Ability of Tahong (Perna viridis), Kuhol ((Helix

pomatia), and Talaba (Crassostrea gigas) to de-

lead simulated waste water preparations
Judilynn N. Solidum 1, Zharama M. Llarena 2
1
Associate Professor, College of Pharmacy, University of the Philippines, Manila
2
MS Pharmacy, College of Pharmacy, University of the Philippines, Manila
Manila, Philippines
graloheus@yahoo.com

Abstract- Shells of Tahong (Perna viridis), Kuhol ((Helix pomatia), and Talaba (Crassostrea gigas) are considered wastes and are
normally dumped in open garbage areas or back to the aquatic system. Many studies showed that family of oysters, mussels, and snails
are used as bio-indicators or used in accumulation monitoring studies of metals like Cd, Pb, Cu and Hg since their soft tissues absorb it.
Their shells, as they may contain chitin or chitosan may also have the ability to absorb heavy metals.As the waste shells are being
reused/recycled, waste materials in the environment are decreased. Vectors of diseases that swarm waste areas will be diminished and
thus safeguard public health.In general, this study aimed to assess the ability of shells of Perna viridis (tahong), Crassostrea gigas
(talaba) and Helix pomatia (kuhol) to de-lead simulated lead-contaminated waste water. Specifically, this study aimed to compare the
ability of shells of Perna viridis (tahong), Crassostrea gigas (talaba) and Helix pomatia (kuhol) to de-lead simulated lead-contaminated
waste water. It also aimed to assess if concentrations of lead in waste water affect the absorption of lead by the shell samples and if
variances in types of shells affect absorption of lead in waste water. Tahong, Talaba and Kuhol absorbed lead in simulated waste water.
However, the inherent amount of lead in the different shells affected its ability to absorb it. Talaba and kuhol, due to its inherently high
lead concentrations failed to absorb lead from simulated waste water as the amount of the heavy metal increased. Statistical analysis
showed that the concentrations of the simulated waste water affected the absorbance of lead by chitin and chitosan from different shells.
Also, chitin and chitosan extracted from different kinds of shells affected lead absorption in varied concentrations of simulated waste
water. Tahong shells showed the most efficient absorption of lead from simulated waste water preparations. Using the different shells in
the differently concentrated SWW, the percentage of lead removed ranged from -15.899% to 63.539%. Flameless Atomic Absorption
Spectrophotometer was used to quantitate the lead levels in simulated waste water, and chitin and chitosan extracted from the three
waste shells used in this study.As there is promise to clean the waste waters from lead pollutants using Tahong, Talaba and Kuhol, it is
recommended that other shell wastes be studied for similar objectives as this study. Likewise, pH of waste water, time of contact,
temperature during treatment and agitation must be looked into to optimize the de-leading ability of these shells. Further absorption of
other heavy metals is suggested to be analyzed as well.

Keywords- Tahong; Talaba; Kuho;, lead; simulated waste water

I.INTRODUCTION
A.Background of the Study
According to sources, the marine ecosystem, and
wastewater are largely contaminated with heavy metals and
oysters, mussels and snails are widely utilized as sentinel
organisms for examining the concentration of certain
contaminants in its immediate aquatic milieu [1]. Bivalves
were reported to extensively accumulate heavy metal
pollutants and act as biological indicators for bioaccumulation
of heavy metals [1,2].
Studies showed that generally, metal accumulation
by a number of marine organisms is preferred due to their
restricted ability to release these contaminants directly after
their uptake, as well as effective physiological inactivation
mechanisms, such as intracellular compartmentalization, or
metal inactivation through affinity to metallothioneins [3].
Factors that can influence the uptake and retention of heavy
metals by marine organisms in their soft tissues include
physicochemical, environmental and biological determinants.
The metal uptake rate by an organism may depend on the
elements chemical form, which may differ from site to site.
[4].
Lead as a contaminant is pervasive, and non-
biodegradable. Being such it is considered a global problem.
The general free +2 ion which is more toxic than inorganic
complexes, are released in the environment and are still being
excreted through mining, smelting, atmospheric deposition
including coal combustion emissions, hair dye products, boat
fuels, vinyl and plastics, and lead-based paint. A study
approximated that about 5.9 million metric tons of lead was
utilized in gasoline and accumulated as residue in the
environment at 4-5 million metric tons. Since lead is fairly
stationary, it will continue to reside in the area where it was
deposited for an ample number of years [5,6].
Mussels, oysters and clams are the most commonly
used bivalve groups for accumulation monitoring studies of
metals like Cd, Pb, Cu and Hg [7,8,9,10]. Pihan and Gomot
(2000) utilized kuhol (Helix pomatia) to assess the intensity of
heavy metal contamination covering the waste dump site
before and after remediation process. With the aid of snails,

