MAKALAH PENELITIAN LPB JATIM 2023

Utilizing Crab Shell Waste as an Alternative Coagulant and Antibacterial Agent to

Mitigate Water Contamination in Surabaya

[Kenzie Alexander Wang], [Mabelle Josephine Putri Dharma]

Guidance Counselor: [Ratna Kusuma Dewi]
[ SMP Cita Hati Christian School ] – [Surabaya & East Java]
[sse_kenzie.wang@bchati.sch.id];[sse_mabelle.dharma@bchati.sch.id] [Environmental
Science]
A. Research Abstract improve the quality of water in
Surabaya. This research concludes
Each year, Indonesia produces 6.4
that the optimal dose for reducing river
million tons of waste from crab shells.
water turbidity in Surabaya as much as
However, within this frequently
250ml, is established at 150 mg, along
underestimated and excessive waste,
with an equally effective dose for
crab shells contain a substance called
antibacterial activity, underlining the
chitin. Chitin, extracted from crab
efficacy of this groundbreaking solution.
shells, yields the antibacterial
polysaccharide known as chitosan.
B. Research Background
Beyond its antibacterial properties,
Indonesia holds a significant
chitosan derived from crab shells plays
fisheries potential, especially in crab
a vital role in water purification through
catches (Luthfiyana et al., 2022). The
its coagulation properties. The
crab industry is highly valuable and
production of chitosan involves stages
popular, but the generated waste has a
such as; demineralization,
negative impact on the environment.
deproteinization, and deacetylation.
Crab shells produce processing waste
Chitosan has the power to clarify water,
that accounts for 50-60% of the whole
mitigating pollution and elevating water
weight (Kurniawan, 2021). Crab shells
quality. Chitosan proves that it can be
can adversely affect the environment
an alternative of coagulant that is more
due to mass disposal as waste,
biodegradable, eco-friendly and
potentially altering marine ecosystems
sustainable to replace alum (aluminum
and causing marine pollution (Ali &
sulfate) in water treatment. This study
Meriatna, 2021). Crab shell waste can
aims to utilize crab shell waste as an
result in environmental pollution as it
alternative coagulant and antibacterial
easily decomposes due to microbial
agent to mitigate water contamination
activity (Asni et al., 2014). These shells
in Surabaya, through this step we can

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contain various valuable components
like chitin, protein, and calcium
carbonate (CaCO3) minerals (Fajri et
al., 2019). Chitin, one of the main
components of crab shells, possesses
antibacterial properties and can be
used in various applications. C. Statement of the problem:
Furthermore, chitosan extracted from Statement of the problem, is a
chitin also has coagulant properties problem statement that encompasses a
that can serve as an effective brief exposition of the problem that a
coagulation agent in water purification research study is trying to solve. And in
processes (Meicahayanti et al., 2018). this research, the statement of the
This study aims to explore optimal problem is explained as follows;
methods for harnessing the benefits of 1. What steps or phases must
crab shell waste. This includes chitin be implemented to
extraction for antibacterial use and synthesize chitosan from
chitosan extraction as a coagulation crab shells?
agent in water purification. In this way, 2. How can chitosan from
the research has the potential to crab shells act as a water
reduce the negative impact of crab clarifying coagulant and an
shell waste on the environment while antibacterial agent?
creating added value from this waste. D. Literature Review
This research holds significant Kepiting dan limbah cangkangnya:
implications in the context of
Crabs, as a significant group of
environmental conservation in
Crustacea, play a pivotal role in
Surabaya, which has a vast marine
Indonesia's fisheries exports. According
area and substantial fisheries potential.
to data from the Central Statistics
By having a positive environmental
Agency in 2017, Indonesia is
impact, this study can be a crucial step
experiencing consistent year-over-year
towards the sustainability of natural
growth in crab exports, with exports
resources and environmental protection
reaching 110,000 tons in 2015 (Badan
in Indonesia.
Pusat Statistik, 2017). The majority of
these exported crabs are in the form of
Picture 1. Crab shell waste frozen crab bodies without heads and
shells, generating a substantial quantity

