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Agricultural-Use Bioplastic Seedling Bag Made from Cassava (Manihot esculenta) Starch
and Deacetylated Chitosan Extracted from Black Tiger Shrimp (Penaeus monodon) Shells

Baldovino, Carla Marie V. Ed. D, Tan, Harvy Angelo D.

Region IV-A CALABARZON Division of Quezon Quezon National High School
harvyangelo.tan@gmail.com

Abstract: The introduction of plastic to industries had made many reliant on using it, especially
in agriculture, which uses seedling bags at the cost of the environment. In Lucena City, were
both agriculture and aquaculture thrive, waste management becomes difficult. Testing
degradation occurred by burying experimental and control films underneath for three weeks with
results tallied after each week for 21 days. Growth stimulation was tested using bioplastic and
commercial seedling bags on mung beans with the same period and data collection as
degradation test. Economic value was compared to commercial product as well. Based on the
results, set-up with more cassava starch degraded at an average of 14%, faster than other
experimental set-ups, on average, with commercial bioplastics not having degraded at all. For
growth stimulation, experimental group with the most chitosan, set-up C, made mung beans to
grow faster than other set-ups with an average height of 16.46cm. Set-up A had the lowest
growth stimulating effect on the post-hoc analysis, while set-up E, without fertilizer lags behind.
Thus, bioplastic seedling bag made from cassava starch and chitosan is more effective than
commercial seedling bags in both degrading to mitigate solid waste problems and growth
stimulation, but the cost of the experimental product is less than that of commercially available
plastics. On the other hand, these conventional products do not offer the ecological benefits the
study’s product have, which is more pressing now especially since the world faces numerous
environmental issues.

Keywords: Chitosan, Cassava Starch, Bioplastic, Agricultural Seedling Bag, Black Tiger
Shrimp

1. Introduction
When the first commercially available
plastic was made, many industries
immediately had an impression on its ease
of use and versatility. Along with this,
people began to realize that this material
takes a long time to degrade, most of which
takes over one hundred years to decay, and
some even reaches upwards of a thousand
years (LeBlanc, 2019). Today, its effects can
be seen in the daily lives of all living things.
Because of the numerous problems society
faces nowadays, the United Nations (2015)
set 17 Sustainable Development Goals,
which is projected to be accomplished for
the year 2030. One of these goals, the 12 th
goal, is to ensure sustainable consumption
and production patterns, in which this study
is contributing towards in many aspects like
lessening waste in industries and producing
materials that benefits producers without the
expense of the environment.
One of the industries that have an evident
impact was agriculture. Farmers needed a
quick and easy solution to what could have
been strenuous work and impossible.
Overall, agriculture alone contributes $5.87
billion USD annually according to the
Transparency Market Research cited on a

report by Grossman (2015) on their use of
plastic.
The Philippines alone contributes 2.7
million tons of plastic waste every year
according to the Ocean Conservancy Charity
and the McKinsey Centre for Business and
Environment (2015). Moreover, since
agriculture accounts for 10% of the
country’s economy (Brown, Ebora, and
Decena 2018), which means the amount of
plastic waste this industry produces is
extensive, and can affect lives.
Alongside the issue of excessive waste on
this industry, it also currently lacks support
in forms like that of government aid and
newer technology. Fertilizing the plants is a
way of local farmers to increase the yield,
but to do this, they have to buy more
chemicals. Worse yet, these fertilizers can
have adverse effects on the plant and pollute
the air, water and soil as studied by
Chandini, Kumar, and Prakash (2013).
Because of these environmental and
economic factors, the researcher looked into
emerging technologies and found that
bioplastic made from waste materials can be
possible, with added benefits as well. In the
coastal city of Lucena, a thriving agriculture
and aquaculture industry has brought about
economic activity. Crustaceans from the
area is a popular delicacy. Black Tiger
Shrimp (Penaeus monodon), as stated by
Vila (2017) in a report, is the most cultured
shrimp in the country. This is further
magnified by the numerous polyculture
farms locally, having both shrimp and fish.
Aquaculture operators that are involved in
this industry accommodates up to 232
hectares in 131 of the farms with Penaeus
m. Chitosan, which can be extracted from
this, is also a natural plant defense activator
and it can be utilized as a growth stimulator,
based on testing by Sabreen, Ibraheim and
Mohsen (2015).
On the other hand, the Philippine
Statistics Authority (2019) reported that
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from April to June 2019, cassava (Manihot
esculenta) production dropped by 7.2
percent, from 809.88 thousand metric tons
during the same quarter of 2018 to 751.46
thousand metric tons this quarter. Though it
seems like the industry is looking to be
declining now, the sector is still large
enough that it can still be a viable
component on this study, especially because
it has plenty of well-established use cases
for making plastics like many of the studies
the researcher based this process on. Since it
is starch, and plastics are primarily made of
polymers, the chemical structure of this
material provides great versatility.
The growing environmental issues of our
society and the lack of investment in local
sectors motivated this study. As mentioned
earlier, the study also aims to contribute to
the Sustainable Development Goals of the
UN. The researcher decided to create a
product that not only lessens waste across
industries while presenting benefits for its
usage and preventing adverse effects
towards our environment.
1.2 Objectives of the Study
The study generally aims to create a
bioplastic seedling bag made for agricultural
purposes.
Specifically, the study wanted to assess:
1.2.1 To determine the degradation of
experimental product and control
commercial bioplastic with varying
concentrations of chitosan and cassava
starch.
1.2.2 To find out if the product made
with different amounts of chitosan and
cassava starch has growt h stimulating
effects for the plant it is used on
compared to commercial products.
1.2.3 To ascertain the differences in the
set-ups and trials made between time
intervals.
1.2.4 To compare its economic value to
available agricultural-use seedling bag.

