J Appl Phycol (2008) 20:6366

DOI 10.1007/s10811-007-9181-8

Heavy metal chelation by non-living biomass of three color

forms of Kappaphycus alvarezii (Doty) Doty
K. Suresh Kumar & K. Ganesan & P. V. Subba Rao

Received: 18 April 2007 / Accepted: 26 April 2007 / Published online: 15 June 2007
# Springer Science + Business Media B.V. 2007

Abstract Water pollution by toxic heavy metals is a

burning environmental problem and has presented a
challenge to humans. Removal of heavy metals using
non-living biomass of seaweeds could be a potential
solution to this problem. In the present investigation,
biomass of three color forms of Kappaphycus alvarezii,
viz. brown, green and pale yellow, were studied in the
laboratory for their heavy metal chelating capacity using
cadmium, cobalt, chromium and copper. Amongst the four
concentrations used (25, 50, 75 and 100 mg L1) maximum
chelation of Cd, Co and Cu was recorded at 25 mg L1
concentration. The highest amount of Cr was chelated at
100 mg L1 by all the three color forms. The pale yellow
form showed maximum chelation for all four metals
studied. Further, chelation in all the color forms was found
to be: Cd 5.370.5915.840.32 %, Co 21.190.1332.32
0.62 %, Cr 65.380.2788.090.51 % and Cu 59.53
0.3790.280.89 %. All the three color forms of K.
alvarezii serve as an excellent biodetoxifier as they all
chelated considerable amounts of heavy metals.
Keywords Biosorbent . Chelation . Color forms .
Heavy metals . Kappaphycus alvarezii

Introduction
Heavy metal pollution is an environmental problem of
worldwide concern. Some industrial processes result in the
K. Suresh Kumar : K. Ganesan : P. V. Subba Rao (*)
Marine Biotechnology and Ecology Discipline,
Central Salt and Marine Chemicals Research Institute,
Bhavangar,
364 002 Gujarat, India
e-mail: pvsubbarao@csmcri.org

release of heavy metals into natural water systems, leading

to increasing concern about their toxic effect as environmental contaminants. Lead (Pb), copper (Cu), cadmium
(Cd), zinc (Zn) and nickel (Ni) are among the most
common pollutants found in industrial effluents. Even at
low concentrations these metals can be toxic to living
organisms, including humans. Conventional methods for
removing heavy metals (precipitation, chemical oxidation/
reduction, ion-exchange, reverse-osmosis, membrane separation, etc.) are often ineffective and costly (Marques et al.
1991; Volesky 1994; Dey et al. 1995; Kapoor and
Viraraghavan 1995; Lu and Wilkins 1996; Zhao and
Duncan 1997). Recently, biological removal processes for
removing heavy metals from aqueous wastes have been
attracting considerable attention. These include new
approaches such as the use of marine algal biomass for
biosorption of heavy metals (Fourest and Roux 1992;
Mattuschka and Straube 1993; Volesky 1994; Volesky and
Holan 1995). Algae are of special interest in the search for,
and development of, new biosorbent materials due to their
high sorption uptake and their ready availability in
practically unlimited quantities in the seas and oceans
(Feng and Aldrich 2004). Biosorption properties of a few
algae are accredited to their cell-wall polysaccharides like
alginate and fucoidan, which have a high affinity for
divalent cations (Fourest et al. 1994; Puranik et al. 1999;
Khoo and Ting 2001; Chen et al. 2002; Davis et al. 2003).
The non-living biomass of Sargassum species, Macrocystis
pyrifera, Kjellmaniella crassiforia, Undaria pinnatifida and
Ulva species are known to effectively remove Cd, Cu, Zn,
Cr and Ni (Seki and Suzuki 1998; Yang and Volesky 1999;
Davis et al. 2000; Suzuki et al. 2005). Kappaphycus
alvarezii, a potential carageenophyte, is reported to occur
in brown, green, pale yellow and red color forms in field
cultivation (Dawes 1992; Hurtado-Ponce 1995; Suresh

