KALUMPANG SEEDS BIOETHANOL

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Physico-Chemical Properties of Kalumpang

(Sterculia Foetida) Seeds as Bioethanol

RESEARCH PLAN
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RATIONALE

A looming energy crisis has been a feature in the Philippines for years. This

leaves the government to rethink its current energy policy including those for renewable

energy and biofuels (Kritz, 2015). According to Coates (2012), biofuel is a transport fuel

that can be produced from plants or animal origin and are utilized as substitutes or

extender for fossil fuels.

Increasing use of Philippine indigenous plant species grows and augments the

value and economic importance that can seek solutions to the current facets of The

Philippine energy problems (Cainta Plant Nursery, 2013). The presence of starch and

saccharides in Kalumpang (Sterculia foetida) seeds makes it feasible substance in

bioethanol production (Gray, Zhao, Emptage, 2006).

However, the produced biodiesel from Kalumpang was not as efficient compared

to the commercial biodiesel (Rao, Ramesh, Kumar, 2015). Also, the challenges to an

industrial reality are multiple and need an investment in research and development (Lima

& Natalense, 2012).

The aim of this research is to contribute new ideas to the body of knowledge and

to support an idea about the feasibility of Kalumpang stock yield as a bioethanol. This

research is relevant to the whole community since it serves as one of the building blocks

of human society. All of the procedures can help and contribute as an idea in making a

bioethanol. The expected product is less toxic that can reduce the amount of carbon

monoxide released by the vehicles thus it may improve the air quality.
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This research will examine the physico-chemical properties of Kalumpang

(Sterculia foetida) seeds as a potent bioethanol through chemical reactions and

performing T-test as statistical tool.

Literature Review

The concern for diminishing limited natural resources that are used to power

demanding industrial society is energy crisis. This occur naturally but it takes millions of

years to fill the stores. Government and concerned citizens took the responsibility to use

renewable resources as an alternative and to properly apply the natural supplies through

increased conservation (Rinkesh, 2009).

According to the Department of Energy, the country compared it internationally

and it is 40% ahead with the use of renewable energy resources such as wind, water, and

solar energy. According to Corpuz (2009), the Philippines also boast its alternative fuel

production globally.

As stated by Bello (2013), the country’s developments on oil production has

deteriorated since circa 1990. Since then, the price of the oil is honestly reached $139 per

barrel. Even the price of unleaded gasoline has arrived beyond ₱56 and ₱49 in diesel. The

Philippines is consuming over 120 million barrel per year but 90% of it is import.

However, energy crisis is a wide and complex topic, it is worthy to try to

understand. Most people don’t feel connected when it talks about energy crisis unless the

price of the gas change, goes up and falls. The energy crisis is a serious topic to make on

because it is ongoing and getting worse despite of too much effort. This will not resolve
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without determination and broad understanding about general-to-specific causes and

specific-to-general solutions (Rinkesh, 2009).

Increasing prices of petroleum fuels and decreasing availability of conventional

and non-renewable energy sources gave way to the development and production of

alternative fuels such as biofuels (Abacan, Acio and Convento, 2007). According to

Coates (2012), biofuel is a generic term for transport fuel that can be produced from

renewable material of plant or animal origin and are utilized as substitutes or extender for

fossil fuels. The production of the biofuels in the Philippines became a trend as the

country became the first to have an ethanol refinery in South East and to be a global

leader in renewable energy usage and production in Asia (Corpuz, 2009). Through this,

research and development in biofuels to supplement or substitute petroleum fuels in the

Philippines focus on these two major classification, the biodiesel and bioethanol.

Bioethanol can be produced, from the raw materials having sucrose and starch which are

recognized as the first generation feedstock, using the fermentation bioprocess. Although

the supply of these raw materials will be limited soon, therefore the lignocellulosic

material can support the production of bioethanol in the near future so this material is

recognized as the second generation feedstock (Lima & Natalense, 2012).