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of waste from crab heads and shells. attract and bind with negatively
Johnson and Peniston (1982) assert charged particles. This electrostatic
that shrimp and crab shells constitute a interaction leads to coagulation and
processing waste that can make up flocculation, making it effective in
50-60% of the total weight. In 2015, the removing impurities, pathogens, and
potential waste from crab shells suspended particles from liquids,
amounted to 66,000 tons, underlining therefore it can be an alternative of
the environmental pollution mitigation coagulant that is biodegradable and
potential through the utilization of crab eco-friendly. A coagulant is a substance
shell waste. Chitin, a homopolymer used to clump particles in a liquid,
composed of crucial in water treatment and industrial
Beta-(1,4)-N-acetyl-D-glucosamine processes (Kurniawan, 2021).
microfibrils approximately 3 nm in Indonesia faces a pressing issue with
diameter, is stabilized by hydrogen contaminated water, particularly in
bonds between amino and carboxyl Surabaya's rivers, which suffer from
groups (Gooday, 1994). Its monomeric toxic waste, waterborne diseases, and
unit, 2-acetamido-2-deoxy-D-glucose ecosystem degradation (Meicahayanti
(N-acetylglucosamine), is represented et al., 2018). Alum, the commonly used
by the molecular formula coagulant, can contaminate water, soil,
(C8H13NO5)n (Horton, 2002). Chitin harm ecosystems, and even affect
shares structural similarities with human health. Therefore, an alternative
cellulose, differing primarily in the coagulant and antibacterial agent are
substituent group at carbon atom 2, required. Utilizing crab shells by
featuring acetyl amide (CH3CONH₂) extracting chitin and converting it into
groups (Muzzarelli, 1985). This chitosan with coagulation properties
biopolymer is soluble in concentrated presents a solution for Surabaya's
mineral acids like HCl, HNO3, and water pollution problem. Other than
H2SO4, with deacetylation degrees of that, Chitosan also has antibacterial
up to 10% (Hartati, 2002). Chitosan, a properties. Chitosan is a biopolymer,
chitin derivative, consists of long-chain possessing a specific structure with
polymers with glucosamine monomers, positively charged amino groups
enabling interactions with negatively (-NH2) along its carbon chain. In an
charged molecules due to its positively aqueous environment, chitosan is
charged amino groups (Goosen, 1997). sensitive to high ion strength. Therefore
Chitosan has coagulation properties it can also reactivate stems from its
due to its positive charge, enabling it to highly reactive amino groups, allowing

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it to interact with the negatively charged
cell walls of bacteria. These
interactions occur in electronegative
regions on the bacterial cell surface.
Furthermore, the -NH2 groups, with
their free electron pairs, can complex
with Ca2+ ions present in the bacterial
cell wall, forming coordinate covalent
bonds. Gram-negative bacteria, with
their outer lipopolysaccharide layer, are
particularly susceptible to the
antimicrobial effects of chitosan due to
their highly negative polarity. Thus, A. The process or utilizing crab shells as a
chitosan is present as an alternative of coagulant and antibacterial agent:
coagulant and anti-bacterial that is eco
1. Deproteination
friendly, biodegradable and
sustainable, and able to help the water In the deproteinization stage, 50 grams of

pollution problem in Surabaya. pulverized crab shell is mixed with a 3.5%

sodium hydroxide (NaOH) solution in a
A. Research Methodology
1:10 weight ratio. Then the solution is
Methodology represents a subjected to controlled heating at 65°c for
meticulously structured systematic 2 hours, and agitated facilitated by a
framework used in research. It magnetic stirrer. Then, the composite is
serves as a strategy for subjected to filtration through filter cloth
researchers to effectively collect and meticulously washed with AquaDES
data and attain the predefined until neutral pH equilibrium. Then, the
goals of their study. In this blend undergoes a drying process in an
research, the methodology is oven set at a precise temperature of 100
shown through the depiction of a degrees Celsius for an extended period of
flowchart following. 24 hours.

4th picture. Research flow diagram.