2. Materials and Methods
The following materials and procedures
had been used in the production of the
agricultural-use bioplastic.
2.1 Raw Materials
2.1.1 Shrimp Shells
Shrimp shells (Peneaus monodon), which
is the source of chitin, will undergo a
deacetylation process. These were gathered
from varying local establishments in Lucena
City, Quezon Province.
Every year, some 6 million to 8 million
tons of waste crab, shrimp and lobster shells
are produced globally — about 1.5 million
tons in Southeast Asia alone. (Chen and
Yan, 2015).
2.1.2 Cassava Starch
Cassava starch (Manihot esculeta) is a
perennial plant cultivated as a crop for its
edible starchy tuberous roots. It is widely
used in studies involving bioplastic
(Suhartono, 2018; Suryanto and
Wahyuningtiyas, 2018; Barbosa et al. 2017;
Akpan and Ogunrinola, 2018)
The researcher acquired this material by
buying it from a local supplier in the market
of downtown Lucena City.
2.1.3 Mung Beans
The study will utilize the mung beans
(Vigna radiata (L.) Wilczek) to assess the
growth stimulation effect of the bioplastic to
be produced. This was sourced from a local
market in downtown Lucena City.
2.2 Chitosan Extraction
To extract chitosan, the following three
steps must be done. First, demineralization,
which reduces mineral substances. Second
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was deproteinization, where proteins
residues are removed. And finally,
deacetylation, which diminishes the
presence of acetyl groups in the substance.
This process was based on study by
Domard, Percot, and Viton (2003) and
Buntong, Huon, Soeung, Sophal, and Tong
(2012).
2.2.1 Demineralization
Shrimp shells dried prior was submerged
in hydrochloric acid for 15 minutes at
ambient room temperature at 1:16 solid-to-
liquid ratio. Shells were rinsed right after.
2.2.2 Deproteinization
1M sodium hydroxide was prepared and
shrimp shells was added at room
temperature for 16h. Chitin was the resulting
material.
2.2.3 Deacetylation
Chitin still on the container was then
heated at 100̊C for 2 hours.
2.3 Bioplastic Production
Bioplastic production was partially based
on studies by Esfandiary, and Samzadeh-
Kermani (2016) and Larotonda, Matsui,
Soldi, and Laurindo, (2004) which used a
traditional method.
A cassava starch:water:glycerol mixture
was prepared, varying the starch
concentration having 200mL of water and
2mL of glycerin. The mixtures were
submitted to heating to 55ºC while stirring.
On a separate beaker, chitosan was
dissolved in 300mL acetic acid (0.1 M) and
stirred for 2 hours at room temperature.
Aqueous 4% PVA was dissolved in 200 mL
of hot distilled water under constant stirring
and then added to Chitosan solution
followed by stirring for 1 hour.

Starch/water/glycerol mixture were added
directly into Chitosan/PVA solution and
stirred for magnetically stirred for 3 hours.
The mixtures were casted into petri dishes
and glass plates, depending on the size
needed and put in a dry oven at 60 ˚C until
peelable.
For degradation, a 15cm x 15cm x 1mm
experimental film was required, weighing at
4g each, while the control group used
commercial bioplastic that weighed the
same. A 5cm x 5cm x 1mm and 9cm high
bag was required for assessing growth
stimulation. Positive control group using
commercial fertilizer was used and a
negative control group with no fertilizer.
2.4 Bioplastic Assessment
2.4.1 Degradation
For testing degradation, each
experimental and commercial bioplastic film
was buried underneath soil in their
respective trials and set-ups. Burial lasted
for three weeks, then the bioplastic was
checked for any signs of reduction at the end
of each week. The weight was tallied every
end of the week and each result at the end of
the week was statistically analyzed. Testing
was parallel to the process of Adamcová,
and Vaverková (2014).
2.4.2 Growth Stimulation
To assess the growth stimulation effect,
bioplastic and control groups was used on
mung beans. For control groups, a positive
control group received commercial nitrogen
fertilizer, and a negative control group
received no fertilizer, only receiving water
like the rest of the groups. Experimental
groups used bioplastic seedling bags with
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the corresponding ratio on the table. To see
the plant growth characteristics, a random
sample of the plants from each plot were
taken after 7, 14 and 28 days after sowing to
investigate the plant height (cm) with each
set-up having three replicates. This was akin
to the testing done by Sabreen, Ibraheim and
Mohsen (2015). Plant was watered and
given sunlight everyday similarly across the
trials.
2.4.3 Economic Value
Cost of production and estimated retail
price was calculated using traditional
formulas. Commercial seedling bag value
was compared to seedling bag tested in
growth stimulating effects of the product.
Only main materials were accounted for in
calculating the cost of production with and
retail price had a 25% profit margin.
2.5 Data Analysis
This research study aimed to test the
degradation and growth stimulation using
pre-post test design. The tallied results went
through a stati3tical treatment using
Analysis of Variance (ANOVA) for
assessing degradation, and growth
stimulation in order to analyze the variance
between the results. For this analysis, the
tallied results at the end of each week were
used and interpreted. After that, Tukey HSD
Post-hoc analysis was employed for tests
with ANOVA in order to see where the
significant difference lies between each set-
up. Treatment was conducted using IBM
Statistical Package for Societal Services 25
(SPSS version 25).
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3. Results

