JST Vol. 21 (1) Jan. 2013 - Pertanika Journal - Universiti Putra ...

Eugene Ng, Y. J., Yap, C. K., Zakaria, M. P. and Tan, S. G.
Fig.1: Map showing the transplantation sites for Perna viridis
Note: A- Sungai Melayu B- Kampung Pasir Puteh
and vigorously shaken to produce homogeneity. All individuals of P. viridis were dissected
into byssus, total soft tissue and shell. Half gram of the samples was digested in concentrated
nitric acid (AnalaR grade, BDH 69%) in a hot-block digestor first at a low temperature (40°C)
for 1 hour before the temperature was increased to 140°C for 3 hours. The digested samples
were later diluted in 40ml double distilled water (DDW). The sample was then filtered through
Whatman No. 1 filter paper and the filtrate was stored in an acid-washed pill box until metal
determination. For the analyses of the total heavy metal concentrations in sediment samples, the
open digestion method was used. About 1g of each dried sample was digested in a combination
of concentrated nitric acid (AnalaR grade, BDH 69%) and perchloric acid (60%) in the ratio
of 4:1. On top of that, the geochemical fractions of Cd, Cu, Fe, Ni, Pb and Zn in the sediments
were obtained by using the modified Sequential extraction technique (SET), as described by
Yap et al. (2002). The first fraction was EFLE (Easily, freely, leacheable or exchangeable).
About 10g of the sample was continuously shaken for 3 hours with 50ml 1.0M ammonium
acetate (NH 4CH 3COO) at pH 7.0 at room temperature for this fraction. The second fraction
was ‘Acid-reducible’. The residue from fraction 1 was continuously shaken for 3 hours with
50ml 0.25M hydroxylammonium chloride (NH 2OH.HCL) acidified to pH 2 with HCL at room
temperature, and this was followed by the third fraction which was ‘Oxidisable-organic’.
The residue from fraction 2 was first oxidized with 30% H 2O 2 in a water bath at 90–95°C.
After cooling, the metal released from the organic complexes was continuously shaken for 3
hours with 1.0M ammonium acetate (NH 4CH 3COO) acidified to pH 2.0 with HCL at room
temperature. ‘Resistant’ was the last fraction where the residue from fraction 3 was digested
in a combination of concentrated nitric acid (AnalaR grade, BDH 69%) and perchloric acid
(AnalaR grade, BDH 60%), as performed in the open digestion method.
78 Pertanika J. Sci. & Technol. 21 (1): 283 - 298 (2013)