1.

INTRODUCTION

The production and consumption of petroleum oil increases

constantly; nowadays consumption is about 75 million barrels of
crude oil daily and in the coming years it will increase by 7%
annually. About 36% of world energy comes from petroleum oil and
22% from

gas. This dependence on fossil fuels has many

disadvantages, for example increasing pollution and increasing

costs. To reduce the use of petroleum oil as fuel, alternative
energies need to be developed; biofuels, biogas and bioethanol are
now the most promising alternatives in energy generation. Biodiesel
is produced in some countries and used efficiently either alone or in
blends with mineral diesel in cars and transport vehicles.
In line with the environmental, public health and economic
worries, the demand for cheap and alternative renewable natural
energy resources has risen. One such alternative being looked at
today is biodiesel. Biodiesel is produced by chemically reacting a
vegetable oil or animal fat with an alcohol such as methanol or
mono-alkyl esters of long chain fatty acids derived from a renewable
lipid stock (Ma and2 Hanna, 1999; Van Gerpen, 2005; Wang et al.,
2006). It is also biodegradable and nontoxic and typically produces
about 60% less net carbon dioxide (CO 2) emissions than petroleumbased diesel (Biodiesel, 2007).
One source of oil is Jatropha curcas .Being an agricultural
country, the Philippines must maximize the resources that it has
1

been richly endowed. Jatropha can be grown in various areas in the

country since it can be planted on any kind of soil and grows well
under tropical and subtropical climate. J. curcas has been found as a
highly promising biodiesel source because of its ubiquity, hardiness
and the quality and quantity of the oil that can be extracted from its
seeds (Ginwal et al, 2005).
It is a plant with many attributes, multiple uses and
considerable potential. The plant can be used to prevent and/or
control erosion, to reclaim land, grown as a live fence, especially to
contain or exclude farm animals and be planted as a commercial
crop. It is a native of tropical America, but now thrives in many parts
of the tropics and sub-tropics in Africa/Asia (Gubitz et a., 1999;
Kumar and Sharma, 2008; Openshaw, 2000; Martnez-Herrera et al.,
2006). The wood and fruit of Jatropha can be used for numerous
purposes including fuel. The seeds of Jatropha contain viscous oil,
which can be used for manufacture of candles and soap, in
cosmetics industry, as a diesel/paraffin substitute or extender. This
latter use has important implications for meeting the demand for
rural energy services and also exploring practical substitutes for
fossil fuels to counter greenhouse gas accumulation in the
atmosphere. These characteristics along with its versatility make it
of vital importance to developing countries (Kumar and Sharma,
2008).
Authority-Davao Office entrusted 14 provenances of Jatropha
curcas L. from diverse origins for research purposes to the Science
2

Department of Mindanao State University General Santos City. To

date, three of these have yet to be characterized. This paper aims to
provide the proximate composition and free fatty acid content of
three (3) J. curcas provenances from Asian and Mexican origin. This
will be done to evaluate the properties of J. curcas industrial
application and as a good source of biofuel.

1.1 Objectives
This study sought to:
a) Determine the proximate composition of Jatropha curcas
seeds ;
b) Determine the free fatty acid content of Jatropha curcas
oil from 3 different provenances;
c) Compare the properties of 3 Jatropha curcas oils from
other oils accepted for potential uses as biodiesel and
for other industrial applications

1.2 Significance of the Study

The unstable supply of oil and triggered several countries to
develop alternative sources of energy such as wind, nuclear and
geothermal power-generating facilities (Horton, 1973). However, for
oil-dependent countries that do not have established powergenerating facilities cutting back on oil consumption is difficult. The
potential solution for these countries is the use of biodiesel.
3

The use of biodiesel is being highly promoted because of its

social, economic and environmental impact. Biodiesel has attracted
significant attention because of its renewability, biodegradability
and for being a cleaner substitute to petrodiesel (Antolin et al,
2002). The production of feedstock will generate employment and
additional income among those that will be involved. Idle lands not
planted with other crops or forest trees can be used (Villancio 2006).
Domestic production of curcas oil will also result to a decreased
dependence on oil imports and thereby saving the hard-earned
revenue of the people.
Entrepreneurs who desire to understand the J. curcas industry
and the current and future business opportunities in this market,
and companies in the biofuel industry will benefit most from this
study.

And also, investment banks and financial investment

companies keen on funding ventures in the biofuel industry will be

provided with valuable information from this study.
This study evaluated three different provenances of Jatropha
curcas from Asian and Mexican origin to determine proximate
analysis and fatty acid profile. These properties were evaluated for
potential use as a biodiesel and for other industrial purposes.

1.3 Scope and Limitation

The study focused on the characterization of 3 Jatropha curcas
provenances coming from Asian and Mexican origins. Free fatty acid
content and proximate composition were determined such as the
4

crude fat, crude protein, carbohydrate, crude fiber, ash, and

moisture. The moisture, ash, crude fiber and crude fat analysis of
Jatropha curcas seed kernels were conducted in ECPC Research and
analytical Laboratory, Alabel, Sarangani Province. For the crude
protein analysis and free fatty acid content, samples were analyzed
in DOST Region XI.

II. REVIEW OF RELATED LITERATURE

2.1 Jatropha curcas

2.1.1 Taxonomy and botanical description
Jatropha is a drought-resistant tropical bush or small tree from
the family of Euphorbioceae. It has a productive life span of 35 to 50
years. Botanist Carl Von Linne first classified and named the plant in
1753. He named it Jatropha curcas; Jatropha coming from the Greek
words jatros meaning doctor and trophe meaning nutrition. The
name was brought about by the many uses of Jatropha as a
medicinal plant. Commonly known as physic nut or purging nut,
Jatropha is a non-edible oil-yielding perennial shrub that has green

leaves with a length and width of 6 cm to 15 cm, and can reach a

height of up to 5 meters.
The genus Jatropha contains approximately 170 known
species.