Table 1. Degradation in Mass Loss of the Bioplastic Film after Three Weeks in grams (g)

Posttest
Pre- Week 1 Week 2 Week 3 Percent
Set-ups Difference Difference
test Trial Trial Trial Trial Trial Trial Trial Trial Trial
1 2 3 1 2 3 1 2 3
A (2:3 3.79 3.82 3.83 3.57 3.64 3.68 3.36 3.45 3.51
Chitosan- -0.56 14%
Cassava) 3.813333333 3.63 3.44*
Experimental

B (1:1 3.89 3.92 3.89 3.74 3.83 3.8 3.59 3.74 3.68
Chitosan- -0.33 8.25%
Cassava) 3.9 3.79 3.67*
4g
C (3:2 3.95 3.93 3.93 3.88 3.85 3.87 3.84 3.79 3.81
Chitosan- -0.19 4.75%
Cassava) 3.936667 3.866667 3.813333*
D 4 4 4 4 4 4 4 4 4
Ctrl

(Commercial 0 0%
Bioplastic ) 4 4 4*
* The data used for calculating difference.

Table 2. Height of Mung Bean Plant with Bioplastic and Control Groups over Three Weeks in centimeters
(cm)

Posttest
Pre-
Set-ups Week 1 Week 2 Week 3
test
Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3 Trial 1 Trial 2 Trial 3
A (2:3 4.25 4.32 4.3 8.4 8.55 8.34 13.43 13.47 13.31
Chitosan-
Cassava) 4.29 8.57 13.40333333
Experimental

B (1:1 5.67 5.59 5.78 9.38 9.08 9.22 14.91 15.15 14.87
Chitosan-
Cassava) 5.68 9.226666667 14.97666667
C (3:2 6.63 7.12 6.39 10.84 11.11 10.78 16.19 16.87 16.32
Chitosan- 0
Cassava) 6.713333333 10.91 16.46
D 5.48 5.39 6.1 8.94 8.76 9 14.22 14.63 14.55
(Commercial
5.656666667 8.9 14.46666667
Ctrl

Fertilizer)
E (No 3.24 2.83 3.27 7.47 7.13 7.56 12.39 12.3 12.56
Fertilizer) 3.113333333 7.386666667 12.41666667

Table 3. Cost of Production between Experimental and Commercial Seedling Bag in Philippine Pesos

Commercial Bioplastic
Experimental Bioplastic
Allied Botanical Corporation (2019)
(Est.) Cost of
Production
Php 580.00 Php 10.00
Production 10 10
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Capacity
(Est.) Retail Price Php 700.00 (Php 70.00 each) Php 32.00 (Php 1.6 each)
Return of
Investment
25% 540%

Table 4. One-way ANOVA of the Significant Difference in the Degradation between Experimental
Bioplastic and Control Films after One Week

Std. Decision
N Mean F-Calculated F-Tabulated Conclusion
Deviation (=0.05)
A (2:3
Chitosan- 3 .1900 .02000
Cassava)
B (1:1
Chitosan- 3 .1133 .02082
Cassava) There is a significant
C (3:2 difference in the
Chitosan- 3 .0633 .01155 80.046 4.07 Reject H0 degradation rate as affected
Cassava) by the chitosan-to- cassava
ratio after one week.
D
(Commercial 3 .0000 .00000
Bioplastic)

Total 12 .0917 .07383

Table 5. Tukey HSD Post-hoc Multiple Comparison Test on the Level of Significance between the
Degradation of Experimental Bioplastic and Control Films after One Week

Mean p-value Decision

Set-ups Conclusion
Difference (Sig.) (=0.05)
There is a significant difference in the degradation
B .07667 .001 Reject H0
A between these set-ups.
(2:3
There is a significant difference in the degradation
Chitosan- C .12667 .000 Reject H0
between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .19000 .000 Reject H0
between these set-ups.
There is a significant difference in the degradation
A -.07667 .001 Reject H0
B between these set-ups.
(1:1
There is a significant difference in the degradation
Chitosan- C .05000 .018 Reject H0
between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .11333 .000 Reject H0
between these set-ups.
There is a significant difference in the degradation
A -.12667 .000 Reject H0
C between these set-ups.
(3:2
There is a significant difference in the degradation
Chitosan- B -.05000 .018 Reject H0
between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .06333 .005 Reject H0
between these set-ups.
D There is a significant difference in the degradation
A -.19000 .000 Reject H0
(Using between these set-ups.
Commercial There is a significant difference in the degradation
Bioplastic) B -.11333 .000 Reject H0
between these set-ups.
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There is a significant difference in the degradation

