1

Chapter 1
The Problem and Its Scope
Rationale

Industrialization proves itself to be both a boon and a bane to the society and
to the environment where it thrives. The rise of different industries and the onset of
large-scale manufacturing have contributed much to the ease at which people operate.
However, this advancement may have also brought with it some detrimental effects--
one such concern is the increasing environmental pollution from industrial wastewater
particularly in developing countries. One of the major classes of water pollutants are the
heavy metals which are of particular concern as they bring harmful effects to
humans.
1
Their intentional or accidental dispersal to the environment also poses a
threat to other living systems especially the to the aquatic biota. Notable sources of
heavy metal-contaminated water are the different kinds of industries such as metal
plating facilities, mining operations, and tanneries.
2

Due to their mobility in aquatic ecosystems and their toxicity to higher life
forms, heavy metals in surface and ground water supplies have been prioritized asmajor
inorganic contaminants in the environment.
3
The need to exert efforts in removing
heavy metals from aqueous solutions is now being addressed using conventional
methods which involve the isolation of the said contaminants. These processes include:
coagulation, flocculation, chemical oxidation and reduction, ion exchange, physical
adsorption, filtration, electrochemical treatment, and evaporation. However, these
procedures have significant fallbacks as they are prone to incomplete removal, high
energy requirement, and toxic sludge production, and are very expensive.
2
Thus,
2

alternative methods for metal separation from aqueous solutions were developed, and
one such alternative is Bioadsorption.

Bioadsorption refers to the removal of metal or metalloid species, compounds,
and particulates from a solution by a biological material.
4
Agricultural waste products
and microorganisms are usually used as the sorbents. The use of lobster
(Panulirusversicolor)shells as Bioadsorbents has been suggested because of their
adsorptive capacity ascribed to the presence of chitosan. It is produced by alkaline N-
deacetylation of chitin, which is widely found in the exoskeleton of shellfish and
crustaceans. The adsorption behavior of chitosan for heavy metal removal is attributed
to: (1) its high hydrophilicitydue to a large number of hydroxyl groups, (2) large number
of primary amino groups with high activity, and (3) the flexible structure of its polymer
chain, making suitable configuration for adsorption of metal ions.
1

Aside from being natural adsorbents, lobster shells are used owing to the fact
that they are of less significance to the community as they are mainly considered as
food left over and generally thrown as wastes. Furthermore, several crustacean
biomasses had also been investigated for heavy-metal ion binding potential such as
shrimp and crab shells. However, very few, if any, studies have been conducted on
lobster biomass. These premises led the researcher to study the capacity of lobster
(Panulirusversicolor) shells as Bioadsorbent for Lead, a useful but toxic metal. It is well
established that lead is toxic to humans at high dosages, and that levels of exposure
encountered by at least some of the population are high enough to constitute a hazard
3

to health.
4
The fact that Lead ions are widely found in industrial wastewater aggravates
the threat that society is facing. One significant characteristic of lead which made the
researcher choose this metal as the contaminant is that Lead (II) can form more stable
metal-adsorbent complexes than Copper, Nickel, Zinc, and Cadmium can.
5
Simply put,
this study aims to highlight the feasible uses of an alternative and affordable adsorption
method in separating hazardous species such as Lead from aqueous environments
though the utilization of biomass such as Panulirusversicolor shells as adsorbents.

Significance of the Study

Lead, a useful but toxic metal is an integral part of the economy. Its uses span
from the production of lead-acid batteries, ballast keels of sailboats, electrodes in
electrolytic devices, fusible alloys, and solders for electronic devices to high-rise
building construction.
5
However, one fallback of this metal is that a considerable
amount of its input is not used for production purposes; a significant portion will be
discharged as waste, which will then pollute aquatic systems. Along with the beneficial
purposes come the environmental and health hazards brought by this metal, especially
when it is not disposed properly. For instance, the Australian Lead Education
Abatement Design Group Inc. reports an incident that happened in the Philippines in
1996 where the Marilao River in Bulacan, Philippines turned black with toxic waste
water discharged by a nearby industrial plant. Water samples from the river were found
out to have severe lead contamination, which reached about 1900 times Australian
allowable standards. Moreover, blood lead levels from the resident children were
4

examined and were found out to be three times higher than the Australian standard of
10 micrograms Pb per deciliter of blood.
7

The incident serves as a reminder about the dangers of water pollution though
heavy metal contamination as its adverse effects not only deteriorate the environment,
but also put peoples health at high risks. This study is conducted to determine the
feasibility of using Panulirusversicolor shells to bind and isolate lead in aqueous
solutions so as to provide an alternative means of water purification.

This study will thus benefit prospective researchers as it will provide insights
about using throw-away organic species as adsorbent materials. The information that
will be generated by this study could also be a significant addition to the existing body of
knowledge which posits that certain waste biological wastes can be used to alleviate an
environmental problem.

This study will also give insights to those working in the water purification and
treatment industry about a new alternative way of treating heavily contaminated water to
make it into a potable one. With the use of the bioadsorption process, these facilities will
be prompted to utilize less costly and equally effective means of treating contaminated
water.

The findings of this study may also give an idea to the local residents living near
bodies of water about a method which they could use in purifying water, making it free
5

of heavy metal contaminants. They may also use this as a springboard in experimenting
with other Bioadsorbents ability to purify water.

According to the World Health Organization (2011), one of the population groups,
which are at the highest risks of Lead exposure are the children. A recent report
suggests that even a blood level of 10 micrograms per deciliter can have harmful effects
on children's learning and behavior (CDC, 2011)
8
. As such, this research may also
benefit local civilians. They may use the knowledge generated by the study to remediate
environmental contingencies which may prove harmful to everyones health, especially
children.

Background of the Study


In this section, the causes, effects, and concept of water pollution are discussed.
Information on heavy metals, with Lead being the highlight, are presented. A discussion
of the two processes by which this study has its grounds on-- adsorption and
Bioadsorption-- follows. After which, properties of ideal Bioadsorbents and the
characteristics of Panulirusversicolorshell which made it suitable to be used in the study
are cited. This section also talks about the process of determining Lead concentration in
aqueous solutions using atomic absorption spectroscopy. Lastly, this section discusses
the possible isotherms by which lead adsorption through lobster shells could be
described.


6

Water Pollution
One thing that distinguishes the Earth from the other planets in the solar system
is the presence of water in its liquid form. Liquid water is in no way less unique as it can
naturally renew and cleanse itself by allowing pollutants to settle out (through the
process of sedimentation), or break down by diluting the pollutants to a point where they
are no longer in harmful concentrations.
9
However, this capability of water diminishes as
an increasing number of pollutants are incorporated into it. It also involves a
considerable amount of time for the pollutants to degrade. This poses an alarming form
of environmental contamination called water pollution. Water pollution occurs when
energy and other materials are released; thereby degrading the quality of wateralso
includes all other materials which cannot naturally be broken down by water.
9

Water pollution is classified into two according to where the pollutants come
from. These classifications are point sources which include factories, waste water
treatment facilities, septic systems and other sources that are clearly discharging
pollutants into water sources; and non-point sources which include run-off sediments,
fertilizers, chemicals and animal wastes from farms, fields, construction sites, and
mines.
9
Significant examples of water pollutants from either point or non-point sources
are the trace elements and heavy metals, like Lead, which are very toxic and harmful to
the environment, to humans, and to other biotic components upon reaching certain
concentration levels.


7

Heavy Metals
Heavy metals are a subset of elements that exhibit metallic properties,
particularly the transition metals and the representative elements. They are potentially
toxic even at very low concentrations.