It was spread as a valuable plant in Africa and Asia by

Portuguese traders (United Nation, 1996). It is multipurpose and

deciduous, reported to be cultivated in drier sites of central and
western India. Jatropha usually grows below 1400 meters of
elevation from sea level and requires a minimum rainfall of 250mm,
with an optimum rainfall between 900-1200mm (Boswell 2003). The
plant shows articulated growth, with amorphological discontinuity at
each increment. The branches contain latex. Normally, five roots are
formed from seedlings, one central and four peripheral. A tap root is
not usually formed by vegetatively propagated plants. Leaves five to
seven

lobed,

hypostomatic

and

stomata

are

of

paracytic

(Rubiaceous) type (Kumar et al 2008).

The trees are deciduous, shedding the leaves in dry season.
Flowering occurs during the wet season and two flowering peaks are
often

seen. In permanently humid regions, flowering

occurs

throughout the year. The plant is monoecious and flowers are

unisexual; occasionally hermaphrodite flowers occur (Dehgan and
Webster, 1979). A flower is formed terminally, individually, with
female flowers (tricarpellary, syncarpous with trilocular ovary)
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
6

flowers. Ten stamens are arranged in two distinct whorls of five each
in a single column in the androecium, and in close proximity to each
other. In the gynoecium, the three slender styles are connate to
about two-thirds of their length, dilating to massive bifurcate stigma
(Dehgan and Webster, 1979). Each inflorescence yields a bunch of
approximately 10 or more ovoid fruits. With good rainfall conditions
nursery plants may bear fruits after the first rainy season, and
directly sown plants after the second rainy season. Three, bivalved
cocci is formed after the seeds mature and the fleshy exocarp dries.
The seeds mature about 34 months after flowering. Under optimal
conditions, jatropha usually flowers about 3 to 6 months after the
seeds have been sown. The time from flower induction to fruit
maturation is 90 days. The female flowers produce fruits that are
first green, and turn yellow when ripening. Later the yellow fruit hull
turns brown nd black when they dry (The Jatropha Handbook: From
Cultivation to Application. 2010).
Because of the vast semi-wild distribution of J. curcas in
different parts of the world there should be a considerable amount
of

genetic

variation among

the species. However, sufficient

information on such aspect is lacking and so far, only few records

exist of provenance trials made to examine the genetic information.
Genetic factors are strong determinants of seed quality and
potential. If the nature of these various provenances would be
known, it could provide elaborate criteria for selection of prominent

traits in both laboratory and nursery for better performances in the

field (Heller, 1996; Ginwal et al., 2004)

2.1.2 Possible Uses

All parts of J.curcas plant have their own uses. Like many
other Jatropha species, J. curcas is a succulent tree that sheds its
leaves during the dry season. It is well adapted to arid and semi-arid
conditions and often used for erosion control. The leaves are used in
traditional medicine against coughs or as antiseptics after birth, and
the branches are chewing sticks (Gbitz et al, 1999). The latex
produced from the branches is useful for wound healing and others
medical uses.
Each fruit contains 2-3 oblong black seeds which can produce
oil. The seed kernel oil contained 30-50% (w/w) oil (Makkar et al.
1997). These seeds contain viscous oil, which can be used for
manufacture of candles and soap, in cosmetics industry, as a
diesel/paraffin substitute or extender. This latter use has important
implications for meeting the demand for rural energy services and
also exploring practical substitutes for fossil fuels to counter
greenhouse

gas

accumulation

in

the

atmosphere.

These

characteristics along with its versatility make it of vital importance

to developing countries (Kumar and Sharma, 2008).

2.2 Proximate Analysis

The etymology of 'proximate' is the Latin word proximatus,
meaning 'to come near' or 'close'. Hence, many of the proximate
analyses are prefaced by the word 'crude', i.e., crude protein, fiber,
fat, carbohydrate, ash, and moisture reported as the percentage
composition

of

the

product

(http://www.aescl.missouri.edu/Prox.html.)
Gubitz et al reported that analysis of Jatropha curcas
seeds shows that it contains; moisture 6.62; protein 18.2; fat 38.0;
carbohydrates 17.30; fibre 15.50; and ash 4.5% (Gubitz et al, 1999).
The oil content is 35 to 40% in the seeds and 50 to 60% in the
kernel (Gubitz et al, 1999). The oil contains 21% saturated fatty
acids and 79% unsaturated fatty acids (Gubitz et al, 1999). It has
also been found that there are some chemicals element in the seeds
which possess poisonous and purgative properties and render the oil
non edible for human consumption. It is also been stated that
technologies are now available, whereby it could be possible to
convert Jatropha oil into an
edible oil which could prove to be a boon for developing countries
(Gubitz et al, 1999). The oil is obtained from decorticates seeds by
expression or solvent extraction and is known in trade as Jatropha.
In general, the oil is reported to be mixed with groundnut oil for
adulteration. This indicates the possibilities of obtaining edible oil
from Jatropha oil base (Gubitz et al, 1999).
9

2.2.1 Moisture and Ash Content

Moisture content is a factor in the quality control of biodiesels.
A recent contribution to the literature (He et al., 2007) notes the
paucity of information on the moisture content of biodiesels. It is
well known that biodiesels have higher moisture contents than
mineral diesels and as biodiesels usage increases moisture content
will become an increasingly important factor in their quality control.
Ash is a measure of the amount of metals contained in the
fuel. High concentrations of these materials can cause injector tip
plugging, combustion deposits and injection system wear. The ash
content

is

important

for

the heating value,

as heating value decreases with increasing ash content. Ash content

for bio-fuels is typically lower than for most coals, and sulphur
content is much lower than for many fossil fuels.
The ash content is determined by ignition of a known weight
of the food at 600C until all carbon has been removed. The residue
is the ash and is taken to represent the inorganic constituents of the
oil (AOAC, 1995).

