Eusebio High School

S.Y. 2018 – 2019

In partial fulfillment in the requirement for the subject

Inquiries, Investigations and Immersion

The Effectiveness of Lanzones (L. domesticum) Fruit as an

Ingredient for Bioethanol Fuel Additives

Submitted by:

Bermas, Lira Mae B.

Fernandez, Rodney E.

Laraño, Alvin M.

Roque, Lenard H.

Submitted to:

Mr. Vladimir Lou Victoria

March 2019
 Chapter 1

Introduction

Alternative Fuel is currently an important issue all over

the world due to the efforts on reducing global warming which is

contributed by the combustion of petroleum or petrol and diesel.

Bio fuel has attracted the attention of researches all over the

world due to reducing the environmental impacts of elevated

Carbon Monoxide.

This study will be conducted using lanzones (L. domesticum)

waste biomass in different parameters using fermentation method.

Some researches produced bioethanol fuel from different

plants using the distillation method, but it takes a lot of time.

Bioethanol is non-toxic, biodegradable produced from renewable

resources and contributes a minimal amount of net Greenhouse

Gases such CO2, SO2, and NO emission to the atmosphere.

The aim of this study is to produce additives for bioethanol

fuel from lanzones (L. domesticum) fruit. The researchers want to

test the effectivity of the additives produced from lanzones (L.

domesticm).

Statement of the Problem

2
 The researchers want to know if the lanzones (L. domesticum)

fruit can be used as an ingredient for additive for bio-fuel and

if it is effective as an eco-friendly fuel compared to other fuel

in terms of its production.

The researchers also aim to answer the following questions:

1. How many milliliters (mL) of additives can be produced from:

a. 2-kg of lanzones fruit

b. 5-kg of lanzones fruit

c. 8-kg of lanzones fruit

2. Is there a significant comparison between the fuel

efficiency before adding the additives and after adding the

additives?

3. Is there a significant difference between the additives for

bio-ethanol fuel from lanzones (L. domesticum) fruit mixed

with other bioethanol to other gasoline in terms of Cetane

Rating?

Hypotheses

Ho1: The production of additives for bioethanol fuel from

lanzones (L. domesticum) fruit is zero.

Ho2: The difference between the additives for bioethanol fuel

from lanzones fruit mixed with other bioethanol to other gasoline

is zero.

3
Scope and Delimitation

The focus of this experimental research is to know if

lanzones (L. domesticum) can produce a substance for additive for

bioethanol fuel. The lanzones (L. domesticum) that will be used

by the researchers will be sourced out from the Pasig City Mega

Market where it is bounty and cheaper. Other materials and tool

will be provided by the researchers themselves and will be

submitted to Department of Energy – Renewable Energy Bureau. The

research experiment proper will be conducted in Department of

Energy – Renewable Energy Bureau. This research is limited only

in terms of number of fruit (disregarding the maturity and the

size), budget and allotted time process. Some calculations,

selections, and assumptions were made to give an alternative to

the kinds of fuels such as Diesel, petrol and were made as a

consideration of a proper, legal, and realistic research.

Significance of the Study

The researchers aim that this research will be helpful to

the following individuals and sectors:

Transportation Companies

- Additives for bioethanol fuel production can help the

companies that administer transportation. Bioethanol fuel

4
 is also eco-friendly giving a better performance for the

engine because it is a renewable energy.

Local Farmers

- Farmers (which the researchers have focused on giving

higher income) that are planting lanzones (L. domesticum)

that will be used for the future production of the said

additive for bioethanol fuel will be benefited in the way

of giving them a higher income rate.

Future Researchers

- Through this research the researchers wanted to help

future researchers to have an idea regarding lanzones (L.

domesticum) benefits.

Botanist

- The production of the bioethanol fuel, if increased, can

be another milestone in the field of botany giving help

to the community (it will not create another sub-branch

in the said field) and creating an eco-friendly renewable

fuel for the people.

Conceptual Framework
The Effectiveness of Lanzones (L.

domesticum) Fruit as an Ingredient for

Bioethanol Fuel Additives

5
 Dependent Variables
Independent Variables
 Fermentation
o Fermentation
of different
 grounded lanzones
pH parameter
(L. domesticum)
o Fermentation
fruit
of different
temperature
o Fermentation
in different
period
 Filtration
 Distillation

 Density / Specific Gravity

 Distillation
 Cetane Numbers

Ho1: The production of additives for bioethanol fuel from

lanzones (L. domesticum) fruit is zero.

Ho2: The difference between the additives for bioethanol fuel

from lanzones fruit mixed with other bioethanol to other

gasoline is zero.

