26

SAMPLE RESEARCH

FIRST PLACE WINNER

REGIONAL SCIENCE FAIR COMPETITION
REGIONAL OFFICE
DEPED, CANDAHUG, PALO, LEYTE
OCTOBER 2-3, 2010

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EXTRACTION OF ETHANOL FROM CORN (Zea maize) STALKS

A Research Paper Presented to

REGIONAL SCIENCE FAIR 2010

September 11-12, 2010
DepED, Candahug, Palo, Leyte

JANELLE S. SARVIDA, II-MG

RESEARCHER

SEPTEMBER 2010

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TABLE OF CONTENTS

Title Page

Cover Page

Table of Contents i

Acknowledgement ii

Abstract iii

CHAPTER I – INTRODUCTION 1
Background of the Study 1
Objectives 2
Null Hypothesis 2
Significance of the Study 2
Definition of Terms 3
Scope and Delimitations 4

CHAPTER II – REVIEW OF RELATED LITERATURE 5

CHAPTER III – METHODOLOGY 11

Materials and Equipment 11
Treatments and Variables 11
General Procedure 11
Data Gathered 13
Data Processing 14

CHAPTER IV – RESULTS, ANALYSIS AND INTERPRETATION OF DATA 16

CHAPTER V – CONCLUSION AND RECOMMENDATIONS 18

Conclusion 18
Recommendations 18

BIBLIOGRAPHY 20

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ABSTRACT

EXTRACTION OF ETHANOL FROM CORN (Zea maize) STALKS

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Keywords: Corn stalks, ethanol, mechanical extraction, filtration, fermentation and
distillation.
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This study is a preliminary evaluation of the feasibility of extracting ethanol from

corn (Zea maize) stalks mostly left by the farmers in the field after harvest through
mechanical extraction, filtration, fermentation and distillation.

Sugar level determination, a critical factor in ethanol production was done using
a refractometer. Two weekly sampling trials were made to determine the level of
ethanol yield using 10 gms and 20 gms yeast treatment during fermentation.

Ethanol yield per 100 ml juice extract 1 week after extraction (WAE) were: 12%
in 10 gms and 15% in 20 gms yeast treated samples. Correspondingly, 14% in 10 gms
and 22% in 20 gms per 100 ml juice extract yeast-treated samples were recovered 2
WAE. While the control recorded only 2.0% and 2.5% ethanol yield 1 and 2 WAE,
respectively or with an average yield of only 2.5%.

Ethanol production increases through yeast application and as the storage period
lengthens.

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ACKNOWLEDGEMENT

This study would not be possible without the Divine Guidance from our Heavenly

Father and some individuals encountered by the researcher in course of her study:

● Mr. and Mrs. Gonzalo Sarvida (my parents), for the moral and financial support;

and for gathering and preparing the corn stalks for juice extraction;

● Mrs. Ermelinda Floretes and Mr. Romeo Floretes for the big help in making this

study possible and in developing this study; for helping me in providing the

materials needed in this study;

● Mr. Aylmer Arellon (our Research teacher) for the guidance and being my

inspiration in conducting this study;

● Mr. Ketz Throy Arteche and Mr. John Romuelle Eclipse (MG-III) for

accompanying me in conducting this study and the helping me in preparing the

materials needed;

● Mr. Hilario Alberto Aguirre (MG-III) for allowing me to ride with them with the

use of their car going to D.A, Regional Integrated Agricultural Research Center,

Brgy. Anquiana, San Jorge, Samar last August 6, 2010 for the juice extraction;

● Department of Agriculture, Regional Integrated Agricultural Research Center,

Brgy. Anquiana, San Jorge, Samar for the extraction of juice from harvested corn

stalks with the use of their Micro-mill and for determining the sugar level with

the use of their Refractometer;

● Mrs. Sadullo for allowing me (with the company of Mrs. Floretes and other MG-
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III students) to use her room (DOST 2) for the distillation process.

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CHAPTER I

INTRODUCTION

Background of the Study

As the Philippines increases its population and strives to develop its resources its

demand for fuel and energy also remarkably intensifies thus spending its dollar reserve

in importing fossil fuel worth millions of dollars just to ease the energy crisis it is facing

today. Consequential to this unbalanced trade is the dramatic increase of prices of its

basic commodities, considered a big burden to most of its people who can hardly meet

both ends in their daily survival.

The country’s high dependence on imported fuel is limiting its financial resources

that could be otherwise spent on viable developmental projects to improve the health

and lifestyle of the Filipinos.