C -.06333 .005 Reject H0
between these set-ups.
*. The mean difference is significant at the 0.05 level

Table 6. One-way ANOVA of the Significant Difference in the Degradation between Experimental
Bioplastic and Control Films after Two Weeks

Decision
N Mean Std. Deviation F-Calculated F-Tabulated Conclusion
(=0.05)
A (2:3
Chitosan- 3 .3700 .05568
Cassava)
B (1:1
Chitosan- 3 .2167 .05686
Cassava) There is a significant
C (3:2 difference in the
Chitosan- 3 .1233 .01528 44.486 4.07 Reject H0 degradation rate as affected
Cassava) by the chitosan-to- cassava
D ratio after two weeks.
(Commercial 3 .0000 .00000
Bioplastic)

Total 12 .1775 .14530

Table 7. Tukey HSD Post-hoc Multiple Comparison Test on the Level of Significance between the
Degradation of Experimental Bioplastic and Control Films after Two Weeks

Mean p-value Decision

Set-ups Conclusion
Difference (Sig.) (=0.05)
There is a significant difference in the degradation
B .15333 .007 Reject H0
A between these set-ups.
(2:3
There is a significant difference in the degradation
Chitosan- C .24667 .000 Reject H0
between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .37000 .000 Reject H0
between these set-ups.
There is a significant difference in the degradation
A -.15333 .007 Reject H0
B between these set-ups.
(1:1
There is no significant difference in the degradation
Chitosan- C .09333 .086 Accept H0
between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .21667 .001 Reject H0
between these set-ups.
There is a significant difference in the degradation
A -.24667 .000 Reject H0
C between these set-ups.
(3:2
There is no significant difference in the degradation
Chitosan- B -.09333 .086 Accept H0
between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .12333 .024 Reject H0
between these set-ups.
There is a significant difference in the degradation
A -.37000 .000 Reject H0
between these set-ups.
D
(Using There is a significant difference in the degradation
B -.21667 .001 Reject H0
Commercial between these set-ups.
Bioplastic)
There is a significant difference in the degradation
C -.12333 .024 Reject H0
between these set-ups.
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*. The mean difference is significant at the 0.05 level

Table 8. One-way ANOVA of the Significant Difference in the Degradation between Experimental
Bioplastic and Control Films after Three Weeks

Decision
N Mean Std. Deviation F-Calculated F-Tabulated Conclusion
(=0.05)
A (2:3
Chitosan- 3 .5600 .07550
Cassava)
B (1:1
Chitosan- 3 .3300 .07550
Cassava) There is a significant
C (3:2 difference in the
Chitosan- 3 .1867 .02517 55.693 4.07 Reject H0 degradation rate as affected
Cassava) by the chitosan-to- cassava
D ratio after three weeks.
(Commercial 3 .0000 .00000
Bioplastic)

Total 12 .2692 .21882

Table 9. Tukey HSD Post-hoc Multiple Comparison Test on the Level of Significance between the
Degradation of Experimental Bioplastic and Control Films after Three Weeks

Mean p-value Decision

Set-ups Conclusion
Difference (Sig.) (=0.05)
There is a significant difference in the degradation
B .23000 .004 Reject H0 between these set-ups.
A
(2:3 There is a significant difference in the degradation
Chitosan- C .37333 .000 Reject H0 between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .56000 .000 Reject H0 between these set-ups.
There is a significant difference in the degradation
A -.23000 .004 Reject H0 between these set-ups.
B
(1:1 There is no significant difference in the degradation
Chitosan- C .14333 .050 Accept H0 between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .33000 .000 Reject H0 between these set-ups.
There is a significant difference in the degradation
A -.37333 .000 Reject H0 between these set-ups.
C
(3:2 There is no significant difference in the degradation
Chitosan- B -.14333 .050 Accept H0 between these set-ups.
to-Cassava
ratio) There is a significant difference in the degradation
D .18667 .013 Reject H0 between these set-ups.
There is a significant difference in the degradation
A -.56000 .000 Reject H0 between these set-ups.
D
(Using There is a significant difference in the degradation
B -.33000 .000 Reject H0 between these set-ups.
Commercial
Bioplastic) There is a significant difference in the degradation
C -.18667 .013 Reject H0 between these set-ups.
*. The mean difference is significant at the 0.05 level
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Table 10. One-way ANOVA of the Significant Difference in the Height of Mung Bean Plant
between Experimental Bioplastic and Control Groups after One Week

Std. Decision
N Mean F-Calculated F-Tabulated Conclusion
Deviation (=0.05)
A (2:3 Chitosan-
3 4.2900 .03606
Cassava)
B (1:1 Chitosan-
3 5.6800 .09539
Cassava) There is a significant
C (3:2 Chitosan- difference in the growth
3 6.7133 .37207
Cassava) 82.084 3.48 Reject H0 stimulation as affected by
D (Commercial the chitosan-to-cassava ratio
3 5.6567 .38657
Fertilizer) after one week.
E (No Fertilizer,
3 3.1133 .24583
Water only)
Total 12 5.0907 1.31671