The focus of this study is to produce an additive for

bioethanol fuel from lanzones (L. domesticum) fruit through the

process of fermentation and distillation. The lanzones after the

6
said two processes will undergo filtration. If the expected

amount of lanzones was produced, the fuel’s density and Cetane

rating will be tested.

Definition of Terms

Bioethanol

Conceptual: in this study, it is a renewable fuel made from

natural resources like plants’ seeds, fruit or crude extract

Operational: an ethanol produced from plants such as sugar cane

or corn, used as an alternative to gasoline. (Gable, 2017)

Cetane Numbers

Conceptual: in this study, it is a rating that measures the

quality of fuel.

Operational: is a measurement of the quality or performance

of diesel fuel. (Barsamian, 2009)

Distillation

Conceptual: in this study, it is a process of separating a

component from a mixture

Operational: it the action of purifying a liquid by a process of

heating and cooling. (Gil et al, 2007)

Fermentation

7
Conceptual: in this study, it is the chemical breakdown of a

substance by bacteria, yeasts, or other microorganisms.

Operational: a metabolic process that produces chemical changes

in organic substrates through the action of enzymes. (Urry et al,

2014)

Filtration

Conceptual: in this study, it is the process of separation of

liquid material from a solid material

Operational: are any of various mechanical, physical or

biological operations that separate solids from fluids by adding

a medium through which only the fluid can pass. (Helmenstine,

2018)

Lanzones

Conceptual: in this study, it is an endemic fruit found in

Philippines

Operational: is yellowish-white, occurring in bunches on a single

stem, ellipsoid or globose, 2 to 4 centimeters long, with bitter

seeds that are surrounded by a translucent pulp. (Santiago and

Stuart, 2015)

8
 Chapter 2

Review of Related Literatures and Studies

Lanzones (Lansium domesticum)

Lansones is a tree growing to a height of 4 to 15 meters.

Leaves are alternate, 20 to 40 centimeters long, with 5 to 7

leaflets, oblong to oblong-elliptic, 7 to 18 centimeters in

length, and pointed at both ends. Flowers are small, yellow and

borne on spikes, solitary or fascicled on the trunk or larger

branches. Fruit is yellowish-white, occurring in bunches on a

single stem, 2 to 4 centimeters long, with bitter seeds that are

surrounded by a translucent pulp (arillus), the pulp occurs in

five sections with one well-developed seed. The outer skin is

thin and tough, abundant in a milky juice. The pulp occurs in

five sections with one well-developed seed. It is believed that

Lanzones came from Camiguin, it is cultivated from Luzon

specifically the province of Quezon and Laguna, and it grows

freely in the Mindanao and Basilan, it can also be found in Indo-

China, in Malay Peninsula and Archipelago, in general

cultivation. The fresh peeling yielded a volatile oil, a resin,

and some reducing acids. The resin is believed to be nontoxic and

protective to the stomach against alcohol. The fruit's skin is

used to treat diarrhea, and in the Philippines the dried skin is

9
burned as a mosquito repellent. The skin, especially of the

langsat variety, can be dried and burned as incense.

Bio-Diesel Extraction from Rambutan Seeds

The researchers decided on using rambutan seeds, because

they live in Southeast Asia, and Rambutan is a common fruit in

there, and it is unique and easy to get. They converted rambutan

seeds into oil first through Soxhlet Extraction (or Soxhlet

Extractor- it is used when the desired compound has a limited

solubility in a solvent, and the impurity is insoluble in that

solvent) and then they have converted it into their final

product, biodiesel through transesterification.

Rambutan is also favorable in making oils because it has a

39% oil percentage in its seeds, higher than its rivals, other

native to Southeast Asia fruits like Lychee and more. The

researchers also said that Rambutan is always coined with the

term “exotic superfruit”, it has a lot of benefits when eaten.

The researcher’s plan is to use Soxhlet Extraction to

extract the oil from rambutan seeds. The researchers chose

Soxhlet Extraction because it is a more efficient method compared

to the use of mechanical press.

As Soxhlet Extraction uses the theory of washing the sample

repeatedly with the use of solvent, it could obtain more bio-oil.

10
Hence, Soxhlet can isolate desired oil, which has limited

solubility in the solvent and the impurity is insoluble in a

solvent. Hence, making the bio-oil have higher purity.

The researchers also said that to achieve efficient Soxhlet

Extraction, the seeds are to be grinded into small pieces and

dried to remove moisture content. After extracting the oil, they

proceeded with transesterification to produce their desired

biodiesel (Naeem et al., 2016).

