Production of Biopesticide From Need Seed

JIMMA UNIVERSITY
JIMMA INSTITUTE OF TECHNOLOGY
SCHOOL OF CHEMICAL AND BIO ENGINEERING
Title: Production of Biopesticide from Neem Seed

A Research Submitted to Jimma University, Jimma Institute of Technology School of
Chemical and Bio Engineering in Partial Fulfillment for the Requirement of the Degree of
Bachelor of Science in Chemical Engineering
By group- 2 peer group -12
Name Id.no.
1. Dawit Dugasa……………………..00952/05
2. Damesa Angasa…………………..00930/05
3. Areki Hailu……………………….00726/05
4. Mekonen Wakjira……………....02437/04
5. Hawa Shafi……………………...01279/05
6. Yerusalem Berhanu……………02128/05
June, 2017
Jimma, Ethiopia
Biopesticide production from neem seed extract
JIMMA UNIVERSITY
JIMMA INSTITUTE OF TECHNOLOGY
SCHOOL OF CHEMICAL ENGINEERING
Title: Production of Biopesticide from Neem Seed

By group- 2 peer group -12
Name Id.no.
1. Dawit Dugasa……………………..00952/05
2. Damesa Angasa…………………..00930/05
3. Areki Hailu……………………….00726/05
4. Mekonen Wakjira……………....02437/04
5. Hawa Shafi……………………...01279/05
6. Yerusalem Berhanu……………02128/05
Name of Advisor Signature Date
Mr. Samuel Gessese
June, 2017
Jimma, Ethiopia
Declaration
We hereby declare that this thesis is based our original work except for citations which have been duly
acknowledged .we also declare that it has not been previously or currently submitted for any other
department at Jimma University institute of technology or other institutes.
By
Name signature date
1. Dawit Dugasa……………………………………...
2. Damesa Angasa………………………………….
3. Areki Hailu……………………………………….
4. Mekonen Wakjira……………............................
5. Hawa Shafi……………………............................
6. Yerusalem Berhanu…………………………….
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ACKNOWLEDGEMENT
We would like to express our immense gratitude to the school of Chemical Engineering for assigning
us on the research topic “production of Biopesticides from neem seed”.
We would like to express our appreciation to our advisor, Mr. Samuel Gessese to complete this
research under his elegant supervision and guidance.
Finally we thank my parents, without their blessing we could not have completed this BSC.
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Abstract
Due to the toxic effects of chemical pesticide and other agro chemicals on human beings and
livestock, residual toxicity, environmental problems, pest out-breaks and drastic effects on beneficial
insects the use chemical pesticide and other agro chemicals are getting reduced /being banned
globally. But Biopesticides that are extracted, for example, from neem seed can tackle these problems
to make it more eco-friendly, economically viable and socially acceptable for the farmers.
Biopesticides are certain types of pesticides derived from such natural materials as animals,
plants, bacteria, and certain minerals. The key insecticidal ingredient found in the neem tree is
azadirachtin, the potent insect and mite killer, anti-feedant, and growth retardant isolated from
the kernel of neem seeds. The general objective of this research is to produce biopesticide from neem
tree seed by the method of solvent extraction. The neem seeds were crushed in to 0.25-0.5 mm sizes
for easy grinding. Sample drying was carried out in sunlight for 24hr to obtain easily crushable
material. The maximum particle sizes of the ground mixed sample were 0.5 mm. The effects of
particle size, solvent, temperature and time on the yield were investigated. The optimum results were
obtained at 0.25mm particle size, 69oC temperature and with yield of 41.78% at the time of four hour.
Under these condition hundred percent of water soluble and stable biopesticide was obtained by
mixing with the neem oil and liquid detergent. Investigation on the technical and economic feasibility
of the work for biopesticide production was performed and results from the feasibility study indicated
that the proposed work was feasible with rate of return (RR) 31% and with the payback period of
estimated to be 3 years.
Key words: neem, Azadirachtin, biopesticide, neem oil
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Table of Contents
ACKNOWLEDGEMENT ........................................................................................................ II
LIST OF TABLES ...............................................................................................................VIIII
LIST OF FIGURES .......................................................................................................... VIIIIII
CHAPTER ONE ........................................................................................................................ 1
1. INTRODUCTION ................................................................................................................. 1
1.1 Background……………………………………………………………………………...1
1.2. Statement of Problem……………………………………………………………………2
1.3. Objective………………………………………………………………………………...3
1.3.1 General objective…………………………………………………………………….3
1.3.2. Specific objectives…………………………………………………………………..3
1.4. Significance of the study………………………………………………………………..3
1.5. Limitation……………………………………………………………………………….Error!
Bookmark not defined.
CHAPTER TWO ....................................................................................................................... 4
2. LITERATUREREVIEW ....................................................................................................... 4
2.1 Definition of pesticide…………………………………………………………………...4
2.1.1. Biopesticide…………………………………………………………………………4
2.1.2 Chemical Pesticides ................................................................................................... 5
2.2. Neem……………………………………………………………………………………6
2.2.1 Reviews on Chemical constituents of Neem ............................................................. 7
2.2.2 Specification of Azadirachtin in neem ...................................................................... 7
2.2.3 Importance of neem ................................................................................................... 7
2.2.4 Bioactive compounds in neem oil ............................................................................. 7
2.5 Factors affecting biopesticide production……………………………………………...8
2.5.1 Temperature, time, particle size and type of solvent used......................................... 8
2.5.2. Seasonal and maturity variations .............................................................................. 9
2.5.3. Geographical variation ............................................................................................. 9
2.5.4. Other factors affecting yield ..................................................................................... 9
2.6. Biological effects of neem on insects………………………………………………….10
2.6.1 Insect growth regulation .......................................................................................... 10
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2.6.2 Feeding deterrent ..................................................................................................... 10

2.6.3 Ovipositor deterrent ................................................................................................. 10
2.7. Methods of bio pesticide production…………………………………………………..11
2.7.1 Mechanical extraction method................................................................................. 12
2.7.2 Steam pressure extraction method ........................................................................... 12
2.7.3 Solvent Extraction method ...................................................................................... 12
CHAPTER THREE ................................................................................................................. 14
3. MATERIALS AND METHODS ......................................................................................... 14
3.1 Materials and equipment’s……………………………………………………………..14
3.2 Description of process flow diagram…………………………………………………..17
3.3. Experimental procedures and method………………………………………………...19
3.4. Sample analysis………………………………………………………………………..20
3.4.1. Determination of moisture content of the seeds ..................................................... 20
3.4.2. Size reduction and sieve analysis of the seeds ....................................................... 20
CHAPTERFOUR ..................................................................................................................... 21
4. RESULTS AND DISCUSSIONS ........................................................................................ 21
4.1. Moisture Contents……………………………………………………….…………….21
4.2. Soxhlet extraction……………………………………………………………………..22
4.2.1. Percent yield of soxhlet extractor ........................................................................... 22
4.3 Stability testing at 50 degree centigrade……………………………………………….24
4.4. Observation and Discussions………………………………………………………….25
CHAPTER FIVE ..................................................................................................................... 26
5. MATERIALAND ENERGY BALANCE ........................................................................... 26
CHAPTER SIX ........................................................................................................................ 32
6. SIZING AND DESIGN OF MAJOR EQUPMENT ............................................................ 32
6.1 Sizing of equipment’s…………………………………………………………………..32
6.2. Design of centrifuge…………………………………………………………………...35
CHAPTER SEVEN ................................................................................................................. 48
7. COSTING AND ECNOMICS ............................................................................................. 48
7.1purchased Equipment cost……………………………………………………………...48
7.2. Estimation of capital investment cost…………………………………………………49
7.2.1. Fixed capital investment (FCI) estimation ............................................................. 49
7.2.2. Total production cost (TPC) estimation ................................................................. 49
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7.3. Economic evaluation…………………………………………………………………..51

7.3.1. Net income, Payback time and return on investment ............................................. 51
7.4. Plant layout, environmental impact and site location…………………………………51
7.4.1 Environmental impact analysis ................................................................................ 52
7.4.2 Plant layout .............................................................................................................. 53
CHAPTER EIGHT .................................................................................................................. 55
8. CONCLUSION AND RECOMMENDATION ................................................................... 55
8.1. Conclusion……………………………………………………………………………..55
8.2. Recommendation………………………………………………………………………56
REFERENCES ........................................................................................................................ 57
APPENDICES ......................................................................................................................... 59
APPENDIX A……………………………………………………………………………..59
APPENDIX B……………………………………………………………………………..60
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LIST OF TABLES
Table 4.1: Moisture content determination of Neem kernel…………………24

Table 4.2: yield of soxhlet extractor for different particle size...................... 24
Table 4.3: characteristics of Neem Oil……………………………………....25
Table 5.1: summary of material balance……………………………………..30
Table 6.1: specification of components of SBDC…………………………...40
Table 7.1: purchased costs of the equipments………………………………..48
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LIST OF FIGURES
Fig 3.1. Block flow diagram of biopesticide production………………………. 15

Fig 3.2. Process flow sheet of biopesticide production………………………... 19
Fig 3.3.Neem seed……………………………………………………………….17
Fig 3.4. Peeled off neem kernel………………………………………………….17
Fig 3.5.Neem oil………………………………………………………………….18
Fig 3.6.Final product……………………………………………………………..18
Fig 4.1.Effect of time and particle size on the yield……………………………..25
Fig 4.2.Effect of temperature on the yield………………………………………..24
Fig 6.1.Conical-cylindrical SBDC, with its main components…………………..36
Fig 6.2.Dimension specification of solid bowel decanter centrifuge…………….37
Fig 6.3.Slippage force on the beach of SBDC…………………………………...41
Fig 7.1.Plant layout of biopesticide production………………………………….61
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LIST OF ABBREVIATIONS
DDT Dichloro Diphenyl Trichloroethane
EPA Environmental Protection Agency
FAO Food and Agricultural Organization
HPLC High Performance Liquid Chromatography
NPEO Nonyl Phenol Ethoxylate
O/W Oil in Water
SLS Sodium Lauryl Sulphate
SBDC Solid Bowel Decanter Centrifuge
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CHAPTER ONE
1. INTRODUCTION
1.1 Background
The use of chemical pesticide and other agro chemicals are getting reduced /being banned
globally because of their toxic effects on human beings and livestock, residual toxicity,
environmental problems, pest out-breaks and drastic effects on beneficial insects. But Bio
pesticide that are extracted, for example, from neem seed can tackle pests to make it more eco-
friendly, economically viable and socially acceptable for the farmers. Neem (Azadirachtin indica
A. Juss.Meliaceae) is thought to have originated in India. The Neem tree contains a thousand of
chemical components. It is remarkable the occurrence of the so-called “terpenoids”, which are
very rare. They appear in Neem in more than one hundred types. By far the most active yet
studied is the Azadirachtin.
Biopesticide is a 'biological pesticides ', include several types of pest management intervention:
through predatory, parasitic, or chemical relationships. The term has been associated historically
with biological control and by implication the manipulation of living organisms.This paper is
aims to produce bio pesticide from neem seed by the solvent method of extraction. In the solvent
method extraction the hexane was used for dissolving the neem oil but not water. It has an
advantage on currently used pesticides. While the conventional pesticides kill insects and
plagues but also other organisms affecting the biological cycle; the bio pesticides from Neem kill
only those which are harmful. This reduces or eliminates any adverse effect. Bio pesticides are
certain types of pesticides derived from such natural materials as animals, plants, bacteria and
certain minerals. For example, canola oil, baking soda and Neem oil have pesticidal applications
and are considered as Biopesticide.
Neem oil is a broad spectrum botanical insecticide, miticide and fungicide agent derived from
the seeds of the Neem tree (Azadirachta indica). Neem oil is powerful and safe pest fighter. It
has many modes of action with the primary role of disrupting an insect’s metamorphosis. It
makes a loss of appetite in some insects and interfering with reproduction and maturation. It is
nontoxic to humans, birds, earthworms, and animals. The toxic effects of the synthetic pesticides
are well known. It is important to note from the toxicological as well as economical point of
view that only 20–50 g of active ingredient is sufficient to treat one hectare for satisfactory
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reduction in pest population (Ghimeray et al,2009)).Neem oil is cheap and contains various
active ingredients but difficult to spray. Hence we have successfully developed an O/W
emulsions which contain 5% of neem oil using various surfactants which are Sodium Lauryl
Sulfate (SLS), Nonyl Phenol Ethoxylate (NPEO) and surface combining agent Guar Gum
(Gunjan et al, 2002).
1.2. Statement of Problem

