Algal Research 7 (2015) 92–99

Contents lists available at ScienceDirect

Algal Research
journal homepage: www.elsevier.com/locate/algal

Preparation of bead-type biosorbent from water-soluble

Spirulina platensis extracts for chromium (VI) removal
Hyo Won Kwak a, Moo Kon Kim a, Jeong Yun Lee a, Haesung Yun a, Min Hwa Kim a,
Young Hwan Park a, Ki Hoon Lee a,b,c,⁎
a
Department of Biosystems & Biomaterials Science and Engineering, Seoul National University, Seoul 151-921, Republic of Korea
b
Center for Food and Bioconvergence, Seoul National University, Seoul 151-921, Republic of Korea
c
Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Republic of Korea

a r t i c l e i n f o a b s t r a c t

Article history: A bead-shaped biosorbent for removal of Cr(VI) from industrial waste waters/aqueous media was fabricated
Received 2 May 2014 using water soluble Spirulina platensis extract (SPE). The addition of LiCl to DMSO affected the solubility of SPE,
Received in revised form 4 December 2014 and 1 M LiCl/DMSO was the optimal solvent system for the preparation of SPE beads. The shape of the SPE
Accepted 17 December 2014
bead was determined by the concentration of SPE, and the type of coagulant affected the mechanical properties
Available online xxxx
of the bead. The optimum pH for the Cr(VI) adsorption was 2.0. The maximum adsorption capacity of the SPE
Keywords:
beads was 41.12 mg/g at pH 2.0, and the adsorption behavior of Cr(VI) on the SPE beads followed the
Spirulina Brunauer–Emmett–Teller (BET) isotherm. The results indicated that 0.1 M NaOH was the most effective desorp-
Microalgae tion agent. The successive recycling of the adsorption–desorption process was stable for more than five cycles,
Spirulina bead and the recycling efficiency was 70%.
Hexavalent chromium © 2014 Elsevier B.V. All rights reserved.
Biosorption

1. Introduction
Recently, the presence of heavy metal ions in waste water has
become one of the most serious environmental problems [1,2]. Among
heavy metal ions, chromium enters aqueous waste streams from
many industries such as dyeing, mining, metal plating, and metal pro-
cessing. In aqueous systems, chromium exists in various oxidation
states, but trivalent (Cr(III)) and hexavalent (Cr(VI)) are the most
common oxidation states. While Cr(III) is sometimes required in tiny
amounts for biological metabolisms, Cr(VI) is known to be a toxic and
carcinogenic substance. The amount of Cr in waste water is therefore
strictly regulated by regulatory agencies [3]. The removal of Cr(VI)
from waste water is more important than the removal of Cr(III), and
many studies have been performed to find more effective systems for
Cr(VI) removal [4].
Among the various techniques that have been employed for the
removal of Cr(VI), adsorption is highly effective and economical because
low-cost adsorbents can be employed, the adsorption technique
has high process flexibility in design and operation, and the adsorbents
can be reused if the adsorption is reversible [5]. Biosorption is an -
adsorption method that uses biomass, living or non-living, as the
⁎ Corresponding author at: Department of Biosystems & Biomaterials Science and
Engineering, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-921,
Republic of Korea.
E-mail address: prolee@snu.ac.kr (K.H. Lee).
adsorbent. Various types of biomass have been investigated [6,7], and
microalgae biomass was suggested as a candidate for heavy metal
removal [8]. The advantages of microalgae biomass for waste water
treatment are wide availability, low cost, and high heavy metal removal
efficiency [9]. Chlorella, Spirulina, and Dunaliella are the microalgae
species that have been most widely tested for the removal of Cr(VI)
[10–12]. However, the use of microalgae in wastewater treatment
has limitations because of the necessity of separation of microalgae
from the treated water. Industrial filtration and centrifugation are the
techniques currently available for the harvesting of the microalgae,
but unfortunately, these techniques are not cost-effective [13]. In this
regard, immobilization of microalgae into polymeric matrices, such as
alginate, silica gel, and carrageenan has been proposed [14–16]. Upon
immobilization, microalgae could be reused for several cycles, and the
continuous process can be applied in addition to the conventional
batch process. However, one of the limitations of the immobilization
method is that the amount of biomass that can be loaded into polymeric
matrices is limited, resulting in low adsorption efficiency per unit vol-
ume [17].
Several mechanisms have been proposed for the bioadsorption
of heavy metals by microalgae: surface adsorption, adsorption on extra-
cellular biopolymers, biosorption, and adsorption onto extracellular
biominerals [9]. Except for the biologic adsorption, the viability of the
microalgae does not matter. Use of the specific substance (cell wall com-
ponent, extracellular biopolymer, biomineral, etc.) that is responsible
for the heavy metal adsorption would be sufficient.

