Biotechnology Letters 23: 249–251, 2001.

© 2001 Kluwer Academic Publishers. Printed in the Netherlands.

249

Batch studies on the removal of gold(III) from aqueous solution by Azolla

filiculoides

A.P.M. Antunes1 , G.M. Watkins2 & J.R. Duncan1,∗

1 Department of Biochemistry and Microbiology, Rhodes University, P.O. Box 94, Grahamstown 6140, South Africa
2 Department of Chemistry, Rhodes University, P.O. Box 94, Grahamstown 6140, South Africa
∗ Author for correspondence (Fax: +27 (0)46-6223984; E-mail: J.Duncan@ru.ac.za)

Received 29 September 2000; Revisions requested 19 October 2000; Revisions received 24 November 2000; Accepted 27 November 2000

Key words: Azolla filiculoides, batch optimization studies, biosorption, gold(III)

Abstract
Azolla filiculoides removed 86% and 100% of gold(III) from initial metal solutions of 2–10 mg gold l−1 increasing
with increased initial concentrations of gold(III). The biomass gave greater than 95% removal efficiency from
solution at all biomass concentrations measured. Complete removal of gold occurred at pH 2, with 42% removal
at pH 3 and 4, and 63% and 73% removal at pH 5 and 6, respectively. No temperature-dependence removal was
observed.

Introduction enhance the effectiveness and feasibility of the metal

Metal wastes result in a critical loss of non-renewable
resources; thus a significant opportunity exists to min-
imize the decreasing resources by recovering and re-
cycling metal solutions (Darnall et al. 1986, Kuyucak
& Volesky 1988). Biomass, such as plant material
(Sanyahumbi et al. 1998) and fungi (Fourest & Roux
1992), have a high potential to bind and concentrate
metal ions from aqueous solutions, even when the
cells are dead. This phenomenon is termed ‘biosorp-
tion’ (Volesky 1987, Fourest & Roux 1992). Naturally
occurring biosorbents offer a potential alternative to
existing methods, not only in the treatment and detox-
ification of wastewaters, but also in the recovery of
valuable metals from dilute solutions (Darnall et al.
1986, Kuyucak & Volesky 1988). An example of such
a biomass is the small aquatic fern, Azolla filiculoides
which, in South Africa, is regarded as a weed because
of its ability to form a dense mat on stream or dam
surfaces. This ‘mat’ leads to eutrophication as the free-
flow of water and oxygen is restricted (Hill 1998). The
use of biosorbents from natural sources such as Azolla
filiculoides may possess a sequestering power supe-
rior to that of commercially available ion-exchange
resins, or the already-in-use activated carbon, and thus
recovery process.
Biorecovery of gold has received considerable
world-wide attention. Gold in mining effluents is gen-
erally at very low concentrations (1–10 mg gold l−1 ).
In addition, gold behaves very differently from other
metals and is not easily removed from solution, al-
though biosorption studies have shown that algae are
able to adsorb gold from aqueous solutions under a
variety of conditions thereby indicating that gold can
be successfully recovered (Greene et al. 1986). The
present study involves the investigation and devel-
opment of an innovative process for the removal of
gold. The advantage of Azolla filiculoides as a biomass
instead of the more expensive ion-exchange resins cur-
rently used is its natural availability and additionally it
provides an impetus to harvest the plant from water
surfaces. Batch study optimization studies were per-
formed to determine the optimal biomass and gold
concentration, pH and temperature for gold removal
by the non-viable biomass.
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Materials and methods