The hazard use of chemical pesticides in agriculture, horticulture, forestry, animal husbandry and
in public health has created some adverse problems. Though these reduction in synthetic
pesticides consumption is should be visualized by all concerned e.g. government, farmers,
pesticide industries, rural development agencies etc. Neem biopesticides offer a reliable,
economic and eco-friendly solution and they have proved to be effective against a number of
pests. The neem-based products or pesticides are included in plant protection schedules of most
of the crops grown in Ethiopia. Synthetic pesticides were considered as an important
component of crop production system and received an overwhelming demand from farmers who
were convinced of high crop yields, pest mortality due to quick knock-down effect, easiness in
application, availability of suitable formulations in local market, recommendations from
government departments etc. in spite of the ill effects of long-term persistence of residues etc.
The guidelines for safe and proper use of pesticides were hardly followed by farmers due to
illiteracy, ignorance of instructions, and inadequate knowledge etc. Thus, overuse or misuse of
these molecules has created some undesirable secondary effects in the agro ecosystems and
environment. Yield in food crops, cotton and vegetables by using non-chemical crop production
techniques including Biofertilizers, Biopesticides, and plant products and Neem-based the
Government did not consider the fate of farmers and did not foresee. In this study we select neem
as Bio- Pesticide because it has proved to be effective and economical, does not pose any threat
to beneficial organisms and human, and eco-friendly. So from the economical point of view and
as an alternative to Chemical pesticides we chose to develop a biopesticide formulation from
Neem seed.

1.3. Objective
1.3.1 General objective

 The general objective of the research is to produce biopesticides from Neem seed.
1.3.2. Specific objectives

 To design the major equipment used in the unit operations.
 To analyze the neem seed samples.
1.4. Significance of the study

This study contributes a way to extract neem biopesticides from neem seed in laboratory scale. It
also indicates the possible way to produce environmentally friend biopesticides and with
optimum cost.

CHAPTER TWO
2. LITERATUREREVIEW
2.1 Definition of pesticide
A chemical or biological substance, designed to kill or retard the growth of pests that damage or
interfere with the growth of crops, shrubs, trees, timber and other vegetation desired by humans,
is called pesticide. Practically all chemical pesticides, however, are poisons and pose long term
danger to the environment and humans through their persistence in nature and body tissue. There
are mainly two types of pesticides: Biopesticides and Chemical pesticides (U.S. Environmental
Protection Agency, 2012).
2.1.1. Biopesticide
Bio pesticides are certain types of pesticides derived from such natural materials as animals,
plants, bacteria and certain minerals. For example, canola oil, baking soda and Neem oil have
pesticidal applications and are considered as Bio-pesticides. Bio-Pesticides are of three types.
And they are as follows.
2.1.1.1 Microbial Pesticides

Microbial Pesticides consist a microorganism (e.g., a bacterium, fungus, virus or protozoa) as the
active ingredient. Microbial pesticides can control many different kinds of pests, although each
separate active ingredient is relatively specific for its target pest. For example, there are fungi
that control certain weeds, and other fungi that kill specific insects. The most widely used
microbial pesticides are subspecies and strains of Bacillus thuringiensis. Each strain of this
bacterium produces a different mixture of proteins and specifically kills one or a few related
species of insect larvae. The target insect species are determined by whether the particular
Bacillus thuringiensis produces a protein that can bind to a larval gut receptor, thereby causing
the insect larvae to starve (U.S. Environmental Protection Agency, 2012).
2.1.1.2 Plant-Incorporated-Protectants
Plant-Incorporated-Protectants are pesticidal substances that plants produce from genetic
material which are added to the plant. For example, scientists can take the gene for the Bacillus
thuringiensis pesticidal protein, and introduce the gene into the plant's own genetic material.
Then the plant, instead of the Bacillus thuringiensis bacterium, manufactures the substance that
destroys the pest (U.S. Environmental Protection Agency, 2012).

2.1.1.3 Biochemical Pesticides

Biochemical Pesticides are naturally occurring substances that control pests by non-toxic
mechanisms. Conventional pesticides, by contrast, are generally synthetic materials that directly
kill or inactivate the pest. Biochemical pesticides include substances, such as insect sex
pheromones that interfere with mating, as well as various scented plant extracts that attract insect
pests to traps. Because it is sometimes difficult to determine whether a substance meets the
criteria for classification as a biochemical pesticide, EPA has established a special committee to
make such decisions (U.S. Environmental Protection Agency, 2012).
2.1.2 Chemical Pesticides
2.1.2.1 Organophosphate Pesticides

These pesticides affect the nervous system by disrupting the enzyme that regulates acetylcholine,
a neurotransmitter. Most organophosphates are insecticides. They were developed during the
early 19th century, but their effects on insects which are similar to their effects on humans, were
discovered in 1932. Some are very poisonous (they were used in World War II as nerve agents).
However, they usually are not persistent in the environment (U.S. Environmental Protection
Agency, 2012).
2.1.2.2 Carbamate Pesticides

“Carbamate Pesticides affect the nervous system by disrupting an enzyme that regulates
acetylcholine, a neurotransmitter. The enzyme effects are usually reversible. There are several
subgroups within the carbamates”(U.S. Environmental Protection Agency, 2012).
2.1.2.3 Organochlorine Insecticides

Organochlorine Insecticides were commonly used in the past, but many have been removed from
the market due to their health and environmental effects and their persistence (e.g. DDT and
chlordane) (U.S. Environmental Protection Agency, (2012)).
2.1.2.4 Pyrethroid Pesticides

Pyrethroid Pesticides were developed as a synthetic version of the naturally occurring pesticide
pyrethrin, which is found in chrysanthemums. They have been modified to increase their stability
in the environment. Some synthetic pyrethroids are toxic to the nervous system (U.S.
Environmental Protection Agency, 2012).

2.2. Neem
Margosa (Neem) tree, which is also known as Azadirachtin indica, is one of the best known trees
in India, which is known for its medicinal properties. The main reason behind the popularity of
the Neem is that it is used to treat few of the most common problems that the people face. The
Neem tree (Azadirachtin indica) is among the fastest-growing trees. Ethiopia has also a potential
for the plantation of Neem tree which grows in humid, arid, and hot places having an altitude of
up to 1,500 meters above sea level. The tree is available in many part of the country such as:-
Jijjiga, Mekele, Bahirdar, wolkite, Harar, Diredawa, Adama, Gambella etc. Mainly in Ethiopia
the neem tree bears a fruit starting from March. Neem can grow in tropical and subtropical
regions with semi-arid to humid climates. Neem tree will adapt to a mean annual rainfall of 450-
1200 mm, mean temperatures of 25-35ºC and grow at altitudes of up to 800 meters above sea
level.
There is so much resourceful that almost all of its parts are used in some form or another. From
toothpastes to oils, from cosmetics to medicines, Neem oil is used as an important ingredient. Its
healing properties are simply awesome. It flourishes very well in the tropical area conditions
where it provides luxurious shade, firewood and also used for afforestation. It produces large
quantities of seeds that are hardly used. The importance of neem as bio-pesticide was realized by
the modern scientific community, as early as 1959, when a German scientist in Sudan found that
neem was the only tree that remained green during a desert locust plague. Literatures confirm
that neem can effectively get rid of over 200 pest species that affects plants. The pesticidal
characteristics of neem is largely attributable to Azadirachtin found in the neem extracts which is
a growth regulator and as well as a powerful feeding. Azadirachtin is non-volatile and an insect
cannot prevent it by smell but has to taste it, in order to respond to it. A taste of azadirachtin
stimulates at least one 'deterrent neuron' in insects which show an anti-feedant response. The
strength of 'deterrent neuron' responses has been correlated with the strength of anti-feedant
responses. Neem oil can also suffocate mites, whiteflies, aphids and other types of soft bodied
insects on contact. So it is clear that neem does not kill on contact, rather it inhibits feeding and
reproduction of the pests. These multiple modes of action make it unlikely that insects and plant
pathogens can develop resistance to neem. Also certain pest such as floral thrips, diamond back
moth and several leaf miners which develop resistance to the inorganic pesticides or that are
inherently difficult to control with conventional pesticides are effectively controlled or managed
with neem (Ahmed, (1989) Econ Bot, 43: 35-38).

2.2.1 Reviews on Chemical constituents of Neem
Melwita E. (2011) studied many bioactive compounds in the Neem tree. Since the pioneering
work by Butterworth Morgan in 1950, hundreds of compounds have been isolated from all parts
of the tree. These compounds possess insecticidal, antifeedant, and growth inhibiting effects
against many species of insects and pests. The fruits and seeds of neem tree are appearing to be
more important. Neem seed contains 40% bitter tasting non edible oil. Extraction of oil from the
seeds gives neem oil which contains important bioactive compounds. Major bioactive
compounds as reported in literatures were Azadirachtin (Azadirachtin A), salanin, nimbin,3-
tigloylazadirachtol(Azadirachtin B), (Azadirachtin D), and Azadirachtin H (Melwita E, (2011)).
2.2.2 Specification of Azadirachtin in neem
“For the purposes of FAO specifications, “azadirachtin” is the collective term applied to a large
group of insecticidal-active limonoid compounds, extracted from seeds of the neem tree
(Azadirachta indica A. Juss.), in which the most active and abundant single compound is known
as azadirachtin A. Azadirachtin, sensulato, is neither completely defined nor quantifiable and so
azadirachtin A is used as a lead or marker compound for the purposes of identification and
quantification”. Empirical formula of Azadirachtin A is C35H44O16 .
2.2.3 Importance of neem
Over thousands of years, Neem has been used by hundreds of millions of people and no hazards
have been documented for normal dosages. (Klaus Ferlow1926). Every part of this fascinating
tree has been used, from ancient to modern times, to treat hundreds of different maladies. While
it is still revered in India for its superior healing properties, recent investigation has dramatically
increased worldwide interest in Neem and many products are now manufactured using this
miraculous herb. More than any other Indian herb, Neem proved useful in helping the body resist
diseases and restore the proper balance to the body’s systems.
2.2.4 Bioactive compounds in neem oil

