PRODUCTION OF BIODIESEL BY TRANSESTERIFICATION OF

DIFFERENT PLANT OILS

PROJECT REPORT
Submitted in partial fulfillment of the
Requirements for the award of the degree of
Bachelor Of Technology
In
Biotechnology
By
G. SINDHU (07241A2320)
M. SWETHA (07241A2324)

DEPARTMENT OF BIOTECHNOLOGY
Gokaraju Rangaraju Institute Of Engineering And Technology
(Affiliated to Jawaharlal Nehru Technological University)
Hyderabad
2011

DEPARTMENT OF BIOTECHNOLOGY
Gokaraju Rangaraju Institute Of Engineering And Technology
(Affiliated to Jawaharlal Nehru Technological University)
Hyderabad

CERTIFICATE
This is to certify that the project entitled
PRODUCTION OF BIODIESEL BY TRANSESTERIFICATION OF
DIFFERENT PLANT OILS
has been submitted by
G. SINDHU (07241A2320)
M. SWETHA (07241A2324)
In partial fulfillment of the requirements for the award of degree of Bachelor
of Technology in Biotechnology from
Jawaharlal Nehru Technological University, Hyderabad.
The results embodied in this project have not been submitted to any other
University or Institution for the award of any degree or diploma.

Internal Supervisor
Ramesh
Asst Professor
Dept of Biotechnology

Head of Department
Dr. D. Sailaja
Professor & HOD
Dept of Biotechnology

CERTIFICATE
This is to certify that Ms G. SINDHU, Ms M. SWETHA, students of final
year
Biotechnology of Gokaraju Rangaraju Institute Of Engineering And
Technology,
affiliated to Jawaharlal Nehru technological university have completed a
project
work entitled PRODUCTION OF BIODIESEL BY
TRANSESTERIFICATION OF DIFFERENT PLANT OILS at our
biotechnology division of laboratory Gokaraju Rangaraju Institute Of
Engineering
And Technology, Bachupally,
Hyderabad.

Head of Department
Examiner
Dr. D. Sailaja
Professor & HOD
Dept of Biotechnology

External

CONTENTS
Acknowledgements
Abstract
Contents
1. Introduction
1.1
1.2
1.3
1.4

What is biodiesel?
Benefits of biodiesel
Biodiesel production
Advantages and disadvantages of biodiesel

2. Biodiesel production from Jatropha Curcus

2.1 Introduction about Jatropha Curcus
2.2 Production of biodiesel by transesterification process
2.3 Materials and methods
2.4 Results and discussions
2.5 Conclusion
3. Biodiesel production from Vegetable oil
3.1
3.2
3.3
3.4
3.5

Introduction about vegetable oil

Methanol transesterification of vegetable oil
Materials and methods
Results and discussion
Conclusion

ACKNOWLEDGEMENT
There are many people who have helped us directly or indirectly
in the successful completion of our project. We would like to take
this opportunity to thank one and all.
First of all we would like to express our deep sense of gratitude
towards our project guide Mr Ramesh, for always being available
whenever we require his guidance as well as for motivating us
through out the project work.
We are also grateful to the Dr. D.Sailaja, (H.O.D) for allowing
us to do the project in GRIET and for her valuable guidance during
our project, which helped me in the successful completion of my
project.
We would like to thank all our friends for their help and
constructive criticism during our project period. Finally, we are
very much indebted to our parents for their moral support and
encouragement to achieve higher goals. We have no words to
express our gratitude and still we are very thankful to our parents
who have shown us this world and for every support they gave us.

Signature
G. SINDHU (07241A2320)
(07241A2324)

Signature
M.SWETHA

WHAT IS BIODIESEL?
Biodiesel is an alternative fuel similar to conventional or fossil diesel. Biodiesel can be
produced from straight vegetable oil, animal oil/fats, tallow and waste cooking oil. The
process used to convert these oils to Biodiesel is called transesterification. This process is
described in more detail below. The largest possible source of suitable oil comes from oil
crops such as rapeseed, palm or soybean. In the UK rapeseed represents the greatest
potential for biodiesel production. Most biodiesel produced at present is produced from
waste vegetable oil sourced from restaurants, chip shops, industrial food producers such
as Birdseye etc. Though oil straight from the agricultural industry represents the greatest
potential source it is not being produced commercially simply because the raw oil is too
expensive. After the cost of converting it to biodiesel has been added on it is simply too
expensive to compete with fossil diesel. Waste vegetable oil can often be sourced for free
or sourced already treated for a small price. (The waste oil must be treated before
conversion to biodiesel to remove impurities). The result is Biodiesel produced from
waste vegetable oil can compete with fossil diesel. More about the cost of biodiesel and
how factors such as duty play an important role can be found here.

WHAT ARE THE BENEFITS OF BIODIESEL?