As mandated by the Philippines Biofuels Act of 2006 (RA 9367) , blending of

biodiesel and ethanol in all locally distributed diesel and gasoline aims to reduce the

country’s dependence on imported fuels with due regard to the protection of public

health, the environment, and the natural ecosystems consistent with the country’s

sustainable economic growth. However, Global Agricultural Information Network (2014)

report points out that the country is nowhere near meeting the E10 standard on a national
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scale, and is falling behind in building new ethanol production capacity. The country

currently can only produce about 17 percent of its ethanol needs, even in falling short of

the E10 standard. As a cause, climate change realities are likely to dampen any dramatic

increase in local biomass output, that’ll results to downgrade of biofuel production in the

year 2017. Another cause is the cultivation for the biomass for fuel will require vast

tracks of land, and is expected to be constrained by a longstanding Philippine agrarian

reform law which limits private land ownership to five hectares (Corpuz, 2016). Based on

the report published by Sy (2015), the Philippines need to have 13 distilleries with annual

capacity of 30 million liters to meet the mandated blend of 10% and meet the annual

quota. Another appeal from the report includes the decline in the number of laborers

joining the force in the production of biomass in industry especially in planting,

cultivating, and harvesting. In general terms, increasing prices of locally produced

biofuels, coupled with declining global oil prices is likely forcing the Philippine

government to rethink its current energy policy, including those for renewable energy and

biofuels (Corpuz, 2016).

Biofuels suddenly becoming unfashionable presents a quandary for Philippine

policymakers. Of the reasonable alternatives the country could pursue in order to tackle

its problem of ludicrously excessive vehicular emissions – the others include alternative

fuels such as liquefied natural gas (LNG) or hydrogen, and hybrid or fully electric

vehicles, but still, biofuels are the most economical option. (Kritz, 2015)

For being dependent on non-renewable resources and working to find an

alternative one to the energy-rich fuels such as gasoline and diesel. Bioethanol can be

produced, from the raw materials having sucrose and starch which are recognized as the
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first generation feedstock, using the fermentation bioprocess. In the Philippines alone, a

total of 11 biodiesel refineries annually supplies aggregate capacity of 585 million liters

throughout the whole country. The Philippines is also the first country in South East

Asia, to have legislation in mandating the blending of biofuel, as suggested by Republic

Act 9367.

Increasing use of Philippine indigenous plant species grows and augments the

value and economic importance that can seek solutions to the current facets of The

Philippine economic problems (Cainta Plant Nursery, 2013). Kalumpang is a spreading

tree, reaching a height more than 30 meters tall, with crowded, digitately compound

leaves at the end of each branch. The putrid smelling flowers whose color ranges from

yellowish to purplish hue has an average length of 2 to 2.5 centimeters in diameter. The

fruit itself is large and smooth that contains an average of 10-15 large seeds (Lee, 2009).

Kalumpang leaves appear in March to April just after its flowering, whereas the flowers

appear on or before March when the tree is leafless. The fruit, however, ripens nearly 9

months after the first appearance of the flowers (Wild, Flora Zambesiaca, 1995). Its leaf

extract is known for the anti-bacterial or antimicrobial property (Singh & Vidyasagar,

2014). The seed extract of Sterculia foetida can be used as insecticide and as an

antifeedant (Wagstaff, 2008). Kalumpang seed can be eaten either raw, roasted, or fried,

and with a pleasant, cacao-like flavor but not bitter taste. The 40 % edible non-drying oil

from the seed has the same properties to the olive oil and can be used in culinary

purposes (Howes, Manandhar, Facciola, Brown, Barwick, Uphof, Dymock, Warden,

Hooper, 2014). About medicinal purposes, Kalumpang is not on the least. Almost of its

parts are aperient including leaves, bark, seeds and the oil of the seeds. Its wood is soft
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and light, and has a property of poor durability that can be used for small project

constructions like boxes, cardage, mats, and bags (Mahakalkar, Upadhye, 2013).