http://dx.doi.org/10.1016/j.algal.2014.12.006
2211-9264/© 2014 Elsevier B.V. All rights reserved.
 H.W. Kwak et al. / Algal Research 7 (2015) 92–99
In the present study, we successfully prepared microalgal Spirulina
platensis extract (SPE) beads as an adsorbent for hexavalent chromium.
We chose S. platensis because it is one of the few commercialized
microalgae and has been used as biosorbent material for heavy metal
ions [18–20]. First, to fabricate the SPE beads, LiCl/DMSO was used as
a solvent system. DMSO is a good solvent for various organic substances
such as synthetic polymers, polysaccharides, and proteins [21–23].
Previously, we found that the addition of LiCl to DMSO can significantly
enhance the solubility of certain proteins, and furthermore, it was also
possible to prepare a highly concentrated protein solution [24]. The
optimal conditions for spherical beads were found by changing the con-
centration of the dope solution. Then, to obtain appropriate beads with
excellent mechanical properties, we attempted to use various coagu-
lants. Finally, we investigated the biosorption potential of SPE beads
for the removal of hexavalent chromium from aqueous solution. This
is the first study to report the fabrication of the biosorbent directly
from the microalgae without any polymeric support.
2. Experimental
2.1. Preparation of SPE
The tablet form of S. platensis was purchased from Cyanotech Corpo-
ration (Spirulina Pacifica®, Purity: 100%, Hawaii, USA). Table S1 of the
Supplementary Material shows the typical composition of S. platensis
biomass, which was used this study. S. platensis was ground to a powder
using a laboratory mill (A 10, IKA Works, Germany) and stored at 4 °C
until use. We used an extraction procedure that has previously been
reported [25]. In detail, a 20 g of sample of S. platensis powder was
soaked in 1 L of distilled water, then shaken continuously for 24 h at 4
°C. The mixture was centrifuged at 5000 rpm for 10 min at 4 °C. The
supernatant was further filtered with nonwoven filter (Miracloth, typi-
cal pore size: 22–25 μm, EMD Millipore, USA) to remove the water-
insoluble fraction to obtain the SPE. SPE was freeze-dried and stored
at 4 °C until use.
2.2. Dissolution rate of SPE in LiCl/DMSO solution
To find the optimum dissolution conditions for SPE in LiCl/DMSO
solvent, the dissolution rate was measured by varying the LiCl concen-
tration. SPE (3.0 g) was added to 10 mL of LiCl/DMSO with various
LiCl concentrations (0.1, 0.5 1.0, 2.0 M) and shaken vigorously for an
hour at room temperature. The soluble portion was separated using
centrifugation (3000 rpm, 30 min at 25 °C), and the dry weight of the
residual insoluble part was measured. The dissolution rate of SPE was
calculated using the following formula:
W1 −W2
Dissolution rate ¼
W1
where W1 is the initial weight of SPE, and W2 is the weight of the residual
insoluble fraction of the solution mixture.
2.3. Preparation of the SPE beads
To prepare the SPE beads, various amounts of SPE (0.5. 1.0, 1.5, 2.0,
2.5, and 3.0 g) were dissolved in 10 mL of 1 M LiCl/DMSO for 3 h at
room temperature. After complete dissolution, the solution was trans-
ferred to a syringe and mounted on a syringe pump (KD Scientific,
USA). The dope solution was dripped into various alcohol coagulants
(methanol, ethanol, isopropanol, and 1-butanol) to study the effect of
the coagulants on the mechanical properties of the SPE beads. The
drip rate was 20 mL/h, the size of the needle was 26 G and the distance
between the needle end and the surface of the coagulant was fixed at
3 cm. Beads were prepared at room temperature. After dripping the
dope solution into the coagulants, the SPE beads were kept in the
93
coagulant for an hour. Then, the SPE beads were soaked in 2% glutaral-
dehyde (GA) in 50 mL of the same coagulant for an hour to improve
the water stability. Finally, the cross-linked SPE beads were washed
with the same coagulant to remove LiCl, DMSO and any residual
unreacted aldehyde groups. Pictures of the solidified SPE beads
were taken using a digital camera (Samsung, Korea). As a control,
raw S. platensis beads were prepared in the same manner using
S. platensis powder without extraction. The as-prepared SPE and
raw S. platensis beads were dried at room temperature and stored
at 4 °C until the adsorption studies.
2.4. Characterization of the SPE beads
The compressive strength of a single SPE bead prepared by various
coagulants was measured using a Material Testing Machine (Lloyd
Instruments, Ltd., UK). After applying 0.05 N to the SPE bead, a compres-
sion curve was obtained. The compression strength was determined
from the strength at rupture. The surfaces of the SPE beads were ob-
served using a field-emission scanning electron microscope (FE-SEM),
(JSM-7600F, JEOL, Korea). The elemental analysis of the SPE bead was
performed by FE-SEM coupled with energy dispersive X-ray analysis
(EDS), (Oxford Instruments X-Max 20, SW AZtec 2.1; Oxford, UK).
Attenuated total reflection Fourier transform infrared spectroscopy
(ATR-FTIR, Thermo Scientific, USA) was used to identify the Cr(VI)
adsorption onto the SPE beads. The samples were examined within
the wavenumber range of 700–4000 cm−1, and 32 scans with 8 cm−1
resolution were used to obtain the spectra.
2.5. Batch adsorption studies
A stock solution of Cr(VI) (1000 mg/L) was prepared in distilled
water using an accurate quantity of potassium dichromate, K2Cr2O7
(Sigma Aldrich, USA). Cr(VI) solutions at other concentrations were pre-
pared from the stock solution by dilution and varied from 25 to
250 mg/L. To determine the optimum pH for the adsorption process,
0.1 g of biomass was added into 100 mL of Cr(VI) solution (100 mg/L).
The initial pH values of the Cr(VI) solutions were adjusted from 1.0 to
5.0 using 1 M H2SO4 or 1 M NaOH. To compare the adsorption capacity
of the raw S. platensis beads and the SPE beads, the adsorption experi-
ments were performed under the same conditions. The equilibrium
adsorption capacity, qe, was determined using the following Eq. (2):
Co −Ce
qe ¼ V ð2Þ
M
where Co and Ce are the initial and the equilibrium concentration of the
Cr(VI) in the testing solution (mg/L), V is the volume of the testing solu-
ð1Þ
tion (L), and M is the weight of the SPE bead (g).
In the biosorption kinetic experiments, 0.1 g of SPE beads was added
to a 100 mL Cr(VI) solution (100 mg/L). The initial pH was adjusted to
2.0, and the samples were taken at different time intervals.
To obtain the adsorption isotherms, varying initial Cr(VI) concentra-
tions ranging from 25 to 250 mg/L were used. The batch adsorption
equilibrium experiments were conducted in 250 mL Erlenmeyer flasks
with 100 mL of the Cr(VI) solution. For all of adsorption experiments,
the flasks were agitated continuously on a multi-stirrer (JEIO Tech,
Korea) at 180 rpm with the temperature controlled at 25 °C up to 24 h.
2.6. Desorption/recycling experiments
Various desorption agents such as distilled water, 0.1 M NaOH, 0.1 M
EDTA, 0.1 M HCl, and 0.1 M HNO3 were used in this study. For the de-
sorption study, after the adsorption experiments, the SPE beads were
recovered from the Cr(VI) solution using nonwoven filter. To remove
the residual Cr(VI) on the surface, the Cr(VI) adsorbed SPE beads were
94 H.W. Kwak et al. / Algal Research 7 (2015) 92–99