2.2.2 Crude Protein

The total protein content is estimated as total nitrogen (e.g.
the Kjeldahl method) after digestion, salt neutralization and titration
of the ammonia released against standard acid. A conversion factor
is applied to calculate the total protein. Protein contains about 16%
Nitrogen. It is not the only thing in food containing Nitrogen, but
10

normally it is the main one. Crude Protein is simply the percentage

of Nitrogen multiplied by 6.25.
The protein content Jatropha oil cake may be used as raw
material for plastics and synthetics fibres. It would also be
advantageous to make use of Jatropha oil as hydraulic oil (Gubitz et
al, 1999). Jatropha oil cake is rich in nitrogen, phosphorous and
potassium and can be used as organic manure. This indicated the
potential of this plant in initiating the process of reduction of surplus
livestock maintain by the rural folk in India, mainly for the purpose
of obtaining cow -dung as manure.
Jatropha oil cakes can, hopefully, replace synthetic fertilizers by
undertaking plantations of Jatropha curcas on wastelands, Jatropha
curcas leaves provide plentiful organic matter and increase the
microbial activity including earthworms which is an indication of
ecological improvement of site (Gubitz et al, 1999). The oil cake is
rich in protein but contains some toxic principle and as such it is
considered unfit for use as cattle feed. But it is reported that the
poisonous principle appears to exist in the alcohol soluble fraction of
the oil. With suitable research it could be possible to convert the
nonedible oil-cake into protein rich cattle and poultry feed on a
massive scale (Gubitz et al, 1999).
2.2.3 Crude Fiber
Fiber (indigestible carbohydrates) stimulates the activity of
bowels, combines heavy metals, cholesterol, bilious acids, and
11

removes them from the organism, and obesity and even cancer.
According to data, up to 13-14 %of crude fiber is found in seeds
(Tarek et al., 2001). Crude fiber contains cellulose, lignin and
hemicelluloses, but not necessarily all of these are present in a
sample (AOAC 1984). There are several methods of fiber analysis,
among other methods of crude fiber, detergent method and the
enzymatic methods each have advantages and disadvantages.

2.2.4 Crude Fat

Fats, usually of animal origin contain saturated fatty acids,
which accounts for its being solid at room temperature. However,
oils, or unsaturated fats, mostly from plant sources, contain
unsaturated fatty acids that are not able to close packing due to
bends or kinks in their molecules.(Holme and Peck, 1983)Fats are
used to describe

those lipids that are solid at a specific

temperature. Crude fat content is estimated by extracting a ground

feed sample with diethyl ether (Association of Official Analytical
Chemists, 1984). The ether soluble components (ether extract) may
include true fats and oils, fatty acid esters, compound lipids and fatsoluble vitamins or provitamins such as the carotenoids, all of which
may have nutritional value. However, ether extract may also contain
significant concentrations of indigestible waxes, resins and essential
oils. There are several methods used for the crude fat analysis:
12

Continuous Solvent extraction or the Goldfisch extraction and the

Soxhlet Method (B.K.K.K Jinadasa, 2010).
Oil content of jatropha kernel was found higher than linseed,
soybean, and palm kernel which is 33.33%, 18.35% and 44.6%,
respectively (Gunstone, 1994). High oil content of Jatropha Curcas
indicated that Jatropha Curcas are suitable as non-edible vegetable
oil feedstock in oleochemical industries (biodiesel, fatty acids, soap,
fatty nitrogenous derivatives, surfactants and detergents, etc).
Currently, Jatropha Curcas can produce 2000 liter/ha oil per annual
(Azam et al., 2005).

2.2.5 Carbohydrates
Total

carbohydrate

consists

of

sugars

(mono

and

oligosaccharides) and polysaccharides (starch and the non-starch

polysaccharides; pectin, soluble and insoluble dietary fibre, e.g.
cellulose and hemicellulose).
In a 1990 report, total carbohydrate was calculated as the
residue by difference from the total of fat, protein, moisture/solids,
ash, and fiber values. A review of collaborative studies of these
parameters was made to determine the likely precision of the
process. The procedure was judged as having poor precision among
laboratories and high variability. Even so, the by difference
method was used in 2002 for the proximate analysis of Nigerian oil
13

seed,and Menezes et al., 2004 averred that most composition

databases contain total carbohydrate data calculated by the
difference method.

2.3 Fats and Oils

Fats and oils are members of the lipid family. Lipids may either
be a solid or liquid at room temperature, depending on their
structure and composition. Normally, oil refers to a lipid that is
liquid at room temperature, while fat refers to a lipid that is solid
or semi-solid at room temperature. Fats and oils primarily consist of
esters of glycerol (mono-, di-, and triglycerides) and low to
moderate contents of free fatty acids (carboxylic acids). Other
compounds such as phospholipids, polypeptides, sterols, water
odorants and other impurities can be found in crude oils and fats.
The structures of mono-, di-, and triglycerides (MGs, DGs, and TGs)
consists of glycerol (a backbone of carbon, hydrogen, and oxygen)
esterificed with fatty acids (chains of carbon and hydrogen atoms
with a carboxylic acid group at one end), Free fatty acids (FFAs) can
can contain 4-24 carbon atoms with some degree of unsaturation
(typically 1-3 C-C double bonds). Fats have more saturated fatty
acids, the compositional building blocks, than oils, which give rise to
a higher melting point and higher viscosity of the former.
Consequently, biodiesel produced from saturated fats have a higher
cloud and gel points than those made from unsaturated oils, making
the former unsuitable to use in cold climates. Food-grade vegetable
14

oils, containing a low FFA level, are currently used for commercial
biodiesel production. Although waste greases such as yellow grease
and brown grease, containing a FFA level of 15% and 33%
respectively, are considered as the attractive feedstocks for
biodiesel synthesis because of its wide availability and low cost
compared to food-grade vegetable oils.