Heavy metals have tremendous affinity for sulfur and disrupt enzyme function
by forming sulfur groups. They can precipitate phosphate biocompounds and catalyze
their decomposition. Due to these toxic properties, heavy metals have been considered
as one of the most harmful elemental pollutants which have significant effects on human
health, aquatic biota, and water toxicity.
1

Moreover, heavy metals are not biodegradable and they tend to accumulate in
living organisms
10
that results in the deterioration of their health. This process, called
Bioaccumulation, can lead to poisoning and other hazards especially when their
concentrations exceed the trace amounts required by the body. They enter the human
body system through food chain, drinking water, and air absorption to the skin.
10
Heavy
metals become toxic when these metals are not metabolized by the body and they
accumulate in soft tissues. (See Figure 3.1)

8


Figure 3.1 Scheme of heavy metal cycle. The heavy metals are moving from the
environment (pollution) to the human body through the food chain.

Although heavy metals are natural components of the Earths crust, their
concentrations in aquatic environments have increased due to mining and industrial
activities and geochemical processes. They can make their way to water sources by the
following contingencies: discrete discharge of industrial and consumer wastes,
percolation of contaminated soil, leaching of wastes from landfills, naturally---since for
some heavy metals, toxic levels can be just above the background concentrations found
in nature, acidic rain breaking down soils and releasing heavy metals into streams,
lakes, rivers, and groundwater.
10


Lead
Lead is a chemical element in the Carbon group with the symbol Pb from the
Latin word Plumbum. It is the most common of the heavy elements, accounting for 13
mg/ kg of the Earths crust.
11
It is a soft and malleable metal, which is also regarded as
a heavy metal. Metallic Lead has a bluish white color after being freshly cut but it soon
9

tarnishes into dull gray after being exposed to air. It has a melting point of 327 degrees
Celsius. Because of its widespread availability, this metal has commonly been used for
thousands of years even reaching as far as the Bronze Age where it was used with
Arsenic and Antimony. Several stable isotopes are found in nature, which include
208
Pb,
206
Pb,
207
Pb and
204
Pb.

Lead is used in the production of lead acid batteries, solder, alloys, cable
sheathing, pigments, and other industrial purposes. Lead pipes were also used in older
water distribution systems and plumbing. Polyvinyl Chloride (PVC) pipes contain some
Lead compounds which can be leached from them and thus contaminate it. This can be
remediated by the addition of lime and the adjustment of the pH to <7, 8-9. It can also
be released from flaking lead carbonate from lead pipes.

Lead is a metal with no known benefit to humans. Too much of it can damage
various systems of the body including the nervous, reproductive, and renal systems.
11
It
can also cause high blood pressure and anemia. Lead accumulates in the bones. Lead
poisoning may be diagnosed from a blue line around the gums. Moreover, Lead is
harmful to the developing brains of fetuses and young children. Exposure to it also puts
pregnant women in danger. Lead interferes with the metabolism of calcium and Vitamin
D. High blood Lead levels in children can cause irreversible learning disabilities,
behavioral problems, and mental retardation. At very high levels, high blood Lead
concentration can cause convulsions, comatose, and even death.
11
Due to its toxicity,
the World Health Organization (2011) established the maximum permissible amounts of
10

lead to be at 5 to 10 micrograms per liter of drinking water.
26
However, this limit is often
times exceeded to a significant amount as in the case of the Marilao River Lead
Contamination incident (1996) in Bulacan, Philippines.

Adsorption
Adsorption refers to the accumulation or binding of molecules at the interface
between two phases. It must be distinguished from absorption which refers to the
process in which a substance penetrates into the actual interior of crystals, of blocks of
amorphous solids, or of liquids. Sometimes, the word sorption is used to indicate the
process of the taking up of a gas or liquid without specifying whether the process
involved is adsorption or absorption.

Adsorption can either be physical or chemical in nature. Physical adsorption
resembles the condensation of gases to liquids and depends on the physical, or Van
der Waals, force of attraction between the solid adsorbent and the absorbate molecules.
No chemical specificity is involved in physical adsorption, contrary to chemical
adsorption where adsorbates are held to the adsorbent, which is usually solid, by
chemical forces. Chemical adsorption frequently involves energy of activation.

Parameters taken into consideration when choosing an appropriate adsorbent
are mainly the sorption capability, regeneration ability, kinetic parameters, price and
market availability.
13
Maximum sorption capability is the most important parameter that
characterizes each sorbent. It is the maximum amount of the adsorbed substance
11

available for the uptake per sorbent unit mass or unit volume (usually in mg/g or meq/g).
The sorption capability is determined experimentally at constant temperature, and the
results are presented as isotherms.

Adsorption plays a critical role in the transport, bioavailability, and fate of
contaminants and naturally occurring trace compounds in both natural and engineered
aquatic systems.
12
It provides the most efficient means of removing contaminants from
solutions to extremely low levels.
12
This study on the Pb
2+
adsorption using lobster
shells does not indicate specific adsorbing mechanism.

Bioadsorption
One such environmental concern which demands necessary attention is the
contamination of aquatic systems with toxic heavy metal ions. Since all heavy metal
ions are non-biodegradable, there is a need for them to be removed from polluted
bodies of water like streams and rivers so that environmental quality standards will be
satisfactorily met.
13


Numerous physiochemical methods of removing heavy metals from aqueous
systems were developed: coagulation, flocculation, chemical oxidation and reduction,
ion exchange, physical adsorption, filtration, electrochemical treatment, and
evaporation.
2
However, these methods have significant disadvantages as they are
prone to high-operating costs, incomplete removal, and production of large quantities of
12

wastes. As an alternative to these methods, a promising and less costly way of
addressing heavy-metal contamination evolved: Bioadsorption.

Bioadsorption is a promising technology for the treatment of heavy-metal
contaminated aqueous systems which is primarily based on metal-biomaterial
interactions. The Bioadsorption process has two phases: the biomaterial as the solid
phase, and the solvent water with dissolved metal ions in it as the liquid phase.
14


Bioadsorption is the capability of active sites on the surface of biomaterials to
bind and concentrate heavy metals from even the most dilute aqueous solutions. The
process of metal ion binding is comprised of many physicochemical processes like ion
exchange, complexation, microprecipitation, and electrostatic interactions.
13

The advantages of this process include low-cost and ease of operation, and
selectivity against the alkaline materials when compared to the conventional methods of
heavy-metal removal. In Bioadsorption, agricultural by-products are utilized as the
adsorbents which exhibit the capability to uptake water contaminants brought by human
activities.

This study utilizes the mechanisms governing Bioadsorption and the effectiveness
it entails in removing lead ions found in aqueous environments.


13


Bioadsorbents
Adsorbents that come from biological materials are known as Bioadsorbents,
examples of which are non-living microorganisms such as bacteria, fungi, yeast, and
algae. Extra-cellular polysaccharides secreted by microorganisms are also
recommended as surface active agents for heavy metal removal.
13
Other examples of
Bioadsorbents are tobacco dust, coconut shell powder, chitin-rich wastes, activated
charcoal, and nutshells.

Plant biomaterials can also serve as Bioadsorbents. Residual products of wheat,
corn, rice plant and other farm crops may serve such purpose. The plant origin
biomaterials contain hemicellulose, lignins, extractives, lipids, proteins, simple sugars,
water hydrocarbons, and starch whose functional groups participate in heavy metal
removal through the mechanism of complexation. Those functional groups include
carbonyl, carboxylic, amine, and hydroxyl groups which may also uptake metal
adsorbates by ion-exchange processes.