2. Demineralization

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The second phase involves
demineralisation. The outcome of the
deproteinization step is combined with 1 N
hydrochloric acid (HCl) in a 1:10 ratio and
subjected to heating at 70ºC for 2 hours
B. Examination
while employing magnetic stirring.
The jar test is a method employed to
Following this, the mixture is filtered and
assess the coagulation capacity of a
rinsed until achieving a neutral pH,
substance and to determine the optimal
utilizing AquaDES. Subsequently, the
operational parameters (dose) in the
mixture is subjected to a 24-hour drying
context of water and wastewater treatment
process in an oven set at 100ºC.
processes. This method involves the
measurement and documentation of
parameters such as turbidity ratios before
and after the introduction of chitosan, as
well as the dosage of coagulant required
for specific wastewater volumes. This test
3. Deacetylation
enables precise quantification of the
In the final step, known as deacetylation,
requisite coagulant volume for river water
the demineralized mixture is blended with
clarification. The jar-test procedure
a 50% sodium hydroxide (NaOH) solution
emulates coagulation and flocculation
in a 1:20 ratio. This mixture is stirred and
mechanisms, targeting the elimination of
heated to 120°C, for a duration of 4 hours
suspended solids and organic
using a magnetic stirrer. After Subsequent
compounds, potential sources of turbidity,
to the heating process, the mixture is
odor, and taste disturbances. When
subjected to filter and thorough washing
executed with precision, it yields pivotal
until it reaches neutral pH level. Then the
insights for optimizing
resulting filtrate is subsequently subjected
coagulation-flocculation and purification
to drying in an oven maintained at a
processes, supplying empirical data for
temperature of 100°C for a period of 24
fine-tuning essential parameters, including
hours. This controlled process yields chitin
coagulant dosage, controlled mixing
extracted from crab exoskeletons,
(flocculation), and settling duration.
characterized by its coagulating and
antibacterial properties.
The Jar-test procedure employed in this
research is outlined as follows

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 ● Formation of 250ml water samples
are introduced into chemical
glassware.
● Chitosan is added in varying doses
of 0.05 grams, 0.10 grams, 0.15
grams, and 0.20 grams, each
subjected to three replicates.
● Then, the samples are settled for a
period of 24 hours, during which
turbidity differentials are monitored.
These observations are then
compared to those of alum, which
serves as the control variable.
Picture 5. Testing as a Coagulant
of microorganisms susceptibility to
antimicrobial agents. As an initial step, all
apparatus are subjected to sterilization
procedures utilizing an autoclave. In this
experiment, filter paper was immersed in
AquaDes solution containing chitosan at
four different doses: 50mg, 100mg,
150mg, and 200mg, along with 50mg of
aluminum-sulfate (alum). Additionally,
water from the river was applied to the
bacterial medium using oasis needle,
which in this case, was sodium agar that
had been solidified and placed in a Petri
dish. Then, small circular filter papers that
had been soaked in AquaDes with varying
doses of chitosan and 50mg of alum were
placed inside the petri dish on top of the
bacterial media.

Furthermore, to assess the chitosan's

Picture 6. Test preparation
antibacterial activity, we employed the disk
diffusion method. The disk diffusion
method is a common approach for testing
the sensitivity of microorganisms to
antimicrobial agents. This involves using
filter paper disks containing antimicrobial
agents, which are placed onto an agar B. Research variable
medium previously inoculated with
microorganisms. Following incubation, the This study encompasses three distinct
appearance of a clear zone in the vicinity variables: the independent variable, the
of the disc signifies the manifestation of dependent variable, and the control
microbial growth inhibition attributable to variable. The independent variable, in this
the presence of the antimicrobial context, is the processed crab
substance. This technique is characterized shell-derived chitosan, known for its
by its expeditiousness, simplicity, and is of coagulant and antibacterial properties.
paramount significance in the assessment This independent variable exerts influence

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on or serves as the causal factor for
alterations in the dependent variable,
which is the turbidity of river water.
Furthermore, the chitosan also plays a
role in disturbing the cellular membranes
of bacteria present in the aqueous
medium.