2.4 Free Fatty Acid (FFA)

One measure of fat quality is the free fatty acid (FFA) content.
Fats are normally composed of three fatty acids linked to glycerol
via ester bonds. FFA are produced when those fatty acids are freed
by hydrolysis. Therefore the presence of high levels of FFA indicates
the fat was exposed to water, acids, and/or enzymes. Fats should be
processed to contain as low a moisture level as is feasible so that
hydrolysis does not occur during subsequent storage.
An important consideration in the feedstock selection for
biodiesel production is the content of free fatty acid (FFA) in the oil.
To be used as a feedstock, the JCO should contain a low percentage
of FFA so that the oil can directly be utilized in a transesterification
reaction with methanol in the presence of an alkaline catalyst
( Azhari et al 2003 ).
In the biodiesel production, to obtain Jatropha curcas methyl
esters (JME), the jatropha curcas oil (JCO) was subjected to a
chemical reaction termed transesterification. In that reaction, the
JCO was reacted in the presence of an alkaline catalyst with
15

methanol to give the corresponding methyl esters. Berchmans and

Hirata reported that the alkaline base catalyzed transesterification
depended on several basic variables. One of them was the level of
FFA in the feedstock should be less than 1%. Therefore, to be used
as a feedstock in biodiesel production, the JCO should contain a low
percentage of free fatty acids (FFA) so that the oil can directly be
utilized in a transesterification reaction with alcohol as an excess
reactant in the presence of an alkaline catalyst. Otherwise, the
saponification shall occur and the separation of products shall be
exceedingly difficult, and as a result, the yield of biodiesel product
would be low.

Some researchers also (Marchetti and Errazu)

reported that if the feedstock has a high amount of free fatty acids,
much higher than the maximum amount suitable to be used with
basic homogeneous catalyst, high amount of soap would be
produced

simultaneously

with

the

transesterification

reaction.

Therefore, to avoid this reaction, alternative technology should be

used for example with a homogeneous acid catalyst, solid resins,
and enzymes or in supercritical process. Chung et al. stated that
due to the corrosion problem, these homogeneous catalyst-based
processes involved elaborate process steps for removal of FFA and
water from the feedstock and catalyst from the products.
In many cases, jatropha curcas oil (JCO) quality deteriorates
gradually due to improper handling and inappropriate storage
condition. It was known that improper handling of the oil would
cause the moisture content of the JCO to increase. In addition,
16

exposing the oil to open atmospheric air and sunlight for long time
would cause the concentration of FFA to increase significantly. The
FFA content of the oil would vary and depend on the quality of
feedstock. Furthermore, other researchers have worked with raw
materials having higher FFA levels using alternative processes,
which included pretreatment step to reduce the FFA of these raw
materials.
Increased levels of FFA in fats have been shown to reduce the
digestibility and thus energy content of fats. On the average, each
increase of 10 percentage units in FFA results in a corresponding
reduction in digestible energy of 1.3 and 1.5 percentage units in
weanling and growing pigs, respectively, (Powles, et al. 1995.
Journal of Animal Science 61:149). A common source of vegetable
fat used in blended feed fats is acidulated soapstock. This byproduct of edible oil refining has very high FFA since it was
intimately exposed to water and acid during its production. High
levels of FFA should be considered when estimating energy content
of fats for feeding.

The acidity of fats is also often expressed

directly in terms of percent Free Fatty Acids (FFA). The assumption

usually made in the calculation is that the acids have a molecular
weight equal to that of an oleic acid. It was known that improper
handling of crude Jatropha curcas oil (CJCO) would cause the water
content increase. In addition, exposing the oil to open air and
sunlight for long time would affect the concentration of FFA increase
significantly to high level of FFA above 1%. The FFA amount of CJCO
17

will
vary and depend on the quality of feedstock. The FFA and moisture
contents have significant effects on the transesterification of
glycerides with alcohol using catalyst (Goodrum, 2002). The high
FFA content (>1% w/w) will happen soap formation and the
separation of products will be exceedingly difficult, and as a result,
it has low yield of biodiesel product. The acid-catalyzedesterification
of the oil is an alternative (Crabbe et al., 2001), but it is much
slower

than

the

base-catalyzed

trans-esterification

reaction.

Therefore, an alternative process such as a two-step process was

investigated for feedstock having the high FFA content (Ghadge and
Raheman, 2005; Veljkovic et al., 2006)

18

3. METHODOLOGY
3.1 Seed Samples
Hereunder is the list of the three (3) Jatropha curcas
provenances that were entrusted to the researcher by the Science
Department of Mindanao State University General Santos City.
Indicated are the names and the country of origin.
Name

Country

1.) Tubao-PhilBio

Philippines

2.) India-D1

India

3,) Mexican-PhilBio

Mexico

3.2 Seed Collection and Sample Preparation

The seed samples of 3 Jatropha curcas provenances were
collected within the Science department area of MSU-GSC, Fatima.
Collected samples were air-dried for 7 days and stored in sealed
glass containers. The hard seed coat was removed by pounding in a

19

mortar and pestle. The collected kernels were then pulverized using
a Waring blender.