Biodiesel has many environmentally beneficial properties. The main benefit of biodiesel
is that it can be described as carbon neutral. This means that the fuel produces no net
output of carbon in the form of carbon dioxide (CO2). This effect occurs because when
the oil crop grows it absorbs the same amount of CO2 as is released when the fuel is
combusted. In fact this is not completely accurate as CO2 is released during the
production of the fertilizer required to fertilize the fields in which the oil crops are grown.
Fertilizer production is not the only source of pollution associated with the production of
biodiesel, other sources include the esterification process, the solvent extraction of the
oil, refining, drying and transporting. All these processes require an energy input either in
the form of electricity or from a fuel, both of which will generally result in the release of
green house gases. To properly assess the impact of all these sources requires use of a
technique called life cycle analysis. Our section on LCA looks closer at this analysis.
Biodiesel is rapidly biodegradable and completely non-toxic, meaning spillages represent
far less of a risk than fossil diesel spillages. Biodiesel has a higher flash point than fossil
diesel and so is safer in the event of a crash.

BIODIESEL PRODUCTION

As mentioned above biodiesel can be produced from straight vegetable oil, animal
oil/fats, tallow and waste oils. There are three basic routes to biodiesel production from
oils and fats:

Base catalyzed transesterification of the oil.

Direct acid catalyzed transesterification of the oil.
Conversion of the oil to its fatty acids and then to biodiesel.

The Transesterification process is the reaction of a triglyceride (fat/oil) with an alcohol to

form esters and glycerol. A triglyceride has a glycerine molecule as its base with three
long chain fatty acids attached. The characteristics of the fat are determined by the nature
of the fatty acids attached to the glycerine. The nature of the fatty acids can in turn affect
the characteristics of the biodiesel. During the esterification process, the triglyceride is
reacted with alcohol in the presence of a catalyst, usually a strong alkaline like sodium
hydroxide. The alcohol reacts with the fatty acids to form the mono-alkyl ester, or
biodiesel and crude glycerol. In most production methanol or ethanol is the alcohol used
(methanol produces methyl esters, ethanol produces ethyl esters) and is base catalysed by
either potassium or sodium hydroxide. Potassium hydroxide has been found to be more
suitable for the ethyl ester biodiesel production, either base can be used for the methyl
ester. A common product of the transesterification process is Rape Methyl Ester (RME)
produced from raw rapeseed oil reacted with methanol.
The figure below shows the chemical process for methyl ester biodiesel. The reaction
between the fat or oil and the alcohol is a reversible reaction and so the alcohol must be
added in excess to drive the reaction towards the right and ensure complete conversion.

The products of the reaction are the biodiesel itself and glycerol.

A successful transesterification reaction is signified by the separation of the ester and

glycerol layers after the reaction time. The heavier, co-product, glycerol settles out and
may be sold as it is or it may be purified for use in other industries, e.g. the
pharmaceutical, cosmetics etc.
Straight vegetable oil (SVO) can be used directly as a fossil diesel substitute however
using this fuel can lead to some fairly serious engine problems. Due to its relatively high
viscosity SVO leads to poor atomisation of the fuel, incomplete combustion, coking of
the fuel injectors, ring carbonisation, and accumulation of fuel in the lubricating oil. The
best method for solving these problems is the transesterification of the oil.
The engine combustion benefits of the transesterification of the oil are:

Lowered viscosity
Complete removal of the glycerides
Lowered boiling point
Lowered flash point
Lowered pour point

PRODUCTION PROCESS
An example of a simple production flow chart is proved below with a brief explanation of
each step.

Mixing of alcohol and catalyst

The catalyst is typically sodium hydroxide (caustic soda) or potassium hydroxide
(potash). It is dissolved in the alcohol using a standard agitator or mixer. Reaction. The
alcohol/catalyst mix is then charged into a closed reaction vessel and the oil or fat is
added. The system from here on is totally closed to the atmosphere to prevent the loss of
alcohol. The reaction mix is kept just above the boiling point of the alcohol (around 160
F) to speed up the reaction and the reaction takes place. Recommended reaction time
varies from 1 to 8 hours, and some systems recommend the reaction take place at room
temperature. Excess alcohol is normally used to ensure total conversion of the fat or oil to
its esters. Care must be taken to monitor the amount of water and free fatty acids in the
incoming oil or fat. If the free fatty acid level or water level is too high it may cause
problems with soap formation and the separation of the glycerin by-product downstream.
Separation
Once the reaction is complete, two major products exist: glycerin and
biodiesel. Each has a substantial amount of the excess methanol that was used in the
reaction. The reacted mixture is sometimes neutralized at this step if needed. The glycerin
phase is much more dense than biodiesel phase and the two can be gravity separated with