Neem tree has been known to contain many bioactive compounds. Since the pioneering work by
Butterworth Morgan in 1950, hundreds of compounds have been isolated from all parts of the
tree. These compounds possess insecticidal, antifeedant, and growth inhibiting effects against
many species of insects and pests. The fruits and seeds of neem tree are appearing to be more
important. Neem seed contains 40% bitter tasting non edible oil. Neem oil, extracted from neem
seeds, contains important bioactive compounds. Major bioactive compounds as reported in

literatures are Azadirachtin (Azadirachtin A), salanin, nimbin, 3-tigloylazadirachtol

(Azadirachtin B), 1-tigloyl-3-acetyl-11-hydroxymeliacarpin (Azadirachtin D), and Azadirachtin
(H). Many other compounds are present in smaller quantities in neem seed. Chemically they are
belonging to highly oxygenated triterpenes classed as tetranor terpenoids or Limonoids which
represent the extreme examples of oxidation of triterpenes in nature. Limonoids possess medium
polarity. They can be dissolved in solvents of medium to high polarity such as methanol, ethanol,
diethyl ether, dichloromethane, and ethyl acetate. Meanwhile, their solubility in water is very
low and they are not soluble in non polar solvents such as hexane and petroleum ether.
Limonoids have polarities that are very close to each other due to their close similarity in
structure. In general Azadirachtin is very labile when exposed to air, moisture and sunlight. This
may be due to presence of C-C pie bonds. Reports in the comprehensive physico- chemical
properties of azadirachtin are very limited. Its melting point is 174 degree centigrade. The
stability of azadirachtin is substantially decreased by the presence of protic solvents, in particular
those solvents having acidic or basic functional groups, specifically water, acids and bases .For
this reason polar solvent like SLS and NPEO have been selected in this study.
2.5 Factors affecting biopesticide production

Factors that determine the composition and yield of the oil for biopesticide obtaining are
numerous. In Some instances it is difficult to segregate these factors from each other, since many
are interdependent and influence one another (Terblanche, 2000). These variables may include
seasonal and maturity variation, geographical origin, genetic variation, growth stages, part of
plant utilized and postharvest drying and storage (Marotti 1994).
2.5.1 Temperature, time, particle size and type of solvent used

The yield of oil can be affected by temperature. This was because of rising the temperature, both
the diffusion coefficient and the solubility of the oil in solvents is enhanced, thus heat treatment
improves the extraction Neem oil. The higher extraction temperatures the easier to break the
molecule inside the seed; as a result, the yield also gets high. But the lower boiling solvents
appropriate, because at the high temperature the active ingredient in the neem may be affected.
Therefore N-hexane has high capacity to dissolve non-polar compounds in the oil. Therefore,
based on the findings hexane is a better solvent for Neem oil extraction.Partcle size also have
effect on the yield. The smaller particle size results in higher yield of oil than larger particle size
because of that smaller particle size have high surface area for extraction to be undertaken.

2.5.2. Seasonal and maturity variations

These two factors are interlinked with each other, because the specific antigenic growth stage
will differ as the season progresses. There are variations in the chemical profile of essential oils
from various plants collected during different seasons. The essential oils yields varied
considerably from month-to-month and was also influenced by the micro-environment (sun or
Shade) in which the plant was growing. Results obtained by Badi (2004) also indicated that
timing of harvest is critical to both yield and oil composition.
2.5.3. Geographical variation

There are many reports in the literature showing the variation in the yield and chemical
composition of the essential oil with respect to geographical regions (Uribe- Hernandez
1992).Chalchat et al. 1995 reported variations in the yield and chemical profile of essential oils
collected from different geographical locations, respectively. Such differences could be linked to
the varied soil textures and possible adaption response of different populations, resulting
indifferent chemical products being formed, without morphological differences being observed
in the plants. (Hussainet al.,2008).
Altitude seems to be another important environmental factor influencing the essential oil content
and chemical composition. Climatic factors such as heat and drought were also related to the
essential oil profiles obtained (Uribe-Hermandezet al., 1992). Moreover, the preference of the
plant for these conditions suggest that genetic make-up of the plant, rather than the soil-type in
which it is growing, should have a greater influence on the chemical profile of the oil
produced(Abdullah 2009).
2.5.4. Other factors affecting yield

Other factors which affect the growing plants thus leading to variations in oil yield and
composition, include part of plant used; post harvest drying; length of exposure to sunlight;
availability of water, height above sea level, plant density, time of sowing and the presence of
fungal diseases and insects. The oil composition and yield may also change as a result of the
Harvesting methods used, the isolation techniques employed the moisture content of the plants at
the time of harvest and the prevailing extraction conditions (Abdullah 2009).
Postharvest drying of material is an accepted practice in the production of oils. Drying methods
include exposure to natural air in the shade, sun-drying, as well as drying by blowing warm air
over the material. Postharvest drying is thought to improve oil yield and accelerate distillation,
by improving heat transfer, in addition to providing increased loading capacity, due to loss of
plant moisture. Further advantages include the reduction of microbial growth and the inhibition

of some biochemical reactions in dried material. However, some amount of the oil may be lost
during such post harvest treatment due to volatilization and mechanical damage to oil glands.
Essential oil components (including terpenoids) are usually present in the free form, but may also
be bound to sugar moieties, usually mono- or disaccharides (Abdullah 2009).
2.6. Biological effects of neem on insects

The action of neem products as pest control agents can be manifested at different levels and in
different ways. This is a very important point to be noted since the farmer would be used to the
“knock-out’’ effect of chemical pesticides. Neem extracts do not exhibit this type of effect on
pests but affect them in several other ways (Vijayalakshmi et al, 1995).
2.6.1 Insect growth regulation

Regulation of the insect growth is a very interesting property of neem products which is unique
in nature, since the products work on juvenile hormones. The insect larva feeds and as it grows,
it sheds its old skin. This particular shedding of old skin is the phenomenon of ecdysis or
moulting and is governed by an enzyme, ecdysone. When the neem components, especially
azadirachtin, enter the body of the larva, the activity of ecdysone is suppressed and the larva fails
to moult, remains in the larval stage and ultimately dies. If the concentration of azadirachtin is
not high enough, the larva will die only after it has entered the pupal stage. If the concentration is
lower still, the adult emerging from the pupa will be 100% malformed, and absolutely sterile
(Vijayalakshmi et al, 1995).
2.6.2 Feeding deterrent
The most important property of neem is feeding deterrence. When an insect larva sits on a leaf, it
will want to feed on it. This particular trigger of feeding is given through the maxillary glands.
Peristalsis in the alimentary canal is thus speeded up, and the larva feels hungry and starts
feeding on the surface of the leaf. If the leaf is treated with a neem product, because of the
presence of azadirachtin, salanin and melandriol, there will be an anti-peristaltic wave in the
alimentary canal which produces something similar to a vomiting sensation in the insect.
Because of this sensation, the insect does not feed on the neem-treated surface. Its ability to
swallow is also blocked.
2.6.3 Ovipositor deterrent

Another way in which neem controls pests is by preventing the females from depositing eggs.
This property is known as Oviposition deterrence, and comes in very handy when the seeds in
storage are coated with neem kernel powder and neem oil. The seeds or grains obtained from the

market may already be infested with some insects. Even these grains could be treated with neem
seed kernel extract or neem oil. After this treatment, the insects will not feed on them. Further
damage to the grains will be halted and the female will be unable to lay its eggs during the egg-
laying period of its life cycle. The use of neem products does not give immediate results, unlike
chemical insecticides. Some patience is required after the application of neem products. Besides
its insecticidal and nematicidal properties, neem is also a promising agent for control of plant
diseases. It has also been demonstrated to possess anti-fungal properties. One of the problems
with the use of chemical pesticides has been their impact on “non-target” species. Often they
have proven harmful to various other species in the ecosystem that could be beneficial. However,
neem extracts are devoid of these effects. Neem leaves and seed kernels, when incorporated into
potting soil containing earthworms, increased the earthworm population by 25%. Neem products
have been proven to be remarkably benign to spiders and also other insects such as bees that
pollinate crops and trees, ladybug beetles that consume aphid, and wasps which act as parasites
on various crop pests. Neem products have to be ingested to be effective. Those insects which
feed on plant tissues, therefore, easily succumb. However, natural predators like spiders feed
only on other insects while bees feed on nectar. Hence they rarely come in contact with
significant concentrations of neem products (Vijayalakshmi et al, 1995).
2.7. Methods of bio pesticide production

Quality of neem oil based biopesticide depends on the type of extraction. Production of neem oil
based biopesticide includes the collection of raw materials for the extraction and selection of
extraction method. Neem oil is extracted from neem leaf and need seed. Neem seed is widely
used in the extraction process instead of neem leaf as the oil content is found to be more in seeds
than in the leaf. Traditional methods of extracting Neem oil neem oil can be extracted
traditionally at home using cold pressed extraction by hand and around 100 to 150 mgs of oil for
every 1 kilogram of Neem seed. To press Neem oil by hand, the kernels of the Neem seed should
be crushed in a mill or pound in a mortar. Add a small amount of water until the mixture forms a
firm paste that can be kneaded. Knead the paste until oil drops form on the surface and press
firmly to extract the oil. The kneading and pressing should be continued in turn until the
maximum amount of oil is removed. The oil content of the seed kernel is about 45%, even
though preparation of the oil at home possible, but this traditional method of processing Neem
oil was not effective on percent yield.

2.7.1 Mechanical extraction method

The mechanical extraction method is the process of extracting oil without involving solvent. The
mechanical extraction has several advantages compared to the other methods, such as simple
equipment and low investment, low operating cost, and the oil does not undergo solvent
separation process, etc. Mechanical pressing extraction consists of grade wise separation of
seeds. Grading of seeds is done according to the amount of oil content in the seeds and with sizes
as well. Firstly, the fruits are collected in a drum, and the kernels are separated to obtain the
seeds. Later the seeds are woven dried and then feed into the oil extracting machine in case of
mechanical pressing method. The neem oil is obtained by pressing it mechanically and collected
in a drum. Thus filtration is done to remove the various unwanted particles left in the extracted
oil in order to obtain pure neem oil .Usually the quality and quantity of the oil obtained by
mechanical extraction process are affected by various operating conditions such as pretreatment
of the Neem seeds, extraction pressure, and storage condition.
Mechanical extraction is the most widely used method to extract Neem oil from Neem seed.
However, the oil produced with this method usually has a low quality, since it’s turbid and
contains a significant amount of water and metals contents
2.7.2 Steam pressure extraction method

Steam pressure extraction method, the neem seeds after the drying process is feed into the steam
boiler. This process makes the extraction process easier. The seeds get swollen by steaming thus
the oil in squeezing becomes easy. The process of steaming is accompanied by increasing of
pressure in the boiler which drives the oil out from the seed without any pressing. In some
industries, the left seed's kernels after the steam boiling is pressed to further extraction of oil up
to 98% leaving just the outer layer of the seeds. The same filtering process is followed as done in
the mechanical pressing method.
2.7.3 Solvent Extraction method
Solvent Extraction is a process which involves extracting oil from oil-bearing materials by
treating it with a low boiler solvent as opposed to extracting the oils by mechanical pressing
methods (such as expellers, hydraulic presses, etc.). The solvent extraction method recovers
almost all the oils and leaves behind only 0.5% to 0.7% residual oil in the raw material. In the
case of mechanical pressing the residual oil left in the oil cake may be anywhere from 6% to
14%. The solvent extraction method can be applied directly to any low oil content raw materials.