978-1-4244-5089-3/11/$26.00 2011 IEEE

pollutant accumulation was modified. Crassostrea gigas is the
molluscan that is in extensive use at present, particularly for
toxicity assessment in coastal sediments [3].
Chitin and chitosan are nitogenous polysaccharides
which consist of acetylglucosamine and glucosamine units.
[11]. In 1970, EPA ordered fishing companies to stop
dumping shells of seafoods (lobsters and crabs in particular)
into the ocean. Since then studies related to chitin started.
With the prospect of chitin being present in tahong, kuhol and
talaba shells, recycling of the said wastes may prove fruitful.
Chitin was discovered to be useful in cleaning water sources
of organic contaminants, minerals, metals and oils. Chitin and
its derivative chitosan is natural, non-toxic, non-allergenic,
anti-microbial and biodegradable [12]. Chitin may even be
used as part of moisturizers, face, hand and body creams. It is
also said to have an antifungal activity [13]. In one laboratory
testing previously conducted, chitosan confirmed to be
effective in the treatment of urban stormwater [11].
B. Objectives
In general, this study aimed to assess the ability of
shells of Perna viridis (tahong), Crassostrea gigas (talaba)
and Helix pomatia (kuhol) to de-lead simulated lead-
contaminated waste water. Specifically, this study aimed to
compare the deleading ability of shells of Perna viridis
(tahong), Crassostrea gigas (talaba) and Helix pomatia
(kuhol) to de-lead simulated lead-contaminated waste water. It
also aimed to assess if concentrations of lead in waste water
affect the absorption of lead by the shell samples and if
variances in types of shells affect absorption of lead in waste
water.
C.Statement of the Problems
The following are the problems stated for this study:
Can the shells of Perna viridis (tahong), Crassostrea
gigas (talaba) and Helix pomatia (kuhol) de-lead
simulated lead-contaminated waste water?
Are the deleading abilities of Perna viridis (tahong),
Crassostrea gigas (talaba) and Helix pomatia (kuhol)
to simulated lead-contaminated waste water
comparable to one another?
Do concentrations of lead in waste water affect the
absorption of lead by the shell samples?
Do variances in types of shells affect absorption of
lead in waste water?
D.Significance of the study
This study will help find good use for shells of Perna
viridis (tahong),Crassostrea gigas (talaba) and Helix pomatia
(kuhol) as deleading agent for waste water. As the waste
shells are being reused/recycled, waste materials in the
environment are decreased. Vectors of diseases that swarm
waste areas will be diminished and thus safeguard public
health.
E.Scope and Limitations of the study
This research will deal only with the analysis of
chitin and chitosan of shells of Perna viridis (tahong),
Crassostrea gigas (talaba) and Helix pomatia (kuhol) from
Cavite, Philippines as deleading agent for simulated lead-
contaminated waste water. Flameless Aromic Absorption
Spectrophotometry is used for lead analysis. The presence of
glucosamine is assessed qualitatively.
II. METHODOLOGY
A.Research design
This study follows an experimental design. Simulated
waste water was used to assess the ability of the different
shells of bivalves Perna viridis (tahong) and Crassostrea
gigas (talaba), and gastropod, Helix pomatia (kuhol).
Instrumental analysis using Flameless Atomic Absorption