2.7. Analysis of the Cr ions

100

The concentration of the Cr(VI) ions was determined spectrophoto-

metrically (540 nm) using a microplate reader (Synergy™ H1, BioTek,
80
USA) after complexation with 1,5-diphenylcarbazide (Sigma Aldrich,
SPE Solubility (%)

USA) [26]. The total Cr ion concentration in aqueous solution was

60
analyzed by inductively coupled plasma optical emission spectrometry
(ICP-OES), (iCAP 7400™, Thermo Fisher Scientific, USA), and the Cr
(III) concentration was determined by deduction from the total Cr
40 ions and Cr(VI) ion concentrations.

2.8. Adsorption isotherm models

20
In this study, the equilibrium data were analyzed using three iso-
therm models, Langmuir, Freundlich, and Brunauer–Emmett–Teller
0 (BET) isotherm models.
0.1M 0.5M 1.0M 2.0M The Langmuir model was used for modeling the adsorption of heavy
Concentration of LiCl salt metals on homogeneous surfaces without any interaction between the
adsorbed ions. The Langmuir isotherm can be defined using the following
Fig. 1. Effect of the LiCl concentration in DMSO on the dissolution of SPE. Three grams of formula [9]:
SPE dissolved in 10 mL of solvent (n = 4). The error bars indicate the standard deviation
of the data.
qm KL Ce
qe ¼ ð4Þ
1 þ KL Ce
agitated with distilled water on a multi-stirrer for 10 min at 180 rpm
and this washing process repeated three times. where qe is the equilibrium metal ion concentration on the adsor-
The beads were then soaked in 100 mL of desorption agent, and the bent (mg/g), Ce is the equilibrium metal ion concentration in the solu-
mixtures were shaken overnight. The relative desorption efficiency was tion (mg/L), qm is the monolayer biosorption capacity of the adsorbent
calculated as: (mg/g), and KL is the Langmuir biosorption constant (L/mg), which is
related to the free energy of biosorption.
In contrast, the Freundlich model describes the adsorption of metal
Desorbed Cr ions by desorption agent
Desorption efficiency ¼ ions on heterogeneous surfaces. This isotherm can be described as
Adsorbed Cr ions
 100: ð3Þ follows [9]:

To study the recycling efficiency, five cycles of adsorption–desorp-
tion experiments were performed using 0.1 M NaOH as a desorption
agent. The adsorption and desorption experiments were performed
during 24 h of contact time and 180 rpm of agitation speed. After each
cycle of experiments, the SPE beads were washed three times with
distilled water to ensure neutral conditions for the next adsorption–
desorption cycle.
1=n
qe ¼ K f Ce ð5Þ
where Kf is a constant related to the biosorption capacity, and 1/n is an
empirical parameter related to the biosorption intensity, which varies
with the heterogeneity of the material.
In addition to the Langmuir and Freundlich models, the BET model
was used to describe the equilibrium metal biosorption in a batch
system. This model assumes uptake of the metal ions in homogeneous

5 wt% 10 wt% 15 wt %

20 wt% 25 wt% 30 wt%

Fig. 2. Effect of the concentration of SPE dope solution on the shape of the SPE beads coagulated with methanol.
 H.W. Kwak et al. / Algal Research 7 (2015) 92–99 95

100 3. Results and discussion

3.1. Preparation and characterization of the SPE beads

80
S. platensis has a high protein content that can be extracted using an
aqueous solution. The final yield of SPE was 30.77% of the initial
S. platensis powder. The main component of SPE was protein, and
60
the protein was approximately 60% of the total SPE, which was consis-
Force (gf)

tent with our previous results [28]. To fabricate the SPE into various
forms such as beads, macro- and nanofibers, or films, one must find an
40 adequate solvent system. In this study, we first checked the solubility
of SPE in LiCl/DMSO (Fig. 1). By increasing the concentration of LiCl in
DMSO to 1 M, the solubility of SPE was increased, and the complete dis-
20
solution of SPE with a 30% concentration (w/v) was achieved at 1 M of
LiCl in DMSO. However, further increase in LiCl up to 2 M decreased
the solubility of SPE, which might be due to the salting-out effect of pro-
tein at high concentrations of salt [29]. Therefore, for the preparation of
0 solutions with high concentrations of SPE, we used 1 M LiCl/DMSO as
Methanol Ethanol 1-Butanol 1-Propanol the solvent for SPE.
Coagulant Fig. 2 shows the shapes of the SPE beads prepared from different SPE
concentrations. From the 5 and 10% (w/v) SPE solutions, the coagulated
Fig. 3. Effect of the coagulant on the compressive strength of the SPE beads (n = 5). The SPE had an irregular shape due to the low viscosity of the SPE solution.
error bars indicate the standard deviation of the data. The dropping droplets cannot maintain their shape upon collision with
the surface of the coagulant, resulting in scattering of the droplet. As the
concentration of the SPE solution increased, spherical-shaped beads
could be formed at concentrations of 20 and 25% (w/v). At these con-
multilayers. This isotherm is expressed using the following equa- centrations, the viscosity of the SPE droplets was sufficient to withstand
tion [27]: the surface tension of the coagulant and maintain their shape. When the
concentration of SPE was increased to 30% (w/v), tails were formed be-
cause of the high viscosity of the SPE solution. The average bead diame-
    
Ce 1 B−1 Ce ter of the as-prepared spherical SPE beads was approximately 2 mm, but
¼ þ ð6Þ
ðCs−CeÞ BQ BQ CS the diameter was reduced to 1 mm after drying.
During the heavy metal removal process, the mechanical properties
of the adsorbent are important because the adsorbent needs to with-
where CS is the saturation concentration of the solute (mg/L), Q is the stand the mechanically harsh environment of the reactor caused by
amount of solute adsorbed per unit weight of adsorbent when forming agitation or stirring. During stirring, beads could collide with each
a complete monolayer on the surface (mg/L), and B is a constant other or with the impellor, resulting in the destruction of the bead.
representing the energy of interaction with the surface. We have previously reported that the type of coagulant can affect the

(a) (b)

(c) (d)

Fig. 4. FE-SEM images of dried SPE beads using various coagulants: (a) methanol, (b) ethanol, (c) 1-propanol, and (d) 1-butanol with magnification: 200× and insert panel: 500×.
96 H.W. Kwak et al. / Algal Research 7 (2015) 92–99

mechanical properties of the protein beads [24]. Figs. 3 and 4 show Raw S.plantensis 1642cm
-1
1536cm
-1
932cm
-1

SPE Amide I Amide II Cr-O stretching

the compressive strength and surface images of SPE beads prepared
SPE bead
using different coagulants. The methanol- or ethanol-coagulated SPE
Cr(VI) sorbed SPE bead
beads have much better compressive strength than the propanol- or
butanol-coagulated beads. In general, the coagulation mechanism of
the polymer is the exchange of the solvent and the non-solvent. The sol-

Absorbance
vent will diffuse out and the non-solvent will diffuse in. In the case of
SPE beads that coagulated in propanol or butanol, a flat region could
be observed on the surface of the bead. This flat region is the result of
temporary adhesion between insufficiently coagulated SPE beads in
the coagulant, which indicates a slow solvent exchange speed. In addi-
tion, cracks were observed in these beads, and these cracks may act as
weak points during real applications. However, the methanol- and
ethanol-coagulated SPE beads exhibited denser and smoother surfaces
and had better compression strength than the propanol or butanol co- 4000 3500 3000 2500 2000 1500 1000
agulated SPE beads. In further studies, the methanol-coagulated SPE Wavenumber (cm )
-1

beads were used.