It can also be used to extract pre-pressed oil cakes obtained from high oil content materials
(ARPN Journal of Engineering and Applied Sciences Extraction of Neem Oil)
The process solvent extraction is basically a process of diffusion of a solvent into oil-bearing
cells of the raw material resulting in a solution of the oil in solvent. Various solvents can be used
for extraction. However, after extensive research developed with neem oil as the internal phase
which was used as pesticide and consideration of various factors, such as commercial economics,
edibility of the various products obtained from extraction, physical properties of the solvent
especially its low boiling point etc. food grade n-hexane is considered to be the best and it is
exclusively used for the purpose. In a nutshell, the extraction process consists of treating the raw
material with hexane and recovering the oil by distillation of the resulting solution of oil in n-
hexane called miscella. Evaporation and condensation from the distillation of miscella recovers
the n-hexane absorbed in the material. The n-hexane thus recovered is reused for extraction. The
low boiling point of hexane (67°C / 152°F) and the emulsifier (detergent) is added to the neem
oil to get the biopesticide. Therefore, in this research solvent extraction method is used for
biopesticide production. As many literatures for extraction they were used petroleum ether
.However, it is desirable to avoid pre-extraction of neem seed kernels with petroleum
ether and avoid extraction in a soxhlet with polar solvents at high temperature, since,
azadirachtin is not quite stable at high temperatures (ARPN Journal of Engineering and
Applied Sciences Extraction Of Neem Oil).

CHAPTER THREE
3. MATERIALS AND METHODS
Here the main raw material was neem seed from which natural Biopesticides have been
prepared. Neem oil extract was prepared from Neem seed first. During the preparation of the
neem oil, three different processes may be followed, the first one is the solvent extraction
process and the second one is the cold pressed extraction process and the third mechanical
extraction. But we use solvent extraction method. A biopesticide formulation was prepared from
neem oil in the form of emulsion. The experiments of production of Biopesticide from neem seed
were carried out in the laboratory of school of Chemical Engineering at the Institute of
Technology, Jimma University.
3.1 Materials and equipment’s
 Neem seed kernels :as raw material

 Detergent (liquid soap) : as additive (surfactant)
 Hexane (300ml): as a solvent
 Soxhlet extractor :for extracting oil
 Beaker: as a container for sample
 Stirrer ;for stirring solution
 Mortar and pestle: for crushing
 Mass Balance :for sample measuring
 Bottle: for containing the sample
 Sieves: to sieve the crushed sample to the particle size of 0.25mm and 0.5mm

Fig 3.1 Block flow diagram of biopesticide production


3.2 Description of process flow diagram
1. collection
Naturally Ripened fruits were collected from Wellega. Fruits with yellowish color should be
harvested. Fruits can harvested by shaking tree branches as well as by plucking fruits and if
possible spread a cloth, tarpaulin under the tree.
Fig 3.3 .Neem seed
2.SeedCleaning: This was done to remove foreign materials such as sticks, stems, leaves, bad
seeds, sand and dirt, to ensure that the oil produced is not contaminated and of high quality
3. Depulping and dehuling: Depulping, the separation of seeds from fruits, was followed by
Dehulling, the process of removing the outer seed coat. Dehulling was done to ensurehigh
extraction efficiency as seed coats contain little or no oil. To achieve this, seed pulps were
soaked in warm water to soften the outer seed coat and the seed coats and fruit were then peeled
off by hand.
Fig 3.4peeled off neem kernel

4. Drying: This was done to remove the moisture content of the seeds so as to ensure high
extraction efficiency. The drying of the seeds was done using sun dryer operated at an ambient
temperature of for about 3 days.
5.GrindingWe ground the seed by mortar and separate the size by sieve, the sizes were 0.25
and0.5 mm.
6. Extraction
300ml of normal hexane was poured into round bottom flask. 45g of the sample powder of neem
seed was placed in the thimble and was inserted in the centre of the extractor. The Soxhlet was
heated to 69.9oC.This was allowed to continue for one, two and three hours. The experiment was
repeated by placing the same amount of the sample into the thimble again by varying particle
size.
7. Evaporation
Oil-solvent mixture obtained was heated and evaporated at 680C in the rotary evaporator.
Solvent-free oil was obtained and the solvent was recovered
8. Blending (mixing)
The oil free solvent obtained above were mixed and stirred with detergent and biopesticide
which is milky (white) color was formed.A beaker was prepared and 20ml of neem oil was
added.4ml of liquid detergent was added and mixed well for 10 minute. Finally the milky color
of biopesticide was obtained as the following figure 3.6.
Fig3.5 .neem oilFig 3.6.Final product

3.3. Experimental procedures and method

Solvent extraction method which involves the extraction of oil from neem seed by treating it
with the low boiling solvent hexane was used in this research. And the experimental procedures
were discussed as follows:
 Neem seeds were depulped by the aid of water. This step ensures easily removal of
outer cover of the seeds, which may reduce the effect of the final product of the
process.
 After depulping, the seeds were dried in a dryer until then moisture content in the seeds
in reduced significantly or until the constant mass of the sample after drying is obtained.
 The dried seeds kernels were grinded in the mortar and sieved by using 0.5mm and
0.25mm size of sieves
 The grounded neem kernels were sent to the next step, the solvent extraction in the
soxhlet extractor. A polar organic liquid such as hexane is chosen for extraction, in
which oil is readily soluble and a hexane-oil solution is obtained.
 After that evaporation was carried out with this solution to evaporate the hexane content
in the solution, and hence to obtain solvent-free oil as the product and by using the
condenser the hexane vapor was recovered. In this way, a solvent free oil yield was
obtained. This oil was blended with an emulsifying agent detergent or liquid soap was
added in a 1: 5 ratio, so that a surfactant based oil biopesticide was formed which can be
diluted with water.

3.4. Sample analysis
3.4.1. Determination of moisture content of the seeds
30g, 40g, and 50g of the cleaned seed sample was weighed and dried in sunlight and the weight was
measured for every 2hr. The procedure was repeated until a constant weight was obtained. The
percentage moisture in the kernel was calculated using the following:
Moisture % = ………………..equation 3.1
Where: W1 = Original weight of the sample before drying; W2 = Weight of the sample after drying.
3.4.2. Size reduction and sieve analysis of the seeds

The moisture was removed by placing the sample in sunlight for three days. The dried Neem kernel
was crushed by using mortar and pestle. The sample was sieved with set of sieves sizes arranged in
ascending order 0.25mm, 0.5mm.This is because to investigate the effect of particles size on yield of
oil.

CHAPTERFOUR
4. RESULTS AND DISCUSSIONS
4.1. Determination of Moisture Contents

The seed was collected on April, 2017, after four months it was decorticated and by taking 30,
40, and 50 grams of Neem kernel, the moisture content of the sample was obtained (table4.1)
using equation 3.1.
Table 4.1: Moisture content determination of Neem kernel
Dryingtime(hour) %moisture content
0 2 4 6 8 10 12
Sample weight
30 28.8537 28.744 28.713 28.682 28.624 28.624 4.59
in grams
40 38.49 38.321 38.108 38.108 38.048 38.049 4.88
50 48.152 48.102 48.048 47.825 47.321 47.320 5.36
The moisture content of the seed kernel of sample with 30, 40, and 50gms was 4.59, 4.88 and
5.36%, respectively. Thus, the average moisture content of the three samples will be 4.94%.

4.2.Soxhlet extraction
4.2.1.Percent yield of soxhlet extractor

Table 4.2: Total %yield for soxhlet extractor for different particle size, temperature and time
using n-hexane as a solvent with initial sample of 45gm.
Cake wt,
Particle size(mm) Extraction times(hrs)
%yield
1 2 3 4
Temperature (0c) 36 47 60 69
0.5 Cake(gm) 28.8 28.2 27 26.9
%yield 36 37.33 40 40.22
0.25 Cake(gm) 28.4 27.3 26.3 26.2
%yield(gm) 36.89 39.33 41.56 41.78
The maximum extraction yield of Neem oil was 41.78% at particle size 0.25mmfor the extraction
time of 4hours and the minimum yield obtained was 36% at maximum particle size (0.5mm) and
minimum extraction time of one hour.
Effect of extraction time on percent yield of oil
Percent yield of Neem oil can be affected by extraction time, temperature, solvent type, particle
size and other components in the seed. Extraction time plays a great role on the percentage yield
of Neem oil using n-hexane as a solvent. Figures4.1 (a), (b) show that as the contact time
increases the oil yield also increases this continues till transfer of oil from the kernel powder to
the solvent attains zero. In other word, when the maximum amount of extractable oil is obtained,
the oil yield level remains invariable even by extending the reaction time. So that in the soxhlet
extraction the maximum oil yield could be finding at an extraction time of 4hours and above. As
shown graph of Figure 4.1, extracting the oil beyond four hours is wasting time because using n-
hexane as a solvent can find maximum yield at this time. The extraction rate is fast at the
beginning of the extraction but gets slow gradually. The reason is that when the kernel powder is
exposed to the fresh solvent, the free oil on the surface of seed is solubilized and oil gets
extracted quickly inducing a fast increase in the extraction rate. Furthermore, since the oil
concentration is low in the solvent at the beginning of the extraction process, the oil diffuses
quickly from the kernel to the liquid phase due to the difference in concentration (driving force)

of the oil. As the time passing by, the concentration of oil increases in the solvent resulting in a
decrease in the diffusion rate
Fig 4.1: effect of time and particle size on the yield
Effect of particle size on percent yield

As we can observed from the figure above. Particle size plays a great role on the yield of Neem
oil. Smaller particle size gives high yield. While samples with large particle size deliver low
yield. That means less oil is extracted from the larger particles (>0.5 mm) compared to the small
size of the particles. The reason is that larger particles with smaller contact surface area, have
more resistant to solvent entrance and oil diffusion towards the solvent. Therefore, less amount
of oil will be transferred from inside the larger particles to the surrounding solution in
comparison with the smaller ones. Thus, an increase in particle size will decrease the oil yield.
Nevertheless, we know that when the particle is too small (very fine particle size) i.e.,
below<0.25 mm, the extracted oil become small in its amount, even though the contact surface
area for small particle is supposed to be significantly higher than that for the larger particles.
This may be due to the agglomeration of the fine particles which reduces the effective surface
area available for the free flow of solvent towards inside the solid particles.
Effect of temperature on oil yield
The quantity of oil can be affected by temperature. From the experimental result, for instance,
let’s look at Figure 4.2 line (a, light blue color) for particle size from 0.25mm % yields of Neem
oil using hexane at a temperature of 30, 40 ,50 and 60 o C are 20.8, 24.4, and 29.8%,
respectively. This result will show us increasing temperature will raise yield. Based on the
findings, for both solvents, the yield was found to enhance with increasing temperature. This was
because of rising the temperature, both the diffusion coefficient and the solubility of the oil in to
both solvents is enhanced, thus heat treatment improves the extraction Neem oil. The higher
extraction temperatures the easier to break the molecule inside the seed; as a result, the higher
the yield obtained.