glycerin simply drawn off the bottom of the settling vessel. In some cases, a centrifuge is
used to separate the two materials faster.
Alcohol Removal
Once the glycerin and biodiesel phases have been separated, the excess alcohol in each
phase is removed with a flash evaporation process or by distillation. In others systems,
the alcohol is removed and the mixture neutralized before the glycerin and esters have
been separated. In either case, the alcohol is recovered using distillation equipment and is
re-used. Care must be taken to ensure no water accumulates in the recovered alcohol
stream.
Glycerin Neutralization
The glycerin by-product contains unused catalyst and soaps that are neutralized with an
acid and sent to storage as crude glycerin. In some cases the salt formed during this phase
is recovered for use as fertilizer. In most cases the salt is left in the glycerin. Water and
alcohol are removed to produce 80-88% pure glycerin that is ready to be sold as crude
glycerin. In more sophisticated operations, the glycerin is distilled to 99% or higher
purity and sold into the cosmetic and pharmaceutical markets.
Methyl Ester Wash
Once separated from the glycerin, the biodiesel is sometimes purified by washing gently
with warm water to remove residual catalyst or soaps, dried, and sent to storage. In some
processes this step is unnecessary. This is normally the end of the production process
resulting in a clear amber-yellow liquid with a viscosity similar to petrodiesel. In some
systems the biodiesel is distilled in an additional step to remove small amounts of color
bodies to produce a colorless biodiesel.
Product Quality
Prior to use as a commercial fuel, the finished biodiesel must be analyzed using
sophisticated analytical equipment to ensure it meets any required specifications. The
most important aspects of biodiesel production to ensure trouble free operation in diesel
engines are:

Complete Reaction
Removal of Glycerin
Removal of Catalyst
Removal of Alcohol
Absence of Free Fatty Acids

ADVANTAGES OF BIODIESEL

It is made from renewable resources.

It performs just as well as the normal diesel fuel.
It causes less pollution as compared to diesel-powered engines.
It is relatively less inflammable compared to the normal diesel.
It can be mixed with normal diesel fuel.
It is biologically degradable and reduces the danger of contamination of soil and
underground water during transport, storage and use.
It contains no sulphur, the element responsible for acid rain.
There are no extra costs for the conversion of engines in comparison to other
biological fuels.
It is suitable for catalytic convertor.
Engines last longer when using it.
Its refineries are comparitively simpler and environmental-friendly in design than
typical petrochemical refineries.
It produces 78% less carbon dioxide (CO2) than normal diesel fuel.
It has a higher cetane and lubricity rating than pure petroleum-based diesel fuel,
which improves engine efficiency and operating life cycle.

DISADVANTAGES OF BIOFUELS

Growing crops for biofuel absorbs the carbon that biofuels emit, but it does not
absorb the fossil fuel emissions created in planting, fertilizing, treating,
harvesting, transporting and processing these crops before they can be converted
into fuel. There are also considerable carbon emissions from the coal or gas
required to heat the required raw materials in the manufacturing process. Its
production can also lead to environmental destruction. Brazil, for example,
produces ethanol from sugar cane but to do so is cutting down the Amazon rain
forest, thus causing great damage to the environment.
Too much concentration on running vehicles on plant oil would set up a direct
competition between feeding the cars and feeding the people. This would not
increase our self-reliance but would increase our food and energy vulnerability.

BIODIESEL PRODUCTION FROM

JATROPHA CURCAS

JATROPHA CURCUS

INTRODUCTION
Jatropha curcus is a drought-resistant perennial, growing
well in marginal/poor soil. It is easy to establish, grows
relatively quickly and lives, producing seeds for 50 years.
Jatropha the wonder plant produces seeds with an oil
content of 37%. The oil can be combusted as fuel without
being refined. It burns with clear smoke-free flame, tested
successfully as fuel for simple diesel engine. The byproducts are press cake a good organic fertilizer, oil
contains also insecticide.
It is found to be growing in many parts of the country, rugged in nature and can survive
with minimum inputs and easy to propagate.
Medically it is used for diseases like cancer, piles, snakebite, paralysis, dropsy etc.
Jatropha grows wild in many areas of India and even thrives on infertile soil. A good crop
can be obtained with little effort. Depending on soil quality and rainfall, oil can be
extracted from the jatropha nuts after two to five years. The annual nut yield ranges from
0.5 to 12 tons. The kernels consist of oil to about 60 percent; this can be transformed into
biodiesel fuel through esterification.
Family: Euphorbiaceae Synonyms: Curcas purgans Medic. Vernacular/common names:
English- physic nut, purging nut; Hindi - Ratanjyot Jangli erandi; Malayalam Katamanak; Tamil - Kattamanakku; Telugu - Pepalam; Kannada - Kadaharalu; Gujarathi
- Jepal; Sanskrit - Kanana randa.
Distribution and habitat
It is still uncertain where the centre of origin is, but it is believed to
be Mexico and Central America. It has been introduced to Africa
and Asia and is now culti-vated world-wide. This highly droughtresistant spe-cies is adapted to arid and semi-arid conditions. The
current distribution shows that introduction has been most successful in the drier regions
of the tropics with annual rainfall of 300-1000 mm. It occurs mainly at lower altitudes (0500 m) in areas with average an-nual temperatures well above 20C but can grow at
higher altitudes and tolerates slight frost. It grows on well-drained soils with good
aeration and is well adapted to marginal soils with low nutrient content.
Botanical Features
It is a small tree or shrub with smooth gray bark, which

exudes a whitish colored, watery, latex when cut. Normally, it grows between three and
five meters in height, but can attain a height of up to eight or ten meters under favourable
conditions.
Leaves
It has large green to pale-green leaves, alternate to subopposite, three-to five-lobed with a spiral phyllotaxis.