Kalumpang’s oil can be used for lightning and as an illuminant. Its sterculic oil is proved

to resist belly fat, obesity and insulin resistance. It is been tested in an obese mice (Coy,

Gouch, Perfield, 2012).

Researchers’ in the University of Malaya, Malaysia (2012), conducted a study

that proves the feasibility of Kalumpang’s fatty acids as one of the non-edible feedstocks

for biodiesel production. As the paper suggested, the produced Kalumpang methyl ester

passed the limits of ASTM D6751 and EN 14214 specification, hence proving the

researchers’ claim. Zero waste management system was deemed feasible in Sterculia

foetida biodiesel production. After the raw seeds of Kalumpang undergoes hydrolysis

and fermentation, the results shown and proved the researchers’ claim in the study. In the

case of the blended bioethanol for example, a research from the India’s Department of

Mechanical Engineering indicates that the engine runs smoothly but the efficiency

decreases (Rao, 2015).

PROBLEM

This study aims to determine the feasibility of Kalumpang (Sterculia foetida)

seeds’ physico-chemical properties in producing bioethanol (C2H5OH) through hydrolysis

and alcoholic fermentation.

Specifically, this research seeks answers to the following questions:

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1. How may the Kalumpang seeds be evaluated and examined in terms of its

physic-chemical properties:

1.1 Physical Properties

a. Bulk density,

b. Particle size?

1.2 Chemical Properties

a. pH stability,

b. Volatility,

c. Lower Flammability Limit?

2. Is there a significant difference between the processed Kalumpang seeds through

oxidation and turbidity in terms of:

a. Sodium dichromate (Na2Cr2O7) solution reaction,

b. Lucas’ Reagent?

HYPOTHESES

1.1 A. Ho : Bulk density doesn’t affect the flow property in terms of viscousity of

powdered Kalumpang seeds on alcoholic fermentation.

Ha : Bulk density affects the flow property in terms of viscousity of powdered

Kalumpang seeds on alcoholic fermentation.

B. Ho : Particle size doesn’t affect the rate of chemical reaction in terms of

speed of powdered Kalumpang seeds on alcoholic fermentation.

Ha : Particle size affects the rate of chemical reaction in terms of speed of

powdered Kalumpang seeds on alcoholic fermentation.

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1.2 A. Ho : There is no significant difference between the pH stability of

Kalumpang bioethanol and commercial ethanol in pH level of 4.5.

Ha :There is a significant difference between the pH stability of Kalumpang

bioethanol and commercial ethanol in pH level of 4.5.

B. Ho : There is no significant difference between the volatility of Kalumpang

bioethanol and commercial ethanol with an average of 78 to 100 oC boiling point.

Ha : There is a significant difference between the volatility of Kalumpang

bioethanol and commercial ethanol with an average of 78 to 100 oC boiling point.

C. Ho : There is no significant difference between the Lower Flammability Limit

of Kalumpang bioethanol and commercial ethanol in chromatography test.

Ha : There is a significant difference between the Lower Flammability Limit of

Kalumpang bioethanol and commercial ethanol in chromatography test.

2. A. Ho :There is no significant difference between the Kalumpang bioethanol and

commercial ethanol through oxidation analysis in terms of Sodium dichromate

(Na2Cr2O7) solution reaction.

Ha :There is a significant difference between the Kalumpang bioethanol and

commercial ethanol through oxidation analysis in terms of Sodium dichromate

(Na2Cr2O7) solution reaction.

B. Ho :There is no significant difference between the Kalumpang bioethanol and

commercial ethanol through turbidity in terms of Lucas’ reagent.

Ha : There is a significant difference between the Kalumpang bioethanol and

commercial ethanol through turbidity in terms of Lucas’ reagent.