Spectrophotometry (FAAS) was utilized to measure lead
concentration removed from the simulated waste water and
lead levels inherent in chitin and chitosan from the different
shells.
B.Collection and Preparation of Samples
Bivalves, Perna viridis (tahong) and Crassostrea
gigas (talaba), and gastropod, Helix pomatia (kuhol), were
collected from Cavite coastal area. The shells were separated,
cleansed and dried. The shells were pulverized.
C.Extraction of chitin and chitosan
To 100 g of pulverized tahong shells, 50 ml of 1-10%
sodium hydroxide solution was added. The solution was
heated from 80 to 100 C in a water bath to remove the
proteins. The solution is decanted. The liquid portion is dried
over low fire using water bath.
To the de-proteinized residue, 50 ml of 1-10%
hydrochloric acid was added at room temperature to
demineralize the tahong shells. The solution was decanted.,
The liquid portion was discarded. A 1% sodium hypochlorite
solution was used to decolorize the chitin.
The same procedure as above was followed for
chitosan except that 50% sodium hydroxide will be used
employing 90-120C temperature [14].
D.Preparation of Simulated Waste Water (SWW)
Lead acetate solutions were prepared to serve as
simulated waste water at 2%,6% and 10%.
E.Deleading of simulated waste water
Ten (10 g) was weighed for each of the ground shells
for both chitin and chitosan extracted. To one hundred (100
mL) of 2%, 6% and 10% lead acetate solution, chitin and
chitosan extracts from the shell samples were added. The test
solutions were made to stand overnight in covered beakers. TABLE II MEAN LEAD CONCENTRATION (ppm) OF SIMULATED
WASTE WATER (SWW)
These were filtered the next day, keeping both the filtrates and
the residues for analysis. Simulated waste water (SWW) Lead concentration
concentration
2% 36.6516
6% 66.6968
10% 76.6516
F.Digestion of the samples
Five grams of the sample were digested in 10 mL
concentrated nitric acid in an open glass container overnight, After addition of chitin and chitosan to separate
at room temperature, and the following day at 80 C for 5 SWW preparations it was shown that there were lowering of
hours. It was cooled to room temperature, and the volume was lead concentrations relative to the control SWW except for 6%
adjusted to 50 mL with distilled water and then analyzed using SWW using Talaba chitin and 50% SWW using both chitin
flameless atomic absorption spectroscopy [15] and chitosan from talaba and kuhol (Table III )
TABLE III MEAN CONCENTRATION OF LEAD (ppm) IN SWW
Five mL of concentrated fuming nitric was added to PREPARATIONS AFTER TREATMENT WITH SAMPLE SHELLS
100mL filtrate then covered with a watch glass. Samples were
heated in a hotplate and evaporated to 50 mL without boiling. Shell Lead in 2% Lead in 6% Lead in 10%
The sides of the beaker and the watch glass were washed with SWW SWW SWW
Tahong 22.3529 39.2459 70.1357
freshly distilled water after which 3 mL of nitric acid was Chitin
added. The solution was evaporated to around 40 mL. The Chitosan 18.914 28.4465 61.4480
samples were filtered after cooling, then diluted to 50mL Talaba 33.0317 88.2956 84.8574
volume using distilled water and transferred to a high density Chitin
polyethylene container. Samples were stored at 28-30C then Chitosan 13.3635 51.8552 88.2353
Kuhol 22.9563 50.2866 79.9698
analyzed using Flameless Atomic Absorption Spectroscopy. Chitin
[adapted 15]. Chitosan 18.19 63.3937 88.8386