Fig. 6. FTIR spectra of raw S. platensis, SPE, SPE beads, Cr(VI) adsorbed on SPE beads.
3.2. Cr(VI) adsorption onto SPE beads

To find out whether the SPE beads can adsorb the chromium, the
existence of chromium on the SPE beads was verified using both EDS
and ATR-FTIR. Fig. 5 shows FE-SEM images and EDS data from the SPE
beads after Cr(VI) removal. The EDS result indicated that chromium
was adsorbed on the SPE bead. Fig. 6 shows the ATR-FTIR spectra
of the SPE bead before and after the Cr(VI) removal. The original
SPE bead showed characteristic protein peaks; both amides I (1700–
1600 cm− 1) and II (1600–1500 cm−1) were observed. Compared to
the original SPE beads, a new peak at 933 cm−1 appeared after Cr(VI)
removal. The new peak is due to Cr\O stretching vibrations from the
adsorbed chromate ion [30]. These results indicate that chromium
could be adsorbed successfully on the SPE bead.
3.3. Removal mechanism of Cr(VI) by SPE beads
Various bioadsorption mechanisms have been proposed, but the ion
exchange mechanism has become the dominant mechanism [31].
Positively or negatively charged ions will be adsorbed on the oppositely
charged surface of the bioadsorbent. The pH of the solution is well
known to be one of the most important factors that affect the metal ad-
sorption process [9]. The pH of the solution not only affects the charge of
the metal ions but also affects the charge of the metal binding site of the
adsorbent. To determine the best pH for Cr(VI) ion biosorption, the ef-
fect of pH on the Cr(VI) removal by the SPE beads was investigated in
the pH range from 1 to 5. Fig. 7 shows the equilibrium Cr(VI) uptake
from a 100 mg/L initial Cr(VI) concentration solution at each pH by
the SPE and S. platensis beads. Clearly, pH 2 was the optimal condition
for the highest Cr(VI) uptake. Generally, the isoelectric points of
microalgal proteins are found near pH 3 [32], which means that the
SPE beads will have positive net charges below pH 3 and negative net
charges above pH 3. In addition, Cr(VI) forms a different chromate ion

50

(a) SPE beads

2.0
cps / eV S. plantensis beads
40
1.8

1.6

30
Q eq (mg/g)

1.4

1.2 Cr
Pt
S N
C O Pt S Cr Pt
1.0 20

0.8

0.6
10
0.4

0.2

0
0.0
0 2 4 6 8 10 1 2 3 4 5
keV
pH
(b)
Fig. 7. The effect of the initial pH on the equilibrium Cr(VI) ion biosorption capacity of raw
Fig. 5. FE-SEM image with magnification: 20,000× (a) and EDS spectrum of Cr(VI) S. platensis and SPE beads (C0: 100 mg/L, adsorbent dose: 1.0 g/L, temperature: 25 °C, agita-
adsorbed on the SPE bead (b). The arrows indicate the weight percentage of Cr(VI). tion rate: 180 rpm). The error bars indicate the standard deviation of the data (n = 4).
 H.W. Kwak et al. / Algal Research 7 (2015) 92–99 97

Total Cr 2.5 60
100 Cr (VI)
Cr (III)
pH 2.4 50
80
Concentration (mg/l)

2.3
40
60

qe (mg/g)
pH
2.2
30
40
2.1 Experiment
20
Langmuir
20
2.0 Freundlich
10 BET
0 1.9
0 5 10 15 20 25
0
Time (hrs)
0 50 100 150 200 250
C e (mg/g)
Fig. 8. Concentration of total chromium, Cr(VI) and Cr(III) in the remaining solution after
removal of Cr(VI) by SPE beads and the change of pH during adsorption. (C0: 100 mg/L,
adsorbent dose: 1.0 g/L, temperature: 25 °C, agitation rate: 180 rpm). The error bars indi- Fig. 10. Adjustment of the Langmuir, Freundlich, and BET models to the experimental data
cate the standard deviation of the data (n = 4). obtained from the SPE beads.

