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International Journal of Application or Innovation in Engineering & Management (IJAIEM)
Web Site: www.ijaiem.org Email: editor@ijaiem.org
Volume 4, Issue 5, May 2015
ISSN 2319 - 4847
Evaluation of macroalgal biomass for removal
of heavy metal Arsenic (As) from aqueous
solution
Johnsi Christobel* , A.P. Lipton
Vizhinjam Research Centre of CMFRI, Vizhinjam, Kerala
*Nesamony Memorial Christian College, Marthandam
ABSTRACT
Biosorption capacity of three common marine macroalgae viz., Ulva fasciata (green alga), Sargassum wightii (brown alga) and
Gracilaria corticata (red alga) was evaluated with respect to the removal of the heavy metal, Arsenic (As) from aqueous
solution. The influence of various parameters such as pH (2 to 10), biomass weight (0.5 to 3.0 g/L), initial metal ion
concentration (2 to10 ppm/L) and contact time (30 to 150 minutes) on biosorption efficiencies were determined. Results
indicated that the optimum pH was 6 for the removal of Arsenic by U. fasciata and S. wighti, while it was greater than 6 for
G. corticata. The maximum removal of arsenic was 90.2% at biomass weight of 2g/100ml for S. wightii and G. corticata. The
optimum arsenic (As) adsorption percentage was obtained at 90 minutes of contact time for initial metal ion concentration in
all the three macroalgae biomass. The biosorption isotherms studies indicated that the biosorption of Arsenic follows the
Langmuir and Freundlich models. The maximum biosorption capacity, q max, was 2.21mmol/g in G. corticata. The Freundlich
Arsenic adsorption capacity was in the order of U. fasciata < S. wightii < G. corticata, which also suggested about the
coexistence of the monolayer adsorption as well as heterogeneous surface conditions. The FTIR analysis for surface
functional groups of unloaded algal biomass and arsenic loaded biomass revealed that the existence of amino, carboxyl,
hydroxyl and carbonyl groups on the surface of biomass cells and the possible interaction between metal and functional groups.
SEM micrograph of pretreated and Arsenic sorbed algal biomass showed morphological changes due to the exposure of heavy
metal to the algae. These results led to refer that the macroalgal biomass could form a potential, eco-friendly, cost effective and
safe alternative biosorbent for fine tuning of waste water treatment.
Keywords:-Algal biomass, biosorbent, Arsenic removal, FTIR, SEM Micrograph, isotherms, water treatment
1. INTRODUCTION
Heavy metals released into the environment seriously affect the biological system including human physiology [17].
One to the heavy metals, arsenic contaminates the environment through anthropogenic and natural sources. Plants
absorb arsenic fairly easily, which results in food contamination with high ranking concentrations. The World Health
Organization (WHO) declared that inorganic arsenic is a human carcinogen. Various health effects have been observed
both in children and adults when exposed to high amount of arsenic [19]. Consumption of arsenic contaminated fishes
cause irritation of the stomach, intestines and lungs. Also small corns or warts appear on the palms and torso.
Considering such health hazards, several remediation techniques have been developed to remove the arsenic
contaminated problems. It includes soil removal, washing, physical stabilization and use of chemical amendments.
Almost all these techniques are considered as expensive and disruptive [10].
Biosorption is an emerging, economic and eco-friendly process of removing heavy metals from aqueous solution using
biological materials such as algae [11][12], bacteria [13], yeast [9] and fungi [2]. Macroalgal biomass is one of the
most promising types of biosorbents due to the rigid macrostructure, high uptake capacity as well as the ready
abundance of the biomass [7]. Different macroalgae exhibited different affinities towards the same metal which in turn
depends upon the chemical structure of the macroalgae [31]. Electron microscopic analysis of macroalgae revealed the
presence of different metal binding sites of their cell wall (carboxyl, aminoacid, polysaccharides and sulfhydral groups)
which could play an important role in the biosorption process [33].
Macroalgae viz; Cladophora fascicularis, Chaetomorpha sp, Caulerpa, Valoniopsis and Ulva lactuca (Green algae),
Sargassum wightii, a brown algae and Gracilaria edulis a red alga were the typical examples that showed the potential
as biosorbents for efficient removal of heavy metals such as Cd, Hg, Pb, and Chromium V1 from contaminated water
[1], [22], [21]. Considering the arsenic pollution and possibility of utilizing marine macroalgae, the present study was
initiated to evaluate the biosorption potentials of 3 macroalgae viz; Ulva fasciata, Sargassum wightii and Gracilaria
corticata towards Arsenic (As) from aqueous solution.