Biomolecules found in Bioadsorbents contain ionizable functional groups such
as carboxyl, phosphate, and sulfate groups. Lead ion-adsorption may involve different
binding mechanisms. One possibility is metal-ion exchange where the Pb
2+
ions will
bind to the functional groups via cationic binding. This study of Pb
2+
Bioadsorption using
chitosan-containing lobster shells are based on the premise of Pb
2+
cationic nature and
the presence of organic functional groups in the Bioadsorbent.
14


Spiny Lobster (Panulirusversicolor)
Spiny lobsters, also known as langouste or rock lobsters, constitute the family
Panulirudae of the class Malacostraca, subphylum Crustacea, under the phylum
Arthropoda. Although they superficially resemble true lobsters in terms of overall shape
and having a hard carapace and exoskeleton, the two groups are not closely related.
Spiny lobsters can be easily distinguished from true lobsters by their very long, thick,
spiny antennae, by the lack of chelae or claws on the first four pairs of walking legs,
although the females of most species have a small claw on the fifth pair, and by a
particularly specialized larval phase called phyllosoma. True lobsters have much
smaller antennae and claws on the first three pairs of legs, with the first being
particularly enlarged.
Panulirus versicolor is a species of spiny lobster that lives in tropical reefs. Other
names include painted rock lobster, blue lobster, and blue spiny lobster. In the
Philippines, it is called Banagan. Reaching as long as 30 cm in body length and
weighing 950 g ,this species has a cylindrical carapace armed with blackish spines of
various sizes. Its supra-orbital spine is stronger and more curved than in other species
of the same genus. The frontal plate is armed with two pairs of spines, the anterior
slightly larger. Its ground color on carapace is dark blue, decorated with irregular white
lines. Its dorsal surface of abdomen is colored with a central white band and a marginal
dark blue on the posterior margin of each segment. The peduncles of first antenna,
walking legs and swimmerets have longitudinal white lines. The tail fans are bluish
15

fringed with white lines. This species is mostly found in rocky areas and sheltered edges
of protected reefs washed by strong currents of clear water.
25








Figure 3.2 Spiny Lobsters (Panulirusversicolor)

Chitosan
Chitin and its deacetylated form, chitosan, are two biopolymers that come from
crustacean shells such as shrimp, lobsters, and crabs which have the ability to fix a
great variety of heavy metals. In this study, chitin and chitosan are the active sites of
bioadsorption. The strong affinity of metal ions for these sorbents is explained by the
relatively high proportion of nitrogen sites. Chitin and chitosan are nitrogenous
polysaccharides that are made up of acetylglucosamine and glucosamine units. In fact,
these two polymers have exactly the same basic chemical structure: (1 4)-2-
acetamido-2-deoxy--D-glucan and (1 4)-2-amino-2-deoxy--D-glucan,
respectively. (See Figure 3.3)

16


Figure 3.3 Scheme of chemical deacetylation of chitin to produce chitosan

Chitosan is produced at an industrial level by chemical deacetylation of chitin
using sodium hydroxide (see Figure 2.1), but chitosan can also be produced by
enzymatic deacetylation of chitin using lysozyme, snailase, neutral protease, and chitin
deacetylase. Due to the free amino groups in chitosan, this polymer chelates five to six
times greater amounts of metals than chitin. It does not take up alkali and alkali earth
metal ions but it collects transition and post-transition metal ions from the aqueous
solution. These sorption properties have been used for environmental purposes (uptake
of heavy metals), separation processes (recovery of valuable metals), and analytical
purposes.
10

Atomic Absorption Spectroscopy
This study utilizes the process known as spectrophotometric method in analyzing
the samples. In this kind of method, the radiant energy of a very narrow wavelength
range is selected form a source and passed though the sample solution contained in a
quartz cell. The amount of radiation absorbed at certain wavelength is proportional to
the concentration of the light-absorbing chemical in a sample. It is a technique
characterized by simplicity and selectivity which makes it quite adequate for the
determination of metal ions in aqueous samples.
17

Among the spectrophotometric methods used to determine metal concentrations,
Flame Atomic Absorption Spectroscopy is particularly useful to perform water analysis.
Flame Atomic-Absorption (AA) spectroscopy uses the absorption of light to measure the
concentration of gas-phase atoms. Since samples are usually liquids or solids, the
analyte atoms or ions must be vaporized in a flame or graphite furnace. The atoms
absorb ultraviolet or visible light and make transitions to higher electronic energy levels.
The analyte concentration is determined from the amount of absorption. Applying
the Beer-Lambert law directly in AA spectroscopy is difficult due to variations in the
atomization efficiency from the sample matrix, and non-uniformity of concentration and
path length of analyte atoms (in graphite furnace AA). Concentration measurements are
usually determined from a working curve after calibrating the instrument with standards
of known concentration.

To determine whether adsorption of Pb
2+
ions has occurred, there must be a
difference between the initial and final Pb
2+
concentrations after treating the sample with
the Bioadsorbent. This can be analyzed by employing the method above. Flame Atomic
Absorption Spectroscopy provides a sensitive means of determining 60 to 70 elements.
The detection limit of this method for Pb
2+
is 5 ng/ mL.
1

Adsorption Isotherms
Adsorption is usually described by isotherms, which show how much solute can
beadsorbed by the adsorbent at a given temperature. An adsorption isotherm ...relates
the concentration of solute on the surface of the adsorbent to the concentration of the
18

solute in the fluid with which the adsorbent is in contact
19
These values are usually
determined experimentally, but there are also models to predict them, both for single
metal adsorption and multi-component adsorption. The best known adsorption
isotherms are the Langmuir and Freundlich isotherm models.

The Langmuir isotherm has three assumptions to describe adsorption: (1) The
adsorbent surface is in contact with the solution containing an adsorbate and a strong
attraction exists between the adsorbent surface and the adsorbates. (2) The solute
molecules (adsorbate) can be adsorbed to a particular number of adsorbent sites. (3) In
the adsorption process, there is only one layer of adsorbate molecules to the adsorbent
surface.
22


Figure 3.4 The Langmuir adsorption process. Scheme of the simplest adsorption of
molecules on a homogeneous adsorbent.

The Langmuir isotherm is given by the following equation:
C/qe = 1/b Qo + c/ Qo
Where Qo and b are the Langmuir model parameters, c is the equilibrium solution in
mg/L of Pb
2+
, and qe is the equilibrium amount of Pb
2+
adsorbed into the adsorbent
which is also expressed as mg Pb (II)/ g absorbent. The shape of the Langmuir
isotherm is a gradual positive curve that flattens to a constant value. The values of
19

qe,max and b can be determined by linearization of this equation, followed by linear
regression or by non-linear fitting using the original form of the isotherm equation.

The Freundlich equation is one of the earliest empirical equations used to
describe equilibrium data. This model can be applied to non-ideal sorption. It often
represents an initial surface adsorption followed by a condensation effect resulting from
extremely strong solute-solute interactions. The Freundlich equation does not consider
all sites on the adsorbent surface to be equal. Furthermore, it is assumed that, once the
surface is covered, additional adsorbed species can still be accommodated. In other
words, multilayer adsorption is predicted by this equation. The Freundlich isotherm is
expressed by the equation:
Log (q) = log (k) + (1/n) log (Ce)
Where q refers to the amount of solute adsorbed per unit weight (mg/g) of the adsorbent
used. Ce is the equilibrium solute concentration in the solution (mg/L), k and n are the
constants representing the adsorption capacity (mg/g) and the intensity of the adsorbent
respectively. In a plot log (q) versus log (Ce), the values of k and n can be obtained
from the slope and the intercept. The higher the k and 1/n, the higher the adsorption
capacity.





