CHAPTER 3

Research Methodology

This chapter consists of the research methods and procedures that will be done in order

to reinforce banana pseudo-stem nanocellulose fiber for the development of bioplastic bubble

wrap (BPBW) to address the problem of packaging wastes and promote green commerce.

RESEARCH DESIGN

An experimental type of research design will be employed in this study. The study will

also involve a single factorial experimental design in which the proportion of banana

pseudostem NCF to PVA and glycerol will serve as the determining factor in the experiment.

The study will also determine the lignocellulosic profile of the banana pseudostem as the source

for the nanocellulose to be synthesized with the selected resin and plasticizer.

The study will evaluate the effect of varying the proportions of the banana pseudo-stem

NCF, PVA, and glycerol in producing the optimal NCF for developing BPBW. The comparison

analysis will be utilized to determine the difference between the BPBW’s physical and chemical

properties and the commercially available petroleum-based Vermatex bubble wrap. The process

will be performed three times for validity and reliability of results. The figure on the next page

shows the schematic diagram of the research flow process.

Reinforced Banana Pseudo-stem Nanocellulose Chapter iII: Research Methodology
Fiber for the Development of Bioplastic Bubble Wrap
Abarintos, M.L.B., Bacquian, M.J.F., Banta, J.B., Castillo, K.A.C.

Figure 3.1 Research Flow Process

Reinforced Banana Pseudo-stem Nanocellulose Chapter iII: Research Methodology
Fiber for the Development of Bioplastic Bubble Wrap
Abarintos, M.L.B., Bacquian, M.J.F., Banta, J.B., Castillo, K.A.C.
MATERIALS AND REAGENT

The raw material, which is banana pseudostem, will be procured locally from the banana

farm in Nasugbu, Batangas. The reagents such as Sodium Bicarbonate, Acetic Acid, and

glycerol, sodium hydroxide, silicon oil and litmus paper, will be bought from the DKL Laboratory

Supplies. The remaining reagents, Sulfuric Acid and Hydrochloric acid will be acquired from

Belman Laboratories.

The equipment for testing such as the analytical balance and graduated cylinder(for

specific gravity testing) , Shore Instron Durometer, type D(for hardness test), Model 5585H

Instron Universal Testing Machine(for tensile and elongation strength), test tubes(for thermal

stability), and Fourier Transform Infrared (FTIR) Spectrometer(FTIR Spectral Analysis) will be

borrowed and done from the DOST, UPLB, or Batangas State University Laboratories.

EXPERIMENTAL PROCEDURE

Extraction of Fibers from the Banana Pseudostem

The banana pseudostem will be placed through the decorticator where the banana

pseudostem will be shredded to expose and extract the fiber. It will be done in a safe and

controlled environment of the DOST Facility. After the extraction, the fibers will sun dried before

storage.

Retting of the Fibers

1. Soaking the Fiber in Sodium Bicarbonate Solution

In a container, the extracted fibers will then be soaked in a 12% wt Sodium Bicarbonate

solution for 120 hours. This solution percentage and time will be the most ideal for the best

tensile strength(Bakri, 2018). After that time, it will be strained and washed with clean water to

rinse the solution away, then will be sun dried again. The researchers will do this step in the

Batangas State University Laboratory.

Reinforced Banana Pseudo-stem Nanocellulose Chapter iII: Research Methodology
Fiber for the Development of Bioplastic Bubble Wrap
Abarintos, M.L.B., Bacquian, M.J.F., Banta, J.B., Castillo, K.A.C.

Degumming of the Banana Pseudostem Fiber

After the retting of the fibers, the researchers will proceed to the degumming. In the

degumming process, the researchers will coordinate with the DOST for the small-scale natural

degumming machine. The acquired materials will be cleaned again with clean water.

Acid Hydrolysis

After degumming, the fibers will then undergo acid hydrolysis to extract the

nanocellulose. The researchers will utilize 72% wt sulfuric acid for this experiment to accord with

Chen’s(2015) work. The fibers will be soaked in the acid for 30 minute at 373 K and then diluted

with boiling water to reach 33% wt concentration and will be continued to soak at the condition

for 60 more hours. After the time, it will be cooled down then strained after. It will then be sun

dried again.

Plasticization and Resin Reinforcement

The researchers will then go again to the DOST Office Advanced Manufacturing Center

for plasticization wherein 50 grams of the powdered nanocellulose will be dissolved in a 1%

acetic acid as the film solvent. Pure glycerol will be added to the solution. It will be then agitated

until it becomes homogeneous and then sundried for 48 hours in an aluminum casting film. The

dried film will be then carefully peeled out. This method is based on the study of Aguila, Alvar, &

Sandoval(2017). After plasticization, the film will be soaked in pure polyvinyl alcohol. There will

be a total of three setups in this experiment where in the nanocellulose:glycerol:PVA ratios are

50:25:25, 70:15:15, and 90:5:5.