in solution that is highly dependent on the pH of the solution [33].
Under acidic conditions such as pH 2, HCrO− 4 is the dominant chromate
ion along with CrO2− 4 and CrO2−
7 . Therefore, at pH 2, the SPE beads will
have a positive net charge and there will be strong electrostatic attrac-
tions between the positive SPE beads and the negative HCrO− 4 . Howev-
er, at a value of the pH above 3, the net charge on the SPE beads becomes
negative, so the biosorption capacity decreases significantly due to
electrostatic repulsion. The biosorption at pH 1 also decreased com-
pared to the biosorption at pH 2 because H2CrO4 dominates at this pH,
and H2CrO4 does not exhibit any charge. At the same pH, the SPE
beads had a higher Cr(VI) adsorption capacity than the raw S. platensis
beads, which indicates that some portions of S. platensis are not involved
in the heavy metal adsorption and were removed during the extraction
procedure.
In addition to the ion exchange mechanism, an adsorption-coupled
reduction mechanism has also been proposed to explain the removal
of Cr(VI) by various types of biomass adsorbents. Park et al. [34] exam-
ined 16 different biomass adsorbents and described the Cr(VI) removal
through indirect reduction followed by an adsorption process or direct
reduction by the electron donor of the biomass. Golder et al. [35]
50
40
30
qe (mg/g)
found that dead Spirulina biomass can reduce Cr(VI) to Cr(III). To
check the reduction of Cr(VI) to Cr(III), we examined the change in
Cr(VI) and Cr(III) concentration during the adsorption process at pH 2
(Fig. 8). With the increase of contact time, the concentration of total
Cr and Cr(VI) decreased while the Cr(III) concentration in the remaining
aqueous solution increased, indicating that reduction of Cr(VI) has oc-
curred during the adsorption process. The pH of the solution also in-
creased slightly with the increase in contact time, consistent with
previous report [36]. The increase in pH is due to consumption of pro-
tons during the protonation of the amine groups of the SPE bead and
the reduction of Cr(VI) to Cr(III). Therefore, we conclude that the
removal of Cr(VI) by SPE beads takes place by an adsorption-coupled
reduction mechanism similar to other biomass adsorbents.
3.4. Biosorption isotherm models
To study the adsorption isotherm, the Langmuir, Freundlich and BET
models were applied to the equilibrium adsorption results obtained by
changing the initial Cr(VI) concentration (C0) from 33 to 250 mg/L.
When C0 was increased from 33 to 250 mg/L, the Cr(VI) uptake by the
SPE beads increased from 24 to 49 mg/g (Fig. 9).
Fig. 10 shows the Langmuir, Freundlich, and BET isotherms obtained
by fitting equilibrium data to Eqs. (4), (5), and (6). The values obtained
for the meaningful parameters and constants of each model are given in
Table 1. The coefficients of determination (R2) were found to be 0.989 in
the Langmuir, 0.946 in the Freundlich model, and 0.999 in the BET
model. These values were sufficiently high to suggest that both the
Langmuir and Freundlich models were able to describe the biosorption

of Cr(VI) ions well. However, when comparing the R2 values, the BET

20
Table 1
Isotherm constants and correlation coefficients for the biosorption of Cr(VI).

10 Parameter Value RMSE (root mean SSE (sum of square R2

square errors) errors)

Langmuir isotherm 0.0567 0.0499 0.989

Q (mg/g) 50.530
0
KL (L/mg) 0.101
0 50 100 150 200 250
Freundlich isotherm 3.176 0.2244 0.946
C 0 (mg/l) n (L/mg) 4.390
Kf (mg/g) 15.755
BET isotherm 0.1628 0.0144 0.999
Fig. 9. The effect of the initial concentration of Cr(VI) on the equilibrium Cr(VI) ion
Q (mg/g) 52.826
biosorption capacity of SPE beads (adsorbent dose: 1.0 g/L, temperature: 25 °C, agitation
B (g/mg) 9.443 × 105
rate: 180 rpm).
98 H.W. Kwak et al. / Algal Research 7 (2015) 92–99

100 4. Conclusions

In this study, we successfully prepared SPE beads as an adsorbent for

80 the removal of Cr(VI). Due to the high content of protein, the SPE could
be fabricated directly into beads without any polymeric additives. The
Desorption efficiency (%)

Cr(VI) removal mechanism of the SPE beads was adsorption-coupled

60 reduction, which initially adsorbs Cr(VI) followed by reduction into
the less toxic Cr(III). The SPE beads could be regenerated more than
five times, still having 70% of the initial capacity. We have found that
40 the optimum removal of Cr(VI) by SPE beads could be performed
under acidic conditions. We expect that the SPE beads could be used
in the treatment of the waste solution from metal finishing, where acidic
20 waste solutions are produced.
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.algal.2014.12.006.
0
Distilled water 0.1M NaOH 0.1M EDTA 0.1M HCl 0.1M HNO3
Acknowledgments
Desorption agent
This work was supported by ‘Technology Development of Marine
Fig. 11. Relative desorption efficiency of the various desorption agents (n = 3). The error Industrial Biomaterials’ of the Marine Biotechnology Program funded
bars indicate the standard deviation of the data.
by the Ministry of Oceans and Fisheries, Korea.