Flowers
The petiole length ranges between 6-23 mm. The
inflorescence is formed in the leaf axil. Flowers are formed
terminally, individually, with female flowers usually slightly
larger and occurs in the hot seasons. In conditions where
continuous growth occurs, an unbalance of pistillate or
staminate flower production results in a higher number of
female flowers.
Fruits
Fruits are produced in winter when the shrub is leafless, or it
may produce several crops during the year if soil moisture is
good and temperatures are sufficiently high. Each
inflorescence yields a bunch of approximately 10 or more
ovoid fruits. A three, bi-valved cocci is formed after the
seeds mature and the fleshy exocarp dries.
Seeds
The seeds become mature when the capsule changes from
green to yellow, after two to four months.

Flowering and fruiting habit

The trees are deciduous, shedding the leaves in the dry
season. Flowering occurs during the wet season and two
flowering peaks are often seen. In permanently hu-mid
regions, flowering occurs throughout the year. The seeds
mature about three months after flowering. Early growth is
fast and with good rainfall conditions nursery plants may

bear fruits after the first rainy season, direct sown plants after the second rainy season.
The flowers are pollinated by insects especially honey bees.
Ecological Requirements
Jatropha curcas grows almost anywhere , even on gravelly,
sandy and saline soils. It can thrive on the poorest stony soil.
It can grow even in the crevices of rocks. The leaves shed
during the winter months form mulch around the base of the
plant. The organic matter from shed leaves enhance earthworm activity in the soil around the root-zone of the plants,
which improves the fertility of the soil.
Regarding climate, Jatropha curcas is found in the tropics and subtropics and likes heat,
although it does well even in lower temperatures and can withstand a light frost. Its water
requirement is extremely low and it can stand long periods of drought by shedding most
of its leaves to reduce transpiration loss. Jatropha is also suitable for preventing soil
erosion and shifting of sand dunes.
Biophysical limits
Altitude: 0-500 m, Mean annual temperature: 20-28 deg. C,
Mean annual rainfall: 300-1000 mm or more.
Soil type: Grows on well-drained soils with good aeration.

PRODUCTION OF BIODIESEL BY TRANSESTERIFICATION OF

JATROPHA OIL USING IMMOBILIZED
Pseudomonas fluorescens

ABSTRACT

Transesterification of vegetable oils is an important reaction that produces

fatty acid alkyl esters, methyl and ethyl esters which are excellent substitutes
for diesel fuel. Biodiesel prepared by catalyzed mild tranesterification has
become of much current interest for alternative fuel production. In the
present study the ability of a commercial immobilized Pseudomonas
fluorescens to catalyze the transesterification of Jatropha oil and methanol
was investigated. The cell of P. fluorescens was easily immobilized within
the sodium alginate during batch process. The important parameters like
temperature, pH, reaction time and amount of beads was studied. From the
study it was found that maximum yield of biodiesel was obtained at the
optimum conditions of at 40 , pH of 7.0, reaction time 48hrs and amount of
beads 3g. the physical properties of the products were analyzed and the
results were compared with the other sources of oils like used vegetable oils.

INTRODUCTION
For more than two centuries, the worlds energy supply has relied heavily on nonrenewable crude oil derived liquid fuels. Out of which 90% is estimated as to be
consumed for energy generation and transportation. It is also known that emissions from
the combustion of these fuels such as CO2, CO, NOx and sulfur containing residues are
the principal causes of global warming. On the other hand, known crude oil reserves
could be depleted in less than 50years at the present rate of consumption. Thus, increased
environmental concerns, tougher clean air act standards necessitates the search for a
viable alternative fuels, which are environmentally friendly. Oil seed crops such as palm,
soyabean, sunflower, peanut, olive etc are by far the largest group of exploitable
renewable biomass resource for liquid fuel and energy generation.
The attractive features of bio-diesel fuel are:
- It is a plant derived, not petroleum derived, and such its combustion does
not increase current net atmospheric levels of CO2, a greenhouse gas.
- It can be domestically produced, offering the possibility of reducing
petroleum imports.
- It is biodegradable.
- Relative to conventional diesel fuel, its combustion products have reduced
level of particulates, carbon monoxide, sulfur oxides, hydrocarbons etc.
- Vegetable oils can be used in diesel engines as they have high octane
number and calorific value very close to diesel.
Transesterification of vegetable oils is an important reaction that produces fatty acid
alkyl esters that are valuable intermediates in oleo chemistry, and methyl and ethyl esters
which are excellent substitutes for diesel fuel. Transesterification as an industrial process
is usually carried out by heating an excess of the alcohol with vegetable oils under
different reaction conditions in the presence of an inorganic catalyst. The most commonly
used catalysts are alkali hydroxides and alcoholates. Transesterification is also possible
under acidic conditions, but this process requires higher reaction temperatures.
Chemical methanolysis using an alkali catalysis process gives high conversion
levels of triglycerides to their corresponding methyl esters in short reaction times, the
reaction has several drawbacks: it is energy intensive, recovery of glycerol id difficult,
the alkaline catalyst has to be removed from the product, alkaline wastewater requires
treatment, and free fatty acids and water interee with the reaction.
Pseudomonas species immobilized with sodium alginate gel can be used directly as
a whole cell biocatalyst. In the present study, Pseudomonas fluroscence cells
immobilized within sodium alginate gel as a whole cell biocatalyst was utilized for
biodiesel fuel production from Jatropha oil.

