Acknowledgement

First and foremost, we would like to thanks God who helps us and being with throughout our
endeavor. Next, we would like to express our deepest gratitude to our advisor Mr. Abelneh.
for his advice, comment and support, suggestion for better compile of our project. Last but
not least, we would like to express our thanks and loves to our families for their support and
giving moral to accomplish this senior essay.
 CHAPTER ONE
INTRODUCTION
1.1 Background
The word “soap” came from the Latin word “Sapo.” it is believed that the name derived from
Mount Sapo in Rome[1]. The first production of soap happened around 2800BC in ancient
Babylon[2]. The Babylonians combined wood ashes with animal and plant fat, and got a
substance that was effective for cleaning. "The cold process method" is the most popular soap
making process today. Some soap makers use the hot process, which was much more
significant in past centuries. Soap is the term for a salt of a fatty acid or for a variety of
cleansing and lubricating products produced from such a substance. Soap is the combination
of fatty acids and alkalis obtained by reacting various animal and vegetable fats and oils with
caustic soda or potash[3]. The soap making reaction is called saponification. Soap prepared
from caustic soda is hard while soap from caustic potash is soft. Both soaps are readily
soluble in hot water. However, they dissolve very slowly in cold water forming a turbid
solution owing to slight decomposition. The soap itself was never actually "discovered", but
instead gradually evolved from a crude mixture of alkaline and fatty acid [4]. Cleanliness is a
very important thing due to the increasing number of diseases caused by bacteria and germs.
The soap and soaps industry is very essential in people's day to day life. The soap and soap
industry includes companies that are primarily engaged in manufacturing soap, synthetic
organic soaps, inorganic alkaline soaps, and crude and refined glycerin from vegetable oils
and animal fats. It is an international industry, and during the early years of 1990, world
demand for its products has increased 1 to 3 percent every year[5]. It has been made for more
than 2500 years and the first recorded manufacture of soap was in 600BC by the Phoenicians.
Actually, they used soap for soap medicinally but in the nineteenth century, it began to spread
in the Western world as cleaning soap. The invasion of non-native species is one of the major
drivers of environmental and socioeconomic changes around the world. Over the past 30-50
years, non-native Prosodic species have emerged as a major invasive plant of the arid and
semi-arid lands of eastern Africa[6]. Invasive plants are naturalized plants that produce a
large number of offspring, have the ability for long-distance dispersal, and thus have the
potential to spread over a considerable area [7]. Soap is a common cleansing agent well
known to everyone. Many authors defined soap indifferent ways and regarded it as any
cleaning agent, manufactured in granules, bars, flakes, or liquid form obtained from reacting
salt of sodium or potassium of various fatty acids that are of natural origin (salt of non-
volatile fatty acids). Soap can also be said to be any water-soluble salt of fatty acids
containing eight or more carbon atoms. Several things affect the soap-making process and the
quality of this soap produced[8]. The characteristics of this soap depend on the quality of oil,
and the amounts of the caustic soda and water used to make it. The speed of the reaction
between the oil and the caustic soda is influenced by the free fatty acid content of the oil, the
heat of the components before mixing, and how vigorously the mixing is to be done. Free
fatty acid contents, vigorous mixing, and heat speed up the given soap-making process. Soaps
are produced for a variety of purposes ranging from washing, bathing, medication, etc. The
cleansing action of the soap is due to the negative ions on the hydrocarbon chain attached to
the carboxylic group of the fatty acids [9]. The affinity of the hydrocarbon chain to oil and
grease, while the carboxylic group to water is the main reason soap is being used mostly with
water for cleaning purposes. Household uses for soaps include washing, bathing and other
types of housekeeping, where soaps act as surfactants, emulsifying oils to enable them to be
carried away by water. Soap usually comes in a solid, molded form, called a bar, based on its
typical shape. The chemical composition is different for the installation of soap, but the same
mechanism, it adversely affected by hard minerals in the water [10]. Therefore, this study
aims to provide a means to use the most invasive. At the beginning of the second half of the
twentieth century past sales exceeded sales of soaps, soap world, where replaced in the field
of laundry and household cleaning and industrial, with the exception of templates (parallel)
soaps are still used to clean the body, which began shampoo replaced by now. The needs for