20

Review of Related Studies

Numerous studies about the different means of remediating water pollution had
been conducted by various researchers. This section presents a few of such
studies,which are also the grounds from which the idea of this research is based.

Mustafiz, et al., (2007) conducted a study on lead metal adsorption in a multi-
component solution consisting of Lead and arsenic using Atlantic cod fish scales upon
varying the pH of the solution. It was found out that the utility of fish scales as
Bioadsorbents associated with high Lead uptake. Elemental analysis of the scales was
also part of the their study where they were able to determine the functional groups
present in the scales and even quantify the number of sites possible for cation
exchange. The adsorptive behavior of the cod fish scales were attributed to the
predominant carbonyl and amide groups found in them.
6

Kamari A., and Wang Ngah, W.S (2008) ascertained the adsorption of Lead and
Copper ions onto a sulfuric acid- modified chitosan. The chitosan they used was
isolated from shrimp scales. The pH of zero point charge, the pH at which a solid has no
net charge when submerged into an electrolytic solution, of natural chitosan and
modified chitosan are 4.15 and 3.37 respectively. They cited that varying values of the
pH of zero point charge varies according to the sources of chitosan. In their
conclusions, they have chosen the pH 6.00 and pH 5.00 as the optimum pHs for Lead
21

and Copper adsorption systems in order to avoid the formation of lead and copper
hydroxides which will affect the adsorbing capacity of the modified chitosan.
18

In the study of Kumar and Gayathri (2009), Bael tree leaf powder was used as
bioadsorbent for lead.The experiments showed that highest removal rate was 84.93% at
a solution with pH 5, setting the contact time at 60 min and initial concentration of 50
mg/L. The effect of contact time was also investigated and it was found out that the
maximum percent lead removal was attained after about 60 min of shaking time at
different initial concentrations. The increasing contact time increased the Pb
2+

adsorption and it remained constant after equilibrium reached in 30 min for different
initial concentrations.
24
This study became the basis of the researcher that the time of
immersion has a significant effect on the lead uptake.

Bamgmose, J.T., Adewuyi S., et al. (2010), carried out an experiment to evaluate
the kinetics and capacity of chitosan to trap lead and cadmium ions in aqueous
solutions a 25 degrees Celsius. Their results show that the adsorption process is
concentration-driven withhigh capacity of chitosan for the adsorption of these metal
ions. The lead and cadmium adsorption kinetic behavior could not be described
usingthe Langmuir isotherm over the whole concentration range but Freundlich isotherm
conforms to theexperimental data. The sorption of the metal ions wasexamined at time
intervals under conditions of vigorous agitation.Contact time was varied between 2 to 12
h. The sorption kineticswas done at pH 4.5.
23

22

A study conducted by Futalan, Kan, et al (2011) showed that the adsorption
data of Copper and Lead in binary systems best fit the Freundlich isotherm while data
for the adsorption of Nickel follows the Langmuir isotherm. The experiment in binary
systems showed Lead having the highest preference for adsorption to the adsorbent,
Chitosan on Bentonite (CHB), followed by Copper, then Nickel.
16

A study conducted by Sibel Tunali Akar, Asli Gorgulub, Burcu Anilanb (2011),
et al, showed that Bioadsorption of lead(II)onto S. albusbiomass has a pH-dependent
profile and lead(II) Bioadsorption was higher when pH ortemperature was increased. As
much as 88.5% removal of lead(II) is also possible in the multi-metal mixture containing
zinc and cadmium.The Langmuir isotherm better fits the Bioadsorption data and the
monolayer Bioadsorption capacity was3.00104 mol g1 at 45 degrees Celsius.
17

Yan-Hui Li,Qiuju,Du Xianjia Peng, et al (2011), conducted a study which showed
that at pH greater than 4.4, the adsorption capacity of E. prolifera, a sea anemone,
increases rapidly with increment as a result of the ionization of the Bioadsorbents
COOH- groups. However, at pH greater than 6.00, hydroxides of lead already start to
precipitate. The study concludes that the optimum pH at which the Bioadsorbent
uptakes Pb (II) is at 5.0. They also found out that that a decrease in the particle size
(lesser than 89 nm) of the Bioadsorbent would lead to a larger surface area, thus
increasing adsorptive capacity. In the study,after a rapid increase in the removal
percentage of Pb
2+
with increasing adsorbent dosage from 0.02 to0.08 g, it only has a
slight change as the adsorbent dosage increases from 0.08 to 0.17 g. The minute rise in
23

the Pb2+removal percentagemay be attributed to the attainment of equilibrium between
adsorbate and adsorbent under the experimental conditions.
19

Liu L., Hongping L., et al (2011) used sesame leaf as bioadsorbent for Lead ions
in aqueous solutions. It was found out in the experiments that the adsorptive behavior
was significantly affected by initial pH, where the optimum pH for maximum Lead
adsorption to occur was pH 5. The immersion time needed for equilibrium to occur was
180 minutes. The monolayer saturation adsorption capacities of lead ions were 279.86,
283.01, and 298.76 mg Lead/ gram adsorbent at temperatures 293, 303, and 313 K
respectively.
26

Bioadsorption studies have also been conducted in the Philippines. Fabio, J.F.
and Varquez, M.F ( unpublished, 2009) studied the lead-binding capacity of Bangus
(Chanoschanos) fish scales whose maximum binding capacity amounted to 99.7 mg
Pb/ g adsorbent at 100 ppm Pb
2+
concentration with 0.1 g adsorbent. The optimum pH
was determined to be pH 4. The data best fitted with the Freundlich equation.
27
Arcillas J.R., and Comaingking J. (unpublished, 2013) conducted a study on the
bioadsorption of hexavalent chromium onto Napier grass (Pennisetum purpureum).
Results showed that an initial concentration of 25 ppm showed the highest percentage
removal after treatment of 2.0000 grams of bioadsorbent. The agitation time which
yielded the highest percentage removal was 120 minutes. The adsorprtion of Cr (VI)
onto napier grass did not conform to both Langmuir and Freundlich isotherms.
28

24

The above studies serve as the bases on which this investigation on the Lead-
binding potential of spiny lobster shells has its grounds. As posited by the previous
researches, it is inferred by the experimenter that the pH which shall optimally be
maintained in the course of this study is between ph 4-5 and temperature should be
held constant at 25 degrees Celsius. The researcher has also used the basic premises
of the previous studies in conducting this investigation.

Statement of the Problem
The study aims to investigate the Lead (II) adsorption potential of powdered spiny
lobster shells specifically with the following parameters:
1. The effect of initial Pb
2+
concentration on lead uptake at constant pH temperature,
agitation time, and mass of bioadsorbent.
2. The effect of the mass of bioadsorbent on lead uptake at constant pH, temperature,
initial Pb
2+
concentration, and agitation time.









25

Methodology

An experimental design on the use of spiny lobster shells as an adsorbent of
Pb
2+
was in this study. In the Pb
2+
adsorption experiment, two parameters were varied--
- bioadsorbent mass and initial Pb
2+
concentration. The schematic diagram that follows
illustrates the flow of the whole experiment.

