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TUMID VENUS CLAM (Gafrarium tumidum) AS AN ADDITIVE FOR STARCH-

BASED BIOPLASTIC: EXTRACTION AND CHARACTERIZATION OF CHITOSAN

A Research Proposal

Presented to the Faculty


Of the Department of Chemical Engineering
School of Engineering and Architecture
Saint Louis University

In Partial Fulfillment
Of the Requirements for the Degree
Bachelor of Science in Chemical Engineering

by:

Engr. Ailine Marie L. Orduña (Research Promoter)


Mark Olivier B. Manzano
Nicole Anne T. Borromeo
Donna Teresa C. Caampued
Nicolle Julia P. Cleto
Jhoisy Dhae M. De Guzman
Mia Lyan Beryll B. Luis
Vay-Ann G. Tayco
Trishia Paulyne N. Villanueva

May 2019
Saint Louis University
SCHOOL OF ENGINEERING AND ARCHITECTURE
Department of Chemical Engineering

INDORSEMENT

The Research entitled “TUMID VENUS CLAM (Gafrarium tumidum) AS AN


ADDITIVE FOR STARCH-BASED BIOPLASTIC: EXTRACTION AND
CHARACTERIZATION OF CHITOSAN” prepared and submitted by

MARK OLIVIER B. MANZANO


NICOLE ANNE T. BORROMEO
DONNA TERESA C. CAAMPUED
NICOLLE JULIA P. CLETO
JHOISY DHAE M. DE GUZMAN
MIA LYAN BERYLL B. LUIS
VAY-ANN G. TAYCO
TRISHIA PAULYNE N. VILLANUEVA

In partial fulfillment of the requirement for the degree BACHELOR OF SCIENCE IN


CHEMICAL ENGINEERING has been examined and hereby recommended for
proposal defense.

Engr. Ailine Marie L. Orduña


Promoter/Lead Author

Members of the Panel

Engr. Vera Lee D. Mendoza, PhD Engr. Maria Nimfa P. Del Rosario
Evaluator Evaluator

Engr. Lilibeth R. Ramos Engr. Emerson Gapuz


Evaluator Undergraduate Research
Coordinator

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ABSTRACT

One major problem involving the use of plastic is its non-

biodegradability that cause a significant threat to the environment. Hence,

alternatives called bioplastic could be a long-term solution to worldwide

plastic pollution. The aim of this study is to extract and characterize chitosan

from wastes seashells and utilize it as an additive to bioplastic. Through the

pre-experiment conducted, Tumid Venus Clam shell was identified to have

the highest chitosan content among the other waste seashells that was

tested. The characterization proves that the extracted material from Tumid

Venus Clam is indeed chitosan; it has low viscosity, 62.92% DDA (degree of

deacetylation) and is 78.70% pure. The starch-based bioplastic with chitosan

as an additive have been successfully produced using modified process of

previous studies presented in the methodology. The researchers focused to

study the biodegradability of the produced bioplastic. The results showed that

by adding 10% weight chitosan to bioplastic had increase the rate of

biodegradability to 70.64% within 1 week in a natural soil.

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TABLE OF CONTENTS

TITLE PAGE i
INDORSEMENT PAGE ii
ABSTRACT iii
TABLE OF CONTENTS iv
LIST OF TABLES v
LIST OF FIGURES vi
CHAPTER 1: THE PROBLEM AND ITS SETTING 1
Background of the Study 1
Statement of the Problem / Statement of the Objectives 5
Theoretical and Conceptual Framework 6
Scope and Delimitation 12
Constraints of the Study 13
Significance of the Study 23
Operational Definition of Terms 24
CHAPTER 2: RESEARCH DESIGN AND METHODOLOGY 26
Research Design and Methodology 26
Population and Locale of the Study 26
Data Gathering Tools 27
Data Gathering Procedure 27
Treatment of Data 33
CHAPTER 3: REVIEW OF RELATED LITERATURE 34
CHAPTER 4: PRESENTATIONS, ANALYSIS, RESULTS AND DISCUSSION 41
Characterization of Chitosan Extracted 41
Biodegradation Test 44
CHAPTER 5: CONCLUSIONS AND RECOMMENDATIONS 52
BIBLIOGRAPHY 56
APPENDICES 61
APPENDIX A: Pretest Computations 62
APPENDIX B: Pretest Results 65
APPENDIX C: Mineral and Protein Test Computations 66
APPENDIX D: Mineral and Protein Test Data 68
APPENDIX E: Documentation 70