producing quality soap from locally source Phytolacca dodecandra require the assessment of
the extraction as will be demonstrated in this studies. This will provide vital information on
the presence of liquid-liquid extraction that can separate the clear juice (Saponione) to the
user. In Ethiopia, toxic plants, berries of Phytolacca dodecandra are being commonly used for
washing clothes and to control fresh water snails. Since the discovery of Endod in 1965, there
have been extensive studies on the chemistry, toxicity, and epidemiology of Lemma toxins,
together with cultivation of the Endod plant[11]. There are many types of soaps, depending
upon the usage. There are hard and soft. Hardness of soap is often achieved through the
addition of hardening agents, so many natural soaps tend to be softer. They are further
categorized into two. Those are;
 Cleansers and
 Soaps
 Cleansers: Those are often made with mild abrasives and they are formulated to eliminate
heavy oil or solid particles and hard-to-remove stains. The cleansers come in many different
types depending on the type of abrasives they contain.
Soaps: Dish soaps are made to remove tough grease and release the solid dirt particles in the
foam that is produced by the soap.
.
1.2 Statement of Problem
In our country the raw material for soap production is basically imported from other
countries which is soap noodles with higher cost, even though the sanitation activity of the
society in our country is poor. Additionally, the price of soap is increasing from day to day
due to the increase in its high demand to the society. In our country, the scarcity of soap is a
chronic problem and also it is not easily accessible to societies. Many people in Ethiopia
specifically in the rural areas, are inhabited with low-income and cannot afford soap, instead
many of them are intending to use Endod for cleaning purpose. This in turn makes the
societies vulnerable to diseases like, diarrhea and cholera, etc. The soap helps in improving
sanitation and hygiene to address stunting, diarrhea, and trachoma. Around 60 to 80 percent
of communicable diseases are attributed to limited access to inadequate sanitation and
hygiene services.
So, to alleviate those problems we have intended to prepare antimicrobial solid soap from
domestic raw materials. We are majorly focus on endod since it is traditionally used by
almost all Ethiopians for hygiene and sanitation purpose and used as a medicine for a skin
attacked by fungal diseases. The Hot process is takes place within a few minutes to a few
hours when compared with other process and it is more suitable for laboratory or industrial
preparation, yields a more chemically pure powder; by-products and excess starting materials
are separated. So that, the oil used to formulate ant fungi bar soap will be extracted from
endod fruits using soxhlet which is used to extract the endod juice from endod fruits and use
Hexane as a solvent.
1.3 Objectives
1.3.1 General Objective
Formulation of ant fungi bar soap from Endod (Phytolacca dodecandra) fruit by using
additives such as sodium chloride, potassium hydroxide and hydrogen peroxide
1.3.2 Specific Objectives
a) Preparation of Raw materials
 b) Extraction of oil from phytolacca dodecandra fruit by using soxhlet extraction
method and Hexane as a solvent
c) Characterization of the extracted oil based on their chemical and physical properties
d) To formulate highly effective ant fungal bar soap by using oil extracted from endod
fruit through hot process soap making method
e) To study the effects of operating conditions (temperature, concentration of lye,
reaction time) on the quantity and quality of the products by using hot process soap
making.
Scope of the study
Our study will extend in collection of Phytolacca dodecandra fruit from East Hararghe up to
production of Anti-fungal bar soap. The fruit will be collected, clean and we measure the
initial moisture content. Drying and grinding to appropriate size. Extraction of oil from
Phytolacca dodecandra fruit the study also further analysis the laboratory experimental work
to determine the pH value, solubility, viscosity and odor of bar soap.
Significances of the study
The aim of this project is to produce bar soap which can be highly used on the infected body
parts of that caused by fungus and other microbial, and additionally to wash cloth both in
hard and soft water with natural based soap from the Endod fruit oil. By doing so the first
beneficial are farmers who are going to prepare a farm of this plant, because raw materials
suppliers for the industries that are engaged in the production of Endod fruit based bar soap.
The soaps are also sold for the purpose of dish washing and cleaning floors industrial use in
textiles, and commercial buildings, hospitals& clinic, and hotels, similarly railway coaches,
road vehicles, aircrafts, milk bottling plants and food preservation equipment are washed with
the help of soaps. Used to minimize the cost of importing / import substitution soap and to
introduce the technology of extracting oil from Phytolacca dodecandra (endod).
 CHAPTER TWO