64

Materials and methods

The three color formsbrown, green and pale yellowof
Kappphycus alvarezii (Doty) Doty were obtained in April
2006 from the cultivation farm of Port Okha (L 2228.528 N;
L 069 04.322 E), on the northwest coast of India.
Brown color form: Plants are dark brown in color, branches
are thick and robust, profusely and irregularly branched, the
branches are tapering towards the tips. The basal part of
the branch is very thick; plant normally measures up to
2025 cm.
Green color form: Plants are dark green in color, branches
are thick and profusely branched with fragile tips; plants are
smaller than the brown form and measure up to 20 cm.

filtered (0.22 m pore size; Millipore) and subjected to

further analysis.
Instrumentation
Heavy metal content in all the filtrates was quantified using
inductively coupled plasma optical emission spectroscopy
(ICP-OES; Perkin Elmer Optima-2000 DV). The difference
between the amount of metal present in the filtrate and the
total amount of metal present initially in the flask yielded
the amount of metal chelated by the biomass. The
percentage chelation for each metal was estimated. Means
and standard deviations were calculated.

a
Co

Cr

Cu

80
60
40
20

Pale yellow form: Plants are light yellowish in color,

smaller than the other two color forms, less branched.
Branches very fragile and break even at moderate water
current; the plant measures up to 1416 cm.

0
25

50

75

100

Metal concentrations (ppm)

b
Cd

100

Chelation (%)

The collected samples were thoroughly washed with

seawater to remove epiphytes and dirt particles and then
dried in the shade for 2 days. Samples were then brought to
the laboratory and oven-dried at 80C for 3 h to constant
weight before being pulverized (size 2 mm) in a grinder.
This non-living biomass was used for further experiments.

Cd

100

Chelation (%)

Kumar et al. 2007). In an earlier study the three living color

forms (brown, green and pale yellow) of K. alvarezii were
examined in the laboratory for their biosorption ability to
uptake Cd, Co and Cr (Suresh Kumar et al. 2007). The
present work is aimed at studying the chelation of Cd, Co,
Cr and Cu from aqueous solution by the brown, green and
pale yellow color forms of non-living biomass of K.
alvarezii in the laboratory.

J Appl Phycol (2008) 20:6366

Co

Cr

Cu

80
60
40
20

Heavy metal chelation

0
25

50

75

100

Metal concentrations (ppm)

c
Cd

100

Co

Cr

Cu

80

Chelation (%)

Chelation of heavy metals was studied by adding a 1 g

sample of each color form to an Erlenmeyer flask
containing 100 mL aqueous solution of Cd, Co, Cr and
Cu at four concentrations, i.e., 25, 50, 75 and 100 mg L1
prepared using analytical grade cadmium sulfate, cobalt
sulfate, potassium dichromate and copper sulfate. Controls
for each color form without addition of heavy metals were
also maintained. The pH was adjusted initially to 4.5 and
was found to be 5.3 at the conclusion of experiment. All
experiments were conducted in triplicate. To avoid heavy
metal contamination, glassware was soaked in 10% HNO3
for 24 h and rinsed with deionized water prior to use. Flasks
were incubated for 72 h on a shaker at room temperature
(331C) and the flask contents were subsequently syringe

60
40
20
0
25

50

75

100

Metal concentrations (ppm)