model is a more appropriate model than the Langmuir and Freundlich
models.
3.5. Desorption and recycling studies
From the point of view of the economical use of the bioadsorbent, it
would be beneficial if the biosorbent could be used repeatedly. There-
fore, the biosorbent should be regenerated by inducing desorption of
the adsorbed metal ions from the biosorbent. Fig. 11 shows the desorp-
tion efficiency of Cr(VI) from the SPE beads after treatment with various
desorption agents. Among the suggested desorption reagents, 0.1 M
NaOH was the most efficient. The desorption mechanism by alkali treat-
ment has a mechanism opposite to the adsorption: By soaking the
Cr(VI)-adsorbed SPE beads in 0.1 M NaOH, the surface charge of the SPE
beads becomes negative, resulting in desorption of the negative chromate
ion. To study the biosorption recycling efficiency of the SPE beads, the ad-
sorption–desorption experiments were performed for five repeated cy-
cles, and 0.1 M NaOH was used as desorption agent. Fig. 12 shows the
adsorption capacity of the SPE beads after each cycle. After the fifth
cycle, the SPE beads can still adsorb 28 mg/g Cr(VI) ions, and the recycling
efficiency was calculated to be 70% of the first cycle.
50
40
30
Q eq (mg/g)
References
[1] S.E. Bailey, T.J. Olin, R.M. Bricka, D.D. Adrian, A review of potentially low-cost
sorbents for heavy metals, Water Res. 33 (1999) 2469–2479.
[2] M.A. Hashim, S. Mukhopadhyay, J.N. Sahu, B. Sengupta, Remediation technologies
for heavy metal contaminated groundwater, J. Environ. Manage. 92 (2011)
2355–2388.
[3] K.R. Krishna, L. Philip, Bioremediation of Cr(VI) in contaminated soils, J. Hazard
Mater. 121 (2005) 109–117.
[4] M. Owlad, M.K. Aroua, W.A.W. Daud, S. Baroutian, Removal of hexavalent
chromium-contaminated water and wastewater: a review, Water Air Soil Pollut.
200 (2009) 59–77.
[5] F. Fu, Q. Wang, Removal of heavy metal ions from wastewaters: a review, J. Environ.
Manage. 92 (2011) 407–418.
[6] J. Wang, C. Chen, Biosorbents for heavy metals removal and their future, Biotechnol.
Adv. 27 (2009) 195–226.
[7] M. Kesaano, R.C. Sims, Algal biofilm based technology for wastewater treatment,
Algal Res 5 (2014) 231–240.
[8] V.K. Gupta, A. Rastogi, Biosorption of lead from aqueous solutions by green algae
Spirogyra species: kinetics and equilibrium studies, J. Hazard Mater. 152 (2008)
407–414.
[9] S.K. Mehta, J.P. Gaur, Use of algae for removing heavy metal ions from wastewater:
progress and prospects, Crit. Rev. Biotechnol. 25 (2005) 113–152.
[10] Z. Aksu, U. Acikel, T. Kutsal, Investigation of simultaneous biosorption of copper(II)
and chromium(VI) on dried Chlorella vulgaris from binary metal mixtures: applica-
tion of multicomponent adsorption isotherms, Sep. Sci. Technol. 34 (1999) 501–524.
[11] G. Donmez, Z. Aksu, Removal of chromium(VI) from saline wastewaters by
Dunaliella species, Process Biochem. 38 (2002) 751–762.
[12] S.V. Gokhale, K.K. Jyoti, S.S. Lele, Kinetic and equilibrium modeling of chromium (VI)
biosorption on fresh and spent Spirulina platensis/Chlorella vulgaris biomass,
Bioresour. Technol. 99 (2008) 3600–3608.
[13] E.M. Grima, E.H. Belarbi, F.G.A. Fenandez, A.R. Medina, Y. Chisti, Recovery of
microalgal biomass and metabolites: process options and economics, Biotechnol.
Adv. 20 (2003) 491–515.
[14] L. Travieso, R.O. Canizares, R. Borja, F. Benitez, A.R. Dominguez, R. Dupeyron, V.
Valiente, Heavy metal removal by microalgae, Bull. Environ. Contam. Toxicol. 62
(1999) 144–151.