LITERATURE REVIEW

2.1 World production trends of soap

Worldwide sales for soap exports by country totaled US$11 billion in 2019. The global soap
market size is projected to reach USD 55.29 billion by 2027, exhibiting a CAGR of 5.0%
during the forecast period. Rapid transmission of the COVID-19 infection is set to play a
central role in fueling the growth of this market, states Fortune Business Insights in its recent
report, titled "Soap Market Size, Share & Industry Analysis, By Product Type (Bath & Body
Soaps, Kitchen Soaps, and Laundry Soap), Form (Solid and Liquid), Application (Household
and Commercial), Distribution Channel (Hypermarkets/Supermarkets, Pharmacies, Online
Channels, and Others), and Regional Forecast, 2020-2027". According to the World Health
Organization (WHO), the coronavirus spreads through respiratory droplets and physical/body
contact[12]. Therefore, to avoid spreading the virus through these routes, the WHO along
with many other bodies such as the Centers for Disease Control and Prevention (CDC) has
recommended regular washing of hands using soap and water. Comprehensive washing of
hands kills the virus and this practice is currently seen as the most effective way of
preventing transmission among people, especially given that a vaccine for the infection is
many months away. Thus, the demand for soaps in all shapes and forms is slated to
experience a meteoric rise as the COVID-19 pandemic intensifies and worsens during the
current year [13].
Table 1: the world’s largest exporters of soap
Rank Country Export value(millions of
USD)
1 United states 558
2 German 543
3 Indonesia 524
4 Malaysia 456
5 China 367

2.1.1 Soap Production in Ethiopia

The industrial sector of Ethiopia is small and highly import-dependent. Ethiopia's
requirement of soap is largely met through import. The average import of soaps is 57% while
the domestic production covers 43%. Data obtained from the Ethiopian Revenues and
Customs Authority on the import of soap for nine years starting from 2003 up to 2011 shows
that the imported quantity during the nine years was highly erratic, which ranges from the
lowest 165.7 tons in the year 2007 to 2,313.6 tons in the year 2006. The demand for the year
2012 for soap was estimated to be 922 tons. According to the Central Statistical Agency on
the survey of Medium and Large Scale and Electricity Industries in 2011, 43 companies are
engaged in the Manufacture of soap and soaps, cleaning and polishing, perfumes and toilet
preparations, Manufacture of chemical products. [14].
2.2 Endod Fruit oil Description
The essential oil from Phytolacca dodecandra, a traditional herb of Ethiopia, will be studied,
including the chemical composition and antimicrobial activity. The essential oils were
obtained by steam distillation.
The essential oils from P. dodecandra fruit were yellowish and the viscosity of oil is range
from 0.0488 ± 0.0002pas at temperature range from 25-50.[15]
2.2.1 Physical Characteristics of Phytolacca Dodecandra
Endod is a perennial plant. It is a climbing plant with hanging branches growing up to 10 m.
usually, in a year twice it used to provide fruits from December – February and then June –
July. Often, it is found in highlands of Ethiopia from 1600m-3000 m above the sea level.
General Uses Parts of the Endod plant have been used as a soap and as traditional medicine
for centuries in Ethiopia. Endod is known as traditional soap in rural Ethiopia and rids clothes
of parasites such as lice. It is also used in traditional medicine against dandruff and other skin
diseases. People of all ages are familiar with the plant and its soaps and medicinal uses.
Endod is considering being associated with poor people. Common medicinal uses include
treatment of skin itching (ringworm), abortion, gonorrhea, leeches, intestinal worms, anthrax
and rabies.[16].
2.2.2 Chemical property of Phytolacca Dodecandra Fruit
The endod plant provides material of special in tersest to organic chemists involved in
saponin chemistry, partly because of the high percentage by weight of crude saponins in dried
berries, and partly because of the chemical complexity of the materials. Most chemical work
on ended until now has concentrated on the saponins of dried berries [17].
The leaves, fruit and root contains numerous saponine (triterpenoid glycosides). These
compound cause hemolysis red blood cells the aglycones of the glycosides are mainly
composed of oleanolic acid (66%), bayogenin (15%), hederogenin (9%), and 2-
hyderoxyoleanoic acid (6.5%). On a dry weight basis the fruit pulp contains 25% saponins,
and the non saponin fraction contains a lipid fraction (palmitic acid, oleic acid, stearic acid
and a non saponifiable bright orange, wax material), 12 sugars, starches, pectin’s and gums
and a water insoluble fraction. The saponins have become important as they have strong
molluscicidal activity against a range of water snail. In Ethiopia different cultivars yielded
between 1050kg/ha to 2750kg/ha of dried fruit, with a saponins content of 20-25% ([18].