3.3 Proximate Analysis

3.3.1 Moisture and Ash Content
The moisture content of Jatropha curcas (after drying) were
measured using the LJ16 Moisture Analyzer, Metler Toledo. The
moisture free samples were ignited at 550oC in a muffle furnace
until a uniformly grayish to white or reddish to blue residue is
acquired. With this process, water and other volatile components
were vaporized while the organic constituents are evolved as other
form of oxides and until weight is constant

3.3.2 Crude Fat

Determination of crude fat percentage was done through
soxhlet method. The soxhlet apparatus was set up with anhydrous
diethyl ether, approximately 175 mL (the empty flask was dried to
constant weight in an oven before using for the extraction). The
samples (2g) were removed after extraction and air dried to remove
the excess ether. The filter paper with the sample was dried in an
oven at 1050C until the weight is constant. The weight of the filter
paper and the samples were then recorded. Total percentage of fat
was calculated using the formula:
% Crude Fat = Corrected weight of the fat x 100
Weight of the dry sample

20

3.3.3 Crude fiber

Analysis of crude fiber was done using manual filtration
method. The defatted sample was boiled in 0.64 N H 2SO4 and then in
1.56 N NaOH to eliminate soluble components. Between successions
the sample was washed with boiling water several times then finally
with ethanol, for the ignition process, the sample was placed in a
gooch crucible with asbestos matting. Ignition was done at 550 oC for
20 minutes to oxidize all carbonaceous materials leaving only the
fibers. The sample was cooled, placed in a dessicator and weighed.
%Crude Fiber =

Weight of crucible @ 550oC

Weight of dry sample

x 100

3.3.4 Crude protein and Free Fatty Acid Composition

Samples were sent to DOST Region XI to determine its free
fatty acid contents and crude protein analysis.

21

4. RESULTS AND DISCUSSION

The proximate composition (crude protein, crude fiber, crude

fat, carbohydrate, ash, and moisture) of three J. curcas provenances
seed kernels of different origins are shown in Table 1. It shows the
mean average composition of three provenances of Jatropha curcas.
Results on the moisture content is comparable to the moisture
content reported by Akintayo, 2004 which is 5.54% for J. curcas. The
percent ash of J. curcas seeds, India D1, Tubao- Philbio, Mexican
Philbio are 4.92%, 4.57% and 4.51% respectively. This indicates that
low ash content of these provenances can be a good source of
biodiesel stated by AOAC, 1984 . The seeds are very rich in fat
content with Tubao-PhilBio

having the highest value of 37.03% and

22

Mexican-Philbio having the least value of 35.86%. These seeds can

be good sources of vegetable oil for human consumption and for
industrial applications such as in cosmetics and the food industry as
mentioned by Islam et al.
The values of the Crude protein for the three samples are
16.71%, 16.64%,15.7% respectively, with India D1 having the
highest and Tubao-PhilBio having the least. However these three
varieties Jatropha curcas can be said to be considerably low in
protein and cannot compete favorably with some other good
sources of protein that were reported to contain 28.4% and 31.85%
protein respectively by (Amoo et al., 2004). The crude fibre contents
of these seeds are considerably high within the range of 12.93% 13.88%. This value is comparably much higher with the crude fiber
reported for Jatropha catharica (1.60%).
Carbohydrate constitutes a major class of naturally occurring
organic compounds. They are essential for the maintenance of plant
and animal life and also provide raw materials for many industries.
Carbohydrate contents of Mexican-Philbio (21.52%), Tubao-PhilBio
(21.18%)

and India D1 (19.35%)

observed is lower to the

carbohydrate content reported for J. curcas of Egyptian origin with

30.11% but higher than the carbohydrate content of J. carthica
(6.45%).
The data revealed that the values between the chemical
compositions of J. curcas are relatively close to each other (Figure 1)

23

which mean that there are slight differences between the chemical
compositions of seeds from different provenances used in the study.

Table 1: Proximate Composition (%) of Three Jatropha curcas

Provenances

Composition

Tubao-

India D1

Phibio
Moisture

6.6

MexicanPhibio

6.38

5.69

24

Ash

4.57

4.92

4.41

Crude fat

37.03

36.76

35.86

Crude fiber

12.93

13.88

13.83

Crude protein

15.7

16.71

16.64

Carbohydrates

21.18

19.35

21.52

Figure 1 Results of proximate composition of three Jatropha

curcas Provenances

4.1 Free Fatty Acid Content in Jatropha curcas oil

As

reported

by

Berchmans

and

Hirata,

an

important

consideration in the feedstock selection for biodiesel production is

the content of free fatty acid (FFA) in the oil. Therefore, to be used
as a feedstock in biodiesel production, the Jatropha curcas Oil
should contain a low percentage of free fatty acids (FFA) not higher
25

than

1%

so

that

the

oil

can

directly

be

utilized

in

transesterification reaction with alcohol as an excess reactant in the

presence of an alkaline catalyst. Otherwise, the saponification shall
occur and the separation of products shall be exceedingly difficult,
and as a result, the yield of biodiesel product would be low.
Figure 2 shows that Tubao- Philbio has 0.16% of free fatty acid,
Mexican-Philbio has 1.03% and India-D1 has 0.19% value of FFA.
Based on these results, the Tubao-Philbio provenance would yield
the highest amount of biodiesel and would require the less energy if
utilized

in

transesterification

reaction.

The

Mexican-Philbio

provenance however exceeded the 1% FFA standard, this indicates

that the oil from this provenance would require processing such as
esterification to neutralize and remove the FFA (Agbogun 1983).