MATERIALS
The non-edible crude Jatropha oil was purchased commercially and was stored at 4 to
avoid rancidity of the vegetable oil. Its quality characteristics were determined according
to the standard methods of fats and oils published by the association of oil chemists,
, acid value of 19.635mg KOH, saponification value
which has the density of 0.92g/
of 187gm KOH and free fatty acid of 17.25mg KOH/g and it was used throughout the
experimentation.

P. fluorescens, was obtained from Microbial Type Culture Collection and Gene Bank.
The culture was maintained on nutrient agar medium. After three days incubation at 25
the agar slants were stored at 4 .The liquid medium for the growth of inoculums for
bacteria was nutrient agar medium composed of 1.0 g/l of beef extract, 2.0 g/l of yeast
extract,
5.0 g/l of NaCl.

INOCULA PREPARATION
Inocula were grown aerobically in 250ml Erlenmeyer flasks containing the above
mentioned medium at 25 in an environmental shaker at 200rpm for 24h.

Active cells were centrifuged in a clinical centrifuge (1200rpm), washed with sterile
water, and were used as inoculum.
IMMOBILIZATION OF Pseudomonas fluorescence CELLS BY ENTRAPMENT
The sodium alginate entrapment of cells was performed according to the standard
method. Alginate solution with a concentration range of 0.5 10% was used or the cell
immobilization and was prepared by dissolving sodium alginate in boiling water and
autoclaved at 121 for 15min. Both alginate slurry and cell suspension was mixed and
stirred for 10min to get uniform mixture. The alginate solution was extruded drop by
drop into a cold sterile 0.2m CaCl2 solution through a sterile 5ml pipette from 5cm height
and kept for curing at 4 for 1h. the beads were hardened by resuspending into a fresh
CaCl2 solution for 24h at 4 with gentle agitation. Finally these beads were washed with
distilled water to remove excess calcium ions and unentraped cells. When the beads are
not being used, they are preserved in 0.9% sodium chloride solution in the refrigerator.

METHANOLYSIS OF JATROPHA OIL

Methanolysis reactions were conducted at stoichiometric molar ratio of oil/methanol; oil
and methano were poured into the reaction flask and heated to the reaction temperature
with constant shaking using magnetic stirrer for 48h.

In subsequent experiments, in which the effect of molar ratio of oil/methanol was

investigated, the volume of the oil is kept constant and the volume of methanol is varied.
Around 3ml of hexane is added to the reaction mixture to increase the solubility of the
reactants.
The appropriate amount of immobilized whole cells based on oil weight was added to the
flask. After 48h of reaction time, the reaction was stopped and the cells were removed
from the reaction mixture by filtration.

The produced ester and byproduct glycerol were separated using separate funnel. The
biodiesel and glycerol were collected and stored.

EFFECT OF TEMPERATURE
Temperature is one of the importance parameter for the production of biodiesel because
the rate of reaction is strongly influenced by the reaction temperature. The effect of
temperature on biodiesel production from Jatropha oil using immobilized cells of P.
fluorescens was studied by conducting experiments at different temperatures 25, 30, 35,
40, 45 keeping initial cell concentration of 3g of beads, substrate concentraton of 50ml
of oil (1:3 molar ratio oil/methanol) with n-hexane (3ml), reaction time of 48h and pH of
7.0 were fixed constant.

Temperature
Yield

25
40%

30
55%

35
65%

40
70%

45
60%

Percentageyeild
80
70
60
50
40
Percentageyeild

30
20
10
0
0

10

20

30

40

50

60

Maximum yield of 70% biodiesel was obtained at temperature 40

and was found
decreasing after 40
due to denaturation of enzyme. The optimum temperature of the
maximum yield of biodiesel was fixed as 40 .

EFFECT OF PH
pH of the reaction media is the another important parameter affecting the yield of
biodiesel. In this experiment, the pH of the reaction media was varied from 5.5 to 7.5. the
effect of pH on biodiesel production of Jatropha oil using immobilized cells of P.
fluorescens was studied by conducting experiments at different pH 5.5, 6.0, 6.5, 7.0 and
7.5.
pH
Yield

5.5
40%

6.0
50%

6.5
60%

7.0
70%

7.5
55%

PercentageYeild
80
70
60
50
40
PercentageYeild

30
20
10
0
0

The yield of biodiesel was found to be increasing and a maximum 70% was obtained at
pH 7.0 and the yield of biodiesel was found to be decreasing when the pH was increased
beyond 7.0.