Fig. 1ac Heavy metal chelation by three color forms of Kappphycus

alvarezii. a Brown, b green, c pale yellow

J Appl Phycol (2008) 20:6366

Results and discussion

The chelating efficiency of the non-living biomass of the
three color forms of K. alvarezii (Fig. 1) revealed that the
maximum amount of Cd, Co and Cu were chelated (%)
when the heavy metal concentration was 25 mg L1, i.e.,
the brown form chelated Cd 5.370.59, Co 21.190.13,
Cu 59.530.37; the green form chelated Cd 8.870.61, Co
28.540.13, Cu 70.870.50; and the pale yellow form
chelated Cd 15.840.32 , Co 32.320.62 , Cu 90.280.89.
In the case of Cr, maximum chelation was observed at
100 mg L1 concentration, i.e. brown 68.790.20, green
77.210.42 and pale yellow 88.100.15.
Chelation decreased with increasing concentration of Cd,
Co and Cu, but increased with increasing concentrations of
Cr. Furthermore, at 100 mg L1 concentration, no chelation
of Cd or Co was found in the brown color form, and no
chelation of Cd in the green color form. Of the four metals
used, the highest chelation was recorded with Cr followed
by Cu, Co and Cd in the brown and green forms. However,
a different trend was observed in the pale yellow form,
where maximum chelation of Cu was noted, followed by
Cr, Co and Cd. This color form chelated highest amount of
heavy metals, followed by the green and brown forms. The
pale yellow form of K. alvarezii is known to chelate 65.28
0.5190.280.89 % of copper and 80.860.8688.10
0.15 % of chromium. Lee et al. (2000) demonstrated 57%
chromate chelation by a red algae Pachymeniopsis species.
A brown seaweed, Padina species, is known to chelate
copper (0.80 mmolg-1; Kaewsarn, 2002). Dried biomass of
Sargassum wightii is also reported to chelate maximally
cadmium metal followed by lead, copper and zinc,
indicating the affinity range for heavy metal ions (Kumar
and Kaladharan 2006). Brown seaweed biomass of Ecklonia maxima, Macrocystis angustifolia and Laminaria
pallida are known to sequester copper, zinc and cadmium
ions at concentrations likely to be encountered in waste
water, i.e., 0100 mg L1 (Stirk and Van Staden 2000).
Ascophyllum nodosum out-performed a commercial ion
exchange resin (DUOLITE GT-73) by accruing 100 mg
Cd2+/g biomass (Holan et al 1993). Hashim and Chu (2004)
and Tsui et al. (2006) have observed that brown seaweeds
exhibit better metal chelation properties than their red
counterparts. In contrast, in the present study, the red
seaweed K. alvarezii has demonstrated more chelation of
heavy metals. In particular, a notable increase in chelation
of Cr (68.8088.08%) was recorded by increasing the
concentration of Cr in the aqueous solution.
Conclusion
The non-living biomass of all three color forms of K.
alvarezii, a carageenan yielding red seaweed, proved to be

65

an efficient biosorbent material, especially for chelation of

Cr and Cu from aqueous solution. This is in contrast with
results obtained by Vieira and Volesky (2000), who opined
that red marine algae containing carageenan do not have
outstanding metal-sorbing properties despite having potential binding sites.
The greater chelation of heavy metals by the pale yellow
form of K. alvarezii in comparison with that by the other
two forms (brown and green) might be attributed to the
cumulative inherent physiological characteristics, especially those related to accumulation of pigments and carageenan at specific binding sites. Since the experiments
conducted here were preliminary in nature, further study is
very much necessary to understand the mechanism and
kinetics of uptake of heavy metals, which may broaden the
scope to use K .alvarezii color forms for bioremediation.
However, the present investigation shows that all three
color forms of K. alvarezii exhibit excellent heavy metal
chelating capacity, thus proving them to be potential
biodetoxifiers.
Acknowledgments The authors are grateful to Dr. Pushpito Ghosh,
Director, Central Salt and Marine Chemicals Research Institute,
Bhavnagar, Gujarat, India, for his constant support and encouragement. They also appreciate the Discipline Co-ordinator of the Marine
Biotechnology and Ecology Discipline, for providing research
facilities, and profusely thank the Department of Biotechnology
(Sanction No: BT/ PR 3309 / PID / 03 / 139 / 2002), New Delhi,
India for providing financial assistance.

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