2.3 Physical and Chemical properties basic raw materials used for the production of
soap in tabular form

Table 2 physical and chemical properties of raw materials [19].

Raw materials Chemical property Physical property
Hydrogen peroxide (H2O2) Colorless liquid, less volatile Highly unstable, act as both
and more viscous than water oxidizing and reducing agent
Potassium hydroxide (KOH) Reactive acid, highly Clear solution, white and
corrosive and combustible granular solid, odorless
Sodium chloride (NaCl) Corrosive to base metals White in color, conduct of
Doesn’t undergo combustion electricity. Soft enough to be
and chemicals have cut with knife
hygroscopic quality

2.4 Processing Technology for Production of Soap

Soap is a water-soluble salt of fatty acids that contain more than eight carbon atoms. The
cleaning properties of soaps depend on their chemical properties as an anionic surface active
agent or surfactant. A soap molecule has a long hydrocarbon chain with a carboxylic acid
group on one end, which has an ionic bond with a metal ion, usually sodium or potassium.
The hydrocarbon end is nonpolar, which is soluble in non-polar substances and the anionic
end is soluble in water. This ability is seen from the molecular structure of soaps, when these
materials are burnt in air, the resulting extraction of KOH from wood ash. Ashes contain
oxides of potassium and sodium that dissolve in water to give hydroxides. The glycerol
which is separated from the crude soap will be sold to lotion and cream manufacturers
There are several different soap making processes that will allow easy production of soap
products [20].
 Cold process soap making
 Melt and Pour soap making method
 Hand milled soap making method
 Hot process soap making
2.4.1 Cold process soap making
This process requires a scale, a large stainless steel pot, measuring cups, kitchen
thermometers, rubber gloves, mold, cardboard, plastic bags, olive oil, coconut oil, shortening,
lye, distilled water, and fragrances. The cold process gets its name from the general low
temperatures that are used to mill this type of soap [21].
2.4.2 Melt and pour soap making method
Out of all of the soap making methods, this is perhaps the easiest for novice soap makers. It
involves simply melting a soap base in a double boiler, adding in fragrance and dyes, and
then placing the soap mixture into a mold[22].
2.4.3 Hand milled soap making method
This method is also called soap re-batching, in which soap is made from soap scraps that can
still be reworked into a new bar of soap. To do this, soap scraps, a mold, a grater, fragrance,
dyes, and other additives are needed. The best type of soap base for this type of soap making
is a cold process soap. To start with, you will want to grate your soap into fine particles. Then
place all of your shavings into a glass bowl that can be placed in a double boiler. As the
double boiler heats up, you will need to stir your grated soap pieces. Add in your vegetable
oils and fragrances at this point, making sure that everything is incorporated. Break up soap
clumps that form. Continue to cook until the soap reaches the string stage. At this point, the
soap will thicken rapidly. Remove the pot from the heat source quickly add in fragrances,
coloring herbs, and optional items in that order. Spoon into molds and allow it to cure [23].
2.4.4 Hot process soap making
This process is named for its boiling pot method of soap milling. For this type of homemade
soap making, you will need oil (palm oil, coconut oil, sesame oil, shea butter, and castor oil),
potassium/sodium hydroxide, water, borax, soap molds, stainless steel pot, stirring stick or
spoon, plastic wrap[24].
Process selection
Selection of hot process over cold process
There are two types of soap. Those are soft soap and hard soap. Soft soap can be made using
either a cold process or a hot process, but hard soap can only be made using a hot process.
 Soap is completely saponified quicker than in Cold Process soap.
 Essential and fragrance oils, super fatting oils, and other additives are added at the
end of the cook and are not affected by the saponification reaction.
 Hot Process soaps are easier to slice and do not crumble.
The cold process may require several days or even months, depending upon the strength and
purity of the ingredients, whereas the hot process takes place within a few minutes to a few
hours. The hot process, more suitable for laboratory or industrial preparation, yields a more
chemically pure powder; by-products and excess starting materials are separated [25].
2.5 Soap Uses and Application
Beyond its cleaning ability, soap has been used in other applications. For example, certain
soaps can be mixed with gasoline to produce gelatinous napalm, a substance that combusts
more slowly than pure gasoline when ignited or exploded in warfare. Soaps are also used in
"canned heat," a commercialized mixture of soap and alcohol that can be ignited and used to
cook foods or provide warmth[26].
Bug repellent: Place a bar of soap in your garden to keep pests away
Rid your house of fleas: place some liquid soap in a bowl mixed with water under a light
and it will attract fleas and they will drowned
Household bug repellent: mix soap and water and place in a spray bottle and spray around
your home to rid your house of spiders and bugs
Drawers: use soap to loosen and quiet dresser drawers or other household drawers by
running the soap along the metal rails
Freshen clothes/luggage: place a bar of soap in a mesh holder to keep stored clothes or
luggage smelling fresh
Car freshener: place slivers of left-over soap in a mesh bag and tie around your rearview
mirror to keep a fresh smell in your car
Removing Wallpaper: mix soap and water and sponge on old wallpaper and it will help in
removing the glue
Remove a light-bulb: remove a broken light-bulb by placing a soap bar on top and then
turning it