Schematic Diagram






















Figure 4.1 Experimental Flow
Preparation of Adsorbent Material
Preparation of Stock Solution
Pb
2+
Adsorption Experiment
Spectrophotometric Analysis
Data Collection
Variation of initial Pb
2+
Concentration
Variation of Bioadsorbent Mass
Testing of Bioadsorbent for Lead
Contamination
Pilot Testing with 30 ppm Pb
2+
initial
concentration
Percent Chitin and Moisture Content
Determination
26

Materials and Equipment


The following materials were utilized in the course of the experiment: powdered
spiny lobster( Panulirus versicolor) shells, rotary sieve shaker, pH- meter, analytical
grade Pb (OAc)
2
.
3H
2
O, Erlenmeyer flasks, distilled water, evaporating dish, rotary
shaker, 1.0 MHCl solution dessicator, Flame Atomic Absorption Spectrometer, 1.0 M
NaOH solution, analytical balance, timer, 500 mL beaker, stirring rod, weigh boats,
volumetric flasks, acid buret, iron stand with clamp, wash bottle, and filter papers.

Sample Description and Preparation of Bioadsorbent

The lobster (Panulirus versicolor) shells were taken from Inday Pinas Sutukil, a
seafood restaurant located in Mactan, Lapulapu City, Cebu. The selection poses a
limitation to this study as the size, age and gender of the spiny lobsters were strictly
taken into account. Only the shells of the spiny lobster were used as bioadsobent
species.

The spiny lobster shells, cut into small pieces, were thoroughly washed with
water so as to remove any water-soluble impurities. After which, they were air- dried for
one week to remove moisture. This step was also important in order to have a more
precise weight of the adsorbent. The bioadsorbents moisture was further eliminated by
oven-drying them at 50 degrees Celsius until the weight of the shells became constant.
The oven temperature must be set at 50 degrees Celsius, since the sample involved is
a biological one. This temperature ensures that the organic matrix found in the shell will
27

not deteriorate. After a constant mass is achieved through oven-drying, the dry mass of
lobster shells was ground using a milling machine and sieved using a rotary sieved
shaker with 850 micrometers mesh size. The smallest possible size of ground lobster
shells was needed so that the surface area of the adsorbent will be maximized, allowing
a higher rate of adsorption. The powdered spiny lobster shells were preserved in
evaporating dishes inside a dessicator.

Preparation of stock Lead (II) Solutions

With 1.830 grams of analytical grade Pb (OAc)
2
.
3H
2
O inside, a 1-L volumetric
flask was filled with distilled water up to the mark in order to make a 1000 ppm
Pb
2+
stock solution. See Appendix C for computation

Determination of Moisture Content and Percent Chitin

The moisture content of the ground lobster shells was determined by obtaining
triplicate samples of 5.00-gram lobster shells and placing them on evaporating dishes of
constant mass. The samples were then oven-dried at 110 degrees Celsius, a
temperature higher than the boiling point of water so as to remove essential water from
the samples. Once constant weight is achieved, the masses of the samples were
measured using the analytical balance.

28

In order to correlate the metal-binding capacity of the spiny lobster shells to the
presence of chitin, an isolation of the said polymer was done first. The procedure will
yield a measurement of how much chitin is there for a given amount of powdered spiny
lobster shell. The European Chitin Society (1997) details how chitin can be isolated from
crustacean shells and this method was adopted in this study.
29
The steps are as follows:

Coarse purification entailed washing the adsorbent thoroughly; air-drying and
oven- drying it. This step was done in the preparation stage of the bioadsorbent in this
study.

The second step involved protein removal. Five grams of the powdered lobster
shells were prepared then added with 50 ml of 2% sodium hydroxide solution. The
mixture was heated at 60-70C for half an hour. The shells were filtered off with a
strainer and the process was repeated. The filtrate should almost be clear and
colorless. Then the shells were washed with demineralized water.

The last step involved calcium carbonate removal where 50 mL of 7%HCl were
added to the deproteinated lobster shells. The mixture was stirred at room temperature
until no carbon dioxide gas escapes anymore. The mixture was filtered off and the
product, chitin, will be oven-dried until constant weight is achieved.



29

Pb
2+
Adsorption Experiment

Prior to experimenting with the variations, a negative control group was tested. In
this case the negative control group consisted of 2.00 grams of the powdered adsorbent
mixed with 50 mL of distilled water. The solutions pH was adjusted to pH 5 using 1.0 M
HCl. The mixture was subjected to agitation using the rotary shaker at 360 rpm for 2
hours enough for dynamic equilibrium to occur. The mixture inside the flask was filtered
through suction-filtration. Any trace of Pb
2+
concentration in the filtrate was then
analyzed using the atomic absorption spectrometer.

Pilot-Testing
Duplicate samples of 30 ppm Pb2+ solutions were placed in individual
Erlenmeyer flasks. This concentration will be obtained by diluting 30 mL of the stock
solution with distilled water and 1.0 M HCl to come up with 1000 mL solutions with pH 5.
Fifty mL aliquots were prepared from this concentration. Two grams of the adsorbent
were added to each of the solutions and were agitated using the rotary shaker at 360
rpm for 2 hours enough for dynamic equilibrium to occur. The mixtures inside the flask
were separated using a strainer and was subjected to suction filtration to separate the
adsorbent from the solution. The Pb
2+
concentration of the filtrates was individually
analyzed using the atomic absorption spectrometer. The absorbance was referred to
the calibration curve and the Pb
2+
concentration of the filtrates will be determined.


30

A.Variation of Initial Pb
2+
Concentration
Duplicate samples of 40, 80, and 120, 170 and 220 ppm Pb
2+
solutions were
placed in individual Erlenmeyer flasks. These solutions of different concentrations were
obtained by diluting 40.00, 80.00, and 120.00, 170.00, 220.00 mL of the stock solution
respectively with distilled water and 1.0 M HCl to come up with 1000 mL solutions with
pH 5. This pH is the one posited by the related studies to be the optimum and will thus
be maintained to deter the formation of lead hydroxides.
Two grams of the adsorbent were added to each of the solutions and were
agitated using the rotary shaker at 360 rpm for 2 hours enough for dynamic equilibrium
to occur. The mixtures inside the flask were separated using a strainer and then
underwent suction filtration to separate the adsorbent from the solution. The
Pb
2+
concentration of the filtrate were individually analyzed using the atomic absorption
spectrometer. The absorbance was referred to the calibration curve and the Pb
2+

concentration of the filtrates was determined.

B. Variation of Adsorbent Mass
Duplicate samples of adsorbents weighing 0.50, 1.00, and 1.50 grams were
obtained using a digital balance and placed in individual Erlenmeyer flasks. Each flask
was added with 50 mL of Pb
2+
solution having a pH 5. The concentration of Pb
2+
will
depend on which concentration in procedure A showed the highest absorbance of Pb
2+
.
The samples were agitated with a rotary shaker with a speed of 360 rpm at room
temperature for 2 hours also. The mixtures underwent suction filtration after being
strained and the Pb
2+
concentrations of the filtrates were analyzed using the atomic
31

absorption spectrometer. The absorbance will be referred to the calibration curve and
the Pb
2+
concentrations of the filtrates were determined.

C. Concentration Analysis of Filtrates
Standard Pb
2+
solutions with concentrations of 0.05, 0.10, 0.30, 0.50, 1.00, and
3.00 ppm and pH 5 were analyzed for their absorbance. Using distilled water as blank
and the standard solutions, the maximum wavelength was identified. A calibration curve
was drawn from their absorbance and will serve as reference to determine the Pb
2+

concentrations of the filtrates. In the same manner, the absorbances of the filtrate
samples were determined. Their absorbance values were based on the calibration
curve.