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2. MATERIALS AND METHODS
2.1 Preparation of biosorbent (macroalgal Biomass)
Three species of macroalgae viz., Ulva fasciata (green alga), Sargassum wightii (brown alga) and Gracilaria corticata
(red alga) available abundantly in the rocky coasts of south west India (Lat 807” to 8022”N, Long 7202” to 77033”E)
were collected and washed gently with distilled water. The algae were shade dried and then oven dried at 60oC over
night. The dried algae were ground into fine particles of 0.5mm size. The algal biomass was preserved in polythene
bags for biosorption experiments.
2.2 Preparation of metal solutions
Sodium arsenate was used for preparing the synthetic metal bearing solution: Arsenic (As) as stock solution of 100
ppm/L was prepared by dissolving Sodium Arsenate (Analytical grade) in 1L of 0.1N Sodium hydroxide solution.
From this, different dilutions such as 2, 4, 6, 8 and 10ppm were prepared using deionised water.
2.3 Batch Biosorption Studies
The Biosorption experiments were conducted in 250ml Erlenmeyer flasks containing 100 ml Arsenic contaminated
aqueous solution. The influence of parameters such as pH (2,4,6,8 and 10), contact times (30,60,90,120 and 150mts),
metal concentrations (2,4,6,8 and 10ppm) and macroalgal biomass (0.5,1.0,1.5,2.0,2.5 and 3 g/100ml) on arsenic
biosorption was investigated by varying any one of the parameters and keeping the other parameters constant. The
experiments were carried out at room temperature of 27±20C. The initial metal ion concentration (Ci) in the aqueous
solution was determined as per APHA (APHA, 2005) using Atomic Absorption Spectrophotometer (AAS). Then the
mixtures were mixed slowly in a rotary shaker [Remi] at an agitation rate of 120 rpm for 30 minutes. After 30mts, the
solid phase was separated by filtration through Whatman No-1 Filter paper. The heavy metal concentration in the
filtrates (Cf) was measured by AA Spectrophotometer.
2.4 Determination of Arsenic ions in the solution
Biosorption experiments were carried out in duplicates and average values were used in the analysis. The percent
biosorption of heavy metal (arsenic) ion was calculated as follows:
Ci-Cf
Biosorption(%) =
x100
Ci(As)
where Ci- initial metal ion concentration of Arsenic (mg/L)
Cf- final metal ion concentration (mg/L).
2.5 Biosorption isotherms
Isotherms were measured by varying the initial metal ion concentrations at the optimum conditions for each metal.
Different biosorption models were used for comparison with the experimental data.
2.6 FTIR Analysis
The pretreated and biosorbed algal biomass samples were analyzed using Fourier Transform Infrared (FTIR)
Spectroscopy. The algal biomass samples were encapsulated in KBr at a ratio of 1:100. The 1K spectrum was
collected using a Perkin-Elmer spectrophotometer within the range 400-4000 cm-1
2.7 SEM analysis
SEM has a high resolution, making higher magnification possible for closely spaced materials. In the present study
surface morphology of control (pretreated) and biosorbed algae were observed using Scanning Electron Microscope
(Hitachi s.54000).
3. RESULTS AND DISCUSSION
Three macroalgae encompassing the green, brown and red ones were used for the biosorption of Arsenic (As) from
aqueous solution and the results of influence of each parameter are discussed below:
3.1 Influence of pH
The influence of pH value on the biosorption of Arsenic on to the three macroalgal biomass was evaluated and the
results are presented in Fig.1. From Fig.1, it is observed that, as the pH was increased from 2 to 6 the biosorption
capacity also increased. The pH is considered as the most important parameter that could affect the biosorption of
metal ions from solutions [8], [16]. This is because, at lower pH, the concentration of positive charge (proton)
increased on the sites of biomass surface which restricted the approach of metal cations to the surface of biomass [14].