26

Figure 2. Free Fatty Acid of 3 Jatropha curcas Provenances of

Asian and Mexican origin

4.3 Statistical Analysis

The researchers wished to assess if the magnitude of the
differences among sample mean measurements (in terms of
moisture, ash, fat, fiber, protein and carbohydrates and free fatty
acid) are large enough to entail rejection of equality of population
means. Hence, a hypothesis testing procedure called Analysis of
Variance was conducted at 5% level of significance.
Based on sample results, there is not enough evidence to
conclude that there is a significant difference in the population
means of the provenances tested for ash, crude fat, crude fiber and
crude protein. There

Figure 2 One-Way Analysis of Variance with =0.05

Proximate

Analysis

value

value

Moisture

0.0147

Ash

0.0690

F crit

Remarks

9.2208

5.1432

Significantly different

4.3124

5.1432

Not significantly
different
Not significantly
Crude Fat

0.3898

1.1066

5.1432
different
Not significantly

Crude Fiber

0.1393

2.7863

5.1432
different

27

Not significantly
Crude Protein

0.2139

2.0162

5.1432
different
Not significantly

Carbohydrates

0.3336

1.3253

5.1432
different

Free Fatty Acid

0.0059

9.0538

4.0661

Significantly different

5. SUMMARY, CONCLUSION AND

RECOMMENDATIONS
From the results of the study the following are the summary,
conclusion and recommendation which answer our main objective.
a. The three varieties of J. curcas are closely related in terms of
moisture, ash, crude fiber, crude protein, carbohydrates and
crude fat contents. Statistically, there is no significant
difference among the three provenances in terms of ash,

28

crude fiber crude protein, carbohydrates and crude fat

however the moisture content slightly differs.
b. The free fatty acid values of the three Jatropha curcas
provenances did not exceed the 1% standard which means it
can be a good source of biodiesel.
c. Based on the result of the study the following evaluation were
made:

J. curcas is a multipurpose species with many attributes and

considerable potential. The present results demonstrated the
chemical composition of J. curcas to know its prospective industrial
applications especially in the oil industry. The three varieties of
Jatropha curcas seeds are closely related in proximate compositions.
From the results of this study the seeds of the different provenances
are a good source of carbohydrate, crude protein, and crude fat. The
plant can be used for medicinal purposes, in cosmetics based on its
proximate composition which determines its potential for industrial
applications (Islam et al). Because of its high crude fat content, it
can be concluded that it can possibly be a good

source of oil which

can be converted into biodiesel. The oil from its seeds is the most
valuable product since it can be converted into biodiesel. Biodiesel
has become more attractive as an alternative to fossil diesel
because of its environmental benefits and the fact that it is made
from renewable resources. J. curcas L. is a promising source of
biodiesel since its seeds contain high amount of oil and the species
29

has good agronomic traits.

provenances

of

J.curcas

Since the seeds of the different

which

are

currently

underutilized/

unexplored in most regions of the world further studies should be

conducted about its chemical composition (e.g. toxicity levels)
because, in this study the phase "crude Protein, crude fat, crude
fiber" does not say anything about the quality of the said
composition. Moreover, there is still lack of scientific evidence to
support claims related to Jatropha high oil yield production
particularly at large scale. But still Jatropha deserves as much
attention as it can receive worldwide so that as many people as
possible can benefit from the obvious advantages.

LITERATURE CITED
30

Adebowale,

K.O.

and

C.O.

Adedire.

2006.

Chemical

composition and insecticidal properties of the underutilized

Jatropha curcas seed oil. Afr. J. Biotechnol., 5: 901-906.

Berchmans, H.J. and Hirata S. 2007.Biodiesel production from

crude Jatropha curcas L. seed oil with a high content of free
fatty acids. Bioresource Technology 99 (2008) 17161721

Chakrabarti, M.H. and R. Ahmad. 2008. Transesterification

studies on castor oil as a first step towards its use in biodiesel
production. Pak. J. Bot., 40: 1153-1157.

Chitra, P., P. Venkatachalam and A. Sampathrajan. 2005.

Optimisation

of

experimental

conditions

for

biodiesel

production from alkali-catalysed transesterification of Jatropha

curcus oil.

ASAE, Vegetable oil fuels: Proceedings of the international

conference on plant and vegetable; 1982

Foidl N, Foidl G, Sanchez M, Mittelbach M, Hackel S. Jatropha

curcas L. as a source for the production of biofuel in
Nicaragua. Bioresour Technol 1996; 58:7782

Goering CE, Schwab AW, Daugherty MJ, Pryde EH, Heakin AJ.
Fuel properties of eleven oils. Trans ASAE 1982;25:147283.
Klass, D. L. Biomass for Renewable Energy, Fuels, and
Chemicals. Academic Press, London. 1998, p. 335

31

Gubitz, G.M., Mittelbach, M., Trabi, M., 1999. Exploitation of

tropical

oil

seed

plant

Jatropha

curcas

L.

Bioresource

Technology 67, 7382

Houfang, Lu., L. Yingying, Z. Hui, Y. Yang, M. Chen and B.

Liang. 2009. Production of biodiesel from Jatropha curcas L.
oil. Comp. Chem. Eng., 33: 1091-1096

Makkar,

H.P.S.,

Aderibigbe,

A.O.,

Becker,

K.,

1998.

Comparative evaluation of non-toxic and toxic varieties of

Jatropha curcas for chemical composition, digestibility, protein
degradability and toxic factors. Food Chemistry 62, 207215

Parawira W. 2010. Biodiesel production from Jatropha curcas: A

review Technol., Vol. 5(14), pp. 1796-1808

Rashid, U., F. Anwar, T.M. Ansari, M. Arif and M. Ahmad. 2009a.