Ethanol is largely used as a motor fuel and fuel additive. It is also considered an

important industrial ingredient and has widespread use as a base chemical for other

organic compounds and ethanol is used in medical wipes and in most common

antibacterial hand sanitizer gels. Ethanol can also be used as an antidote for poisoning

by other, more toxic alcohols, in particular methanol and ethylene glycol. It is found in

paints, tinctures, markers, and personal care products such as perfumes, deodorants

and also used as alcoholic beverages and solvent in cooking, such as in vodka sauce

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(Myers, 2007).

Given this outlook, the researcher was enthused of coming up with this timely

and very relevant study to explore the possibility of ethanol production from harvested

corn stalks which are considered waste products after harvesting. It is therefore

significant to carry out this kind of investigation because in one way or another, this

research project study will lessen the country’s dependence on importing fuel.

Objectives:

1. To determine feasibility of extracting ethanol from corn stalks.

2. To assess the presence of sugar from extracted corn stalk juice.

3. To measure and compare the ethanol yield in the various stages of

fermentation as influenced by the addition of yeast.

Null Hypothesis:

Ho: There is no significant difference of ethanol yield between the control and

yeast-treated corn stalk juice.

Ha: There is a significant difference of ethanol yield between the control and

yeast-treated corn stalk juice.

Significance of the Study

The current energy crisis is one of the major contributors to this research. The
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speedy increase of commodities in the global market confuse consumers where to get

extra income to this added burden. It is therefore a necessity to remedy to the existing

present crisis where consumers are now crabby due to the great effects on various

items. As a way out, this study aims to conduct a test on the feasibility of ethanol in

corn stalks where ethanol is extremely significant nowadays. The largest single use of

ethanol is as a motor fuel and fuel additive, with this study this gives everyone an

overview that after harvesting corn, instead of throwing the stalks which can only add

to the bulk of waste in the environment, these waste can be converted into something

very vital to everyone which is the ethanol. This study will yield information

regarding the use of harvested corn stalks. Consequently, the result of this study would

be very useful because this can be considered as an initial phase to scale up the

production of ethanol from corn stalks, and this study can also be an avenue in solving

the present problem on energy.

Definition of Terms:

Bagasse = corn stalks refuse after crushing.

Corn = plants that yield grain or maize.

Corn stalks = the stem or body of corn plant.

Distillation = is the process in which a liquid or vapor mixture of two or more

substances is separated into its component fractions of desired purity by the application
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or removal of heat.

Extracted Corn Stalks Juice = is the juice collected from corn stalks after

extraction.

Fermentation = is the process of deriving energy from the oxidation of organic

compounds, such as carbohydrates, and using an endogenous electron acceptor, which

is usually an organic compound, as opposed to respiration where electrons are donated

to an exogenous electron acceptor, such as oxygen, via an electron transport chain.

Refractometer = an instrument used to measure the level of sugar.

Micro mill or Juice Extractor = an instrument used to crush the corn stalks to extract

the juice.

Yeast = a fungus that causes alcoholic fermentation

Scope and Delimitations:

This study was conducted at the Department of Agriculture, Regional Integrated

Agricultural Research Center, Brgy. Anquiana, San Jorge, Samar and in the DOST 2

Chemistry laboratory at Samar National School from July-August 2010.

Juice extraction from freshly harvested corn stalks was done in the field using

micro mill juice extractor and the determination of the presence of sugar level was
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made using a refractometer. After fermentation and distillation the researcher

computed only the ethanol yield 1 and 2 weeks after extraction and did not further

attempt determine the percent alcohol content and bioassay of the product.

CHAPTER II

REVIEW OF RELATED LITERATURE

Ethanol is a renewable energy source because the energy is generated by using

a resource, sunlight, which is naturally replenished. Creation of ethanol starts with

photosynthesis causing a feedstock, such as sugar cane or corn, to grow. These

feedstocks are processed into ethanol. About 5% of the ethanol produced in the world

in 2003 was actually a petroleum product. It is made by the catalytic hydration of

ethylene with sulfuric acid as the catalyst. It can also be obtained via ethylene or

acetylene, from calcium carbide, coal, oil gas, and other sources. Two million tons of

petroleum-derived ethanol is produced annually. The principal suppliers are plants in

the United States, Europe, and South Africa. Petroleum derived ethanol (synthetic

ethanol) is chemically identical to bio-ethanol and can be differentiated only by

radiocarbon dating.

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(Klein, Donald W.; Lansing M.; Harley, John (2004). Microbiology (6th ed.). New York:
McGraw-Hill. ISBN 978-0072556780. http://highered.mcgrawhill.com/sites/0072556781/
information_center_view0/ ).

