N. Siahaya et al. / Advances in Biological Chemistry 3 (2013) 549-552
550
habitat of a living sponge. This is in line with the
opinions [3,4,9] that the metal content in aquatic biota
generally increases over time because the metal is ex-
pected, so the presence of a sponge in water Hative cycle
can be used to determine the heavy metals Pb, Cd , Cr,
and Zn in the sponge type callispongia sp, sediments and
water in the waters Hative Ambon bay.
2. MATERIALS AND METHODS
Research materials are: sponges from Hative waters in
Ambon bay; Acetone (Merck), HNO3 (p.A), double dis-
tilled water and Whatman filter paper. Sponge samples
were collected by diving, cleaned and then placed in a
plastic bag and put in the ice box. 0.5 grams of sample
was put in beaker glass, add 5 mL HNO3 and then heated
at 150˚C for 2 hours. After being cooled at room tem-
perature, sample put in 25 mL volumetric flask, match
the volume with double distilled water and filtered with
Whatman paper and solution is ready to be analyzed by
ICP-OES Perkin Elmer 3000. One litre of water sample
was taken at the bottom, and immediately filtered with
filter paper of cellulose nitrate (0.45 μ) after previously
washed with 1N HNO3 and then preserved in HNO3 5%.
250 ml water sample is inserted in a Teflon separating
funnel, then extracted with APDC- NaDDC/MIBK. The
organic phase was extracted again with 5% HNO3 solu-
tion, filtered back, and ready for analysis by ICP-OES
Perkin Elmer 3000.
For sediments, they were taken from the bottom with a
Van Veen Grab sampler, stored in polyethylene bottles
and taken in laboratory put in a Teflon beaker and dried
in an oven at a temperature of 105˚C and after drying,
rinsed 3 times with double distilled water then dried
again. A total of 5 g sample was destructed in Teflon
beaker with solution of HNO3/HCl (1:3) at 100˚C for 8
hours. After that, the solution was filtered, and the fil-
trate is ready to be analyzed by ICP-OES Perkin 3000.
3. RESULTS AND DISCUSSION
3.1. General Situation of Sampling Locations
Sampling was conducted on July 4, 2011, about noon
under cloudy weather in Hative waters (Figure 1) sur-
rounded by population settlement, estuaries, navy com-
plex, harbour, oil depot of Pertamina, and sago plantation.
Callispongia sp, ca 50 grams were taken under physico-
chemical as shown in Table 1. The data clearly shows
that its water quality fits with environmental conditions
in general where sponges grow in tropical and sub-
tropical conditions with vertical distribution on coral
reefs at low tide up into the area of approximately 50
meters [6]. Also, water content, ash, and biomass of Cal-
lispongia sp, represent an integral part of metal content
in a biological sample [9].
Accumulation of Pb, Cd, Cr and Zn Callispongia sp
can be seen in Table 2 as a function of organ as skeleton
and tissue. Also determined were water and sediment
around sponge. Logically before entering into cellular
level of sponge, metals will be firstly existed in water
and sediment It seems obvious that each element shows a
different partitional pattern where cadmium as the least
amount remain largely in sediment (86.4 %). Oppositely,
chromium was almost all adsorbed by cellular sponge. It
was also clear that majority of elements have stayed in
cellular level.
As for zinc as an essential element, its highest con-
centration revealed that the source might not only come
from industrial by products but also from excretion of
living system in marine environment. So the total zinc in
this Callispongia sp apparently not an abnormal case
especially if one looks at a fraction remaining in sedi-
ment (54.1%) and consumption level of skeleton and
tissue on zinc. Generally the highest metal concentration
in sediments is influenced by several processes like
sedimentation, flocculation, precipitation, and adsorption
[5,6]. Another important parameter is bioconcentration
factor (BCF) in measuring the capability of organisms to
accumulate metals from environment into its tissue. Ac-
cording to Abdullah et al. 2007, BCF can be estimated
by comparing metal absorption between in sponge tissue
and in water (BCF sw) or sediment (BCF ss).
The ability of organisms to accumulate metals from
the environment into the tissues of the body can be cal-
culated using the bioconcentration factor (BCF). BCF
value can be obtained by comparing the ability of organ-
isms (e.g. sponges) to absorbsi metals from water and
sediment. Therefore there are two BCF values, BCF
sponge-sediment (BCFs-s) and BCF-water sponge (BCFs-
w). BCFs-s is the value of the ratio between the concen-
tration of the metal absorbed into the sponge tissue with
metal concentrations in the sediment, while the BCF sw
is the value of the ratio between the concentration of
metal that accumulates into the sponge tissue metal con-
centrations in water [8] as found in table-3 below.
From the result in Table 3 one can find the highest
value for BCF is Cr either in water or in sediment. and
this indicated that for the case of the location and sponge,
Cr is the most appropriate element to be assigned as a
bioindicator or biomonitor for Callispongia sp compared
to other metals. Some results from several investigations
have used the same method [4,5,7].
To assess the possible chemical bonding occurs be-
tween organic molecules in sponge and metal, an FTIR
analysis was carried out to look at the key functional
groups that may link to bond formation with metals.
FTIR spectrum of Callispongia sp sample can be seen in
Figure 2.
According to Terada et al. (1983) the interactions that
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