CHAPTER 2
PRESENTATION, ANALYSIS, AND INTERPRETATION OF DATA

Data gathered from the preceding experimental procedures are presented in this
section along with their corresponding analyses and interpretation. This section
discusses the results obtained from the determination of the moisture content and
percent chitin of the spiny lobster shells. The effect of varying the initial concentration
and bioadsorbent mass on the lead-uptake of the adsorbent are presented in the form
of tables and graphs. Analyses and interpretations of the results are furnished with
accompanying literature for the purpose of clarity and conciseness.
32


Moisture Content Determination

The moisture content of the bioadsorbent was analyzed for reproducibility
purposes. Triplicate samples of 5.00 g spiny lobster shells were placed on evaporating
dishes having constant mass and were oven-dried at 110 degrees Celsius until constant
weight was achieved. The temperature was such in order to remove any essential water
which are chemically bonded to the structure of the adsorbent material. The percent
moisture with respect to the mass before oven-drying was found out to be 8.5856 %.
See Table 2.1

Table 2.1 Moisture Content Determination
Initial
Bioadsorbent Weight (g)
Final
Bioadsorbent Weight (g)
Percent Moisture (%)
5.0001 4.5718 8.5658
5.0030 4.5705 8.6448
5.0093 4.5812 8.5461
AVERAGE 8.5856 %

The percent moisture obtained can be accounted by the fact that the
bioadsorbent material had already been air-dried for two months before oven-drying in
the course of its preparation.

33

Percent Chitin Determination

This part of the study seeks to find any correlation between the amount of chitin
present in the boadsorbent material and its lead-uptake capacity. Chitin in the spiny
lobster shell acts as an active site for bioadsorption though it may not be the only
species involved. After coarse purification, the lobster shells underwent deproteination
and demineralization by immersion in 2% NaOH and 7% HCl respectively under chitin
was obtained. The immersion was followed with the addition of I
2
in KI solution where
no discoloration of the IKI occurred, implying a positive result that chitin, not chitosan,
was obtained. Chitosan would have rendered the I
2
in KI solution colorless upon
contact. The spiny lobster shell was found out to be composed of 21.7775 % chitin. See
Table 2.2

Table 2.2 Percent Chitin in the Spiny Lobster Shells
Weight of Lobster Shells
before Treatment (g)
Weight of Chitin Obtained
after Treatment (g)
Percent Chitin (%)
5.0138 1.2019 23.9718
5.0528 1.0171 201294
5.0440 1.0709 21.2312
Average 21.7775 %

Curita, as cited by Lee
30
provided a list of selected crustaceans with the
corresponding percentage of chitin and CaCO
3
in their structures. Apparently, there was
34

no entry for any lobster species. However, crab shells, which more or less have the
same structure as that of lobsters shells, were found out to have 15- 30 % chitin and 40-
50 % CaCO
3
. The study further emphasized that the chitin percentage varies from
species to species.

The moisture content and chitin determination are just some of the ways by
which this study characterized the bioadsorbent material. One limitation of this study is
its inability to provide the complete physico-chemical parameters like ash content and
mineral content of the spiny lobster shells.

Pb
2+
Adsorption Experiment

Prior to proceeding with the variations, the spiny lobster shells were examined
first for any lead contamination. Results of the Atomic Absorption Spectroscopy analysis
on the negative controls indicated that there was no lead contamination on the
bioadsorbent sample as no lead ions were detected. A significant limitation on this part
of the experiment is the lack of financial resources which would have allowed the
researcher to carry out his sampling in triplicates. As such, the samples were prepared
only in duplicates.

A pilot test was also conducted in order to find the appropriate starting
conditions. Duplicate samples of 30 ppm Pb
2+
solutions were analyzed using AAS after
treatment of 2.00 grams of bioadsorbent. The idea behind was to treat the solution
35

having the least concentration with the highest dosage of bioadsorbent in order to
ensure a reasonable range for the variation of initial concentration. Apparently, the
lead-ions were almost completely adsorbed by the bioadsorbent during the pilot test,
leaving only 0.07 ppm of lead ions after treatment. That made the researcher increase
the first denomination in the variation of initial concentrations to 40 ppm to ensure that a
notable difference can be obtained from the initial and final concentrations in the sample
variations.

Effect of the Initial Concentration on Pb
2+
adsorption
The effect of varying the initial lead concentrations on lead-uptake by the
bioadsorbent was studied by preparing lead solutions of different concentrations ( 30,
40, 80, and 120 ppm). These were treated with 2.0000 grams of bioadsorbent, agitated
with a speed of 360 rpm, and under a pH of 5.00. Agitation time was set at two hours.
Table 2.3 summarizes the results.
Table 2.3 Effect of varying initial Concentration
Initial Concetration (ppm) Final Concetration (ppm) Percentage Removal (%)
mg Pb
adsorbed
Lead uptake
(Qe)
33.5 0.07 99.79 1.67 0.835
43.5 0.12 99.72 1.99 0.997
83.5 0.17 99.80 3.99 1.995
123.5 1.1 99.11 5.94 2.972
174 7.3 95.80 8.13 4.067
223 20.3 90.90 9.90 4.950


The table shows that the concentration with the highest percentage removal was
83.5 ppm (99.80 %) with a final concentration of 0.17 ppm lead after treatment with 2.00
36

grams of spiny lobster shells. The values show that there was a gradual increase in
percentage removal as the initial concentration increased from 33.5 to 83.5 ppm and
there was a gradual drop when it reached the 123.5 ppm concentration onwards. This
drop in percentage removal maybe attributed to the saturation of active coordination
sites of the bioadsorbent, as posited in the study of Rao in 2007.
31
in that study, a
decrease in the percentage removal of chromium from 94.6 % to 78.6 % when initial
concentration was increased from 25 ppm to 125 ppm. In the given initial
concentrations, an increasing number of coordination sites that are utilized as the
system approaches equilibrium, thereby diminishing the final concentrations. Arcillas
and Comaingking
28
further quote Ghoulipour in explaining that a specific amount of
bioadsorbent offers a finite amount of active sites which can accommodate a certain
amount of heavy metal adsorbate, any excess will remain in the solution once the
saturation point was reached. Figure 2.1 shows the percentage removal plotted against
the initial concentration,


y = -0.0004x
2
+ 0.0526x + 98.295
R = 0.9982 90
92
94
96
98
100
102
0 50 100 150 200 250
Percentage
Rmeoval (%)

Initial Concentration
Percentage Removal v.s. Initial Concentration
Percentage Removal
Poly. (Percentage Removal)
37


Figure 2.1 Percentage Removal vs. Initial Concentration

However, a gradual increase can also be observed in the amount of lead ions
adsorbed as the initial concentration increased. The highest amount of lead adsorbed
can be attributed to the 223.5 ppm concentration. Figure 2.2 shows the linear
relationship between the increase of initial concentration and the lead-uptake (mg Pb
adsorbed per gram of the bioadsorbent).

Figure 2.2 Lead-uptake vs. Initial Concentration

The reason for the very high adsorption may be attributed to the combined
action of the chitin and CaCO
3
in the structure of the bioadsorbent material. Lee, et al
32

conducted a study on the lead uptake of crab shells and asserted that the high
adsorptive capacity was due to the microprecipitation mechanism that happens to lead
y = 0.0222x + 0.1117
R = 0.997
0
1
2
3
4
5
6
0 50 100 150 200 250
Lead uptake (mg
Pb/ g adsrobent)
Initial Concentration
Lead uptake v.s Initial Concetrtion
Lead uptake
Linear (Lead uptake)
38

once it comes into contact with the crab shell particles. In their study, the equilibrium
isotherm showed that at optimum pH 5.5, the crab shells were able to have an uptake of
1300 mg Pb per gram of adsorbent. Their results confirmed that the N-acetyl and
calcium carbonate groups of the crab shells were mainly responsible for the adsorption.
Furthermore, the removal of lead occurred mainly through dissolution of CaCO
3

followed by the precipitation of Pb
3
(CO
3
)
2
(OH)
2
and PbCO
3
on the crab shell surface.
These microprecitipates were then adsorbed by the chitin. The same mechanism could
have had happened in this study as posited by the grayish-white coloration of the
powdered lobster shells after the solutions were filtered.