Sweet sorghum (Sorghum bicolor) is similar to grain sorghum with a sugar-rich

stalk, almost similar to sugarcane. Besides having wide adaptability, rapid growth and

high sugar accumulation and biomass production potential, sweet sorghum, is tolerant

to drought, water logging, soil salinity and acidity toxicity. It has great potential for

jaggery, syrup and alcohol (most importantly Gasohol, which is ethanol blended with

petrol) production. The sugar content in the juice extracted from sweet sorghum varies

from 16-23% Brix.

In recent years, there is an increased interest in the utilization of sweet sorghum

for ethanol production in India as its growing period of about four months, and water

requirement of 8,000 cubic meters (cu m) over two crops are one-fourth that of

sugarcane, which has a growing period of 12 to 16 months and a water requirement of

36,000 cu m per crop. In addition, sweet sorghum is better suited for mechanized crop

production and seed propagation.

According to a pilot study conducted by VSI, sweet sorghum is the best

alternative raw material, which can supplement the use of sugarcane in ethanol

production. At 5,600 liters per hectare per year (over two crops, at 70 tons per hectare

of millable stalk per crop at 40 liters per ton), the ethanol production from sweet

sorghum compares well with the 6,500 liters per ha per crop for sugarcane (at 85-90
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tons per hectare of millable cane per crop at 75 liters per ton).

According to estimates made by National Research Center for Sorghum (NRCS),

Hyderabad-India, the per liter cost of production of ethanol from sweet sorghum is Rs

13.11* (at Rs 500 per ton of stalk), when compared to Rs 12.55* (at Rs 1,600 per ton

of stalk) from sugarcane molasses. However, the increased cost of production of

ethanol from sweet sorghum is more than compensated by grain yield of 1 ton per

hectare (which can be used as food or feed) and the superior quality of ethanol. The

really significant advantage is that the production of ethanol from sweet sorghum is

environment friendly since it uses the non-molasses route. (www.practicalaction.org).

Sugar cane must be crushed to extract the juice. The crushing process must

break up the hard nodes of the cane and flatten the stems. The juice is collected,

filtered and sometimes treated and then boiled to drive off the excess water. The dried

cane residue (bagasse) is often used as fuel for this process. The remaining liquid is

allowed to set into a solid mass known as jaggery, gur, chancaca or panela. (Gur is

used in the rest of this document.) The yield of gur from sugar cane depends mostly on

the quality of the cane and the efficiency of the extraction of juice. High quality cane

has a good juice content with high sugar levels (20%+). Poor quality cane or cane that

has been harvested early may have similar juice content but the sugar levels will be

reduced.

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The efficiency with which juice can be extracted from the cane is limited by the

technology used. The simple three roller crushers used by most artisanal producers will

never extract more than 50 kg of juice from each 100 kg of cane.

Yields

Yields are also improved by careful control of the boiling process. Boiling should

be completed as rapidly as possible and the conditions kept as clean as possible.

Crushing

Most artisanal sugar producers use a simple crusher consisting of three metal

rollers. This is driven by either animal or diesel power. A crusher driven by a single ox

can be expected to process around 50 kg of cane per hour. A 5HP diesel set could

increase this to around 300 kg per hour. Important points to remember during crushing

are, namely: 1) Cane must be crushed within 24 hours of being cut. After this time the

sugar begins to ‘invert’ into different sugars that will not set solid, and; 2) Crushing

efficiency is the most important factor in good sugar yields. Every possible amount of

juice needs to be squeezed from the cane.

Juice Treatment

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Juice should be filtered through a cloth before boiling in order to remove any

solids such as dirt or particles of cane.

Large-scale sugar processors add lime to the juice in order to coagulate

impurities which then settle out. This is rarely done at the artisanal level. The juice is

then neutralized with sulphur dioxide. Small-scale producers add a variety of clarificants

to the juice including wood ash. All of these have the effect of settling out impurities.

Many producers also add ‘Hydros’ (sodium hydrogen sulphate) at the final stages of

boiling. This releases sulphur dioxide into the juice and lightens the color of the final

product. However, high sulphur content often remains in the final product.

Juice Boiling

This is done in large pans over open fires or simple furnaces. The essential

requirement is for clean pans and tools. Sediment settles to the bottom of the pan

during boiling and is dredged out. Scum rises to the top and is skimmed off.

The end point of the boiling process is judged from experience; from the sight

and sound of the boiling juice. Small samples can be removed to see if they set solid

when cooled. For those with access to simple sugar measuring devices, this usually

corresponds to a Brix (sugar content) of 90-95%.

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After removal from the heat, the pans of juice are usually stirred rapidly to

incorporate air and promote an even crystallisation. The cooling juice is then poured

into pots or moulds to set.