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ENGINEERING GOALS

This study will aim to achieve the following:

 Elaborate the idea of the importance of bioethanol derived from the Kalumpang

(Sterculia foetida) seeds that will contribute new ideas to the body of knowledge.

 Examine the physico-chemical properties of Kalumpang (Sterculia foetida) seeds

as a potent bioethanol through chemical reactions.

EXPECTED OUTCOME

 The performance of the physico-chemical properties of Kalumpang (Sterculia

foetida) seeds as a bioethanol will be as feasible as the commercial one’s own

properties.

MATERIALS AND METHODS

The research method will be carried out with three (3) replicates through

saccahrification with the aid of invertase at optimal conditions and the hydrolyzed

Kalumpang (Sterculia foetida) starch containing saccharomyces cervisiae will undergo

alcoholic fermentation through zymase to use as carbon source for ethanol production

of Kalumpang (Sterculia foetida) seeds with addition of ethyl alcohol ( C2H6O) after

distillation. 

The 500 g of one week old Kalumpang (Sterculia foetida) seeds will be collected

in the premises of Barangay Baseco, Mariveles, Bataan and will be placed in a rattan

basket. The measuring instruments and laboratory apparatus including the 500mL

Erlenmeyer flask, 10x2 steel wool, 1000mL vapor recycler beaker with 12 inch spiral
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glass tube, digital weighing scale and 10mL evaporating dish will be borrowed from the

Mariveles NHS-Poblacion’s laboratory property. The Saccharomyces cervisiae and the

Carbon Hydroxide (CaOH2) will be bought in the local market of Mariveles, Bataan.

Hydrolysis and fermentation of the seeds will be conducted at Mariveles National

High School – Poblacion. The oxidation test of the produced bioethanol will be tested at

ECHEM Environmental Testing Laboratory, Clark, Pampanga.

The 500 g of Kalumpang (Sterculia foetida) seeds will be will be sun-dried for

six hours at 9 A.M. until 4 P.M. to remove the excess moisture and to avoid the formation

of eukaryotic and single-celled organisms. The acquired dried seeds will be milled to 1.0

mm particle size using a 500 mL coffee grinder at 120 rpm to obtain 450 g of Kalumpang

(Sterculia foetida) starch that will be divided into three replicates, 150 g per each . The

milling process will be completed to break down the seeds into fine particles in order to

increase the speed rate in producing sugar solution during invertase. Invertase is an

enzyme that splits sucrose into glucose (C6H12O6) and fructose (C6H12O6) that uses yeast

culture through saccharification.

C6H12O6 +H2O + Invertase 2(C6H12O6)

(Starch) (Water)

The saccharification will be

carried out in 500 mL Erlenmeyer flasks in which the three (3) replicates contains 150 g

Kalumpang seed starch, Saccharomyces cerviciae as the yeast culture and 200 ml

distilled water. The saccharification process will be accomplished to increase ethanol

yield and production rate that remains active in the fermentation at a temperature of 4-

32℃ and pH concentration of 4.0 to 5.0. The three (3) replicates will be left standalone
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for 72 hours at 4-32℃ to allow intervase to broke down the starch of Kalumpang

(Sterculia foetida) into glucose (C6H12O6). The 500mL Erlenmeyer flask will be utilized

with a 10x2 steel wool so that the hydrolysis and fermentation assays will be in anaerobic

process. On a twelve hourly basis, the three (3) replicates’ pH level will be monitored by

addition of Carbon hydroxide (CaOH2) so that the first replicate will attain a constant pH

of 4, the second replicate will attain a constant pH of 4.5 and the third replicate will attain

a constant pH of 5. Thus, making the results vary in terms of ethanol concentration.