As the pH increase, the proton concentration decreases and the biomass surface is more negatively charged. The
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biosorption of the positively charged metal ion increased till reaching their maximum biosorption around pH6 for U.
fasciata (84.6%) and S. wightii (87.1%) and pH8 for G. corticata (85.4%). In accordance with our finding, [24]
reported that the maximum up take of cadmium, nickel and zinc by green algae, Codium vermilara, and Spirogyra
insignis, brown algae viz; Fucus spiralis, Ascophyllum nodosum and red algae such as Asparagopsis armata and
Chondrus crispus was at pH6. The increase of biosorption with increasing pH also be related to both Carboxyl and
sulphonate groups on the cell walls of the biomass and to metal chemistry in solution [27], [30], [5].
% adsorption by
100
Macroalgae
Arsenic adsorption (%)
90
Concentration
80 of
Ulva
Sodium arsenate
70
fasciata
(ppm)
69.5
2 60
76.2
4 50
84.6
6 40
82.1
8 30
72.5
10
20
Sargass Gracilar
um
ia
wightii corticata
75.4
72.1
83.5
75.3
87.1
82.5
82.7
85.4
67.8
76
10
0
2
4
6
8
10
pH values
Fig. 1. Influence of pH on Arsenic adsorption by macroalgae
3.2 Influence of biosorbent dosage
The pattern of influence of macroalgal biomass dosage on the biosorption of Arsenic from aqueous solution is shown in
Fig.2. As observed from Fig.2, a steep increase in the Arsenic uptake took place as the biomass of S. wightii and G.
corticata was increased from 0.5g to 2g/100 ml dosage. The biomass concentration of 2g/100ml has shown maximum
removal of 90.2% of Arsenic from the aqueous solution. In contrast, U. fasciata biomass at concentration 3g/100ml
showed maximum removal of 80.4% Arsenic from aqueous solution. Above the optimum biomass dosage, the
percentage uptake of Arsenic was decreasing gradually with the increase in concentrations. This effect could be
explained by the formation of aggregates of biomass at higher doses which decreases the surface area of biomass
adsorption [15]. These results coincided with the findings of [29] in the uptake of lead from aqueous solution by brown
algae Sargassum myriocystum and biosorption of Pb(II) and Cd(II) from aqueous solution using green alga Ulva lactuta
biomass [25]. Similarly [24] suggested that the electrostatic interactions between cells could function as a significant
factor in the relationship between biomass concentration and metal biosorption.
Fig. 2 Influence of algal biomass on adsorption of Arsenic
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3.3 Influence of Contact time
Contact time was highly influencing the biosorption process. The Fig. 3 indicates the effect of contact time on the
biosorption of Arsenic ions using optimum biomass (2g/100ml) of U. fasciata, S. wightii and G. corticata. These
results indicates that the biosorption of Arsenic metal was rapid during the initial 30 minutes, then it gradually
increased further the equilibrium was attained at 90 minutes in all the three macroalgal biomass used. Increased in
contact time beyond 90 mts did not improve the uptake capacity of biomass. Contrary to that, in G. corticata, the
biosorption percentage was maximum (87.5%) at 60 mts contact time. It was similar to the observation made by [32].
According to them maximum adsorption (90%) of lead, copper, cadmium, zinc and nickel occurred within 60 minutes
by the marine algae, Sargassum sp., Ulva sp., Gracilaria from dilute solutions.
Macroalgae
100
90
Agitation time
80
mts 70
30 60
60 50
90 40
30
120
20
150
10
Ulva
fasciata
79.4
81.1
82.6
68.5
62.2
Sargass Gracilar
um
ia
wightii corticata
79.2
81.2
84.6
87.5
85.1
85.4
75.4
72.3
64.2
65.6
0
30
60
90
120
150
Contact time (minutes)
Ulva fasciata
Sargassum wightii
Gracilaria corticata
Fig. 3. Influence of contact time on Arsenic adsorption by Macroalgae
3.4 Influence of metal concentration
The data presented in Fig.4 showed the effect of metal concentrations on biosorption process. The biosorption of
arsenic had increased with the increasing concentration of arsenic from 2 to 10 ppm in the presence of macroalgae such
as S. wightii and G. corticata. They removed maximum percentage of 75.1% and 77.4% respectively at 10 ppm
concentration. It indicated that biosorption was even favourable for the higher initial metal ion concentration. Similar
results were reported by [29] in biosorption of heavy metal lead from aqueous solution by non living biomass of
Sargassum myriocystune. In contrast, U. fasciata attained its maximum biosorption percentage of 67.2% at 6 ppm
metal concentration. Then the biosorption was decreased with increasing the metal concentrations. These results were
in accordance with observation made by [4] on the biosorption of cadmium and lead from aqueous solution by fresh
water alga, Anabaena sphaerica biomass. This behavior was due to the fact that initially all binding sites on the
biomass surface were vacant resulting in high metal biosorption at the beginning. After that with increasing metal
concentration, the biosorption of metal was decreased because of a few active sites were available on the surface of the
algal biomass.