Table 11. Tukey HSD Post-hoc Multiple Comparison Test on the Level of Significance between the
Height of Mung Bean Plant Tested with Experimental Bioplastic and Control Groups after One
Week

Mean p-value Decision

Set-ups Conclusion
Difference (Sig.) (=0.05)
B -1.39000 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
A
(2:3 C -2.42333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D -1.36667 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
ratio)
E 1.17667 .002 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A 1.39000 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
B
(1:1 C -1.03333 .006 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D .02333 1.000 Accept H0 There is no significant difference in the growth stimulation between these set-ups.
ratio)
E 2.56667 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A 2.42333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
C
(3:2 B 1.03333 .006 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D 1.05667 .005 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
ratio)
E 3.60000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A 1.36667 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

D B -.02333 1.000 Accept H0 There is no significant difference in the growth stimulation between these set-ups.
(Commercial
Fertilizer) C -1.05667 .005 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

E 2.54333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A -1.17667 .002 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

E (No B -2.56667 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Fertilizer,
Water only) C -3.60000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

D -2.54333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
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*. The mean difference is significant at the 0.05 level

Table 12. One-way ANOVA of the Significant Difference in the Height of Mung Bean Plant
between Experimental Bioplastic and Control Groups after Two Weeks

Std. Decision
N Mean F-Calculated F-Tabulated Conclusion
Deviation (=0.05)
A (2:3 Chitosan-
3 8.4300 .10817
Cassava)
B (1:1 Chitosan-
3 9.2267 .15011
Cassava) There is a significant
C (3:2 Chitosan- difference in the growth
3 10.9100 .17578
Cassava) 188.198 3.48 Reject H0 stimulation as affected by
D (Commercial the chitosan-to-cassava ratio
3 8.9000 .12490
Fertilizer) after two weeks.
E (No Fertilizer,
3 7.3867 .22679
Water only)
Total 12 8.9707 1.20009

Table 13. Tukey HSD Post-hoc Multiple Comparison Test on the Level of Significance between the
Height of Mung Bean Plant Tested with Experimental Bioplastic and Control Groups after Two
Weeks

Mean p-value Decision

Set-ups Conclusion
Difference (Sig.) (=0.05)
B -.79667 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
A
(2:3 C -2.48000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D -.47000 .034 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
ratio)
E 1.04333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A .79667 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
B
(1:1 C -1.68333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D .32667 .176 Accept H0 There is no significant difference in the growth stimulation between these set-ups.
ratio)
E 1.84000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A 2.48000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
C
(3:2 B 1.68333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D 2.01000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
ratio)
E 3.52333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A .47000 .034 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

D B -.32667 .176 Accept H0 There is no significant difference in the growth stimulation between these set-ups.
(Commercial
Fertilizer) C -2.01000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

E 1.51333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A -1.04333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
E (No
Fertilizer, B -1.84000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Water only)
C -3.52333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
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D -1.51333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
*. The mean difference is significant at the 0.05 level
Table 14. One-way ANOVA of the Significant Difference in the Height of Mung Bean Plant
between Experimental Bioplastic and Control Groups after Three Weeks

Std. Decision
N Mean F-Calculated F-Tabulated Conclusion
Deviation (=0.05)
A (2:3 Chitosan-
3 13.4033 .08327
Cassava)
B (1:1 Chitosan-
3 14.7400 .49790
Cassava) There is a significant
C (3:2 Chitosan- difference in the growth
3 16.4600 .36097
Cassava) 157.157 3.48 Reject H0 stimulation as affected by
D (Commercial the chitosan-to-cassava ratio
3 14.4667 .21733
Fertilizer) after three weeks.
E (No Fertilizer,
3 12.4167 .13204
Water only)
Total 12 14.2973 1.42962

Table 15. Tukey HSD Post-hoc Multiple Comparison Test on the Level of Significance between the
Height of Mung Bean Plant Tested with Experimental Bioplastic and Control Groups after Three
Weeks

Mean p-value Decision

Set-ups Conclusion
Difference (Sig.) (=0.05)
B -1.57667 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
A
(2:3 C -3.05667 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D -1.06333 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
ratio)
E .98667 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A 1.57667 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
B
(1:1 C -1.48000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D .51333 .084 Accept H0 There is no significant difference in the growth stimulation between these set-ups.
ratio)
E 2.56333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A 3.05667 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
C
(3:2 B 1.48000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Chitosan-to-
Cassava D 1.99333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
ratio)
E 4.04333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A 1.06333 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

D B -.51333 .084 Accept H0 There is no significant difference in the growth stimulation between these set-ups.
(Commercial
Fertilizer) C -1.99333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

E 2.05000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

A -.98667 .001 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

E (No B -2.56333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
Fertilizer,
Water only) C -4.04333 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.