[15] A. Pellon, F. Benitez, J. Frades, L. Garcia, A. Cerpa, F.J. Alguacil, Using microalgae
Scenedesmus obliquus in the removal of chromium present in plating wastewaters,
Rev. Metal. 39 (2003) 9–16.
[16] N. Rangsayatorn, P. Pokethitiyook, E.S. Upatham, G.R. Lanza, Cadmium biosorption by
20 cells of Spirulina platensis TISTR 8217 immobilized in alginate and silica gel, Environ.
Int. 30 (2004) 57–63.
[17] J.M. Guisan (Ed.), Immobilization of Enzymes and Cells, Humana Press, Totowa, NJ,
2006, pp. 15–30.
10 [18] S.V. Gokhale, K.K. Jyoti, S.S. Lele, Modeling of chromium (VI) biosorption by
immobilized Spirulina platensis in packed column, J. Hazard Mater. 170 (2009)
735–743.
[19] C. Solisio, A. Lodi, E. Finocchio, Effects of pH on chromate(VI) adsorption by Spirulina
0 platensis biomass: batch tests and FT-IR studies, Water Sci. Technol. 67 (2013)
1 2 3 4 5 1916–1922.
Regeneration cycle [20] E. Finicchio, A. Lodi, C. Solisio, A. Converti, Chromium (VI) removal by methylated
biomass of Spirulina platensis: the effect of methylation process, J. Hazard Mater.
156 (2010) 264–269.
Fig. 12. Adsorption capacity of the SPE beads after each cycle (n = 3). The error bars [21] M. Strukelj, J. Hedrick, J. Hedrick, R. Twieg, Solvent effects on the preparation of
indicate the standard deviation of the data. novel amorphous poly(aryl ether benzil)s, Macromolecules 27 (1994) 6277–6285.
 H.W. Kwak et al. / Algal Research 7 (2015) 92–99
[22] S. Xu, J. Zhang, A. He, J. Li, H. Zhang, C.C. Han, Electrospinning of native cellulose from
nonvolatile solvent system, Polymer 49 (2008) 2911–2917.
[23] H. Oh, M.K. Kim, K.H. Lee, Preparation of sericin microparticles by electro-
hydrodynamic spraying and their application in drug delivery, Macromol. Res. 19
(2011) 266–272.
[24] H. Oh, J.Y. Lee, A. Kim, C.S. Ki, J.W. Kim, Y.H. Park, K.H. Lee, Preparation of silk sericin
beads using LiCl/DMSO solvent and their potential as a drug carrier for oral admin-
istration, Fiber Polym. 8 (2007) 470–476.
[25] W.L. Chu, Y.W. Lim, A.K. Radhakrishnan, P.E. Lim, Protective effect of aqueous extract
from Spirulina platensis against cell death induced by free radicals, BMC Com-
plement. Altern. Med. 10 (2010) 53.
[26] F.D. Snell, Colorimetric Methods of Analysis, 3rd ed. D. Van Nostrand Company Inc.,
New York, 1949.
[27] E. Oguz, Adsorption characteristics and the kinetics of the Cr(VI) on the Thuja
oriantalis, Colloids Surf. A 252 (2010) 121–128.
[28] B.G. Cha, H.W. Kwak, A.R. Park, S.H. Kim, S.Y. Park, H.J. Kim, I.S. Kim, K.H. Lee, Y.H.
Park, Structural characteristics and biological performance of silk fibroin nanofiber
containing microalgae Spirulina extract, Biopolymers 101 (2014) 307–318.
[29] R.K. Scopes, Protein Purification: Principles and Practice, 3rd ed. Springer, 1994.
99
[30] M. Bolte, Y. Israeli, F. Djouani, A. Rivaton, L. Frezet, R.A. Lessard, Hologram formation
reconsidered in dichromated polyvinylalcohol: polymer cross-linking around chro-
mium (V), Proc. SPIE 5742 (2005) 195–204.
[31] K. Chojnacka, Biosorption and bioaccumulation; the prospects for practical applica-
tions, Environ. Int. 36 (2010) 299–307.
[32] M.A. Devi, G. Subbulakshmi, K.M. Devi, L.V. Venkataraman, Studies on the proteins
of mass-cultivated, blue-green alga (Spirulina platensis), J. Agric. Food Chem. 29
(1981) 522–525.
[33] M. Owlad, M.K. Aroua, W.M.A. Wan Daud, Hexavalent chromium adsorption on
impregnated palm shell activated carbon with polyethyleneimine, Bioresour.
Technol. 101 (2010) 5098–5103.
[34] D. Park, S.-R. Lim, Y.-S. Yun, J.M. Park, Reliable evidences that the removal mechanism of
hexavalent chromium by natural biomaterials is adsorption-coupled reduction,
Chemosphere 77 (2010) 298–305.
[35] M.K. Gagrai, C. Das, A.K. Golder, Reduction of Cr(VI) into Cr(III) by Spirulina dead
biomass in aqueous solution: Kinetic, Chemosphere 93 (2013) 1366–1371.
[36] J. Wu, H. Zhang, P.-J. He, Q. Yao, L.-M. Shao, Cr(VI) removal from aqueous solution by
dried activated sludge biomass, J. Hazard Mater. 176 (2010) 697–703.