Optimization of the Bioplastic Bubble Wrap

After the reinforcement process, the DOST Office Advanced Manufacturing Center will

then utilize the procured products to be converted into bubble wraps.

Reinforced Banana Pseudo-stem Nanocellulose Chapter iII: Research Methodology
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Abarintos, M.L.B., Bacquian, M.J.F., Banta, J.B., Castillo, K.A.C.

ANALYTICAL TESTING

The analytical testing for this study will either be performed at Batangas State University

Laboratories or DOST Laboratory depending on the availability of equipment that are needed for

each test. Each sample will be triplicated in every test to ensure accuracy in obtaining data and

results.

Determination of Cellulose and Hemicellulose of Banana Pseudo-Stem

The determination of the lignocellulosic profile of banana pseudo-stem, specifically the

cellulose and hemicellulose content, will be done using the procedure similar to the study by Li

et. al. (2010). The layered outer bark will be separated from the pseudo-stem by hand then the

remaining layers will be air-dried. The sample that will pass the 40 mesh screen and will be

subjected to the tests. The alcoholic nitric acid solution will be prepared by mixing one volume of

65 wt.% solution of nitric acid and four volumes of 96% purity ethanol. The cellulose content of

the BPS will be determined by following the Kurschner-Hoffner approach, where a gram of

sample will be treated with 25mL of alcoholic nitric acid under reflux at four cycles for 1 hour.

After each cycle, the solution will be removed to be replaced in fresh volume. Holocellulose, on

the other hand, will be determined using the method of sodium chlorite. In this method, the

solution will be prepared by mixing 65mL distilled water, 0.5mL acetic acid, and 0.6g of pure

sodium chlorite. The 2g sample will be subjected to the solution for one hour at 75℃. The test

will be repeated until the sample turns white in color. In order to determine the final content of

holocellulose, the ash content of the sample must be defined as well. The ash content will be

determined by conforming with ASTM D2584 or the ash test where the sample will be subjected

to a temperature higher than 500℃ in air atmosphere. The procedure will utilize a pre-weighed
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crucible which will hold the sample and the ash residue. The ash residue will be weighed and

the ash content will be calculated by:

mass of the ash

%ash= ×100 %
mass of sample

The ash content will be subtracted to both the cellulose and holocellulose content to arrive at

the final cellulose and holocellulose content since the banana pseudo-stem is known for high

ash content.

Determination of Nanocellulose Fiber Characterization and Percent Yield

The NCF will be characterized using an electron microscopy test to ensure that the

extracted fiber is in nanoscale. For the percent yield, the sample banana pseudostem will

initially be weighed using an analytical balance. The extracted nanocellulose-fiber will also be

weighed in the same analytical balance for the determination of nanocellulose fiber percent yield

using the equation used by Landicho et. al. (2012):

MA
% yield= ×100 %
MO

where M A is the mass of the actual extracted nanocellulose fiber and M O is the mass of

the banana pseudo-stem where the nanocellulose fiber was extracted.

Determination of the Characteristics of Bioplastic Bubble Wrap Samples

The BPBW samples with varying NCF-PVA-Glycerol ratio along with the commercially

available bubble wrap (Vermatex) will each undergo the identical tests in order to accurately

gather the data that is needed for the study.

Color

The color of the BPBW and Vermatex will be characterized qualitatively and

determined through visual inspection.

Elongation and Tensile Strength

Reinforced Banana Pseudo-stem Nanocellulose Chapter iII: Research Methodology
Fiber for the Development of Bioplastic Bubble Wrap
Abarintos, M.L.B., Bacquian, M.J.F., Banta, J.B., Castillo, K.A.C.
The elongation of the BPBW and Vermatex will be determined using tensile

strength and elongation test where in the machine called Model 5585H Instron Universal

Testing Machine (UTM) with the crosshead speed of 50mm/min with 50mm as gauge

length will be utilized. The process is done at the DOST Laboratory under the process

defined as ASTM D882. Both the tensile strength and elongation will be tested at break

which means that the samples’ ability to resist breaking under tensile stress will be

tested. The tensile strength and percent elongation at break will be calculated using the

equation used by Aguila et. al. (2015) which is as follows:

load at break
tensile strength( MPa)=
thickness ×width

position at break
% elongation= ×100 %
gauge length

Hardness

The hardness of the materials will be measured using the standard procedure

called ISO 868. The procedure utilizes the Shore Instron Durometer type D where the

sample is placed on a hard-flat surface and the indentor of the said instrument will press

the sample in order to make sure that it is parallel to the surface. The hardness of the

sample will be determined in one second of firm contact with the specimen.