Comparative studies of Bioethanol Production from Different Fruit

Wastes

The depletion of fossil fuel impacts on the increase of

petroleum price and has triggered the finding of alternative and

renewable energy. Ethanol fuel is the most common biofuel

worldwide. Ethanol can be used in petrol engines as a replacement

for gasoline. The production of bioethanol must be increased

using cheaper and ecofriendly. Ethanol has a smaller energy

density than that of gasoline; this means it takes more fuel

(volume and mass) to produce the same amount of work. The first

generation of ethanol production used corn as substrate, later

corn was considered as a feedstock lead to the second generation

of production of ethanol which is used microorganism and

different waste as substrate.

11
 The ethanol produced from fermentation process by different

fruit wastes such as sweet lime, papaya, pineapple, watermelon,

and banana was compared in this study. The ethanol concentration

was estimated by alcohol extermination method/potassium

dichromate K2CR207 method/saccharomyces cerevisiae is used in

fermentation process since it converts sugars with oxygen to give

carbon dioxide. (Akalya et al., 2017)

Essential Chemicals in Selected Fruit Peels from Manila,

Philippines

According to this study by Judilynn N. Solidum, general

aimed to define the phytochemicals present in the peels of the

seven (7) fruits obtained from Manila, Philippines which is the

Rambutan, Lanzones, Pomelo, Longgan, Dalandan, Ponkan, and

Mangosteen. Definitely it aimed to define the presence of total

sugars and alkaloids. Further it intended to obtain the amounts

of total sugars and alkaloids in the fruit peel samples analyzed.

Based on the phytochemical screening was conducted, the 7

selected fruits showed the presence of reducing sugars such as

glycosides. For the part of Lanzones, its pH parameters is 6, and

its glycosides is white fine ppt. For is alkaloids, it resulted

in Mayer's, Brownish yellow solution with brown ppt, Valser’s is

Brownish yellow solution With red ppt, Wagner’s Reddish brown

12
solution with brown ppt, Dragendorff’s is Reddish orange solution

with brown ppt. For Plant acids, Light brown solution; no stable

and dense froth, for its Satd alcoholic solution of cholesteol

resulted Brown solution with brown ppt. with the concentration of

total sugars (ug/mL), the results for Lanzones in 20% decoction

>164, for 1:1 is 145.1, for 1:4 is 110.05, and for 1:9 is 39.4

and lastly for 1:99 is 1.801. Lanzones considered that it has a

highest intensity of carbohydrates and alkaloids and gave the

positive results to alkaloids.

Comparative Analysis of Single and Two Stage Fermentation Systems

under Various Process Conditions

In addition to the increasing need for fuels and energy in

industrial uses the demand for the production of bio-fuels from

lignocellulosic biomass and agro-industrial waste is increasing

Anaerobic fermentation that produces biogas from crop straw is a

potential solution to meet the demands and can help to solve

environmental pollution (such as burning). The process of biogas

fermentation can be divided into Semi-continuous or continuous

from the point of import and export mode. The concentration of

fermentation substrate that is needed depends on whether it is

wet fermentation, high concentration fermentation or dry

fermentation, thus the choice of fermentation process is vital

13
for the operational efficiency and stability of biogas

production. The period of anaerobic fermentation is usually 40-50

days for fibrous raw material under medium temperature conditions

when the TS concentration of the raw material is 20%, and the

period of high solid anaerobic fermentation is usually longer, 2

to 3 months. Or even upwards of 4 months or more.

The anaerobic biological conversion rate of wet fermentation

is higher compared to high solid fermentation, but the load of

raw material for the reactor per unit volume is low, which result

in a low utilization rate of the reactor. The biogas Production

from the single - stage net fermentation was 83.35% higher

compared with the single dry fermentation. This showed that the

net fermentation was more conductive in improving the production

of biogas. This biogas slurry can be added again during the stage

conversion in the wet-dry process which belong to the

replenishment operation. Can achieve reuse of mixed liquid slurry

after fermentation solid-liquid separation. (Du et al., 2017)

Biofuels from Lignocellulosic Biomass

The most promising biofuels are bioethanol, biodiesel, pure

plant oil, and bio methane. Many biofuels generate and has

benefits when compared to fossil fuels. The use of bioethanol

will largely depend on the potent ion of available and

14
continuously regrown biomass, the feedstock sources. Bioethanol

produced from renewable feedstock has great potential to reduce

greenhouse gases emission related to the combustion of fossil

fuels .Lignocellulose is a generic term for describing the main

constituents in most plants namely cellulose, hemicellulose , and

lignin. It is the non-starch based fibrous part of plant

material. The cumulative impacts of fuse concern have increased

the interest in developing biofuels produced from non-food

biomass feedstock from lignocellulose materials include cereal

straw, bagasse, forest. Lignocellulose biomass has an advantage

over other agriculturally important biofuels such as cornstarch,

sugar cane because it can be produced quickly and at

significantly lower cost than food crops.