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LIST OF TABLES

Table 1.1 The roles of each members in the accomplishment of the study 16

Table 1.2 Gantt Chart 19

Table 1.3 Schedule of Outputs 20

Table 1.4 Bill of Materials 20

Table 2.1 List of Materials and Equipment 27

Table 4.1 Degree of Deacetylation Results (DDA) 41

Table 4.2 Solubility Test Results 42

Table 4.3 Viscosity Test Results 43

Table B.1. The results for the pretest for the four shells 65

Table D.1 Mineral Test Data before Demineralization 68

Table D.2 Mineral Test Data after Demineralization 68

Table D.3 Total Calcium removed by Demineralization 68

Table D.4 Spectrophotometric reading of Protein in the sample 68

Table D.5 Concentration of Protein before and after Deproteinization 69

Table D.6 Total Protein Removed by Deproteinization 69

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LIST OF FIGURES

Figure 1.1 Dependent-Independent-Expected Output Paradigm 10


Figure 1.2 Input-Process-Output Research Paradigm 11
Figure 2.1 Summary of Experimental Procedure for Chitosan 28
Extraction
Figure 4.1 Measured weights of bioplastic produced with three 45
different composition of chitosan (0%, 10% and 20%) buried in natural
soil for trial 1
Figure 4.2 Measured weights of bioplastic produced with three 45
different composition of chitosan (0%, 10% and 20%) buried in natural
soil for trial 2
Figure 4.3 Measured weights of bioplastic produced with three 46
different composition of chitosan (0%, 10% and 20%) buried in natural
soil for trial 3
Figure 4.4 Measured weights of bioplastic produced with three 47
different composition of chitosan (0%, 10% and 20%) buried in
compost soil for trial 1
Figure 4.5 Measured weights of bioplastic produced with three 47
different composition of chitosan (0%, 10% and 20%) buried in
compost soil for trial 2
Figure 4.6 Measured weights of bioplastic produced with three 48
different composition of chitosan (0%, 10% and 20%) buried in
compost soil for trial 3
Figure 4.7 Relationship between the percentage weight loss of 49
bioplastic produced with three different composition of chitosan (0%,
10% and 20%) and the burial time in both natural and compost soil for
trial 1
Figure 4.8 Relationship between the percentage weight loss of 49
bioplastic produced with three different composition of chitosan (0%,
10% and 20%) and the burial time in both natural and compost soil for

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trial 2
Figure 4.9 Relationship between the percentage weight loss of 50
bioplastic produced with three different composition of chitosan (0%,
10% and 20%) and the burial time in both natural and compost soil for
trial 3
Figure E.1 The shells: Ostrea edulis, Donax vittatus, Gafrarium 70
tumidum, and Melongena, were obtained from Tondol, Ando,
Pangasinan
Figure E.2 The shells were thoroughly washed with water and sun- 70
dried until no moisture was left.
Figure E.3 40 g of the sample from each shell were taken for the 71
extraction of chitosan. The milled shells were soaked in a 1:10 w/v of
0.68 M HCl for four hours at ambient temperature
Figure E.4 The solution was stirred in the acid until no bubbles and no 71
color change was observed
Figure E.5 The sample was washed thoroughly with distilled water 72
until a neutral pH was measured using a pH paper
Figure E.6 The sample was dried in a hot plate until fine powder will 72
be obtained
Figure E.7 The prepared solution was filtered in order to remove the 72
organic solids present
Figure E.8 Titration method using EDTA was used to determine if 73
minerals are still present in the shells.
Figure E.9 The dried shells from the demineralization process were 73
soaked in a 1:10 w/v 0.62 M NaOH solution for 16 hours at ambient
temperature
Figure E.10 The chitin was dried in a hot plate until fine powder was 73
obtained
Figure E.11 The chitin was deacetylated by soaking it in liquid sosa 74
(1:10 w/v) for 5 hours maintaining a temperature of 90°C.
Figure E.12 The chitosan was washed thoroughly with distilled tap 74

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water followed by distilled water. The washed chitosan was dried in a
hot plate.
Figure E.13 0.2 grams are weighed for each chitosan sample and 75
dissolved in 20 mL 0.1M hydrochloric acid and 25 mL deionized water.
Figure E.14 The solutions are continuously stirred for 30 minutes and 75
another 25 mL of deionized water were added. pH are measured using
pH paper. Then, solutions were titrated with 0.1M sodium hydroxide
solution.
Figure E.15 The prepared solutions are mixed using a magnetic stirrer 76
for 15 minutes with a speed of 1000 rpm. After 15 minutes, the
remaining chitosan which did not dissolve are filtered, dried and
weighed again.
Figure E.16 The viscosity of the chitosan samples was determined 76
using an Ostwald viscometer.
Figure E.17 The extracted chitosan was placed in a 4.5% acidity 77
Silver Swan Vinegar while stirring it with a magnetic stirrer and was
heated on a hotplate with temperature of 50˚C.
Figure E.18 5 grams of corn starch and 1.5 grams of glycerine were 77
added in the chitosan solution
Figure E.19 The mixture was stirred until it became homogeneous 77
and thickened at a temperature of 80˚C.
Figure E.20 After it thickened, it was poured into an aluminum tray 78
and was placed in a hot plate for drying at a temperature of 40˚C.
Figure E.21 After drying, the bioplastic was removed carefully from 78
the aluminum tray and subjected to characterization.
Figure E.22 The bioplastic containing 0%, 10% and 20% chitosan 79
were initially weighed and buried using natural soil for one week.
Three trials for each variation of bioplastic were made. The sticks
served as markers and labels.
Figure E.23 The bioplastic containing 0%, 10% and 20% chitosan 79
were initially weighed and buried using compost soil for one week.

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Three trials for each variation of bioplastic were made. The sticks
served as markers and labels.
Figure E.24 The results of the biodegradation test after the bioplastics 80
containing 0%, 10% and 20% (right to left) chitosan were buried in
natural soil (left) and compost soil (right) for two weeks.
Figure E.25 The bioplastics obtained after one week of burial were 80
weighed to determine how much was degraded.

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