Fig 4.2 effect of temperature on the

yield
Table 4.3: characteristics of Neem Oil

Property Literature value Unit
Odour Garlic -
Specific gravity at 30oC 0.908-0.934 -
Refractive index at 30°C 1.4615-1.4705 -
Iodine value 65 – 80 g/g
Color varies Golden yellow , yellowish brown

- ,
reddish brown
PH 5.7 – 6.5 -
Acid Value 40 mg KOH/g
Saponification value 175-205 mg KOH
4.3 Stability testing at 50 degree centigrade

Procedure
1) 40 ml water was taken in a beaker.
2) Then the water was heated to 50 degree centigrade.
3) 10 ml emulsion was added slowly at the time of stirring in the beaker for well
mixing.
4) Then the sample was kept at 50degree centigrade for 2 hours in a temperature
controlling device.

4.4. Observation and Discussions

In this study oil-in-water emulsion was developed with neem oil as the internal phase which was
used as biopesticide and its stability was examined after 2 hours of constant temperature
controlling operation. After two hours no sign of phase inversion was noticed, so the sample was
considered to be a stable emulsion. The observation was that the sample was found to be
astable one in this study and no breakdown of phase was observed. Also we put it at room
temperature for 1 week and observed no breaking of emulsion. The milky color
biopesticide obtained was tested on bed bugs, weevils and observing that it kills bedbugs in 2
minute. But it took almost 2days to kill weevils and other insects 2-7 days this not indicate the
weakness of biopesticide when compared to other pesticides since it is very systematic to work
on insects and pests by entering their hormones and affect their cycles like to eat, mate, laying
eggs.
Azadirachtin is the most active. It reduces insect feeding and acts as a repellent. It also interferes
with insect hormone systems, making it harder for insects to grow and lay eggs. Neem oil has
many complex active ingredients. Rather than being simple poisons, those ingredients are similar
to the hormones that insects produce. Insects take up the neem oil ingredients just like natural
hormones. Neem enters the system and blocks the real hormones from working properly. Insects
"forget" to eat, to mate, or they stop laying eggs. Some forget that they can fly. If eggs are
produced they don't hatch, or the larvae don't moult. Obviously insects that are too confused to
eat or breed will not survive.

CHAPTER FIVE
5. MATERIALAND ENERGY BALANCE
Note: All material balances had performed and scaled up based on the experimental work in the
laboratory.
Balance on depulping
· Weight of the flesh from the fresh fruit=5% of fresh fruit
· Amount of water for depulping=50% of Kg of fresh fruit
Basis: 1000kg/hr of fresh fruit
Total Material Balance
Accumulation=Output + Consumption – Input – Generation
Since there is no reaction, the generation and consumption terms are zero, no accumulation.
M2 =water
M1=fresh fruit M4=seed (95% M1 +

Depulping
+water (5%M2) water (5%M1
M3=water (95%M2) +flesh (5% M1)

Input = Output
But the amount of water flow rate required for pulping 1000kg/hr fresh fruit will
be500kg/hr=M2.
 Water in stream three = 95% of M2= 475kg/hr of water
 Amount of water leaving with seed (stream 4) =5% of M2 = 25kg/hr of water,
Assume that all water contents are removed from the seed by drying using sun light for 24hours.

Balance on dryer
M4 =seed +shell Dryer M6 =dried seed
M5=water
 Weight of the flesh= 5% of wt. fresh fruit=50kg/hr of flesh
 Neem fruit with the shell=95% of M1=950kg/hr of Neem seed
Balance on decorticating machine

From laboratory work, wt. Neem kernel= 60-55% of Neem seed, let’s take the average value
57.5%.
M6=neem seed M8=neem kernel

Decorticator
M7=shell
 Wt. of shell=42.5% of Neem seed=403.75 kg/hr of shell have been removed,

and can be used as an adsorbent.
 Wt. of Neem kernel=57.5% of Neem seed= 546.25kg/hr Neem kernel send to

the storage tank.
Balance on size reduction Machine

Here by assuming that, there is 1% of Neem kernel loss, then = 5.4625kg/hr of kernel lost
Then, the amount of raw material left to the extraction vessel with particle size ranges from
0.25-0.5mm will be 540.79kg/hr=M9

Balance on Extraction machine
M10=solvent type
M9=crushed kernel
Extractor
M11=kernel+solvent+oil
In the laboratory, for 45 g of Neem kernel, the amount of solvent required should be 2 times.
Thus, 300 ml of solvent was used, therefore, for 540.79kg/hr of Neem kernel 3605.2667lt of
solvent will be required which is 2361.45 kg solvent required.
Therefore M11= wt. of solvent, oil and cake= 2902.24kg/hr
Balance on Centrifuge
From the experimental work, the cake from the extractor was 28.8 gm.Thus, to process 540.79kg
Neem kernel there is 346.1 kg of cake as by product.
M11=output from extractor M13=miscella

Centrifuge
M12=cake
Therefore M13= 2902.24 -346.1kg/hr =2556.1kg/hr
 The amount of miscella is 2556.1kg/hr which contains oil and solvent. This is separated
by evaporation and the solvent will recycled by using the condenser.

M14=solvent
M13=miscella M15=neem oil

Evaporator
From the experimental work, cake from the centrifuge contains 6.1% solvent. For processing
45gm Neem kernel, there is 2.745gm of the solvent loss. Thus, to process 2556.1kg there is a
loss of 155.9kg of solvent.
M14 =2361.45kg -155.9kg =2205.55kg/hr
M15=M13-M14 =2556.1-2205.kg = 350.55kg/hr
Balance on blending
M15= neem oil
Blending
M16 = Detergent and M17=biopesticide
stirring
From the experimental work 5gm of detergent was mixed with 10 gm of neem oil .Thus to
processes 350.55kg it requires 175.275kg/hr of detergent.
M17 = M16 +M15 =175.27kg +350.55kg = 525.82kg/hr

Table 5.1: summary of material balance

Equipment Input type Flow rate (kg/hr) Flow rate (kg/hr)
Output type Capacity (kg/hr)
Pulping  Fresh fruit 100  Water +flesh 525 1500

 Water  Water +seed
500 975
Drying Seed and water 975  Water 25 975

 Seed
950
Decorticator Seed 950  Kernel 546.25 950

 Shell
403.75
Pulverizer Kernel 546.25  Loss 5.4625 546.25

 Crushed kernel
540.79
Extractor  Kernel 546.5  Oil 2902.24 2902.24

 Solvent  cake
2361.45
Centrifuge Oil and cake 2902.24  Miscella 2556.1 2902.24

 Cake
346.1
Evaporator Miscella 2556.1  Oil 350.55 2556.1

 Solvent
2205.55
Blending  Detergent 175.27 Biopesticide 525.82 525.82

 Oil
350.55

Energy balance on evaporator
QSolvent
QmiscellaQOil
Evaporator
Qsupplied
Cpoil=2.053J/g.oc
Cps =2.195J/g.oc
Qsteam+QM=QS+QO
Cpmix= 0.86*2.1956+0.14*2.053= 2.18j/g.oc
Qsteam = MSCpS(T2-T1) +MOCpO(T2-T1) -MMCpmix(T2-T10)
=614.98g/s*2.195J/g.oc*(68oc-25oc)+97.38g/s*2.053J/g.oc(68oc-25oc)-
710.08g/s*2.18J/g.oc*(680c-25oc)
= 138.44j/s energy supply is required

CHAPTER SIX
6. SIZING AND DESIGN OF MAJOR EQUPMENT

6.1 Sizing of equipment’s
Sizing of depulping machine
Assume the density of fruit in the depulping machine is 1200kg/m3and taking the mass flaw rate
from the mass balance. The Operating hour is 8hr per day
Solid to be handled neem seed depulping by water
Density solid 0.667*1000kg/m3+0.333*1200kg/m3=1066.6kg/m3
Mass flow rate of solid 1500kg/hr
Temperature of the solid 25oC
Material of construction carbon steel
Volume flow rate
==1.406m3/hr
v=1.406m3/hr*8hr=11.25m3
Sizing of dryer
Solid to handled dried neem seed
Density of solid 1200kg/m3
Mass flow rate 975 kg/hr
Temperature 25oc
Construction material carbon steel
Q = 0.8125m3/hr
V =8.8125m3/hr*8hr=6.5m3

Sizing of decorticator
Solid to be handled decortized neem seed
Mass flow rate 950 kg/hr
Temperature 25oc
Constriction material carbon steel
Volume flow rate =
==0.779m3/hr
v =0.79m3/hr*8hr= 6.33m3
Sizing of pulverizer (size reduction machine)
Solid to handled crushed neem seed
Mass flow rate 546.25 kg/hr
Temperature 25oc
Constriction material carbon steel
Volume flow rate=
=0.46m3/hr
v =0.46m3/hr*8hr= 3.64m3
Sizing of extractor
Slurry to handled crushed neem seed and solvent (hexane)
Density of slurry (0.186* 1200+0.814*655) kg/m3= 756.37kg/m3
Temperature 65oc

Constriction material stainless steel
Volume flow rate=
=3.84m3/hr
V =3.84m3/hr*8hr=30.7m3
Sizing of centrifuge
Slurry to handled cake, oil and solvent (hexane)
Density of the salary 756.37kg/m3
Temperature 65oc
Volume flow rate=
=3.84m3/hr
V =3.84m3/hr*8hr=30.7m3
Sizing of evaporator
Slurry to handled miscella (solvent and oil)
Density of the salary0.94*906kg/m3+0.06*655kg/m3=890.94kg/m3
Temperature 65oc
Construction material stainless steel
Volume flow rate=
=2.87m3/hr
V =2.87m3/hr*8hr=28.95m3

Sizing of blending
Slurry to handled detergent and oil
Density of the salary0.67*906kg/m3+0.33*725.78kg/m3=846.53kg/m3
Temperature 65oc
Volume flow rate=
=0.62m3/hr
V=0.62m3/hr*8hr=4.96m3
6.2. Design of centrifuge

MaterialSelectionforDesignProcedure
The Solid bowel decanter centrifuge in its effective performance has its own operating
temperature. The temperature of the feed entering SBDC can range from 10 to 90oC, but due to
localized friction within the centrifuge the temperature could exceed this. Any material used
must be able to withstand temperature up to 120oC and be able to operate in acidic or basic
environment. Thus the materials used to construct this plant should have to withstand the high
temperature recommended. Accordingly stainless steel (316) is used to construct and design
Solid bowel decanter centrifuges. Because; this material can withstand a temperature of about
200oC and above with effective wear resistance and suitability for this design.
Conical-Cylindrical SBDC:-developed based on conical and cylindrical bowel designs. This

particular design allows efficient dewatering ability, effective clarification, and good recovery
within one unit. Thus conical-cylindrical SBDC design is the most widely used in oil industry
with a minimum recovery of 90% and above(Leung, “Industrial Centrifugation Technology”,
McGraw-Hill Professional, 1998).As a result we selected a conical-cylindrical SBDC in case of
our project design.