Optimization of alkaline transesterification of rice bran oil for
biodiesel production using response surface methodology. J
Chem. Technol. Biotechnol., 84: 1364-1370

Sonntag NOV. Structure and composition of fats and oils. In:

Swern D, editor. 4th ed., Baileys industrial oil and fat
products, vol. 1, 4th ed. New York: John Wiley and Sons; 1979.
p. 1

Wang, P.S. M.E. Tat and J.V. Gerpen. 2005. The production of
fatty acid isopropyl esters and their uses as a diesel fuel. J.
Am. Oil Chem. Soc., 82: 845-849. (Received for publication 7
March 2009)

32

33

Appendix A

Procedures
Sample preparation
Ash and Moisture Determination
Equipment/ Apparatus

LJ16 Moisture Analyzer *(Metler Toledo)

Set drying time to AUTO. Press ENTER

Set drying to 110oC. Press ENTER.

Open drying cover. Mount aluminum pan on the pan

support.

Tare. Distribute the sample evenly on the pan. Close the dryer.

Press START.

Press STOP after analysis.

Record the moisture loss.

Bring the down the temperature of the furnace to 105oC and

maintain for 20 minutes.

Cool crucible in a dessicator and weigh after 30 minures.

Reheat the crucibles in an oven at 110oC, let cool and weigh.

34

Repeat this operation at constant intervals until constant

weight is obtained.

Record the final weight of the crucibles.

%Ash = weight of the residue

Weight of the dry sample

% Crude Fat (Soxhlet method)

Reagents:
Anhydrous diethyl ether
Equipment/ Apparatus
Filter paper
Soxhlet apparatus
Drying oven
Analytical balance

Weigh in triplicate 2.00 g sample in a pre- weighed filter

paper (15x15), fold, and wrap with another piece of the
same size. Place the sample in the extraction chamber
of the soxhlet apparatus.

Set up the soxhlet flask with enough anhydrous diethyl

ether, approximately 175 mL (the empty flask should be
dried to constant weight in an oven before using for the
extraction). Extract the fat in the sample.

35

Remove the sample and air dry to remove the excess

ether. Dry the filter paper with the sample in an oven at
105oC until the weight is constant.

Record the weight of the filter paper and the sample.

Run the blank using the same size of the filter paper.
Subtract the weight obtained from the weight of the fat
in the sample.
% Crude Fat = corrected weight of the fat x 100
Weight of the dry sample

% Crude Fiber (Manual Filtration Method)

Reagents:
0.64bN H2SO4 (71.12 mL cont H2SO4 per L soln)
1.56 N NaOH (124.8 g NaOH per 2 L soln
0.2 % methyl red (ttitrate 0.2 g methyl red with 7.4 mL 0.1 N NaOH
and make up to 100

mL with of distilled water)

1% phenolphthalein (1 g H2Ph in 90mL ethanol and make up to

100mL with ethanol) antifoam.
Equipments/Apparatus
600 mL beaker
Hot plate
Dress linen
Filter papers

Gooch crucibles
Oven
muffle furnace
dessicator

36

Transfer the defatted sample into a 600 mLbeaker (tall

form).

Add boiling 0.64 N H2SO4 upto the 200 mL mark. Add 1

drop antifoam compound.

Heat the resulting mixture and allow to boill for 10

minutes with constant shaking.

Filter immediately through dress linen.

Wash the residue on the filter paper with boiling water

until the washings are no longer acidic (washings turn
red to yellow with MR indicator)

Return the residue into the beaker using approximately

100 mL hot distilled water. Make up to the 100 mL mark
with the distilled water.

Add boiling 1.56 N NaOH up to the 200 mL mark. Add 1

drop antifoam compound.

Heat the mixture and allow to boil for 10 minutes with

constant shaking

Filter immediately through dress linen

Wash the residue with boiling water until the washings

are no longer alkaline (washing turn pink to colorless
with phenolphththalein indicator
37

Transfer the residue into the prepared Gooch crucible

with asbestos matting by washing the residue with 10
mL ethanol.

Heat the sample at 105oC until the weight becomes

constant.

Ignite the residue in the muffle furnace for about 20

minutes at 550oC until all carbonaceous matter are
oxidized.

Cool, place in a dessicator and weigh.

%Crude Fiber = Weight of crucible @ 550oC

Weight of dry sample

x 100

Appendix B

Documentation
Sample Preparation

38

Moisture and Ash Content Analysis

Constant Weighing

39

Samples in the Furnace

Soxhlet Method for Crude fat and Oil Extraction

40

Appendix C
Raw Data
41

Proximate Composition (%) of 3 Jatropha curcas Provenances

Table 1 Tubao-Philbio

Replicate
No.

Moisture

Ash

Fat

Fiber

Protein

Carbohydra
tes

1
2

6.6
6.59

4.56
4.64

37.2
35.48

12.8
13.42

14.54
16.22

23.3
21.65

12.56
12.92667

16.34
15.7

18.58
21.1766666
7

3
Ave

6.62
4.5
38.4
6.603333 4.566667 37.02667

Table 2 India-D1
Replicate
No.
1
2

Moisture

Ash

Fat

Fiber

Protein

Carbohydrates

6.2
6.43

5.3
4.56

37.7
36.32

13.43
13.62

17.1
16.53

19.27
20.54

6.5

4.9

36.25

14.6

16.5

18.25

Ave

6.376667

4.92

36.75667

13.88333

16.71

19.35333333

Table 3 Mexican-PhilBio

Replicate
No.