D -2.05000 .000 Reject H0 There is a significant difference in the growth stimulation between these set-ups.
 Tan 12

*. The mean difference is significant at the 0.05 level

4 Figure 3. Cost of Production and Estimated
Retail Price Comparison
Weight in grams (g)

0.6
3.5

Mean of Degradation (g)

0.5
0.4
3
0.3
0.2
2.5 0.1
1 2 3 1 2 3 1 2 3 1 2 3
A B C D
-8.32667268468867E-17
a b c d
Week 3 Week 2 Week 1 -0.1
Week 1Set-up
Figure 1. Degradation in Mass Loss (g) in
Experimental Bioplastic and Control Films
after Three Weeks Figure 4. Means Plot Difference of the
Height of Plant (cm)

Degradation of the Bioplastic and

Commercial Films after One Week
20
0.6
15

Mean of Degradation (g)

0.5
10 0.4
0.3
5
0.2
0
1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 0.1
A B C D E
-8.32667268468867E-17
a b c d
Week 1 Week 2 Week 3 -0.1
Week 2 Set-up
Figure 2. Growth in Height (cm) of Mung
Bean Plant in Experimental Bioplastic and
Control Groups Figure 5. Means Plot Difference of the
Price in Philippine Pesos

Degradation of the Bioplastic and

Commercial Films after Two Weeks
700
0.6
600
Mean of Degradation (g)

0.5
500 0.4
0.3
400
0.2
300 0.1
200 -8.32667268468867E-17
a b c d
-0.1
100
Week 3 Set-up
0
Bioplastic Seedling Bag
Cost of Production Estimated Retal Price Figure 6. Means Plot Difference of the
Degradation of the Bioplastic and
Commercial Films after Three Weeks
 Tan 13

M ean of G row th (cm)

4. Discussion
18
16 In the degradation assessment, the most
14
12 effective set-up was revealed to be
10 experimental set-up A, which had a
8
6 chitosan-to-cassava ratio of 2:3, had an
4 average degradation of 0.56g, 14% of its
2
0
total mass initially. In contrast, the third
A B C D E experimental set-up, which had a 3:2
Week 1 Set-up chitosan-to-cassava ratio, yielded the least
degradation among the experimental groups,
Figure 7. Means Plot Difference of the Height
of Mung Bean Plant with Bioplastic and with an average of 4.75% after the testing.
Control Groups after One Week Meanwhile, set-up B lies between the results
of set-ups A and C. This was the group with
18 equal portions of chitosan and cassava starch
Mean of Growth (cm)

16 and ended the testing with 8.25% of its mass

14 having degraded. The control group that
12
10 used commercial bioplastic films did not
8 degrade at any point during the testing
6 process. Additionally, all the data collected
4
2
in testing degradation followed a common
0 trend among the results. These results are
A B C D E observable in Table 1 and Figure 1, where
Week 2 Set-up the differences become more apparent.
In a study by, Hasan, Rahmayani, and
Figure 8. Means Plot Difference of the Height Munandar (2018) on creating bioplastic
of Mung Bean Plant with Bioplastic and from similar materials, stated that the more
Control Groups after Two Weeks usage of starch creates a product that
degrades faster. This is in line with the
M ean of G row th (cm)

results of the degradation test, where higher

18 cassava starch yielded faster than groups
16
14
with lesser concentration of it. This is
12 because of the composition of the starch,
10 which provides the most degradation
8
6 because of the hydroxyl content. Moreover,
4 Barbosa, Cacique, Rios and Wentz (2017)
2
0
conducted a study using chitosan as well in
A B C D E creating bioplastic, and it was found that
Week 3 Set-up many of the resulting characteristics was
Figure 9. Means Plot Difference of the Height caused by the materials used, like acetic acid
of Mung Bean Plant with Bioplastic and influencing the consistency, elasticity and
Control Groups after Three Weeks shape, and the chitosan influences on the
stiffness and thickness. This could explain
as to why set-up C did not degrade as fast as