Separation of Ethanol and Water by Extractive Distillation with

Salt and Solvent as Entrainer: Process Simulation

According to I.D. Gil et al., 2007, this study simulate and

analyze an extractive distillation process for azeotropic ethanol

dehydration with ethylene glycol and calcium chloride mixture as

entrainer. All of this study was developed using Aspen plus

Simulator, version 11.1. Activity coefficient calculation for the

mixture were made with the NRTLE thermodynamic model for

electrolytes system and validated with experimental data that was

15
reported in previous works (Zhigang et al. 2002). The researchers

used dehydration process and used the main extractive column and

the recovery column. In first dehydration column to which an

alcoholic solution, near to the azeotropic composition, is fed on

of the middle stages; and separating agent on one of the upper

stages. As a product, alcohol with a 99.5% molar concentration is

obtained, and as bottom product a mixture of water entrainer is

withdrawn. In second regeneration column, which is vacuum is

applied in order to separate water from the separating agent and

recycle it later to the dehydration process, while water with

some content of ethanol is recycled back to the rectification

step located in an upstream zone of overall. The thermodynamic

model chosen by the researchers fits properly the experimental

vapor-liquid equilibrium data for the mixture studied. Therefore,

the simulation results were supported on a solid thermodynamic

basis, which can describe the mixture behavior in a very accurate

way. This study establish operating condition for an extractive

distillation process using salt and solvent. Reflux ratio has the

greatest effect on energy consumption and it must be operated at

low values together with an entry solvent temperature between 70

and 90oC. Also, it concludes that it can be used to obtain high

ethanol concentration in the overhead product without implying in

important energy consumption.

16
Methods in Determining Diesel Cetane Number

Using the ignition delay principle, ASTM published an

updated ASTM D7170 test method for derived cetane numbers. It is

another way to determine diesel fuel’s cetane numbers

economically and more reliable. The cetane rating is a very

important property of diesel oils which indicates the ignition

quality of diesel fuels, the higher, the better. It is somewhat

analogous to octane for gasoline.

There are four methods in determining the cetane number of

diesel fuel. First is the use of cetane engine. The measurement

is direct once the fuel is setup and the engine is calibrated.

Another method is the IQT or ignition quality tester wherein the

delay of time is measured between the start of fuel injection and

the start of the significant combustion. The third method is the

near infra – red (NIR) analyzer, the obtained absorption spectrum

of diesel is fed to a chemometric model to estimate cetane. The

last method is the cetane indices where the measured cetane

number is estimated based in the formulas in ASTM D976 or ASTM

4737. The overall calculation will be based from the T10, T50,

and T90 distillation points and the gravity or the density of the

fuel. (Barsamian, 2009)

17
 Chapter 3

Introduction

This chapter shows or defines the methods that will be used

to conduct the study. The researchers will state the information

and data needed to elaborate their goal or objectives and

questions that will be answered during the whole research

process. The materials, the lanzones fruit, the equipment that

the researchers will be used for the variable. To see what the

lanzones fruit contains that can be used for bioethanol fuel.

This chapter will also state the research design, the test

that was used, the Gantt Graph, and the researcher's list of

schedules for how they have organized and planned a schedule of

process that researchers followed. The process not just the

methodology but also the researcher's task to achieve the whole

research process at a time table.

Materials, Tools, and Equipment

The following are the possible materials, tools, and

equipment that will be used for this research:

The main material that will be used in conducting this

research is lanzones (L. domesticum) fruit which will be air

dried and grind.

18
 The tools that will be used during the experimentation are

Erlenmeyer flasks, which will be used as a container for the

samples, graduated cylinders and beakers, which will be used for

measuring the extracted and fermented solutions.

The equipment that will be used for this study is blender

machine for the grinding of lanzones fruit.

Experimental Procedure

The researchers will gather the tools and materials that

will be used for the experimentation. It will be peeled, dried

and then ground. The researchers will undergo on different

processes:

Pre-treatment

The ground lanzones will be mixed with hydrochloric acid

(HCl) and will be sterilized at 75 degree Celsius for 15 minutes

and then measure its pH value.

Figure 1.1

19
Fermentation

The pre-treated lanzones solution will be fermented from

different parameter: Fermentation in different pH parameter,

different temperature, and different period.