Moisture

Ash

Fat

Fiber

Protein

Carbohydrates

6.2

4.4

36.35

13.43

16.6

22.02

2
3

5.43
5.43

4.43
4.4

35.74
35.5

13.62
14.6

16.12
17.2

22.66
19.87

Ave

5.686667

4.41

35.86333

13.88333

16.64

21.51666667

Appendix D

Statistical Analysis

42

MOISTURE
Let the following be:

1. Hypothesis:

2. Level of significance:

3. Test Statistic:
4. Critical Region:
5. Computations:
Anova: Single
Factor
SUMMARY
Groups
Tubao-Philbio
India D1
Mexican PhilBio

ANOVA
Source of
Variation
Between
Groups

3
3

Sum
19.81
19.13

Averag
e
6.603
6.377

Varian
ce
0.000
0.025

17.06

5.687

0.198

Count

SS
1.3677555
56

df

MS

P-value

F crit

0.684

9.221

0.015

5.143

43

Within Groups

0.445

Total

1.8127555
56

0.074

6.

7. Conclusion: At 5% level of significance, based on sample

results, there is sufficient evidence to claim that not all
population mean moisture content of the three J. curcas
provenances are equal.
ASH
Let the following be:

1. Hypothesis:

2. Level of significance:

3. Test Statistic:
4. Critical Region:
5. Computations:

Anova: Single
Factor
44

SUMMARY
Groups
Tubao-Philbio
India D1
Mexican PhilBio

Count
3
3

Sum
13.70
14.76

Average
4.57
4.92

Varian
ce
0.00
0.14

13.23

4.41

0.00

ANOVA
Source of
Variation
Between
Groups
Within Groups

SS

df

MS

0.409
0.285

2
6

0.205
0.047

Total

0.694

Pvalue

F crit

4.312

0.069

5.143

6.

7. Conclusion: At 5% level of significance, based on sample

results, we do not have sufficient evidence to conclude that
the population mean ash content of the three J. curcas
provenances differ from one another.
FAT
Let the following be:

1. Hypothesis:

2. Level of significance:

3. Test Statistic:
45

4. Critical Region:
5. Computations:
Anova: Single
Factor
SUMMARY
Groups
Tubao-Philbio
India D1
Mexican PhilBio

Count
3
3

Sum
111.08
110.27

Average
37.03
36.76

Varian
ce
2.15
0.67

107.59

35.86

0.19

ANOVA
Source of
Variation
Between
Groups
Within Groups

SS

df

MS

2.22
6.03

2
6

1.11
1.00

Total

8.25

Pvalue

F crit

1.11

0.39

5.14

6.

7. Conclusion: At 5% level of significance, based on sample

results, we do not have sufficient evidence to conclude that
the population mean fat content of the three J. curcas
provenances differ from one another.

FIBER
Let the following be:

46

1. Hypothesis:

Level of significance:

2. Test Statistic:
3. Critical Region:
4. Computations:
Anova: Single
Factor
SUMMARY
Groups
Tubao-Philbio
India D1
Mexican PhilBio

Count
3
3

Sum
38.78
41.65

Average
12.93
13.88

Varian
ce
0.20
0.39

41.65

13.88

0.39

ANOVA
Source of
Variation
Between
Groups
Within Groups

SS

df

MS

1.83
1.97

2
6

0.92
0.33

Total

3.80

Pvalue

F crit

2.79

0.14

5.14

5.

6. Conclusion: At 5% level of significance, based on sample

results, we do not have sufficient evidence to conclude that
the population mean fiber content of the three J. curcas
provenances differ from one another.
47

PROTEIN
Let the following be:

1. Hypothesis:

2. Level of significance:

3. Test Statistic:
4. Critical Region:
5. Computations:
Anova: Single
Factor
SUMMARY
Groups
Tubao-Philbio
India D1
Mexican PhilBio

Count
3
3

Sum
47.10
50.13

Average
15.70
16.71

Varian
ce
1.01
0.11

49.92

16.64

0.29

ANOVA
Source of
Variation
Between
Groups
Within Groups

SS

df

MS

1.91
2.84

2
6

0.95
0.47

Total

4.75

Pvalue

F crit

2.02

0.21

5.14

48

6.

7. Conclusion: At 5% level of significance, based on sample

results, we do not have sufficient evidence to conclude that
the population mean protein content of the three J. curcas
provenances differ from one another.

CARBOHYDRATES
Let the following be:

1. Hypothesis:

2. Level of significance:

3. Test Statistic:
4. Critical Region:
5. Computations:
Anova: Single
Factor
SUMMARY
Groups
Tubao-Philbio
India D1
Mexican PhilBio

Count
3
3

Sum
63.53
58.06

Average
21.18
19.35

Varian
ce
5.74
1.32

64.55

21.52

2.14

49

ANOVA
Source of
Variation
Between
Groups
Within Groups

SS

df

MS

8.12
18.38

2
6

4.06
3.06

Total

26.50

Pvalue

F crit

1.33

0.33

5.14

6.

7. Conclusion: At 5% level of significance, based on sample

results, we do not have sufficient evidence to conclude that
the population mean carbohydrates content of the three J.
curcas provenances differ from one another.
FREE FATTY ACID
Let the following be:

1. Hypothesis:

2. Level of significance:

3. Test Statistic:
4. Critical Region:
5. Computations:

SUMMARY
50

Groups
TubaoPhilbio
India D1
Mexican PhilBio

Count

Sum

Average
0.1566666
67
1.0266666
67
0.1933333
33

Variance
3.33333E05
0.0006333
33
0.0002333
33

MS
0.7263444
44

Pvalu
F
e
2421.1481 1.9E
48
-09

0.0018

0.0003

1.4544888
89

0.47

3.08

0.58

ANOVA
Source of
Variation
Between
Groups
Within
Groups
Total

SS
1.4526888
89

df

6.
7. Conclusion: At 5% level of significance, based on sample
results, there is sufficient evidence to claim that not all
population mean moisture content of the three J. curcas
provenances are equal.

51

F crit
5.1432
53