other experimental groups. Additionally,
Ginting, Lubis, Sidabutar, and Sirait (2018)
studied that the adding of chitosan to a
starch bioplastic solution resulted in
chitosan acting as the bioplastic thickener.
It was also discovered that experimental
set-up A, though having the fastest
degradation rate, did not have the most
growth stimulating effect on the plant it was
tested on. In fact, the highest growth in a
plant that was tallied was in set-up C, which
had more chitosan in the bioplastic solution
it was tested on. It grew up at an average of
16.46cm, about 1.8cm more than the
following highest growth, which was set-up
B. For positive control group D for growth
stimulation, which used no seedling bag but
with commercial nitrogen fertilizer, very
similar results on the growth stimulation test
of set-up B was observed across all of the
three weeks. Set-up A grew 13.23cm on
average after three weeks, not as much as
the other experimental groups, but still more
than the negative control group E, the set-
up, which used water only, had an average
of 12.42cm. The result of set-up B lies in
between that of A and C, with this group
having an average height of 14.74cm after
three weeks. Also analogous to the
degradation test was that the data collected
in each trial for the set-ups in assessing
growth stimulating effects followed a
common trend. The results for are tallied in
Table 2 along with Figure 2 where it shows
a clear illustration of how the growth of each
trial went.
As observed by Ismail, Malik, Mondal,
and Puteh (2013), when chitosan is used as a
bio-fertilizer, it showed that shrimp waste is
rich in nitrogen, which is commonly used in
agricultural fertilizers. The composition of
its waste is characterized by its high
nitrogen content, which plants can use in
growing. However, according to Kirchhoff
(2019), mung beans fix their own nitrogen,
so they normally do not need nitrogen
Tan 14
fertilizer. Despite the fact that mung bean
produces its own nitrogen, the chitosan in
the experimental bioplastic in this study,
stimulated the growth as evident in the
results. Theoretically, this can have
applications that are more practical
especially on plants that requires a good
amount of nitrogen fertilizer without
polluting the cost of the environment.
Additionally, the study conducted by
Changwei, Qiang, Qing, Xirong and Yide
(2005), where chitosan and chitosan
derivatives were used for mung bean plants
as well, results indicated that all chitosan
and chitosan derivatives increased yield and
quality of mung bean sprouts, which
validates the results of this study even more.
After accounting for the value of the
experimental bioplastic, even after using
waste material, it still falls behind the
commercial plastic in terms production cost,
retail price and . This is understandable
considering the fact that commercial plastic
production has become cheaper, especially
since it is mass-produced and has had
decades of time to develop. As shown in
Table 3 and Figure 3, the profit and
production cost cannot be matched in terms
of percentages. Even if the experimental
bioplastic rakes in Php 140.00, more than
the Php. 27.00 of commercial plastics, the
production cost pulls back the experimental
product. The return of investment is 540%
for the seedling bags available for
purchasing, while it is only 25% for the
product of this study. Though the
commercial product offers more profit at the
end of the day, this does come at the cost of
the environment, and in the end, it had
detrimental effects to lives. Today’s
challenges are affecting how businesses
tackle their markets, how they make
products and how they should be held
accountable in the growing environmental
issues of our time. This is why corporations
are now accounting for environmental costs.

In the case of this study, the results for the
experimental seedling bag boils down to
Php. 0.00, since it not only leaves no lasting
adverse effects on the environment, it
lessens it as well because of the waste
materials used while also helping
agricultural production. In contrast, the
environmental cost is more substantial on
the control group, especially when the tons
of plastic used in agriculture is accounted
for. When this is considered, the
environmental cost stacks up to about the
same as the $5.87 billion USD (Grossman,
2015) they spent in getting these plastic
products. This further illustrates stacks up
to about the how these industries need to
find a way in which they produce the
commodities everybody requires without
trading it for the environment.
Since the calculated F-ratio of 80.05 for
week 1 is greater than the tabulated F-ratio
of 4.07, the null hypothesis stating that the
chitosan-to-cassava ratio will have no
significant effect on the degradation of the
bioplastic is rejected, as seen in Tables 4.
Therefore, the ANOVA verifies that the
chitosan-to-cassava ratio does indeed have
an effect on the degradation of the product.
In order to determine where the significant
difference lies, a Tukey HSD post-hoc test
was conducted as shown in Tables 5, where
the mean difference is significant at 0.05
level. Based on the statistical treatment, the
null hypothesis was rejected every single
time. The mean difference against the
control group (set-up D) and every
experimental group (set-ups A, B and C)
yielded mean differences that are more than
that of the differences between experimental
groups. Despite this, the mean difference
between set-up B and C is less than that of
the other experimental set-ups, which could
entail that as time goes by, these would not
have a statistically significant difference. As
shown in Figure 4, the mean differences are
Tan 15
clearly apart showing its height in set-up A
and lowest in D, the control group.
On the second week of testing
degradation, the F-calculated value of 44.49
is still statistically significant compared to
the tabulated F-ratio of 4.07, therefore
rejecting the null hypothesis again. But, in
the Post-hoc test, and as predicted, the
differences between set-ups B and C were
not statistically significant compared to
others. Although C still degraded less based
on the raw data, statistically, their difference
was not significant like what happened in
the first week. A means plot showing these
statistical results can be seen in Figure 5,
and in Tables 6 and 7.
For the third and final week, the F-
calculated value of 55.69 is greater than the
F-tabulated value of 4.07 as shown in Table
8, meaning the null hypothesis is rejected
again. Post-hoc analysis was done to ensure
where the difference lies, and equivalently
to the second week, the difference between
set-ups B and C were not significant as seen
in Table 9. Despite this, the difference this
time was not as big as the second week. The
means plot difference between groups B and
C were not that largely apart as shown in
Figure 6, but differences elsewhere was
significant.
In assessing the growth stimulating
effects, similar to testing degradation, the
results were tallied and statistically analyzed
at the end of each week. For the first week,
since the tabulated F-ratio of 3.48 is less
than the F-calculated value of 82.084 as
shown in Table 10, the null hypothesis
stating that the chitosan-to-cassava ratio will
have no significant effect on the growth
stimulating effects of the bioplastic is
rejected. Therefore, the ratio between the
used materials does indeed affect the growth
of the plant it is used on. To verify where
the significance lies, a post-hoc test was
conducted and the results were tallied in
Table 11. It shows that all the difference