Picture 7 reveals that in the

In this research, the dependent variables
coagulation-flocculation process
are the turbidity of river water and the
employing crab shell-derived
bacteriological activity within a liquid
chitosan as a coagulant, the
medium mixed with chitosan. Dependent
majority of the dosage of chitosan
variables are those parameters whose
effectively attenuates the turbidity
states are contingent upon or influenced
levels in river water. The most
by the independent variable, which, in the
optimal chitosan dosage was
present study, is represented by the
determined to be 150mg. Prior to
introduction of chitosan.
chitosan treatment, the average
turbidity of the river water was
The control variable is a variable that is
measured at 21.33 NTU, while
manipulated to ensure that the influence
post-chitosan application, this value
of independent variables on the
decreased to 15.5 NTU. The
dependent variable is not affected by
reduction in turbidity in formation
unexamined external factors. In the
water is attributed to the addition of
context of this research, one of the control
chitosan as a coagulant, which
variables includes aluminum sulfate
possesses polyelectrolyte cations
(alum).
that interact with the colloidal
B. Results and discussion
particles present in the formation
water, leading to the formation of
a. As a coagulant alternative:
easily sedimentable flocs. Chitosan
derived from crab shells serves as
The results of the jar test are presented in
evidence of the existence of
the bar graph below:
colloidal particle binding
interactions within the formation
Gambar 7. Persentase penurunan turbidity
water.

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 To assess the effectiveness of membrane, leading to bactericidal
chitosan from crab shells works as effects.
a coagulant, we used this formula:
Gambar 7. diameter zona bening
Turbidity Reduction Percentage = dari alum, kitosan 50mg, 100mg ,
[(Average turbidity before chitosan - 150mg dan 200mg:
Average turbidity after chitosan) :
Average turbidity before chitosan] X
100.

= [(21.33 - 16.7) : 21.33] X 100

= 27.33%

In simpler terms, this formula helps

Image 7, pertaining to the clear zone,
us measure the effectiveness of
concludes that a 150 mg dose of chitosan
chitosan in clearing cloudy water. A
represents the most optimal dosage for
27.33% reduction in turbidity
this antibacterial activity. This substantiates
indicates that chitosan is good at
the potential utility of chitosan as an
making water clearer. This analysis
antibacterial agent. Chitosan's
is crucial for evaluating how well
antimicrobial properties render it highly
chitosan works as a water
effective in inhibiting bacterial growth in
treatment agent.
river water. Chitosan functions by
disrupting bacterial cell walls, interfering
b. As an antibacterial agent:
with membrane permeability, and
sequestering heavy metals commonly
Chitosan is a bioactive substance
found in river water. Tha high molecular
with versatile applications in the
weight polymer of chitosan forms a
fields of pharmacy, agriculture, and
protective layer around microorganisms,
industrial environmental
thereby preventing the proliferation of
management. In its capacity as a
bacteria.
bioactive agent, chitosan exhibits
the ability to inhibit the growth of
C. Conclusion
Streptococcus mutans bacteria.
The chitosan compounds function The exoskeletons of crustaceans,

by disrupting the bacterial cell such as crabs, present an excellent

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 opportunity for reducing water pollution (2019). Penambahan mineral kalsium
in Surabaya by beeing as a dari cangkang kepiting bakau (Scylla
replacement for alum (aluminum serrata) pada pakan terhadap
sulfate) coagulants. This is possible pertumbuhan dan kelangsungan
through a three-stage process that hidup udang galah (Macrobrachium
involves deproteinization, rosenbergii). Depik, 8(3).
demineralization, and deacetylation to https://doi.org/10.13170/depik.8.3.12
produce chitosan. Optimum 090
effectiveness was achieved at a
Kurniawan, Y. (2021). EFEKTIVITAS
dosage of 150 mg, which successfully
LIMBAH CANGKANG KEPITING
reduced the levels of turbidity and
SEBAGAI BIOKOAGULAN DALAM
exhibited antibacterial properties in
PENURUNAN KADAR KEKERUHAN
Surabayan river water.
DAN WARNA AIR BAKU SUNGAI
KAPUAS. Jurnal Teknologi
Lingkungan Lahan Basah, 10(1).
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