EFFECT OF REACTION TIME

Effect of time on biodiesel production from Jatropha oil using immobilized cells of P.
fluorescens was studied by conducting experiments with different periods of 12, 24, 36,
48 and 60h. experiments were carried out at th optimum temperature 40 , pH 7.0,
immobilized cell concentration of 3g beads and substrate concentration of 50ml of oil.

Reaction
time
Yield

12hrs

24hrs

36hrs

48hrs

60hrs

40%

50%

60%

70%

65%

Percentageyeild
80
70
60
50
40
30
20
10
0

Percentageyeild

10

20

30

40

50

60

70

Yield of biodiesel increases till 48h and thereafter decreases. Further increase in the
reactiontime does not have effect on the production of biodiesel.

EFFECT OF AMOUNT OF BEADS

The effect of amount of immobilized beads on production of biodiesel from jatropha oil
using immobilized cells of P. fluorescens was studied by conducting experiments at
different amounts 1, 2, 3, 4 and 5g at constant levels of substrate concentration of 50ml of
oil (1:3 molar ratio of oil/methanol) and n-hexane (3ml) at optimum temperature 40 ,
pH 7.0 and reaction time 48h.

Amount
beads
Yield

of

1gms

2gms

3gms

4gms

5gms

32%

45%

70%

65%

60%

PercentageYeild
80
70
60
50
40
30
20
10
0

PercentageYeild

The percentage of yield of biodiesel increases till 4g and thereafter decreases, so 4g was
chosen as the optimum amount of beads. The maximum yield of biodiesel is 70%.

ANALYSIS
Temperature
pH

OPTIMUM

MAXIMUM
PERCENTAGE YIELD

40

70%

70%

Reaction time

48hrs

70%

Amount of beads

4gms

70%

Transesterification reaction was carried out using jatropha oil and short chain alcohol
(methanol on hexane) using immobilized cells of P. fluorescens. The various parameters
affecting biodiesel yield namely temperature, Ph, reaction time and amount of beads. The
maximum yield of 70% was obtained at optimum values of temperature 40 , pH 7,
reaction time 48h and amount of beads 4g and molar ratio of oil to alcohol 1:4. The
biodiesel produced was analysed for its physical properties and compared with the values
of petroleum based diesel. The specific gravity, flash and fire point, cloud and pour point,
kinematic viscosity are slightly lower than that of diesel, whereas the diesel index is
much higher and the smoke point is slightly lower.

BIODIESEL FROM USED

VEGETABLE OIL

VEGETABLE OIL
Vegetable oil is an alternative fuel for diesel engines and for heating oil burners. For
engines designed to burn diesel fuel, the viscosity of vegetable oil must be lowered to
allow for proper atomization of the fuel, otherwise incomplete combustion and carbon
build up will ultimately damage the engine. Many enthusiasts refer to vegetable oil used
as fuel as waste vegetable oil (WVO) if it is oil that was discarded from a restaurant
or straight vegetable oil (SVO) or pure plant oil (PPO) to distinguish it from biodiesel.

PROPERTIES
The main form of SVO/PPO used in the UK is rapeseed oil (also known as canola oil,
primarily in the United States and Canada) which has a freezing point of -10C. However
the use of sunflower oil, which gels at around -12C,[7] is currently being investigated as
a means of improving cold weather starting. Unfortunately oils with lower gelling points
tend to be less saturated (leading to a higher iodine number) and polymerize more easily
in the presence of atmospheric oxygen.

Material compatibility
Free fatty acids in WVO can have a detrimental effect on metals. Copper and its alloys,
such as brass, are affected. Zinc and zinc-plating (galvanization) are stripped by FFA's
and tin, lead, iron, and steel are affected too. Stainless steel and aluminum are generally
unaffected.

Temperature effects
Some Pacific island nations are using coconut oil as fuel to reduce their expenses and
their dependence on imported fuels while helping stabilize the coconut oil market.
Coconut oil is only usable where temperatures do not drop below 17 degrees Celsius (62
degrees Fahrenheit), unless two-tank SVO/PPO kits or other tank-heating accessories,
etc. are used.