between the set-ups were statistically
significant. Furthermore, the means plot
difference illustrated in Figure 7 show that
all of the set-ups are significantly different
after assessing the growth for the first week.
The results for the second week was
statistically analyzed and tabulated in Tables
12 and 13 for ANOVA and post-hoc
respectively. Since the F-calculated value of
188.198 is greater than tabulated the F-ratio
of 3.48, the null hypothesis is therefore
rejected once again. The post-hoc analysis is
results were different from the week prior,
there was no significant difference between
set-ups B and D. Experimental group B had
a 1:1 chitosan-to-cassava ratio, while D used
commercial fertilizers. From this, it can be
inferred that using conventional fertilizer
equates to using this set-ups solution for
creating bioplastic, but it does not offer the
ecological benefits this study presents. The
means plot difference in Figure 8 shows this
difference more and it the results peak at set-
up C once again for growth stimulating
effects.
Finally, in the last week of testing
growth-stimulating effects, the one-way
ANOVA test showed that the F-tabulated
value of 3.48 is less than the calculated F-
ratio of 157.157, tallied in Table 14. Which
means that there is a significant difference
across the results once more. Post-hoc
analysis was done to see where the
difference lies. Set-up B and positive control
group D showed no significant difference
between the results, similar to last week.
The means plit of this follows the same
trend as last week, which is shown at Figure
9.
5. Analysis
Based on the tallied results for
degradation and growth stimulation, each
group had significant variations from one
another, based on both the ANOVA and
Tan 16
post-hoc tests, which entails potentially
different use cases for each of the set-ups
when used in agriculture. Set-up A degraded
at a faster rate than the other set-ups, but
along with this, it provided the least amount
of growth stimulation for the experimental
groups as well. This could be used when
farmers require a plastic that does not need
a whole lot of fertilizing, similar to mung
bean in this study which it does not require,
but at the same time, provides the peace of
mind of not adversely affecting the
environment the most. When growth-
stimulating effect is more important, using
the solution in set-up C offers the most of
this, while still having degradation,
something the control commercially
available seedling bag does not provide.
Additionally, set-up B demonstrates the
average experience in terms of degradation
and growth stimulation, which is nice to
have when both of these are equally valued.
To determine other properties of the
bioplastic, especially more on technical and
physical terms, testing its tensile strength
using standardized methods like that of
ASTM, along with using an SEM (Scanning
Electron Microscope) to better understand
its composition in a microscopic level. The
researcher recommends assessing multiple
physical characteristics of the bioplastics,
like tensile strength, hardness, and water
solubility among others, also recommends
using more materials in the study, like
different plants to test the growth
stimulating effects on, or diversifying the
products used.
In this study, to prove the effectiveness of
the product, which is the bioplastic seedling
bag in terms of degradation, the researcher
based this on studies that assessed bioplastic
by entombing it underneath soil for a period
of time (Adamcova and Vavercova, 2014).
For growth stimulation, the study by
Sabreen, Ibraheim and Mohsen (2015) was
the basis for testing and measuring the

height of the plant every end of the week.
The results show that there are two main
differences between the control and
experimental groups. First, the experimental
groups made a product that degrades unlike
any commercially available product. This
characteristic is important especially now
where pollution and solid waste problems
are happening all around the world,
eventually affecting climate change as well.
The second differentiating factor is that the
product has growth-stimulating effects on
the growth of plants. This can reduce the
usage of fertilizers on seedling bags with
young plants, which can cause air, water and
soil pollution when used excessively,
according to Chandini, et al., (2013).
The post-hoc results after three weeks
showed that for degradation, the differences
between B and C, the groups with the
average and most cassava starch in the set-
up respectively, is not significant. Similarly,
in growth stimulation, set-up B, and the
positive control group with fertilizer, set-up
D, showed that their difference is not
significant. From this observation, it can be
inferred that amount of chitosan and cassava
starch affects how the bioplastic behaves.
While commercial products does its purpose
for agriculture just right, these do affect the
environment negatively.
6. Conclusion
After thoroughly testing the usage of the
product for agricultural-use, it can be
concluded that the bioplastic seedling bags
is more effective in growing young plants,
and useful in reducing use of harmful
plastics. Since it can be used as both
seedling bag and fertilizer at once, the
widespread application of damaging
products like plastics and chemical-based
fertilizers to the environment can be
mitigated. Along with this, it is also
conclusive to note that the cost of producing
Tan 17
the bioplastic is more than that of
commercially available products, but the
environmental cost these commercial
products imply hindrances its value.
7. Acknowledgement
The researcher would like to recognize
the efforts of the people who made this
study happen in this section. Their notable
actions made all the difference in making
this entire endeavor possible. To his friends,
peers and family, to Quezon National High
School, Mrs. Rea Ethel Gatbonton, his
qualified scientist, Johanna Elyssa Ramos,
his colleague that provided connections to
people and opportunities this study needed,
and most especially to his research adviser
and motivator, Carla Marie V. Baldovino,
Ed. D.
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