Effect of Mass of Bioadsorbent on Adsorption

Variation on the adsorbent dosage was done and its effect on lead-uptake was
studied. Samples containing 0.5000, 1.0000, and 1.5000 of spiny lobster shells were
mixed with 123.5 ppm lead solutions at pH 5, agitated at 360 rpm and 2 hours agitation
time. Table 2.4 presents the results.
Adsorbent Mass
(g) Final Concentration (ppm) Percent Removal (%)
mg Pb
adsorbed
Binding Capacity
(Qe)
0.5002 14.5 88.3 5.45 10.9
1.0004 7.5 93.2 5.8 5.80
1.5 6.1 95.1 5.87 3.91

Table 2.4 Effect of Mass of Bioadsorbent at 120 ppm initial concentration

The highest percentage removal was after treatment with 1.5000 grams of
bioadsorbent (95.1%). The values show that as the amount of bioadsorbent increased,
39

an increase in the lead adsorbed can also be noticed. This can be primarily attributed to
the number of active sites participating in the biaodsorptive process. As the number
active sites increase, so does the number of lead being adsorbed. Figure 2.3 presents a
linear proportionality of this parameter of the lead uptake employing the line of best fit.


Figure 2.3 Percent Removal vs. Adsorbent Mass

Similar trends can be observed in other studies concerning bioadsorption such
as those conducted by Fabio
27
, Wang
3
and many others. Any increase in the adsorbent
dosage gives way to an increase in heavy-metal uptake due to the presence of more
active sites relative to the prior dosages.

Adsorption Isotherms
The Pb
2+
binding capacity of the spiny lobster shells was investigated whether it
will fit the Langmuir and/ or Freundlich isotherms, which give an idea on how these ions
are adsorbed onto the bioadsorbent material. The initial concentration which was
y = 6.8023x + 85.613
R = 0.8709
87
88
89
90
91
92
93
94
95
96
97
0 0.5 1 1.5 2
P
e
r
c
e
n
t
a
g
e

R
e
m
o
v
a
l

Adsorbent mass (g)
Percent Removal v.s. Adsorbent Mass
Linear (Percent Removal)
Linear (Percent Removal)
40

considered was 120 ppm since the 170 and 220 ppm concentrations were only
conducted as a check towards the behavior of the percentage removal with further
increase in initial concentration.

The Langmuir adsorption isotherm describes the formation of a monolayer
adsorbate on the outer surface of the bioadsorbent, and after which, no more adsorption
occurs. This isotherm represents the equilibrium distribution of metal ions between solid
and liquid phases and is valid for monolayer adsorption onto a surface containing a
finite number of identical sites. It is represented by the following equation:

Qe= Qo KL Ce
1+ KL Ce
The Langmuir isotherm parameters are determined by transforming the equation
into linear form:

1/ Qe= 1/Qo + 1/ ( Qo * KL * Ce)

Where Ce= equilibrium concentration of adosrbate ( mg Pb/ L)
Qe= metal uptake ( mg Pb/ g adsorbent)
Qo= maximum monolayer coverage capacity (mg Pb/ g)
KL= Langmuir isotherm constant

41

The linearized Langmuir isotherm is plotted by graphing 1/ Qe v.s. 1/ Ce. The
values of Qo and KL can be computed from the slope and y- intercept of the graph
respectively. See Figure 2.4
Biodorbent
Mass
Equilibrium
Conc. (Ce)
1/ Ce Lead Uptake
(Qe)
1/ Qe
0.5002 14.5 0.068965517 10.9 0.094
1.0004 7.5 0.133333333 5.80 0.178
1.5 6.1 0.163934426 3.91 0.263

Table 2.5 Bioadsorbent Mass, Ce, 1/Ce, Qe, and 1/ Qe values

The table above shows the necessary values needed in order for a linear graph
to be drawn. The resulting graph is shown in Figure 2.4

Figure 2.4 Langmuir Adsorption Isotherm in Linear Form

y = 1.6518x - 0.0284
R = 0.9562
0
0.05
0.1
0.15
0.2
0.25
0.3
0 0.05 0.1 0.15 0.2
1/Qe
1/Ce
Langmuir Model
1/ Qe
Linear (1/ Qe)
Linear (1/ Qe)
42

The values of Qo and KL parameters were computed to be -35.71 mg Pb/ L
and -0.0165 respectively from the equation. The figure above shows that the
bioadsorption behavior of the spiny lobster shells as bioadsorbent mass is increased
somehow conforms to the Langmuir isotherm as indicated by an R
2
value of 0.956 of
the line of best fit.
The Freundlich adsorption isotherm is used to describe the adsorption processes
of heterogeneous surfaces.It is described by the equation

Qe= Kf Ce
1/n

and its linear form is
logQe= log Kf + 1/n log Ce
Where Kf= Freundlich isotherm adsorption constant (mg/g)
Ce= equilibrium concentration of adsorbate (mg/L)
Qe= lead uptake (mg Pb/ g adsorbent)

The Freundlich isotherm is drawn by plotting log Qev.s. log Ce. See
corresponding table and graph.


Biodorbent
Mass
Equilibrium
Conc. (Ce)
Log Ce Lead Uptake
(Qe)
Log Qe
0.5002 14.5 1.161368002 5.275 1.037252815
1.0004 7.5 0.875061263 5.625 0.763254311
1.5 6.1 0.785329835 5.695 0.592546842
43


Table 2.6 Bioadsorbent mass, Ce, log Ce, Qe, an log Qe

Figure 2.5 Freundlich Adsorption Isotherm in Linear form


From the equation of the line, the constants Kf and n, which are parameters of
the sorbent-sorbate system, are determined. Kf is an approximate indicator of
adsorption capacity. From the graph above, it was computed to be 0.53 mg Pb/ g
adsorbent. 1/n is a function of the strength of adsorption in the adsorption process. If n=
1, then the partition between the two phases are independent of the concentration. If 1/n
is below one, it indicates normal adsorption. A 1/n value greater than 1 indicates
cooperative adsorption
25
. In this case the 1/n value is 1.124, which means that the
adsorptive behavior is cooperative in nature. The adsorption data also fits the
Freundlich model with a relatively higher R
2
value of 0.974 compared to that of the
Langmuir isotherm.
y = 1.1277x - 0.2631
R = 0.9747 0
0.2
0.4
0.6
0.8
1
1.2
0 0.2 0.4 0.6 0.8 1 1.2 1.4
Log Qe
Log Ce
Freundlich Model
Linear (Log Qe)
44

The resulting analysis may have yielded adsorption data which show conformity
to the Freundlich isotherm than to the Langmuir model. However, it is important to point
out that the adsorbent-adsorbate systems have not yet reached equilibrium state as
evidenced by the high percentage removal (> 90%) even at concentrations 170 ppm
and 220 ppm of Pb (II). It cannot be claimed whether the adsorption behavior can be
modeled by either of the two isotherms because of the possibility that there may be
more free active coordination sites compared to the number of adsorbate species which
impedes the system from attaining equilibrium.