Cleanliness

Cleanliness is vital to the whole process. Once the juice has been heated,

impurities will speed the ‘inversion’ of sugar and lead to reduced yields. All boiling pans

and tools need to be thoroughly cleaned between uses.

Tools

The tools required are very simple Filtration before boiling is done through a fine

woven cloth. Scum is removed from the boiling juice by a simple perforated scoop on a

long handle. Sediment is removed by scraping a stretched cloth along the bottom of the

pan. Once the pan has been removed from the heat, a simple rake is used to stir the

thickened juice. (www.practicalaction.org, photo from ecofriend.org)

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CHAPTER III

METHODOLOGY

Cork
Test tube
Alcohol lamp
Denatured alcohol
Iron stand, ring and clamp

Materials/Equipment

Corn stalks
Micro mill or juice extractor
Refractometer
Yeast
1000-ml and 250-ml beakers
Distilling flasks

Treatments and Variables

T0 = 0-gm yeast; corn stalk juice only

T1 = 10-gm yeast treated corn stalk juice
T2 = 20-gm yeast treated corn stalk juice

General Procedure

Gathering and Preparation of Materials. Freshly harvested corn stalks were

obtained from Brgy. Tagbayaon, Jiabong, Samar. The corn stalks were immediately

cleaned by removing the dried and fresh leaf sheaths. After which the stalks were cut

into pieces by about eight inches (8”) long to facilitate easy extraction, and then the

samples were weighed.

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Extraction. The extraction was done in the Department of Agriculture, Regional

Integrated Agricultural Research Center in Brgy. Anquiana San Jorge, Samar. The micro

mill or juice extractor was cleaned by wiping using a cheese cloth and the strainer were

washed using clean water. After cleaning the micro mill were set up ready for

extraction. Four (4) stalks at a time of about eight (8) inches long were feed in into the

micro mill to extract the juice and the stalks were feed in into the extractor four times

to fully extract the juice or until no more juice coming out from the extractor.

Filtration. The extracted juice was initially strained by passing through a screen

strainer to remove solid particles from corn stalks. After passing through the strainer it

was further processed and filtered using filter paper to remove the fine particles

embedded in the juice.

Measuring the Filtered Juice and Sugar Level Determination. The volume

of the filtered juice was measured using a 250-ml graduated cylinder and its sugar level

was assessed using a refractometer. For the sugar level determination two (2) drops of

corn stalk juice were placed in the lid of refractometer making sure that no bubbles will

appear and it was repeated three times and the mean for the three trials were

computed to assure accurate results.

Fermentation. The corn stalk juice extract were then prepared for

fermentation. The three treatments previously defined were applied: T0 = no yeast

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added; T1 = ten (10) grams yeast; and T2 = twenty (20) grams yeast. The three

treatments were stored at ambient conditions and fermented for a period of one and

two weeks. Note: Fermentation is the natural metabolic process that produces energy

by breaking down carbohydrates (like sugars) in the absence of oxygen. It is catalyzed

by the action of enzymes present in microorganisms like yeasts with ethyl alcohol as the

end product.

Distillation. Two weekly sampling trials were made during fermentation to

determine the alcohol content of the product. During the weekly sampling, distillation of

the fermented corn stalk juice was made by transferring the fermented corn stalk juice

to a distillation process where the ethanol is separated from the remaining stillage

(residue non-fermentable solids and water). Distillation is the process in which a liquid

or vapor mixture of two or more substances is separated into its component fractions of

desired purity by the application of heat. This process can usually produce ethanol

product.

Data Gathered

After the distillation the ethanol recovered from corn stalks was measured

and the percent yield of the ethanol was then computed using the formula:

Percent of ethanol yield = Volume of ethanol (ml)______ X 100

Volume of the extract sample (ml)
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The process was used from the first to second trials both in the control and

yeast-treated samples. In measuring the percent yield of the ethanol, the researcher

used the available graduated cylinder.

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Gathering and Preparation of Materials

FLOW CHART

Cleaning & cutting of corn stalks (30 minutes)

Extraction Procedure

Pressing (60 minutes)

Filtration

Straining & filtering (15 minutes)

1.
Measuring the Filtered Juice and Sugar Level Determination

Refractometer reading (15 minutes)

Treatment Application and Fermentation

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1 & 2 –week storage and sampling

Distillation

Distilling 60 minutes

Measuring of Recovered Ethanol and Determining the Percent Yield

Figure 1. Flowchart of the general procedure of the study.