% adsorption by
Macroalgae
90
80
Concentration of
70
Ulva
Sodium arsenate
fasciata
(ppm)
60
60.1
2 50
63.5
4
40
67.2
6
56.5
8 30
52.3
10 20
Sargass Gracilar
um
ia
wightii corticata
65.8
63.7
66.7
65.2
69.5
71.3
70.2
75.5
75.1
77.4
10
0
2
4
6
8
10
Arsenic concentration (ppm)
Ulva fasciata
Sargassum wightii
Fig. 4.Influence of arsenic concentration on the percentage adsorption of macroalgae
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3.5 Adsorption isotherms
The Langmuir and Freundlich isotherms describe the adsorption phenomena at the solid liquid interface and the
isotherms data were used for the design of adsorption system and to understand the relation between adsorbent and
adsorbate [26].
The results of Langmuir model parameters for arsenic adsorption of macroalgae indicated that the efficacy of
adsorption was maximum in G. corticata (qmax=2.21±0.26 m/mol/g) and minimum in S. wightii (qmax=1.31±0.16). The
correlation coefficient was high in S. wightii (2.9±0.31) and G. corticata (2.12±0.12) but low in U. fasciata (1.1±0.22)
as could be noted from the Table1.
The Freundlich Arsenic adsorption capacity increased in the order U. fasciata<S. wightii<G. corticata which was in
agreement with the findings of Langmuir isotherms (Fig.5 to 7). The adsorption intensity increases in the order U.
fasciata<S. wightii<G. corticata which was also in agreement with Langmuir trends. This observation may imply that
monolayer adsorption as well as heterogeneous surface conditions could co-exist under the applied experimental
conditions and therefore both Freundlich Langmuir isotherms could be used to model the experimental adsorption data
for the three selected marine macroalgae. Among these, G. corticata had shown the highest affinity of arsenic
adsorption making it an extremely promising adsorbent for the removal of arsenic when present at low residual
concentration.
Table 1 Langmuir model parameters for arsenic adsorption of macroalage
Macro algae
qmax(mmol g-1)
b (L mmol -1)
r2
Ulva fasciata
1.82±0.34
1.12±0.22
0.98
Sargassum wightii
1.31±0.16
2.29±0.31
0.97
2.21±0.26
2.12±0.12
0.98
Fig. 5 Freundlich adsorption isotherm of Arsenic by Ulva fasciata
(Arsenic concentration 1.0 g/L, T = 25oC, pH = 6.2, Agitation Time 80 min)
Fig. 6 Freundlich adsorption isotherm of Arsenic by Sargassum wightii
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Fig. 7 Freundlich adsorption isotherm of Arsenic by Gracilaria corticata
3.6 Characterization of Biosorbent
3.6.1 Fourier Transformation Infrared Spectroscopy (FTIR)
Fig. 8 FTIR Spectrum of U. fasciata (control)
Fig. 9 FTIR Spectrum of U. fasciata (Arsenic treated)
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Fig. 10 FTIR Spectrum of S. wightii (control)
Fig. 11 FTIR Spectrum of S. wightii (Arsenic tretaed)
Fig. 12 FTIR Spectrum of G. corticata (control)
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Fig. 13 FTIR Spectrum of G. corticata (Arsenic treated)
The results of FTIR spectra are illustrated in Fig. 8 to 13 of the unloaded biomass (control) and Arsenic (As) loaded
biomass. These results represented the information about the functional groups on the surface of the cell wall of the
biomass and the possible interaction between metals and the functional groups. From these data, it is clear that in
Fig.11 the FTIR spectra of Arsenic loaded S. wightii biomass showed a significant reduction in the Hydroxy (-OH) and
–NH stretching band (3400 to 3315 cm-1). [23] reported that the reduced distance between bands after metal binding
was due to higher symmetry in the cell wall matrix as a result of complexation with the metal ion. It was confirmed by
their studies on the mechanism of adsorption of heavy metal cadmium by Sargassum biomass. The change in band
structure was due to nitrogen-hydrogen band stretching. Similarly, a significant decrease in wave numbers of 2966.31
to 2927.74 and 2964.39 to 2929.67 cm-1 was observed in the C-H stretching band of U. fasciata and G. corticata. It
indicated that significant involvement of C-H stretching band in the binding of Arsenic to macroalgae viz., U. fasciata
and G. corticata.