Zippers: Loosen stuck or rusty zippers by sliding some bar soap up & down the zipper
Detect gas leaks: mix a solution of water and soap and rub along pipes, if it bubbles, you
found your leak.
Lubricate screws/nails: Dip a screw or nail into a bar of soap before drilling or hammering
and it will move through wood more easily/also for saw blades too.
Fabric Marker: use those little left-overs to make fabric for hemming and it washes right
away when complete
2.6 Processes Descriptions on extraction of oil from endod fruit
2.6.1 Preparation for Extraction
The Phytolacca dodecandra fruit will be collected from East Hararghe area, the pre-treatment
involves the preliminary preparations of the Phytolacca dodecandra fruits to make suitable for
extraction. The fruit will be wash with tap water and lean to remove dirty introduced during
preparation. After cleaning the fruit will grind and mill with mortal and pistol in order to
achieve size reduction and so as to increase reactivity and to homogenize for extraction
process.
2.6.2 Extraction of Fruit Oil
Extraction is the withdrawing of an active agent or a waste substance from a solid or liquid
mixture with liquid solvent. The solvent is not or only partial miscible with the solid or the
liquid. By intensive contact the active agent transfers from the solid or liquid mixture in to
the solvent (extract). After mixing the two phases are separated which happens either by
rotary separator. For recovery of the solvent (hexane) and to get the active agent in pure form
a further separation process is necessary. Hexane is poured into the round bottom flask.
Endod fruit is placed in the thimble and inserted in the center of the soxhlet extractor. The
extractor is heated when the solvent is boiling; the vapor rises through the vertical tube into
the condenser at the top. The liquid condensate drips into the filter paper thimble in the center
which contains the solid sample to be extracted. The extract sips through the pores of the
thimble and fills the siphon tube where it flows back down into the round bottom flask. This
is allow for 4 hours the output from soxhlet is mixture of oil and solvent, hexane this mixture
cool for few minute and inter to the rotary evaporator to recover solvent that is hexane the
temperature in the rotary evaporator is the boiling point temperature of solvent(hexane) .after
the required oil was obtained from the recovery process the viscosity, free fatty acid specific
gravity and density of oil was identified. The desired yield on this experiment (soxhlet
extraction) is free fatty acid oil.
2.7 Factors influencing the rate of Endod Fruit oil Extraction
The factors which affect the laboratory extraction of endod fruit thus leading to variations in
oil yield and composition, includes; moisture, contents, samples, particle size, temperature,
reaction time, solvent type and solvent to sample ratio. The parameters are discussed in this
paper as follows:
2.7.1 The effects of temperature in oil extraction
Temperature is increased for fruit oil after pre-treatments such as cleaning, grinding and
milling by drying, sieving and milling again the over -size of endod fruit prior to extraction
and is termed thermal treatment of fruit oil. Better extraction is achieved by heating, which
reduces the oil viscosity and releases oil from intact cells, and also reduces moisture in the
cells. Temperature plays an active role in the fruit treatment for mechanical extraction and
ensures an effective solvent process by heating the solvent which hastens the extraction
process. At the right temperature and moisture content, the individual oil droplets unite to
form a continuous phase and flow out maximizing oil yield.
2.7.2 Effect of reaction times
Oil yield obtained (expressed in percent) was extraction time dependent. In general, the oil
yield increased with increase in extraction time. But after a time the amount of oil would be
lower and constant oil yield would be obtained. See chapter three.
2.7.3 Effect of particle size
Particle size plays a great role on the yield of endod fruit oil. Medium particle size gives high
yield while samples with large particle size and very smaller particle size deliver low yield.
That means less oil is extracted from the larger particles compared to the medium size of the
particles.
2.8 Soap manufacturing process
The length of the hydrocarbon chain ("n") varies with the type of fat or oil but is usually quite
long. The anionic charge on the carboxylate head is usually balanced by either charged
potassium (K+) or sodium (Na+) cations. In making soap, triglyceride in fat or oils are heated
in the presence of a strong alkali base such as’ sodium hydroxide producing three molecules
of soap for every molecule of glycerol, the process is called saponification. The equations
below represent typical saponification reactions;[27]
Soap is produced industrially in four basic steps: saponification, glycerin removal, soap
purification, finishing.
Saponification: a mixture of tallow (animal fat) and oil is mixed &heated with sodium
hydroxide then the soap produced is the salt of a long chain carboxylic acid.
Glycerin removal: glycerin is more valuable than soap, so most of it is removed. Some is
left in the soap to help make it soft and smooth. Soap is not very soluble in salt water whereas
glycerin is soluble; salt is added to the wet soap causing it to separate out.
Soap purification: any remaining sodium hydroxide is neutralized with a weak acid such as,
citric acid and two thirds of the remaining water is removed.
Finishing: additives such as preservatives, color and perfume are added and mixed with the
soap and it is shaped into bars for the market. Soaps are similar in structure and function to
soap and for most uses they are more efficient than soap. In addition to the actual 'soap'
molecule, soaps usually incorporate a variety of other ingredients that act as water softeners,
free flowing agents etc.