The fermentation assays will be carried out in the three (3) replicates through

alcoholic fermentation with the aid of zymase. Zymase is an enzyme complex that

catalyzes the fermentation of glucose into ethanol and carbon dioxide. The glucose will

came from the three (3) replicates after the 74 hours saccharification hydrolysis was

finished. The three (3) replicates will then be left stand alone for another 74 at room

temperature not exceeding 35℃. This will let the zymase produce two (2) moles of

ethanol 2(C2H5OH) and two mole of carbon dioxide 2(CO2). The yeasts will mainly

metabolize glucose and fructose to form pyruvic acid through the stages of the reaction

pathway Embden-Meyerhof-Parnas, nevertheless pyruvic acid generated would be

decarboxylated to acetaldehyde which then experiences dehydrogenation to ethanol.

C6H12O6 + Zymase 2(C2H5OH) + 2(CO2)

(Glucose)

After fermentation is completed, the distillation process can separate ethanol

based on boiling point. The boiling point of pure ethanol is 78 ℃. By heating the solution

at a temperature range of 78 – 100℃ will result in most of the ethanol evaporated, and the

condensing units will be produced with a concentration of 95% ethanol by volume. The
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precipitate will pass through the spiral glass tube from vapor recycler beaker in which the

three (3) replicates will be placed and heated by a bunsen burner in a temperature of 80-

85℃. The distilled mixture will be then combined with 70% solution of isopropyl

alcohol with a proportion of 1mL: 2.5mL. to control the pH level of 4.5.

The researchers will gather information of the 3 replicates by the use of Lucas’

Reagant and Sodium Dichromate (Na2Cr2O7) reaction, and determining the pH level

concentration and the level of flammability.

In Lucas’ test, an alcohol will be treated with Lucas’ Reagent that is basically a

solution formed by the combination of HCl (Hydrochloric Acid) and ZnC12 (Zinc

Dichloride). It will be used as a reagent to test alcohols and classify them in accordance

to their reactivity. The classification of the alcohols is usually done based on the

difference in reaction with concentrated hydrochloric acid. A simple reaction is given

below:

ZnC12

ROH + HCl—>RCl + H2O

About oxidation test, the alcohols will be oxidized with Sodium Dichromate

(Na2Cr2O7). The rate of oxidation varies between primary, secondary and tertiary alcohol.

It is an orange to red colored, crystalline, inorganic compound that emits toxic Chromium

(Cr) fumes upon heating. Sodium Dichromate (Na2Cr2O7) is highly corrosive and is a

strong oxidizing agent. Conducting the two test in ECHEM will give the researchers the

data they needed for classifying an alcohol and in oxidation reactions.

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The changes of pH in all fermentations were determined by pH Paper (Arrhenius

model). The pH will be checked before, during and after the fermentation process. A pH

indicator is a halo chromic chemical compound added in small amounts to a solution so

the pH (acidity or basicity) of the solution can be determined visually. The indicator

causes the color of the solution to change depending on the ph. Indicators can also show

change in other physical properties. The color will be listed from the 3 replicates and the

researchers will write the indicated pH level of the ethanol.

In testing the level of flammability, the cotton balls with two 5 mL of the ethanol

will be lighted and the duration of fire will be recorded separately per set- up, it will be

listed in tabular form. While the percent ethanol of the Kalumpang (Sterculia foetida)

seeds bioethanol will be measured by ethanol- gas chromatography. The ethanol with the

higher duration of the fire will consider its level of flammability bigger than the other

commercial bioethanol.

Statistical Analysis

The t- test will be used for comparing the means of two samples (or treatments)

with the same number of replicates.

After measuring the Kalumpang (Sterculia foetida) seeds bioethanol’s in category

of the alcohol, fire duration, the t- test was used since Kalumpang (Sterculia foetida)

seeds bioethanol fuel will only be compared to commercial bioethanol. The results will

be considered statistically significant at 90% (ᾳ=0.05) confidence to determine significant

differences between the two fuels.