The C-O (carbonyl) group exhibited the peak at 1026 and 1020 cm-1 and c-o-s (sulphated polysaccharide) band
observed at 1255 and 1278 cm-1 in the pretreated samples of U. fasciata and S. wightti were missing in the Arsenic
loads samples due to their predominant interaction of that groups with the Arsenic. Similar to that, in G. corticata the
unloaded sample showed 3, 6 anhydro galactose vibration band at 927 cm-1. It was not found in the arsenic treated
sample indicates the significant involvement of this group in binding of heavy metal arsenic. A significant reduction in
the wave number of non sulphated B_D galacto phranose band (889 to 811cm-1) was observed in the arsenic treated G.
corticata sample suggesting that group involved in the biosorption process.
3.7 SEM Analysis
The SEM micrograph of raw U. fasciata showed folded structures (Fig. 14a). But those folded structures were not
found in the Arsenic loaded image of U. fasciata. Instead of that an apparently smoother surface was observed (Fig.
14b). These results coincided with the findings of [20] on the SEM micrograph of Ulva lactuca, a green alga loaded
with or without the metal of Cu II and Cr III. Similar to U. fasciata, some morphological differences were observed
between the pretreated and arsenic loaded brown alga, Sargassum wighti as could be noticed from Fig.15(a&b). On the
surface of pretreated S. wightii, protuberance and microstructures were observed. [5] studied the raw Sargasum sp. and
suggested that protuberance and microstructure on the surface may be due to the calcium and other salt crystalloid
deposition. In the SEM micrographs of arsenic loaded S. wightii, shrinkun surface was noticed. According to [6] such
morphological changes were due to the binding of Arsenic with alginic acid and sulphated polysaccharides embedded
in the matrix of S. wightti. It was confirmed by the change observed in the hydroxyl and ester sulphate link of FTIR
spectrum of arsenic loaded S. wightti in the present study. SEM micrograph of raw red alga, Gracilara corticata
Fig.16(a) also showed some folded structures similar to those observed in U. fasciata. But the numbers of folded
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structures were fewer and less pronounced. In the arsenic loaded G. corticata, Fig. 16(b) the surface was apparently
smoother with mount like structures. This result coincided with earlier studies by [20] on the heavy metal Cu II and Cr
III binding on the surface of Palmaria palmate, red alga. Electron Microscopic studies on the structure of G. corticata
thallus by [28] revealed the presence of high content of sulphur with metals like zirconium, chromium, zinc, iron and
potassium could produced in the changes in surface morphology of G. corticata.
Fig. 14( a)
EM Micrograph of U.fasciata (control)
Fig. 15 (a)
SEM Micrographs of S. wightii (control)
Fig . 16 (a)
SEM Micrographs of G. corticata (control)
Fig. 14(b)
SEM Micrograph of Arsenic loaded U. fascia
Fig. 15 (b)
SEM Micrographs of Arsenate loaded S. wightii
Fig. 16. (b)
SEM Micrographs of Arsenate loaded G. corticata
4.CONCLUSION
Results of experiments could lead to the conclusion that the dried algal biomass effectively removed the arsenic from
aqueous solution. This has tremendous potential as an economic, effective, safe alternative to existing commercial
adsorbents. It is easy and effective technique for fine tuning of waste water treatment and removal of heavy metals
from the various industrial wastes containing heavy metals
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Acknowledgements
The authors express sincere thanks to Dr. Shakina, Assistant Professor, Dept of Chemistry, Sara Tucker College,
Palayamkottai for her valuable guidance throughout the tenure of this work. The first author gratefully acknowledge
the Director, CMFRI, Cochin and Scientist In charge and staff of Vizhinjam Research Centre of CMFRI, Vizhinjam for
the facilities provided to carry out this work. Thanks are extended to Dr. Sarika, Scientist, Mr. Shine, CMFRI,
Vizhinjam and Dr. K. Paulraj, H.O.D. Dept. of Botany N.M.C.College, Marthandam for their valuable suggestions
during the preparation of MS of this paper.
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