saponification Glycerin removal

Finishing Soap purification

Figure 3 Process flow diagram of soap production.

 CHAPTER THREE
MATERIALS AND METHODOLOGY
3.1 Materials and Chemicals
3.1.1 Chemicals
Chemicals Their Functions
Potassium Hydroxide(KOH) Used for saponification process
Sodium Chloride (NaCl) Used to increase the brightness of the product and change the
product to semi solid

Hydrogen Peroxide (H2O2) Used us bleaching agent

Water (H2O) Used for dilute the ingredients and act as the reaction media
Hexane (C6H14) Used as a solvent

3.1.2 Equipment
Equipment’s Their function
Test tubes Use to measure the liquid ingredients
Soxhlet Used to extract the endod juice from endod
fruits.
Beakers To mix the ingredients
Rotary evaporator Used to separate the solvent from the
 produced oil
Stand To carry the stirrer motor
PH meter To identify the pH of the product weather it
is acidic or basic
Measuring cylinder To prepare solution of potassium hydroxide
and to read the volume
Mixer Used for mixing the ingredients
Thermometer For measuring the temperature of reaction
Heater or Bunsen burner To heat the solution or for saponification
reaction

Analytical balance Measure the required amount of ingredients

3.2 Raw material Preparation

3.2.1 Phytolacca Dodecandra Fruit Oil
1. Phytolacca dodecandra fruit collection: the samples of phytolacca dodecandra fruit
were obtained from east Hrarghe area.
2. Cleaning: the fruit were separated from the chaffs and other impurities. This
Preparation is very important since any impurity in the fruit will eventually reflect on
the oil extracted.
3. Size Reduction: the endod fruit was milled and grinded using mortal and pistol to
provide a greater surface area to facilitate the extraction process.
4. Drying: after the Phytolacca dodecandra fruit had been grinded and milled
thoroughly; they were dried in sun light for 8:00 hour to reduce the moisture content
of the fruit.
5. Weighing: These were done before and after the crushed fruit were dried. The weight
was taken and recorded using electronic weighing balance. The methodology that
going to use for this project experimentation the experiment extraction of oil from
endod fruit will be carried out in laboratory of Haramaya University department of
chemical engineering.
6. Determination of moisture content of the seeds: 2250g of the wet sample will be
weighed and dried in sun light for 8:00 hour and the Weight will be measured. Then
the moisture content will be obtained. The Percentage moisture in the dried fruit will
be calculated using the following:
 Time (hr) 0 8 Weight (gm) 2250 1621
MC% = (M1-M2) / (M1) * 100………………………………. (eqn3.1) Moisture content
(%) = (Initial mass of sample – dried sample mass) / (Initial sample mass) * 100 Moisture
content (%) = (2250 - 1621) / (2250) * 100 = 27.9%
7. Size reduction and sieve analysis of the fruit powder
The sample was vibrating shaker with set of sieves sizes arranged in decreasing order
4mm, 3mm, 2mm, 1mm and 0.5mm. This is because to Investigate the effect of particles
size on yield oil. But it is not advisable to use very small size sieve, because it affects the
produced oil quality, the oil became residue and black in color. To produce anti-microbial
bar soap we follow two experiments.
 The first experiment is extraction of Endod oil from Phytolacca dodecandra fruit
and
 The second is saponification of Endod oil to produce anti-fungi bar soap.
3.3 Extraction Of Endod Oil Procedure Using Hexane As A Solvent
 Pour 240ml of normal Hexane in to round bottom flask, the proportionality of Endod
fruit to hexane is 1:3.
 Place 80g of the sample in the thimble and insert in the center of the extractor. Heat