HOME HEATING
When liquid fuels made from biomass as used for energy purposes other than transport
they are called bioliquids.
With often minimal modification, most residential furnaces and boilers that are designed
to burn No. 2 heating oil can be made to burn either biodiesel or filtered, preheated waste
vegetable oil. These are generally not as clean-burning as petroleum fuel oil, but if
processed at home, by the consumer, can result in considerable savings. Many restaurants
will give away their used cooking oil either free or at minimal cost, and processing to
biodiesel is fairly simple and inexpensive. Burning filtered Waste Vegetable Oil (WVO)
directly is somewhat more problematic, since it is much more viscous, but it can be
accomplished with suitable preheating. WVO can thus be an economical heating option
for those with the necessary mechanical and experimental aptitude, where fire regulations
and insurance policy permit it.
AVAILABILITY
Waste vegetable oil
As of 2000, the United States was producing in excess of 11 billion liters of waste
vegetable

oil

annually,

mainly

from

industrial deep

fryers in potato processing

plants, snack food factories and fast food restaurants. If all those 11 billion liters could be
collected and used to replace the energetically equivalent amount of petroleum, almost
1% of US oil consumption could be offset.]Use of waste vegetable oil as a fuel competes
with some other uses of the commodity, which has effects on its price as a fuel and
increases its cost as an input to the other uses as well.
Pure plant oil (Straight vegetable oil)
Pure plant oil (PPO) (or Straight Vegetable Oil (SVO)), in contrast to waste vegetable oil,
is not a byproduct of other industries, and thus its prospects for use as fuel are not limited
by the capacities of other industries. Production of vegetable oils for use as fuels is
theoretically limited only by the agricultural capacity of a given economy. However,
doing so detracts from the supply of other uses of pure vegetable oil.

METHANOL TRANSESTERIFICATON OF
VEGETABLE OIL

ABSTRACT

The result of the investigation on methyl esters obtained on the basis of used
vegetable oil are given in this paper. Transesterification reaction conditions
that effect yield and purity of the product esters including oil quality, type
and concentration of catalyst, temperature and reaction time were examined.
Methanol transesterification of different oils at 60 with 1-10 % (v/v)
sodium met-oxide was studied, with appropriate percent of sodium metoxide, temperature 60 and 1 hour. Vegetable oil was sufficiently
transesterified and could be used as fuel in diesel engines.

INTRODUCTION

Over 45 million tonnes of greenhouse gases are produced every year from the burning of
diesel in trucks. Of course we are trying to find new creative ways to save our
environment and one way to do this is to use vegetable oil as a renewable source of fuel
for transportation and also the use of heating.
There are so many benefits of using this source to replace fossil fuels and some of these
include reduced air pollution, reduced greenhouse gas emissions, and conservation of
limited fossil fuels. There are two different ways that you can use vegetable oil as a fuel
in engines.
The first way is that you can use straight vegetable oil either waste frying oil or freshpressed oil, however you will need an extra fuel tank and a system for heating and
filtering the oil before it reaches the engine. The reason why you will need this is because
pure vegetable oil is too thick to work in the engine unless the oil is heated up.
The other way is to simply convert the vegetable oil into biodiesel which can be used in a
diesel engine without any modifications. Biodiesel is a fuel source made from vegetable
oil when a chemical reaction occurs between methanol and lye. It can be created either
from using waste vegetable oil from the food industry, or you have the other option of
using it from fresh-pressed vegetable oil.
This is something that is now being made to sell commercially in thousands of countries
all around the world, however with the right equipment and enough time it can also be
made right at home. Some of the toxic air pollutants that are reduced include soot,
particulates, carbon monoxide, and sulphur oxides, however nitrous oxide emissions may
increase slightly.

MATERIALS AND METHODS

Warm up oil:
Pour the oil into the flask and turn the electric heater to low.
We need to warm up the oil for a couple of reasons. First, the heating
will make the oil less viscous so it will stir more easily.
Second, the heat helps the conversion to biodiesel go faster.

Temperature:

Weigh out lye:

Using a thermometer bring the temperature to

between 120 to 130 which is about 50 .
This will feel hot but not too hot to
touch. Instead of electric heater, the flask could be placed in
hot tap water.

Open the lye bottle, add sodium hydroxide

until the balance reads 5.0g.
Be sure to close the bottle of lye as you get
right amount weighed.
Lye will absorb water out of the air making
it heavier and less effective.

Pour methanol into NaOH:

Take the lye in a bottle. Open the methanol
bottle and pour
some methanol into the bottle of lye. Continue
adding methanol until the bottle is about full.

Dissolve NaOH in the methanol:

Close the lid. Tip the bottle back and forth until the
NaOH dissolves.
The NaOH reacts with the methanol to form a
powerful substance called methoxide (C ONa).

The bottle will get warm as the NaOH

dissolves.

Place flask on magnetic stirrer:

The flask can now be placed on the magnetic stirrer. Be sure
to put the magnetic bar in the flask. To keep the
temperature between 120 to 130 , adjust the temperature.

Pour methoxide into oil:

Open the methoxide bottle and pour it into the flask
containing oil. Pour the remaining methanol into the flask
containing the oil and methoxide. This extra methanol helps
to convert more of the oil to biodiesel.

Stir mixture:
Turn on the magnetic stirrer to mix the methoxide, oil and
methanol. The stirrer should not be too fast, the meagnetic
bar may get unstable. Stirring time is normally listed as 2hrs.
To get maximum yield of biodiesel, 2hrs is recommended.