Figure 1.2

Filtration

After the lanzones (L. domesticum) fruit fermented, it will

undergo to filtration process to separate the fermented extract

from any residue.

Figure 1.3

20
Distillation

After the lanzones fermented and filtrated, it will undergo

to distillation process to separate the ethanolic content from

water.

Figure 1.4

The fermented and distilled seed extract will be then send

to Department of Energy – Renewable Energy Bureau for them to

test the distillation properties, its density using the specific

gravity test and its Cetane rating.

Figure 2.1 Figure 2.2

21
Figure 2.1 Setup A contains the mixture of bioethanol fuels with

the additive produced from the lanzones (L. domesticum) fruit.

The DOE – REB laboratory will test its distillation properties,

its density using the specific gravity test and its Cetane

rating.

Figure 2.2 Setup B contains only the bioethanol without the

additive produced from the lanzones (L. domesticum) fruit. The

DOE – REB laboratory will test its distillation properties, its

density using the specific gravity test and its Cetane rating.

Data Gathering Procedure

The said data will come from the results gathered by the

researchers. The researchers will then compare the data gathered

from the laboratories where it is tested.

Statistical Analysis

The researchers will give the Department of Energy

Laboratory the permission to analyse and test the fermented and

distilled seed extract’s density / specific gravity and Cetane

Rating. The researchers will also test the effectiveness of the

fermented seed extract as an additive for the bioethanol fuel.

22
 The researchers will use the formula for t-test for the data

analysis:

where

x−μ
t=
sx

where

s
sx= ¿
√n

μ = Proposed constant for the population mean

x = Sample mean

n = Sample size

s = Sample standard deviation

x
s x = Estimated standard error of the mean ( )
√n

23
 Chapter 4

This chapter includes the data gathered by the research

throughout the whole experimentation process.

The researchers aren’t able to finish the whole

experimentation process that they needed to gather the data

necessary for this research. They’re done the first part of the

experimentation in which they pre-treated the lanzones fruit with

a 12mL diluted hydrochloric acid and sterilized it in 75 degree

Celsius for 15 minutes. The researchers are considering the

following factors that affected their research for not to be able

to finish:

 Financial issues. The researchers are supposed to use a

different plant material for this research and are supposed

to send the said material to a laboratory but the only

laboratory that is available is the University of the

Philippines in Los Baños, Laguna which is too far from their

location and they don’t have enough finance to support them.

 The availability of plant material. The researchers then now

decided to conduct the fermentation themselves but another

problem aroused, the plant material that they needed for the

research is not available for it is seasonal. The

researchers tried to find another source of it but it is

said to take a long time for it to arrive.

24
 Lack of time for the experimentation. The researchers

decided to use another plant material for the additive

production but the problem is the time frame of their study.

Due to the extensions and the problem occurred they only

have at least 2 weeks to finish the research.

 The pH value of the pre-treated solution. Fermentation is an

anaerobic process that breaks down the sugar molecules into

simpler molecules. It is affected by the different factors

like the pH value. pH value affects the shape of the

enzymes. Fermentation by yeast will be faster at a lower pH

values. It is suggested that the best range for fermentation

in yeast is 4.4 – 5.5. (Bloomingdale et. al., 2018) The

researchers are unable to produce a solution that has the pH

value suggested before they proceed to the fermentation.

25
 Chapter 5

This chapter includes the conclusion and recommendation done

by the researchers.

Conclusion

The researchers throughout the whole processes that they've

undergo, therefore conclude that bioethanol fuel additives is a

research that needs a lot of funding, effort and time.

Fermentation is a long process counting a couple or more weeks.

Also it takes lots of time in Distillation processes, before

undergoing to test its number of cetane rating and specific

gravity. The researchers failed to do the process because of

lacking source of variable the main reason why to change its

variable as soon as possible.

Recommendation

The researcher, recommend the following for the future

researchers to achieve best results in their future researches:

 Time Management. The researchers must have a time management

to ensure that they can perform their different test;

26
 experiment and can do their research paper. Time management

is very important.

 Examine, observe keenly whenever conducting experimentations

 Work in an organized and clean space because it doesn’t only

improve workflow by saving steps, it also forces you to

think through the different aspects of your operation, and

how to approach them efficiently and systematically.

 The future researchers should have enough money, or it is

best if the researchers should save money beforehand, for

the future expenses such as materials, equipment and for

laboratory testing.

 Seek help from Research Advisers. The researchers should

seek further help from their research advisers as the

research continues its process, consistent consultation, ask

for advice and guide, in a proper time and proper manner.

Appendices

27
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