Fig 6.1 Conical-cylindrical SBDC, with its main components.
Physical Parameters Design
Factors which affect the separating ability of conical-cylindrical SBDC are determined by the
physical parameters such as the geometric design and the rpm’s at which the unit spins. The
main ones to consider are as follows:
 Centrifugal force
 Suspension volume
 Retention time
 Beach angle
 Clarifying area
 Equivalent clarifying area
The measurements and parameters which an engineer will need to carryout in design of conical-
cylindrical SBDC are the following:
Db =Inner bowel diameter
Dw=Weir diameter (m)
Lcyl=Cylindrical length (m)
Lc = conical length
n = bowel speed (rpm)
α =cone angle

Lcyl
Lc
Fig 6.2.Dimension specification of solid bowel decanter centrifuge
Clarifying Area
This is a parameter which is often used to make one SBD centrifuge appear more effective than
the other. The clarifying area in square meters is the wetted surface of the bowl interior. The
problem with this value is that every manufacturer uses a different formula to calculate it. There
is no common standard formula for this parameter. The results obtained by the various formulae
in use vary significantly, as can be seen in table below where the clarifying area for 73 cm bowl
diameter decanter with a 4:1 bowel length to bowel diameter ratio is calculated.
The results vary dramatically. If the clarifying area is to be compared, it is critical that the same
formula is applied to each conical-cylindrical SBD centrifuge. Of these formulae the one used by
Sokolov (W.J.Sokolov, Berlin, 1971) is the simplest and as long as this formula is used for all
of the cases, it provides a reasonable basis for comparison of the equivalent clarifying area.
Ac = π *Db * Lcyl ……………………………………. (1)
2
Ac= Clarifying area (m )
Db = inner bowel diameter (m)
Lcyl= Cylinder length (m)
In theory, the greater the clarifying area is, the more effective the separation of the centrifuge. In
practice the clarifying area is not a precise measurement and can at best be used as an indication,
with little bearing on the actual performance of the centrifuge. Thus in case of our design we
selected a SBDC by Sokolov of (conical-cylindrical shape)

Ac = 5.50 m2
Rds = 0.126 m
Rdl = 0.132 to 0.155m
Bl: Db = 4:1
Db = 0.73m
Dw =Rds+Rdl …………………………………. (2)
Where:
Ac = clarifying area (m2)
Rds = solid discharge radius, conical section (m)
Rdl = liquid discharge radius, cylindrical section (m)
Bl = bowel length (m)
Dw = weir diameter
Now it is possible to calculate/find the values of Bl, Lcyl, and Dw respectively.
Step 1.Calculating bowel length (Bl).
It is given that the ratio of Bl to Db =4 to 1(i.e. 4:1), and Db =0.73m.By this data;
Bl/Db =4/1, from this
Bl=4/1*Db
=4/1*0.73m
= 2.92m
Step 2.Length of cylindrical section (Lcyl)
From equation (1)
Ac = π *Db * Lcyl, and Db=0.53m from this

Lcyl = Ac/ (π *Db)
= 5.50m2/ (3.14*.730m)
=2.39m
Step 3.weir diameter (Dw)
From equation 2,
Dw = Rds+Rdl
But the value of Rdl is 0.132 to 0.155m; then taking mean of the two values
Rdl = (0.132+0.155)/2
=0.1435m
Thus;
Dw = 0.126m + 0.1435m
= 0.269 m

Centrifugal Force
This is the most obvious parameter which comes to mind when considering the action of a
centrifuge. The maximum centrifugal acceleration, developed inside a centrifuge is a function of
its radius and angular rotational speed. More commonly the term G-force or G-value is used
instead of acceleration. “G--‐force “is defined as the ratio between the centrifugal acceleration
created inside the bowl and the earth gravity acceleration g=9.81m/s2. A centrifuge works like a
gravity settling tank, except instead of operating at 1G, the centrifuge operates at several
thousand G thus reducing the settling time.
A formula for approximating the G-force at the bowl periphery is:
G = (n2*Db)/1800 …………………………………….. (3)
Where, G = G-force
n =bowel speed (rpm)
According to data source by Sokolov (W.J.Sokolov, Berlin, 1971) the variation for bowel
speed, G-force and scroll speed, and volumetric flow rate in case of cane is:
Table 6.1: specification of components of SBDC

Volumetric flow rate(Q) 3_300m3/hr.
Bowel speed (rpm) 500_3500rpm
G-force 400G _3000G
Conveyor/scroll speed(rpm) Exactly lower than bowel speed by a

minimum of 10 rpm, and adjusted as
needed for effective separation.
Based onSokolovthe effective separation is with rotational speed (i.e. bowel speed) of (n=
2500rpm).By this value:

G = (n2*Db)/1800
= (25002*0.730m)/1800
= 2,534.7, say =2,535G
As we observe from the above formula (3), the centrifugal acceleration or G- value will increase
with the bowel diameter and bowel speed.
With particular range of SBD centrifuges, the larger ones generally run at lower G-forces than
the smaller ones, as there are structural limitations to running larger machines at high G-values.
A larger decanter running at the same G-force as smaller one will give batter separation. This
means that when comparing two centrifuges of different diameters but similar bowl speeds, the
larger unit will generate more G-force and can be expected to provide better separation.
BeachAngle
When solids are transported along the beach of a SBD (the conical section), there is a force
acting on the solids in the direction of the liquid pool, named the slippage force (S) as shown in
Figure below. This force depends on the value of the difference between the specific gravity of
the solid and the surrounding medium. This means that the slippage force increases considerably
when the solids pass out of the liquid pool onto the beach where they are surrounded by air.
n (rpm)
-------------------------------------------------------------------------- S
Fig 6.3 Slippage force on the beach of SBDC

Theangleofinclinationcanvaryfrom8to15degree, and is selected depending on its recovery.
For a given set of feed densities the slippage force can be calculated as follows:
S = G * sin α ……………………….. (4)
Where; G = Gravitational force generated by the centrifuge
α = the cone angle of the centrifuge

Centrifuge with a small cone angle generate lower slippage forces than ones with a steep cone
angle. A small cone angle is desirable when the solids do not compact well and have a soft
texture such as in the case of digested sewage sludge.
A low cone angle also is advantageous when dealing with highly compacted solids which require
high torque to convey. A lower cone angle results in a lower wear rate on the scroll.
Steeper cone angles are suited to materials which are conveyed easily by the scroll. They also
result in a greater pond depth. Thus we selected an angle of 10 degree (W.J.Sokolov, Berlin,
1971)
Thus, S = G * sin α
= 2535sin (10o)
= 440.2G.
Suspension Volume
The suspension volume of a decanter can be considered as the total content of the liquid zone in
the bowl. This volume may change in relation to the “weir plate” diameter. The suspension
volume (Vs) consists of two components: the volume contained in the cylindrical section (Vcyl)
and the volume contained in the conical section (Vcn).It can be calculated as follows:
Vcyl = π/4*(Db2 – Dw2) * Lcyl…………………….. (5)
Vcn = π/8*(Db – Dw)/tanα*((Db2 + Db*Dw + Dw2)/3 – Dw2)….. (6)
Vs = Vcyl + Vcn ………………………………… (7)
o
Where; α = angle of inclination or cone angle ( )
Now it is possible to solve the above equations (5, 6, and 7) respectively.
Vcyl = π/4*(Db2 – Dw2) * Lcyl………….5
= 3.14/4*(0.73 m2 - 0.269m2)*2.39m
= 0.785*(0.460539m2)*2.39m
= 0.864 m3

Vcn = π/8*(Db – Dw)/tanα*((Db2 + Db*Dw + Dw2)/3 – Dw2.................6
= 3.14/8(0.73m – 0.269m)/tan10o ((0.73m2 + 0.73m*0.269m + 0.269m2)/3- 0.269m2
= 1.03*(0.801631)/2.93
= 0.2820m3
Now it is possible to solve Vs as:
Vs = Vcyl + Vcn
= 0.864m3 + 0.282m3
= 1.146m3
The effect of the suspension volume in a SBDC shows larger settling volume generally leads to a
better degree of separation. A larger suspension volume results in better separation.
Retention Time
Retention time is a parameter most engineers can relate to quite well. Unfortunately it is quite a
complicated issue in the case of centrifuges. Different phases are discharged, and as there may a
buildup of the solid phase in the machine, the retention time should take this into consideration.
In most case it is possible to deal with quite dense solids, so the solids generally make up a
relatively small percentage of the volume of the feed.
Each manufacturer will also calculate retention time in a totally different manner depending on
how they interpret the flow of fluid through the internals of their centrifuge design. So any
calculation will be an approximation at best. As long as the same approximation is used, this can
still give a sound basis of comparison between different SBDC.
The suspension volume provides a reasonable approximation as the basis for calculating the
retention time of the centrifuge, giving an approximation of the time which the slurry resides in
the centrifuge under the effect of centrifugal forces. The retention time can be calculated as
follows:
Based on general data from material balance on centrifuge and literature:
Let ρ density of miscella =756.37Kg/m3,

Q=M/ρ =(2556.1Kg/hr)/756.37Kg/m3=0.00094m3/sec=0.00094m3/sec*3600sec/hr.
=3.38m3/hr. where
ρ=density of miscella, Q=volumetric flow of the micella
M= miscella
TR= VS/Q ……………………….. (7), Where,
3 3
TR = Retention time (sec), Vs= Suspension volume (m ), Q = Volumetric feed rate (m /h)
Q=3.38m3/hr.Thus
TR = (3600sec/hr*1.146m3)/ (3.38m3/hr.) = 1214.2sec
The longer the retention time, the better the separating efficiency of the centrifuge. As can be
seen, the larger suspension volume leads to a higher retention time. This does, however, not give
any indication whether the available retention time is actually enough to achieve the desired
degree of separation. The actual retention time required for each particular sludge will be
different and is affected by parameters such as:
 Particle size
 Relative densities of the phases
 Viscosity of the liquid phases
 Ratio of phases
For a given feed rate with no change in the speed at which the centrifuge is operated, the
retention time of a centrifuge can be varied by changing weir settings, thereby increasing or
decreasing the suspension volume. This can be achieved by either installing static weir plates of
different diameters, or in more technologically advanced machine by means of an adjustable
weir. This gives such machines much greater flexibility.
A further adjustment which can affect the retention time of the solids in the centrifuge is the
differential speed between the bowl and the scroll. Slowing down the scroll relative to the bowl
means that the solids are conveyed more slowly from the centrifuge. This generally results in a
more compact and dryer cake and a clearer centrate.
Equivalent Clarifying Area

