve the longevity of the

perfume). The solution was shaken and poured into a 50ml bottle.

Yield of oil that obtained was calculated by:

Yield of essential oil =

a m o u n t of essential oil (g) obtained / amount of raw materials (g) used

CYMBOPOGON CITRATUS(Lemon Grass)