Chapter 3
Summary, Conclusions, and Recommendations

Summary
This study entails the determination of some physico-chemical properties of
spiny lobster ( Panulirusversicolor) shells which make it a suitable bioadsorbent for
lead, and more importantly highlights the effects of initial adsorbate concentration and
adsorbent mass on such bioadsorptive activity. Furthermore, the favorability of the
adsorption process was examined using the Langmuir and Freundlich adsorption
isotherm models.
Spiny lobster (Panulirusversicolor) shells were air-dried for 8 weeks and were
further oven-dried at 50 degrees Celsius. These samples were then powdered. Moisture
analysis was conducted. Determination of percent chitin of the bioadsorbent material
was done through coarse purification, deproteination, and demineralization. A liter of
45

1000 ppm lead stock solution was prepared using 1.840 g analytical grade Pb
(OAc)
2
.
3H
2
O and distilled water. Serial dilution was done to the stock solution in order to
come up with 30 (pilot test), 40, 80, and 120, 170, and 220 ppm working lead solutions.
Two grams of the bioadsorbent were added to these working solutions, and with the pH
set to 5, these were agitated at 360 rpm for two hours. The best concentration was
chosen to be used in the next step involving variation of bioadsorbent mass. 0.5000,
1.0000, and 1.5000 g of bioadsorbent were added to 50 mL of the lead solution with 120
ppm concentration. The final concentration for each variation was determined through
Atomic Absorption Spectroscopy with maximum wavelength at 283 nm. The percentage
removal, amount of lead ions adsorbed, and binding capacity of the bioadsorbent were
calculated and their relationship with the varied parameters was analyzed.

Findings
The powdered spiny lobster shells were found out to have 8.59 % moisture and
21.78 % chitin. In the variation of initial concentration, the 80 ppm solution has the
highest percentage removal at 99.80% and the trend truned to a gradual drop starting at
120 ppm with 99.08% removal attributed to the increasing number of coordination sites
involved which are in equilibrium with the lead ions. There is a direct relationship
between the initial concentration and the lead uptake of the bioadsorbent species. As
the initial concentration increased, so did the lead uptake. However, a nonlinear
relationship can be observed in the plot of percentage removal v.s. initial concentration.

46

In the variation of the bioadsorbent mass while keeping all other parameters
constant, the dosage 1.50 g showed the highest percentage removal at 95.10 %.
Increasing the adsorbent mass also increased the lead-uptake activity of the
bioadsorbent.

The adsorption mechanism cannot be determined whether it can be modeled by
either Langmuir or Freundlich isotherms because the system has not yet reached
equilibrium even at 170 ppm and 220 ppm Pb (II) concentrations.

Conclusions
Increasing the initial concentration and bioadsorbent mass results in an increase
in the lead-binding capacity of the bioadsorbent. At 120 ppm lead concetration, a
gradual drop in percentage removal can be observed. Adsorptive behavior of the spiny
lobster shells can be ascribed to the combined action of chitin and calcium carbonate
found in its structures. Microprecipitation of lead ions into Pb
3
(CO
3
)
2
(OH)
2
and PbCO
3

and its adherence to the chitins N-acetyl groups may be accountable for the high
percentage removal.. The mechanism governing lead adsorption onto spiny lobster
shells cannot be modeled by either the Freundlich or Langmuir isotherm due to the
inability of the adsorbent-adsorbate systems to reach equilibrium.

Recommendations
The researcher recommends the following to further improve this study and/ or
use this study as a springboard for future ones related to this:
47


Determine the other physic-chemical parameters of the bioadsorbent such as
ash content, protein content, and mineral content.
Investigate the effect of other parameters excluded in this research such as pH,
agitation speed, time of immersion, particle size, and temperature
Investigate the adsorption capacity of spiny lobster shells with other heavy
metals
Study the desorption capacity of the bioadsorbent
Conduct the experiment in triplicate amounts
Use large scale initial concentrations to ensure equilibrium attainment.













48

References:

1
Stanley Manahan.2005. Environmental Chemistry. CRC Press

2
Ramachandra T.V., Ahalya N., Kanamadi RD., Bioadsorption: Techniques and
Mechanisms Center for Ecological Sciences, Indian Institute of Science, Technical
Report No. 110
http://wgbis.ces.iisc.ernet.in/biodiversity/pubs/ces_tr/TR110/index.htm

3
Ramachandra T.V., Ahalya N., Kanamadi RD., Bioadsorption: Techniques and
Mechanisms. Center for Ecological Sciences, Indian Institute of Science, Technical
Report No. 112

http://wgbis.ces.iisc.ernet.in/biodiversity/pubs/ces_tr/TR112_Ahalya/CESTechnical%20r
eport%20-Metals210607.pdf

4
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54

APPENDIX
Moisture Content Analysis

[( ) ( )]
[( ) ]

TRIAL 1 TRIAL 2 TRIAL 3 AVERAGE
DISH Weight (g) Weight (g) Weight (g) Weight (g)
1 51.1236 51.122 51.122 51.122533
2 49.1016 49.1001 49.1002 49.100633
3 45.812 45.8112 45.8108 45.811333

SAMPLE Wo + Dish Wf + Dish wt. Dish Wf sample % Moisture AVERAGE
1 56.123 55.6938 51.1225 4.5713 8.5658
8.5856
2 54.1036 53.6707 49.100633 4.570067 8.6448
3 50.8203 50.3922 45.8113 4.5809 8.5461

Percent Chitin Determination




TRIAL 1 TRIAL 2 TRIAL 3 TRIAL 4 AVERAGE
DISH Weight (g) Weight (g) Weight (g) Weight (g) Weight (g)
1 51.1394 51.1371 51.1316 51.1312 51.134825
2 44.3278 44.3253 44.3217 44.3215 44.324075
3 44.3236 44.3207 44.3169 44.3173 44.319625


55

Sample Wt. Sample Dish + Chitin Wt. Chitin % Chitin Avergae
1 5.0138 52.3333 1.2019 23.9718
21.7775 % 2 5.0528 45.3387 1.0171 20.1294
3 5.0440 45.3380 1.0709 21.2312

Preparation of Stock Solution
Weight of Pb (OAc)2 3 H2O needed:

()

()
()
()

Serial Dilution of Stock Solution to Make Working Solutions:
M1* V1 = M2*V2
V1= (M2* V2)/ M1
For 30 ppm
V1= ( 30 ppm * 1000 mL)/ 1000 ppm = 30 mL then dilute to 1000 mL
For 40 ppm
V1= ( 40 ppm * 1000 mL)/ 1000 ppm = 40 mL then dilute to 1000 mL
56

For 80 ppm
V1= ( 80 ppm * 1000 mL)/ 1000 ppm = 80 mL then dilute to 1000 mL
For 120 ppm
V1= ( 120 ppm * 1000 mL)/ 1000 ppm = 120 mL then dilute to 1000 mL

Atomic Absorption Spectroscopy Analysis
Negative Control, Pilot Testing, and Variation of Initial Concentration
Sample ID Description
1896.1
1896.2
1896.3
1896.4
1896.5
MC1311-2021-01
MC1311-2012-03
MC1311-2012-03
MC1311-2012-03
Negative Control
Pilot Test ( 30 ppm after treatment)
40 ppm Pb after treatment
80 ppm Pb after treatment
120 ppm Pb after treatment
170 ppm Pb after treatment
220 ppm Pb after treatment
170 ppm Pb before treatment
220 ppm Pb after treatment



57

Variation of Adsorbent Mass
Sample ID Description
1924.1
1924.2
1924.3
At 0.5 g adsorbent dosage
At 1.5 g adsorbent dosage
At 1.5 g adsorbent dosage













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