This is a value which aims to put the effectiveness of a SBD centrifuge into terms which are
easier to visualize. Essentially a centrifuge can be likened to a settling pond in which high G
forces are applied to improve the settling characteristics of the phases. In order to gets an idea of
the relative settling capacity of a centrifuge; one can calculate the Equivalent Clarifying Area or
Sigma value (Σ)
Σ =Ac*G………………………….. (8)
2
Σ = sigma value (m )
2
AC = Clarifying area (m )
G = G-force (m)
This Σ can be seen as the equivalent surface in square meters of a static settling pond required
producing the same separation result as the centrifuge.
Re-substituting the previous formulae gives:
2
Σ = (π* n *Db2*Lcyl)/1800 ……………………...(9)
Thus Σ = (3.14*2500rpm2*0.73m2*2.39m)/1800 = 13,886 m2
As can be seen, the effect of the diameter of the bowl is more pronounced than that of the length
of the bowl. Increased bowl speed and increased length of cylindrical section of a centrifuge will
tend to improve the settling of fine solids, resulting in a clearer liquid phase.
Given two machines generating the same G forces, the one with the larger diameter would tend
to be more effective in achieving separation, assuming that both machines are properly designed
for the given application.
Since 13,886m2 ≈ 13900m2, the design is effective by Sokolov.
Terminal Falling Velocity of Particles
This terminal falling velocity of particles is calculated as follow.
Uo = Q / Σ…………………… (10).Thus,
Uo = (3.38m3/hr) / 13886m2=0.00024m/hr

= (0.00024m/hr)/3600s/hr
= 6.7* 10-8 m/s
Fluid (Centrifugal) Pressure
If the machine is separating solid liquid particles, the fluid pressure is given by:
Pf = ((ρl*ω2 (r12-r22))/2……………………(11)
Where, Pf = pressure of fluid
ρl = density of liquid
ω = angular velocity
r1 and r2= radius of bowel and liquid surface respectively
For design case, the maximum fluid pressure will occur when the bowel is full, (r2= 0).
Again angular velocity;
ω = (2πn)/60……………….. (12)
= (2*3.14*2500rpm)/60
= 262 rad/s, and taking the density of miscella in case of density of liquid (ρ=
756.37Kg/m3) with bowel radius r1=0.73/2=0.37m and radius of liquid surface
r2=0m, also 262rad/sec=4.4m/sec
Pf = ((ρl* ω2 (r12-r22))/2…………………(3)
= (756.37kg/m3*(4.4m/sec) 2*(0.37m) 2)/2
= 2004.7N/m
Wall Thickness
The maximum allowable wall thickness required can be estimated using the equation
Et = (Pt*r1)/Fm*103……………………….. (13)
Where, Et = wall thickness

Pt = total (fluid and self-pressure) which is almost equal to fluid pressure
Fm = design stress
But from table Fm=120N/mm@200 oc, thus
Et = (2004.7 N/m2*0.37m)/ (120N/mm*1000mm/1m)
= 0.0062m =6.2mm and with a corrosion allowance of 2mm ≈ say 8mm, which is
perfectly meet the selected design thickness given in table (6.1).
How Solid Bowel Decanter Centrifuge Works
Solids discharge: Solids/cakes which are sediment inside the bowel discharged due to high
rotational speed of the bowel and G-force.
Solids deposited: These are the settled solids forming an annular of 360 degree, shape around
the bowel, and being conveyed forward by the scroll at the rate relative to the differential speed.
Liquid discharge: is the liquid discharge end of SBDC, this liquid is sent to the processing plant
for further clarifications and treatments.
Cylindrical section: Dewatering is occurred in this section if necessary.
Beach (conical section): This section is designed to exert additional force on the solids
squeezing out the last drops of liquid as we are not only applying centrifugal G-force but also
pushing the solids. It is designed to elevate the solids above the waterline into the discharge
chamber.
Gearbox: SBDC is equipped with a drive system composed of different motors. The electric
motor provides the power required to turn the complete rotating assembly driving the bowel
directly and the scroll through the hydraulics

CHAPTER SEVEN
7. COSTING AND ECNOMICS

7.1purchased Equipment cost
The ultimate purpose for developing such a detailed process design and cost estimate is to
determine the economics of production of biopesticide from neem seed.
Table 7.1: purchased costs of the equipments
No. Equip. type Required Capacity or Size Cost
Dollar Birr
1. Pulping machine 1500kg/hr 11.25m3 8000.99 187303.18
2. Decorticator 950kg/hr 6.33m3 1667 57357.36
3. Pulverizer 546.25kg/hr 3.64m3 236 7915.45
4 Dryer 975kg/hr 6.5m3 1460 5676.78
5. Extractor 30.7m3 31000 1252440.75
6. Centrifuge 30.7m3 16715.68 391314
7. Evaporator 28.95 m3 12154.6 284539
8. Storage tank-1 1900 m3 161245.6 3774759.6
9. Storage tank-2 2. 207 m3 2795.87 65451.3
10. Pumps(3) Flow rates
0.0617m3/min, 13410.25 313933.87
0.0618m3/min, 13425.01 314279.52
and
0.0046m3/min 2819.62 66007.4
Total purchased equipments cost 6720968.47
Source: http//: www. Alibaba.com

7.2. Estimation of capital investment cost
7.2.1. Fixed capital investment (FCI) estimation

Direct cost
A. Costs of equip. + installation + instrumentation +piping+ electricity+ painting (50-60%of
FCI)
I. Purchased equipment cost (PEC) =6720968.47Br.
II. Installation including painting (30% of PEC) =2016290.54Br.
III. Instrumentation and control (6% of PEC) =403258.11Br.
IV. Piping (5% of PEC) =336048.44Br.
V. Electricity (40% of PEC) =2688387.39Br.
B. Building, process and auxiliary (10% of PEC) = 672096.85Br.
C. Service facilities (40% of PEC) =2688387.39Br.
D. Land (4% of PEC) = 268838.74Br.
Direct cost (DC) = A + B + C + D = 15794275.93Br.
Indirect cost
A. Eng’g and supervision ( 20% of DC) = 3158855.19Br.
B. Construction expense and contractor fee (18% of DC) = 2842969.67Br.
Indirect cost (IC) = A + B = 6001824.86Br.
Fixed Capital Investment (FCI) = DC + IC = 21796100.79Br.
Working Capital investment (WCI) = 15% of TCE =1008145.27Br
TCI = FCI + WCI =22,804,246.06Br
From the above two equations and the value of FCI, the WCI = 1008145.27Br.
7.2.2. Total production cost (TPC) estimation

 Total fresh fruit required: 1000kg/hr = 2,400,000 kg/yr
 Price of fruit = 5Br/kg
 The total costs of fresh fruit = 12,000,000 Br/yr
Total fresh n- hexane solvent required
 Total crushed kernel = 540.78kg/hr * 8 * 300 = 1,297,672 kg/yr
From the experimental work, for 0.045kg of crushed kernel, we used 0.3lt solvent. Then, for
1,297,672 kg of kernel we will use 8,651,146.7solvent. Assuming that the recovered solvent can
be used again at least two times.Therefore, total solvent required 4,325,573.35lt/yr. and this costs
216278667.5 Br.

 Price of detergent 20birr/kg and the annual price is 175,27kg/hr

*8hr/day*300days/yr=420648kg/yr
 Price of detergent = 420648kg/yr*20birr/kg = 8412960birr/yr
Hence the total production cost of raw material is = 236,691,627.5birr/yr.
A. Fixed Charges (FC)

1. Depreciation=10% FCI = 2179610.8Birr
2. Local taxes = 2.5% of FCI = 0.025*21796100.79=544902.52Br
3. Insurance = 0.7% of FCI = 0.007*21796100.79Br=152572.71Br
Total Fixed Charge = 688899.27+544902.52+152572.71=1386374.5B.
B. Production cost (PC)
FC = 15% of TPC, TPC = 9242496.67Br.
1. Raw material and inputs = 236,691,627.5birr/yr.
2. Operating labor (15% of TPC) = 0 15*9242496.67=1386374.5Br.
3. Direct supervisor and clerical labor (20%operatin labour) = 277274.9.Br
4. Utilities (15% of TPC) = 0.15*9242496.67Br=1386374.5Br
5. Maintenance and repair (6% of FCI) = 0.06*21796100.79Br=1307766.05Br
6. Laboratory Charge (15% of operating labor) = 0.15*1386374.5=207956.12Br
Total product cost = 241257373.6Br.
C. Plant overhead cost (POC) (10% of TPC) = 0.1*9242496.67=924249.67Br
Manufacturing cost = FC + PC + POC = 243567997.8Br.
General expenses
A. Administration cost (4% of TPC) =0.04*9242496.67=369699.87Br
B. Distribution and selling cost (11% of TPC) = 0.11*9242496.67=1016674.63Br
C.Researchanddevelopment cost (5% of TPC) = 0.05*9242496.67=462124.83Br.
General expense = 1848499.33Br.
Total production cost = Manufacturing cost + General expense = 245,416,497.1Br.
Whole selling price of biopesticide
Price of biopesticide = 200Br/kg
Total income = 200birr/kg*525.82kg/hr*8hr*300days/yr = 252393600birr/yr
Gross income = Total income – total production Cost
= 252393600birr/yr -245,416,497.1birr/yr =6977102.9birr =Net profit before tax
Let take the tax rate in Ethiopia 35% of gross income, tax = 2441986.02birr.
Net profit after tax = Gross income – tax = 4535116.89Birr.

7.3. Economic evaluation
7.3.1. Net income, Payback time and return on investment

Rate of return
Rate of return on investment after tax
== = *100%= 20%
Rate of return on investment before tax= =31%
Payback period = =
3.25yrs
Neem oil extraction using n-hexane is profitable as it is clearly observed from the above cost
estimation. The rate of return on investment 31% implies the plant returns 31% of its total capital
investment in three year. The payback period tells us the plant return its total investment cost in
around three years and then it will become profitable. The income statement and the other
indicators of profitability show that the project is viable. The project can be implemented after
detailed feasibility study has been done.
7.4. Plant layout, environmental impact and site location

The geographical location of the final plant can have strong influence on the success of an
industrial venture. Considerable care must be exercised in selecting the plant site, and many
different factors must be considered. Primarily, the plant should be located where the minimum
cost of production and distribution can be obtained, but other factors, such as room for expansion
and safe living conditions for plant operation as well as the surrounding community, are also
important.
A general consensus as to the plant location should be obtained before a design project reaches
the detailed estimate stage, and a firm location should be established upon completion of the
detailed-estimate design. The choice of the final site should first be based on a complete survey
of the advantages and disadvantages of various geographical areas and, ultimately, on the
advantages and disadvantages of available real estate. The following factors should be
Considered in selecting a plant site: Raw materials availability, Markets, Energy availability,
Climate, Transportation facilities, Water supply, Waste disposal, Labor supply, Taxation and
legal restrictions, Site characteristics, Flood and fire protection, Community factors
Considering the above factors Biopesticide production plant the main raw material is neem tree.
Therefore the Neem tree (Azadirachtinindica) which is the fastest-growing trees in Ethiopia has

also a potential for the plantation of Neem tree which grows in humid, arid, and hot places
having an altitude of up to 1,500 meters above sea level. The tree is available in many part of the
country such as:-Jijjiga, Mekele, Bahirdar, wolkite, Harar, Diredawa, Adama, Gambella etc.
Mainly in Ethiopia the neem tree bears a fruit starting from March. Neem can grow in tropical
and subtropical regions with semi-arid to humid climates. Neem tree will adapt to a mean annual
rainfall of 450-1200 mm, mean temperatures of 25-35ºC and grow at altitudes of up to 800
meters above sea level. We selected Dire Dawa for this biopesticide production plant since the
availability of the main raw material and its location is also good for transportation of product to
other places else.
7.4.1 Environmental impact analysis

Neem oil based biopesticide management system pest is an environmental friendly, low cost, and
healthy method of pest control system. Bio-pesticides are a set of tools and applications that will
help our farmer’s transition away from highly toxic conventional chemical pesticides into an
era of truly sustainable agriculture. Of course bio-pesticides are only a part of a larger
solution; sustainable agriculture is a broad and deep field. But helping farmers move from their
current chemical dependency to organic agriculture and beyond requires tools for the
transition and tools for a new era. Biopesticides can and will play a significant role in this
process. The remain, however, serious questions about the safety of these products from both
a human and ecosystem health standpoint. By definition, green chemistry is about continuous
improvements aimed at reducing or eliminating hazard. Fully defining hazard is difficult. Even
products hailed by green chemistry and regulators alike as safer for human health may
turn out to have unforeseen negative environmental health impacts- for example, Spinosad,
a green chemistry award winning bio-pesticide, is significantly safer for humans than other
treatments but is toxic to bees. We must encourage pest management solutions and regulations to
continuously evolve ensure that multi-disciplinary teams, including green chemists,
environmental health sciences and other sciences, approach these products systemically to both
discover and refine them. They are the most likely source for alternatives to some of the most
problematic chemical pesticides currently in use that are under ever -increasing scrutiny.
Bio-pesticides may also offer solutions to concerns such as pest resistance to traditional chemical
pesticides, public concern about side effects of pesticides on the surrounding environment and
ultimately, on human health. Finally the cake as byproduct is used for fertility of the soil so it
have not effect on the environment.