the Soxhlet at 69oC. While the solvent boiling, the vapor rises through the vertical
tube into the condenser at the top. The liquid condensate drips into the filter paper
thimble in the center, which contains the solid sample to be extracted. The extract
seeps through the pores of the thimble and fills the siphon tube, where it flows back
down into the round bottom flask. Allow this to continue for 4 hours.
 The extract is then removed from the tube. Further extraction is carried out repeatedly
until the required amount of oil is obtained.
 At the end of the extraction, the resulting mixture containing the oil will be heated to
recover solvent from the oil.
3.3 Characterization of physical and chemical property of oil

3.3.1 Determination of PH value Procedure

PH is a measure of how acidic/basic water is. The range goes from 0 - 14, with 7 being
neutral. PH of less than 7 indicate acidity, whereas a pH of greater than 7 indicates a base. PH
is really a measure of the relative amount of free hydrogen and hydroxyl ions in the water
The pH electrode will be standardized with buffer solution and the electrode immersed into
the sample and the pH value will be read and recorded. The pH value of the oil which is
found to be 6.19
3.3.2 Determination of Specific Gravity
The specific gravity of oil is the ratio of its density to a specified reference substance (water).
This ratio is a pure number, containing no units. If the specific gravity ratio for a given
substance is less than 1, that means the material will float in the reference substance. When
the specific gravity ratio for a given material is greater than1, that means the material will
sink in the reference substance. Specific gravity is determined as
S G = Density of oil /Density of water……………………………………. (eqn.3.2)
3.3.4 Determination of Saponification Value
Saponification is the hydrolysis of esters. Oils and fats are the fatty acid esters of the
trihydroxy alcohol, glycerol. The saponification value of an oil is defined as the number of
milligrams of potassium hydroxide required to neutralize the fatty acids resulting from the
complete hydrolysis of 1 g of the sample

Saponification value (SV) =56.11* ( Vo−Vi

m )
N …………..…………….…..(eqn.3.3)

Where: Vo = volume of HCl solution used for the blank test,

Vi = volume of HCl solution for the determination,
N = actual normality of HCl used, and
m = mass of sample
3.3.5 Acid Values
The acid value is defined as the number of milligrams of Potassium hydroxide required to
neutralize the free fatty acids present in one gram of fat. It is a relative measure of rancidity
as free fatty acids are normally formed during decomposition of triglycerides
V∗N
Acid value =56.11* ………………………………………… ……. (eqn3.4)
m

3.4 Formulation of highly effective ant fungal bar soap

 Test plant material and pathogen

Fresh healthy-looking mature leaves of Phytolacca dodecandra were collected from natural
habitats from areas in the vicinity of Sinana Agricultural Research Center (SARC) located at
07° N and 40° 10ʹ E in Bale Zone, Ethiopia. This was done when Phytolacca dodecandra was
bearing flowers.[28].
P. dodecandra plants were collected, identified, processed and extracted using ethanol (95% v/v)
and extracts stored at -4 °C for quality tests.

CHAPTER FOUR
4.1 Work Plan

Table 4.1

S/ Activities
N
1 Finalizing the proposal and getting the approval

2 Raw material preparation and characterization

3 Conducting experiments for production of starch

base
4 Characterizing / testing the bio plastic film produced

5 Data/ result analysis and interpretation

6 Writing the final BSc thesis

7 Final BSc thesis submission and defense
No
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