F. Statistical Analysis
Two-way ANOVA was used to statistically compare After subtracting the levels of lead obtained in the
if there were differences on the uptake of lead from the treated SWW preparation by chitin and chitosan of tahong,
simulated waste water relative to its concentration. talaba and kuhol, generally, the shells were effective in
removing heavy metal lead from SWW. Talaba chitin at 6%
III. RESULTS AND DISCUSSION SWW, Talaba chitin and chitosan at 10% SWW and Kuhol
cjitin and chitosan at 10% SWW showed no absorption of lead
After extraction and pulverization of chitin and by the said shells. Using the different shells in the differently
chitosan from tahong, talaba and kuhol shells, their inherent concentrated SWW,the percentage of lead removed ranged
lead levels were analyzed. Among the three samples, chitin from -15.899% to 63.539%. (Table IV). There was an
and chitosan from tahong showed the lowest lead content increase in the concentration of lead amounts in their
while talaba showed the highest lead level in its chitin and respective SWW preparations. The inherent amount of lead in
kuhol in its chitosan (Tables I). the different shells could have affected its ability to absorb it.
The discrepancy was pronounced with concomitant increase in
TABLE I MEAN LEAD LEVELS IN CHITIN AND CHITOSAN OF lead concentration in SWW. The lead saturation point in
TAHONG, TALABA AND KUHOL SHELLS Talaba and Kuhol might have been reached that instead of
Chitin Lead concentration (ppm) Lead concentration (ppm)
being able to absorb the heavy metal to itself it even gave part
Tahong 3.1427 4.0171 of its lead contents to the SWW. Tahong chitin and chitosan
Kuhol 11.896 13.576 due to its inherently low lead concentration, showed lead
Talaba 12.995 11.875 absorption in all concentration of SWW preparations.

Simulated waste water (SWW) was prepared using
2%, 6% and 10% lead acetate solution. The concentration of
lead (ppm) in SWW was analyzed using FAAS. As expected
the lowest amount was seen in 2% followed by 6% and 10%
SWW. This served as the control for this research (Table II).
Using two way ANOVA it was shown that the
concentrations of the SWW affected the absorbance of lead by
the chitin and chitosan from different shells. Also, chitin and
chitosan extracted from different kinds of shells affected lead
absorption in varied concentrations of SWW (Tables V and
VI).
 SWW. Tahong shells showed the most efficient absorption of
lead from SWW preparations.
TABLE IV MEAN LEAD LEVELS REMOVED FROM THE SWW
PREPARATIONS AFTER TREATMENT WITH DIFFERENT SHELL V. RECOMMENDATIONS
SAMPLES
As there is promise to clean the waste waters from
Shell sample Lead levels Lead levels Lead levels
removed removed removed lead pollutants using Tahong, Talaba and Kuhol, it is
from from from recommended that other shell wastes be studied for similar
2%after 6%after 10%after objectives as this study. Likewise, pH of waste water, time of
treatment treatment treatment contact, temperature during treatment and agitation must be
Tahong Chitin 14.2987 27.4509 6.5159
(39.012%) (41.158%) (8.501%) looked into to optimize the de-leading ability of these shells.
Tahong Chitosan 17.7376 38.2503 15.2036 Further absorption of other heavy metals are suggested to be
(38.395%) (57.349%) (19.835%) analyzed as well.
Talaba Chitin 3.6199 -11.5988 -8.2058
(9.876%) (-17.390%) (-10.705%)
VI REFERENCES
Talaba Chitosan 23.2881 14.8416 -11.5837
(63.539%) (22.252%) (-15.112%)
Kuhol Chitin 13.6953 16.4102 -3.3182 [1] Amiard, J.C., et. al., Influence of ploidy and metal-metal interactions
(37.366%) (24.604%) (-4.329%) on the accumulation of Ag, Cd and Cu in oysters Crassostrea gigas
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(50.371%) (10.041%) (-15.899%) December 2004; 48 (1): 68-74.
[2] Kasiotis, Konstantinos M., Organochlorine Pesticides Residues in
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SHELLS IN VARIED SWW CONCENTRATIONS
[3] Oehlmann, Jorg & Schulte-Oehlmann, Ulrike, Molluscs as
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