CHAPTER THREE

3.0
3.1

MATERIALS AND METHOD

Preparation of sida acuta

Materials and equipment: Seeds and sponges of sida acuta, clean plastic bag,
grinding mill, a sieve.
Method: The seeds and sponges of Sida acuta gathered into a clean plastic bag
have been dried in the oven at 105oC for 24 hours and afterwards ground with a
grinding mill. The ground seeds and sponges were sieved and of particle sizes
ranging from 0.3 to 0.6 mm, allowing for shorter diffusion path, which resulted in a
higher rate of biosorption (Adeyinka, Liang and Tina, 2007). The ground seed and
sponge were mixed at a ratio of 1:1.

3.2

Preparation of Aqueous Solutions

Materials and equipment: CdCl2, k2Cr2O7, distilled water, a pH meter, atomic

absorption spectrophotometer.
Method: Different metal concentration of cadmium and chromium was prepared by
dissolving CdCl2 and K2Cr2O7 with deionized water to have concentration of 2, 4,
6, and 8 mmol/L. Binary metal solutions were prepared by the use of the four
metal concentrations at equimolar ratios. All glassware washed with 0.1 M HCl

before and after each experiment to avoid binding of the metal to it (Hany et al.,
2004). The pH of the solutions was adjusted to the pH of 7. The concentration of
metal ions in solutions was analyzed by Atomic Absorption Spectrophotometer. A
duplicate was analyzed for every sample to track experimental error and show
capability of reproduced results (Marshall and Champagne, 1995).

3.3

Biosorption Experiment

The biosorption studies on evaluation of the Sida acuta mixture for removal of
chromium (vi) ion and cadmium (ii) ion from aqueous solutions was carried-out in
triplicate using the batch biosorption procedure (Basil et al., 2006; Lima et al.,
2007). The batch experimental procedure to determine the effect of metal ion
concentration is described below. An equilibrium contact time of 2 h was used for
metal ion-Sida acuta biomass. A 10 mg of the biomass samples was weighed and
placed in pre-cleaned test tubes in triplicates. Several metal ion solutions with
standard concentrations of 2, 4, 6, and 8 mmol/L were made from HPLC
analytical grade standards of Cr6+ and Cd2+(from k2Cr2O7 and CdCl2 respectively).
The two sets of metal solutions made separately were adjusted to pH 7.0 with
concentrated HCl. 2 mL of each metal solution were added to each tube containing
the biomass and equilibrated for 2 h by shaking at 29C. The supernatants were

analyzed for Cr6+ and Cd2+ using flame atomic absorption spectrometer model
300A. (Horsfall et al., 2005).
3.4

Characterization of Sida acuta

3.4.1 Determination of Surface Area

The specific surface area of sida acuta was estimated using Sears method by
agitating 1.5 g of the sida acuta sample in 100 mL of diluted hydrochloric acid of a
pH = 3. Then a 30 g of sodium chloride was added with stirring and the volume
was made up to 150 mL with deionized water. The solution was titrated with 0.10N
NaOH and the volume, V, needed to raise the pH from 4-9 was then recorded. The
surface area according to this method was calculated by the following equation:
S (m2g1) = 32 V-25
Where, V is the volume of sodium hydroxide required to raise the pH of the sample
from 4-9. This volume was measured in replicate and the average value was taken
for the surface area calculation.

3.4.2 Determination of Chemical Bonds in Sida acuta Mixture

Fourier transform infrared spectroscopy (FTIR) of the adsorbent was done by using
an FTIR spectrometer (Model FTIR 2000, Shimadzu, Kyoto, Japan) (Coates,
2000).

3.4.3 Determination of Bulk Density of Sida acuta Mixture

The method of Okaka and Potter (1979) was used to determine the bulk density.
Using the procedure of Okaka and Potter (1979), 50 g of sida acuta powder was
put into a 100 ml measuring cylinder and tapped to a constant volume and the bulk
density (gcm-3) calculated using the formula:
Bulk density = weight of sida acuta powder (g) / sida acuta powder volume (cm3).