7.4.2 Plant layout

After the process flow diagrams are completed and before detailed piping, structural, and
electrical design can begin, the layout of process units in a plant and the equipment within these
process units must be planned. This layout can play an important part in determining
construction and manufacturing costs, and thus must be planned carefully with attention being
given to future problems that may arise. Since each plant differs in many ways and no two plant
sites are exactly alike, there is no one ideal plant layout. However, proper layout in each case
will include arrangement of processing areas, storage areas, and handling areas in efficient
coordination and with regard to such factors as:
1. New site development or addition to previously developed site
2. Type and quantity of products to be produced
3. Type of process and product control
4. Operational convenience and accessibility
5. Economic distribution of utilities and services
6. Type of buildings and building-code requirements
7. Health and safety considerations
8. Waste-disposal requirements
9. Auxiliary equipment
10. Space available and space required
11. Roads and railroads
12. Possible future expansion
Preparation of the Layout
Scale drawings, complete with elevation indications can be used for determining the best
location for equipment and facilities. Elementary layouts are developed first. These show the
fundamental relationships between storage space and operating equipment. The next step
requires consideration of the safe operational sequence and gives a primary layout based on the
flow of materials, unit operations, storage, and future expansion. By analyzing all the factors that
are involved in plant layout, a detailed recommendation can be presented, and drawings and
elevations, including isometric drawings of the piping systems, can be prepared.

Fig 7.1 plant layout of biopesticide production

CHAPTEREIGHT
8. CONCLUSION AND RECOMMENDATION

8.1. Conclusion
There are different methods of essential oil extraction for the production of biopesticide from
neem seed. In our project we use solvent extraction method by using hexane. The extracted oil is
golden yellow color with smell of garlic and the percentage oil yield obtained was 41.78% with
the particle size of 0.25mm at a temperature of 69oc. The milky color biopesticide was obtained
and tested on bed bugs, weevils and observing that it kills bedbugs in 2 minute. But it took
almost 2days to kill weevils and other insects 2-7 days this not indicate the weakness of
biopesticide when compared to other pesticides since it work on insects and pests by entering
their hormones and affect their cycles like to eat, mate, laying eggs. Investigation on the
technical and economic feasibility of the work for biopesticide production was performed and
results from the feasibility study indicated that the proposed work was feasible with rate of return
(RR) 31% and with the payback period of estimated to be 3 years.

8.2. Recommendation
 To increase the agricultural productivity, to control environmental pollution and hazards

of chemical pesticides, the industries producing pesticide from neem should be installed
in Ethiopia.
 It is advisable to use extracts of neem seed using n-hexane as a solvent for biopesticides
production due to its low boiling point and also its non polarity which extracts the oil.
 When we are going to use neem oil biopesticide we should also be concerned
about the time of application because neem oil, which contains azadirachtin, for which
we used it as a pesticide, breaks down under sunlight and moisture. So it should be
used at afternoon or early in the morning so that tree leaves can absorb all the
portions of the diluted pesticide.

REFERENCES
Ahmed,S.,Bamofleh, M., Munshi, A., (1989) Cultivation of neem (Azadirachta indica) in Saudi
Arabia. Econ Bot, 43: 35-38.
Anonymous., (2006) Neem – Growing neem, organic farming, health, animal health,
environmental use, and home uses, economic potential, patents, new bazaars, research
papers, world neem conference. Neem foundation (Internet) Mumbai, India
ARPN Journal of Engineering and Applied Sciences Extraction Of Neem Oil (Azadirachtin
indica A. Juss) Using N-Hexane And Ethanol: Studies Of Oil Quality, Kinetic And
Thermodynamic.
Bulletin ,1998/15, Sankaram, Akella Venkata Bhavani Andhra Pradesh (IN) Azadirachtin
formulations and a process for preparing them from neem seed/kernel
Gunjan P., Bhojwani S. S., and. Srivastava A. K., (2002). Production of Azadirachtin from Plant
Tissue Culture, Biotechnol. Bioprocess Eng. Vol. 7,185-193.
Ghimeray A. K., Jin C.W., Ghimire B.K and Dong H.C., (2009). Antioxidant activity and
quantitative estimation of azadirachtin and nimbin in Azadirachta Indica A. Juss grown in
foothills of Nepal, African Journal of Biotechnology, 8, pp. 3084-3091
ICAR, (2009). Hand book of agriculture. Sixth Edition, Directorate of Information and
Publications of Agriculture,ICAR, New Delhi –110012
Kumar, R.V., Gupta, V.K., (2002) Thrust on neem is need of today. In: Employment news, July
20 26.New Delhi, India.
Mattie’s Giger, (2011)
Mc Ewen, F.L., (1978) Food production – the challenge of pesticides. Bio Sci, 28: 773-777
Neem A Tree for Solving Global Problems Report of an Ad Hoc Panel of the Board on Science
and Technology for International Development National Research Council National
Academy Press Washington, D.
Neem Foundation, 1997-2

Nebil El-Wakeil, Botanical Pesticide and Their mode of action University, Napaam India
Nigam, S.K., Mishra, G., Sharma, A., (1994) Neem: A promising natural insecticide. Appl Bot
Abstr, 14: 35-46.
Nirmala Devi1andTarun K. Maji2Gauhati University, Guwahati Tezpur University, Napaam

India
Plant Based Pesticides: Green Environment with Special Reference to Silk Worms Dipsikha
Bora, Bulbuli Khanikor and Hiren Gogoi.
Sateesh, M.K., (1998) Microbiological investigation on die-back disease of neem (Azadirachta

indica A.Juss.).Ph.D. thesis. University of Mysore. Mysore, India.
Siddiqui, B.S., Afshan, F., Gulzar, T., et al., (2003) Tetra cyclic Triterpenoids from
the leaves of Azadirachta indica and their insecticidal activities. Hem Pharm Bull (Tokyo),
51: 415-417.
Wilkinson CF (1976) Insecticide Biochemistry and Physiology. Plenum Press. New York.
U.S. Environmental Protection Agency, (2012), About Pesticides,

“http://www.epa.gov/pesticides/about/types.htm”, updated on Wednesday, May 09, 20

APPENDICES
APPENDIX A
Table A1. Material properties for 316 stainless steal
Density 7870-8070 kg/m3 Endurance limit 256-307MPa
Young’s modules 190-205 GPa Fracture toughness 112-278MPa.m1/2
Tensile strength 480-620 MPa Hardness-Vickers 170-220 HV
Compressive strength 170-310 MPa Modulus of rupture 170-310MPa
Bulk modules 134-152 GPa Poisson’s ratio 0.265-0.275
Elongation 30-51% Shape factor 63
Elastic limit 170-310 MPa Shear modulus 74-82GPa
Design stress at 200oC = 120N/mm2
Minimum wall thickness without corrosion allowance,Et = 4mm to 12mm

APPENDIX B
Laboratory equipments and samples photo
B1: Soxhlet B2: rotary evaporatorB3: neem oilB4: Biopesticide product
B5: Killed bedbugs B6: powder of neem seed

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JIMMA UNIVERSITY

JIMMA INSTITUTE OF TECHNOLOGY

SCHOOL OF CHEMICAL AND BIO ENGINEERING

Title: Production of Biopesticide from Neem Seed

By group- 2 peer group -12

JIMMA INSTITUTE OF TECHNOLOGY

SCHOOL OF CHEMICAL ENGINEERING

Title: Production of Biopesticide from Neem Seed

Name of Advisor Signature Date

Mr. Samuel Gessese

Name signature date

JIT ,School of Chemical Engineering Page I

JIT ,School of Chemical Engineering Page II

Key words: neem, Azadirachtin, biopesticide, neem oil

JIT ,School of Chemical Engineering Page III

JIT ,School of Chemical Engineering Page IV

2.6.2 Feeding deterrent ..................................................................................................... 10

JIT ,School of Chemical Engineering Page V

7.3. Economic evaluation…………………………………………………………………..51

JIT ,School of Chemical Engineering Page VI

Table 4.1: Moisture content determination of Neem kernel…………………24

JIT ,School of Chemical Engineering Page VII

Fig 3.1. Block flow diagram of biopesticide production………………………. 15

JIT ,School of Chemical Engineering Page VIII

DDT Dichloro Diphenyl Trichloroethane

EPA Environmental Protection Agency

FAO Food and Agricultural Organization

HPLC High Performance Liquid Chromatography

NPEO Nonyl Phenol Ethoxylate

O/W Oil in Water

SLS Sodium Lauryl Sulphate

SBDC Solid Bowel Decanter Centrifuge

JIT ,School of Chemical Engineering Page IX

JIT ,School of Chemical Engineering Page X

JIT, School of Chemical Engineering Page 1

1.2. Statement of Problem

JIT, School of Chemical Engineering Page 2

1.3.1 General objective

1.3.2. Specific objectives

 To analyze the neem seed samples.

1.4. Significance of the study

JIT, School of Chemical Engineering Page 3

2.1.1.1 Microbial Pesticides

JIT, School of Chemical Engineering Page 4

2.1.1.3 Biochemical Pesticides

2.1.2 Chemical Pesticides

2.1.2.1 Organophosphate Pesticides

2.1.2.2 Carbamate Pesticides

2.1.2.3 Organochlorine Insecticides

2.1.2.4 Pyrethroid Pesticides

JIT, School of Chemical Engineering Page 5

JIT, School of Chemical Engineering Page 6

2.2.1 Reviews on Chemical constituents of Neem

2.2.2 Specification of Azadirachtin in neem

2.2.3 Importance of neem

2.2.4 Bioactive compounds in neem oil

JIT, School of Chemical Engineering Page 7

literatures are Azadirachtin (Azadirachtin A), salanin, nimbin, 3-tigloylazadirachtol

2.5 Factors affecting biopesticide production