RISKS
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Fermentation. Hazards including microbial, chemical and physical hazards coincide

under fermentation assays that will be conducted in the research. These hazards are

caused by uncertain parameters in unconditioned areas and for deeper understanding,

some listed as follows:

Accuracy to research methods may decrease the harm to the researchers. It is

advised to use protective equipments such as eye goggles and laboratory gloves. And for

the variables that will be used in the research, lists for safety precautions are showed

below:

a. Saccharomyces cerevisiae. The yeast was found to cause sensitivities to an

individual when exposed and may trigger allergies and in bad cases, to cause

Candida yeast infection. However, this may occur due to incautious, improper

execution of fermentation processes.

Less exposure to the yeast by wearing protective equipments may reduce

the possibility of hazards. An individual must also be properly educated about the

aforementioned yeast culture.

b. Calcium Hydroxide. Ingestion and inhalation of Calcium Hydroxide may cause

irritation to the respiratory tract and is corrosive to the eyes and the skin.

An individual must not touch or inhale any solutions or powders present in

the laboratory for the sake of his safety. Wearing face mask and gloves are

advised to do so before conducting experimentations in the laboratory.

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c. Laboratory Apparatuses. Equipments and tools left haphazardly could lead to

unwanted accidents and falls.

Checking for any hazards present in the area can keep the researcher on its

safety. Background education about laboratory apparatuses are necessary to

conduct the experiments efficiently.

Distillation. The common accidents, which often occur simultaneously, are fire,

explosion, chemical and thermal burns, cuts from broken glass tubing and thermometers,

absorption of toxic, but non-corrosive chemicals through the skin, and inhalation of toxic

fumes. Less common, but obviously dangerous, is the ingestion of a toxic chemical. Each

of these types is discussed in a general way below:

Accidents in the laboratory are often the result of carelessness and unawareness of

the researchers conducting the experiments. Stay alert and pay constant attention to every

actions executed.

a. Bunsen burner. Not observing the experimentation area while performing a

distillation process may cause fire related injuries such as burn and boils.

Exceeding temperatures causes explosions to laboratory apparatuses that may

harm all the individuals in the vicinity.

There should never be open flames in the laboratory. Specifically, never

heat any organic solvent in an open vessel, such as a test tube, Erlenmeyer flask,

or beaker, with a flame. Never keep volatile solvents in an open beaker or

Erlenmeyer flask. It is researcher’s responsibility to know where the nearest

safety shower and fire extinguisher are located.

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Determining the pH level. In getting the pH level of a water soluble substances by the

use of pH paper may cause harm when its getting more acidic. The pH paper may be

swallowed by someone, especially children, may cause skin infections, and other

infections in other part of the body. Since ethanol is expected to be acidic, so it is a

chemical with corrosive elements. Even though it may not totally affect the human body,

but it may cause some damage in skin tissues.

Before doing the experiment, make sure the appropriate PPEs or the Personal

Protective Equipments is worn. Immediately put the permented ethanol in a safe place

wherein there is no fire and can't be spill out in the floor. All the materials, especially

chemicals, must handle with care. And after using the equipments, there must be

someone who is obligated to put things and materials in place.

Testing the flammability. Fire hazards is evaluated by the use of flammability testing

by cotton balls wherein the commercial ethanol and bioethanol from Kalumpang seeds is

used apply separately. However, it is not intended to directly simulate actual fire

condition. The temperature, humidity, the mass, orientation and shape of the material, air

flow, available oxygen, the source and intensity of the ignition source, heat from radiance

and/or conduction and so on affect the fire from the experiment. It may burn the whole

place by only comparing their flammability. It can also burn your skin by a simple

contact with the fire.

Flammability testing is commonly performed on almost all the materials and may

be able to support combustion. The government suggested to make another test for the

product safety and liability information. When something will be happened during the the
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experiment, the person or group must be aware of the surroundings and don't panic. In the

end, the persons involved must collect all the materials and put it properly to a nice and

safe place where none can drink the ethanol and the children can't reach it.

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