Data Processing

The results were gathered and tabulated. Quantitative analysis was done based

on comparison of ethanol yields between the untreated and yeast treated samples.

Arithmetic mean or mean was used to determine the average of the percent yield of

ethanol.

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CHAPTER IV

RESULTS, ANALYSIS AND INTERPRETATION OF DATA

Table 1 shows the amount and level of sugar extracted from the 2,500 grams of

corn stalks. After the pressing the corn stalks the bagasse weighed 1,920 g and the

volume of extracted corn stalk was 560 ml. Three readings of sugar level were

conducted and a mean of 10.5 was computed. Based on the results of sugar level

determination, it is assumed that ethanol could be extracted from corn stalks as any

plant or grass containing sugar, the presence of ethanol is possible.

Table 1. Amount and Sugar Level of the Sample.

Volum Readings
Weight
Total e of of the
of
MEDIUM weight juice Amount
bagasse
(g) extract of Sugar
(g)
(ml) Level
1st 2nd 3rd MEAN
Corn
2,500 1,920 560 11 10.5 10 10.5
Stalk

A comparative mean ethanol yield 7 DOF and 14 DOF were shown in Table 2 and

Figure 2. The percentage of ethanol recovered were 2.5%, 13.0% and 18.5% from T0,

T1, and T2, respectively. It could be observed that the volume of ethanol recovered

increases as the amount of yeast is increased. Addition of yeast to the corn stalk

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extract speed-up the chemical process of fermentation that make ethanol extraction

feasible.

The length of fermentation period had also influenced on the percentage of

ethanol recovery as reflected in the same table. However, the percent ethanol content

per treatment per sampling period was not determined due to limitations of chemicals

and facilities.

Table 2. Comparison of ethanol yields between the control (untreated) and yeast-
treated samples per 100 ml of Juice 7 and 14 days of fermentation (DOF).

Treatments Ethanol
yield (%)
7 DOF 14 DOF Mean

T0 - Control 2.0 3.0 2.5

T1 - 10-gm yeast 12.0 14.0 13.0
T2 - 20-gm yeast 15.0 22.0 18.5

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Figure 2. Comparison of ethanol yields between the control (untreated) and yeast-
treated samples per 100 ml of Juice 7 and 14 days of fermentation (DOF).

CHAPTER V

CONCLUSION AND RECOMMENDATIONS

Conclusion

The presence of sugar in corn stalk juice indicated that ethanol can be extracted

from freshly harvested corn stalks. The ethanol yield from this otherwise wasted

resource could significantly contribute to the development of ethanol industry and

paints a promising future for the corn farmers as they could be able to develop a new

alternative source of income from their farm by products.

Addition of yeast to corn stalk juice and lengthening the period of fermentation

had significantly increased ethanol yield and/or production.

The impact of the study on the utilization of corn stalk as source of ethanol could

not be understated and offered an avenue to provide the growing demand for ethanol

for particular and the alcohol industry of the country in general.

Recommendations

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To further improve the study the researcher recommends the following:

1. Conduct of confirmatory, replicated studies and statistical analysis on the

level of significance of the treatments and variables used.

2. Application of new and efficient technology on corn stalk juice extraction.

3. Conduct of feasibility study on the large-scale corn stalk juice extraction

and ethanol production to further explore the economic viability of the process.

4. Assessment on the enhancement of the physico-chemical properties that

determines the percent alcohol content of the product and microbial studies to the

effectiveness and efficacy of ethanol from corn stalk should be done.

5. Conduct of exploratory studies on other plants or grasses apart from corn

stalk is also recommended.

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BIBLIOGRAPHY

1. Klein, Donald W.; Lansing M.; Harley, John (2004). Microbiology (6th ed.). New York:
McGraw-Hill. ISBN 978-0072556780. http://highered.mcgraw-
hill.com/sites/0072556781/information_center_view0/

2. Myers, Richard L.; Myers, Rusty L. (2007). The 100 most important chemical compounds:
a reference guide. Westport, Conn.: Greenwood Press. p. 122. ISBN 0313337586.
http://books.google.com/?id=0AnJU-hralEC&pg=PA122.

3. PhD thesis on algae production for bioenergy) (PDF). Murdoch University, Western
Australia. http://wwwlib.murdoch.edu.au/adt/pubfiles/adt-MU20050901.140745/
02Whole.pdf

4. Root Crop Digest Vol. 2 No. 1 1987 PRIS ISSN 0116-4325

5. Roy Robison, Donald B. White, and Mary H. Meyer: Plants in Prairie Communities.
University of Minnesota, 1995.

6. www.practicalaction.org

7. Webster’s Universal Dictionary and Thesaurus 2002

2.

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