Materials

The water fern, Azolla filiculoides, was harvested lo-

cally from dams around Grahamstown, in the Eastern
Cape, South Africa. The plant was thoroughly washed
in deionised water and dried at 37 ◦ C. It was stored
in a cool, dry place for subsequent utilization. All
reagents used were analytical standard and obtained
from Saarchem, South Africa. Aqueous gold solutions
were prepared from hydrogen tetrachloroaurate(III).
All glassware used for experimental purposes was
washed in 2.5 M nitric acid and subsequently rinsed
with deionised water to remove any possible inter-
Fig. 1. The effect of pH on the uptake of gold(III) by Azolla fil-
ferences by other metals. NaOH and HCl were used iculoides. The experiments were conducted in duplicate. Initial
for pH adjustments. Atomic absorption spectrometric metal concentration is 8 mg gold l−1 and the Azolla filiculoides
standards were prepared from 1000 mg gold metal l−1 concentration is 5 g biomass l−1 .
solution.
20 min. For convenience, this concentration of gold
Methods
was utilized for all further experiments. Gold(III) re-
All experimental work was conducted in dupli- moval decreases progressively with increased biomass
cate. Biomass and gold concentrations were ad- concentrations greater than the optimum concentration
justed according to the respective experiment. Sam- but significant removal (95%) was still observed at the
ples (100 ml) placed in 300-ml Erlenmeyer flasks were highest biomass concentration examined.
constantly shaken at 200 rpm at room temperature.
Aliquots (3 ml) were withdrawn and filtered using Effect of initial concentration of hydrogen
cellulose-acetate filters (25 mm, 0.45 µM pore size). tetrachloroaurate(III)
The filtrate was then analyzed for gold using atomic
Due to the low concentration of gold(III) found in ef-
absorption spectrophotometry. The results were ex-
fluents, it was decided to use typical concentrations of
pressed as percentage removal. Control experiments
the metal at 2, 4, 6, 8 and 10 mg gold l−1 . Results
involving the studies of gold(III) solutions in the ab-
show that removal of 86%, 95%, 94%, 98% and 99%
sence of biomass were run to exclude the possibility
were achieved, respectively.
of gold precipitation. In the pH study, the pH was
adjusted every half-hour. In the temperature study,
Effect of pH
the flasks were shaken in a thermostatically-controlled
incubator. pH studies showed a substantial change in the bind-
ing capacity of gold(III) to the biomass, with pH 2
showing optimal removal. The adsorption mechanism
Results
of gold is ionic rather than covalent due to the depen-
dence of gold binding with pH. The results in Figure 1
Gold removal was rapid with the majority being re-
indicate that the gold(III) complex is in the anionic
moved within the first 20 min. The exception was in
form [AuCl4 ]− shown by its preference to bind at pH 2
the pH studies.
(Cotton & Wilkinson 1980). pH affects the protonation
of the functional groups on the biomass as well as the
Effect of biomass concentration
metal chemistry. At pH 3 and 4, probable incomplete
The percentage removal of gold(III) at biomass con- functional group protonation results in decreased up-
centrations of 1 to 9 g biomass l−1 (in intervals of take, while at pH 5 and 6 maximum uptake is achieved
2 g l−1 ), demonstrated that the optimum concentra- after 180 min suggesting a slow equilibrium between
tion was 5 g biomass l−1 with 99.9% removal within [AuCl4 ]− and gold(III) hydroxy species.

Effect of temperature
Contrary to previous biomass studies (Kuyucak &
Volesky 1988), variations in temperature from 10–
50 ◦ C had little effect on the percentage removal of
gold at pH 2, with 100% removal for all temperatures
investigated. The process may be energy-independent
under the experimental conditions, suggesting anionic
binding.
Conclusions
Azolla filiculoides is able to remove gold from dilute
solutions, which is desirable since gold concentrations
in effluents, especially mine effluents, are very low.
The results indicate that the biomass under the opti-
mal conditions may achieve 100% removal, strongly
supporting the possible application of Azolla filicu-
loides biomass in wastewater bioremediation. From
the batch studies performed with hydrogen tetra-
chloroaurate(III) solution, it can be deduced that the
gold is in the anionic form, [AuCl4 ]− , shown by its
preference to bind to the biomass at pH 2 and that
the process of adsorption is independent of the tem-
perature within the range of 10 ◦ C to 50 ◦ C. Further
studies are being undertaken employing wastewater
rather than synthetic solutions.
251
Acknowledgements
The authors would like to gratefully acknowledge the
financial support and funding for this project by the
National Research Foundation and Water Research
Council of South Africa.
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