Separating funnel:
The pear-shaped glassware is called a separating
funnel because it is good for separating liquids that split
into two layers. In the biodiesel process, the methoxide reacts
with the oil and makes two products.
One is glycerin and the other is biodiesel.The glycerin will
sink to the bottom. A separating funnel makes it easier to
drain off the glycerin.

Pour product into separating funnel:

After 2hrs of stirring, the obtained product is poured into
the separating funnel. In a few minutes, the glycerin will
settle down at the bottom of the separating funnel. It will
not be pure glycerin, because some of the sodium hydroxide
will be with it.

Drain off glycerin:

Turn the stopcock so the glycerin drains out.
Glycerin can be saved to be used for other purposes.
This glycerin has some contamination with NaOH
and methanol, but the NaOH is neutralized and the
methanol will evaporate. The NaOH can be neutralized
with a little lemon juice to get it around a pH of 6 or 7.

Add water for washing:

After draining out glycerin, add tap water to the separating
funnel. To get rid of any residues of glycerin and NaOH.
Mix the water and biodiesel using an air pump. After few
minutes the water and biodiesel gets separated.

Drain off biodiesel:

The biodiesel is collected in a beaker. The biodiesel is
obtained in a Liquid gold colour. The biodiesel is dried,
to remove the traces of water. After drying, the biodiesel
is ready to be used in a diesel engine.

CONFORMATION TEST FOR GLYCEROL

1. Check to make sure the absent-minded professor has set up a pan of boiling water
for you. This should be in an "on" fume hood as some chlorine gas will be
produced.
2. Set up three test tubes: blank, glycerol std, and your experimental.
3. Add one ml of water to the blank; one ml of 10% glycerol to the standard; and
one ml of your fermentation mixture to the experimental tube.
4. To each, add one ml of Clorox; wait three minutes.
5. Add 4 drops of concentrated (12M) HCl to each tube.
6. Place the three tubes in the boiling water for a minute to drive off
any chlorine gas.
7. Add 200 L (0.2 mL) 5% -naphthol*
8. Add 4 mL of concentrated H2SO4 (take care to not get any of this on
your fingers or clothes!)
9. Shake the tubes carefully to mix; prevent splashing!
10. An emerald green color indicates the presence of glycerol.

CONFORMATION TEST FOR BIODIESEL

Clarity Testing Finished Biodiesel for Water

This is a visual inspection of the finished biodiesel. Water free biodiesel will be clear.
The common method of testing is to put a sample in a jar and if you can read a newspaper
through the biodiesel then it passes the test. For larger batches, the ability to see the
bottom of a drum of biodiesel clearly is often used. This is a good test, however it does
not detect water that is dissolved in the biodiesel. Water dissolved in biodiesel will be
evenly distributed on a molecular level throughout the biodiesel. There is some debate as
to the effect of dissolved water on certain types of diesel engines.
One important fact is that hot biodiesel will hold more dissolved water than cold
biodiesel. This results in hot or warm biodiesel passing the test and cold biodiesel failing
the test. It is important to pass the test when the biodiesel is slightly colder than the
expected operating temperature.

Cloud Point Testing

This test is performed by placing a sample of finished biodiesel in the freezer and
watching it for cloudiness. The temperature at which the biodiesel first turns cloudy is the
cloud point. This test is important for winter fuel. It is related to the "Gel Point Test". The
cloud point is near the temperature at which your filter will clog from frozen biodiesel.

Gel Point Testing

In this test you place a sample of finished biodiesel in a freezer and record the
temperature at which the biodiesel starts to gel. This is an important test for winter fuel.
Simply put, you don't drive on biodiesel at temperatures below the gel point.

The Methanol Test from JTF or the 27/3 Test

Neither Triglycerides or Diglycerides are soluable in dry methanol, but Monoglycerides
and Biodiesel will dissolve in dry methanol. If significant Triglycerides or Diglycerides
present will fall to the bottom of the test.
To perform the test add 1 part biodiesel to 9 parts methanol, by volume. On infopop
many use 3 mL of biodiesel and 27 mL of methanol and call the test either the Jan

Warnqvist test or the 27/3 test. Testing has shown that for the test to appromimate ASTM
level conversions the original 25ml of biodiesel and 225ml of methanol need to be used.
Triglycerides and Diglycerides will form a precipitant on the bottom. Only a liquid
precipitant that collects together to form a bead or bubble that rolls arond on the bottom
of the test is significant. If you have a liquid precipitant after about 5 minuites, then your
biodiesel is under converted.
Your methanol should be dry and you perform this test at room temperature, The test has
been found to be temperature sensitive. Hot biodiesel straight off the processor can give
false readings as can using cold methanol stored outside in winter.

The pHLip Test

The pHLip test is a commercial test that was designed as a "Firewall" for bad fuel in the
biodiesel distribution chain. It is a simple test consisting of a vial of red liquid, to which
you add your sample of biodiesel, flip end over end 10 times and wait 10 minutes. It will
tell you if your biodiesel is high quality or if it fails for total glycerine, soap content, free
glycerine, oxidation and catalyst contamination.