Density

The density or specific gravity of the material can be determined using the

procedure ASTM D792/ISO 1183. The mass of the sample will be measured using the

analytical balance and the volume will be determined by submerging the sample in the

graduated cylinder with 20mL of water. The change in level of water in the graduated

cylinder is the volume of the sample. Using the mass and volume obtained, the specific

gravity will be calculated the same way as Aguila et. al. (2015) did in their study which is:

mass of sample
Sp .Gr=
volume of sample∗density of water
Reinforced Banana Pseudo-stem Nanocellulose Chapter iII: Research Methodology
Fiber for the Development of Bioplastic Bubble Wrap
Abarintos, M.L.B., Bacquian, M.J.F., Banta, J.B., Castillo, K.A.C.
Water Absorption

The water absorption will be determined using the standard ASTM D570 where

the sample will be dried in the oven for 5 hours and stored in the dessicator. It will be

submerged in a graduated cylinder with distilled water for three days. The moisture

uptake can be determined by weighing the samples in specific intervals as cited in

Landicho et. al. (2012).

Biodegradability

The biodegradability of each sample will be determined using two methods:

Composting/Soil Burial and Marine Floating Condition and Degradation.

Composting/Soil Burial

Composting or soil burial will be accomplished using a laboratory simulation

where a piece of sample will be buried in a loam soil enclosed in a container. The plastic

will be monitored and weighed each month for a 3-month duration to measure the weight

loss which is considered as the degradation rate. (Landicho et. al 2012)

Marine Floating Condition and Degradation

For the marine floating condition and degradation, seawater will be utilized and

stored in a closed container where the samples will be submerged for three months. The

change in weight, gain or loss, will be monitored each month to conclude marine floating

condition and the rate of degradation. (Aguila et. al. 2015)

The analytical balance will be utilized in weighing the specimen for both methods.

The rate of degradation can be determined using the equation used in a study by

Landicho et. al (2012):

MD
% degradation= ×100 %
MO

where M D is the mass difference and M O is the original or initial mass of the specimen.

Thermal Stability
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The thermal stability of the samples will be determined using the standard ASTM

D2307 where the samples will be cut into pieces and stored in a silicon oil bath at 180℃.

It will be set aside for a few minutes with cotton dipped in the oil bath which serves as

the stopper. Afterwards, the red litmus will be subjected to the litmus paper. The reaction

of the litmus will indicate whether the sample underwent decomposition which is when

the litmus paper turns blue. The initial time and the time it started decomposing will be

recorded and the difference of the two will determine how long the sample can sustain

heat. Longer time before it decomposes means a higher heat stability (Tosoh Polyvin

Corp., 2005). The thermal stability percentage can be calculated the same way Aguila

et. al, (2015) did in their study which is:

time reading
% thermal stability= ×100 %
60 mins
80 hrs ×
1 hr

Chemical Resistance

The chemical resistance, for this study, will be measured in accordance with the

ASTM D543 which uses two methods: reaction to bases and reaction to acids.

Reaction to Acids

The test for the reaction of the sample to acids will utilize 2M of hydrochloric acid

(HCl) where the samples are subjected for 24 hours at room temperature.

Reaction to Bases

For the reaction of the samples to bases, the same method will be applied but,

instead of HCl, the test will utilize sodium hydroxide(NaOH).

The percentage of chemical resistance for both methods will be determined using

the similar equation used by Aguila et. al, (2015) in their study which is:

% chemical resistance= 1 ±( M 2−M 1

M1 )
× 100 %
Reinforced Banana Pseudo-stem Nanocellulose Chapter iII: Research Methodology
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Abarintos, M.L.B., Bacquian, M.J.F., Banta, J.B., Castillo, K.A.C.

where M 1 is the initial mass of the sample before it is subjected to the test and M 2 is the

mass of the sample after it is subjected to the test.

STATISTICAL TREATMENT

The data that will be obtained from the experiment and tests in laboratories will be

tallied, tabulated and interpreted using the different statistical tools. Whereas, the findings in

this study will be statistically analyzed using T-Test and Variance Test Analysis.

Moreover, One Way Analysis of Variance (ANOVA) will be used for the evaluation of the

effect of different level content of reinforced banana cellulose fiber in its properties (mechanical,

chemical, physical).

The T-Test will also be utilized in this study wherein the comparison of the average

performance between the reinforced bioplastic bubble wrap and petroleum based (Vermatex)

are differentiated.