Algal Research 9 (2015) 178185

Contents lists available at ScienceDirect

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

Review article

Harvesting microalgae by magnetic separation: A review

Shi-Kai Wang a,c, Amanda R. Stiles b, Chen Guo a, Chun-Zhao Liu a,
a
b
c

National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China
Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, USA
University of Chinese Academy of Sciences, Beijing 100049, PR China

a r t i c l e

i n f o

Article history:
Received 11 November 2014
Received in revised form 3 March 2015
Accepted 10 March 2015
Available online xxxx
Keywords:
Microalgae harvesting
Magnetic nanoparticles
Magnetic separator

a b s t r a c t
Magnetic separation has been utilized for the removal of microalgae for nearly forty years. Due to its advantages
compared to traditional harvesting methods, magnetophoretic harvesting of microalgal cells has received much
attention in recent years. In this context, synthesized magnetic particles for microalgae harvesting are summarized in this review. In addition, the particlecell interaction and factors inuencing the separation process are
discussed as well as the feasibility of its scale-up applications using a magnetic separator. Furthermore, the downstream techniques including the extraction of desired products and the reuse of the culture medium and magnetic particles are also assessed. Finally, the current challenges are outlined and future directions to achieve efcient
and economic magnetic harvesting of microalgae are discussed.
2015 Elsevier B.V. All rights reserved.

Contents
1.
2.
3.

Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Characteristics of algal broth . . . . . . . . . . . . . . . . . . . . . .
Magnetic particles for the separation of microalgae . . . . . . . . . . .
3.1.
Naked magnetic particles . . . . . . . . . . . . . . . . . . . .
3.2.
Surface functionalized magnetic particles . . . . . . . . . . . .
3.3.
Summary . . . . . . . . . . . . . . . . . . . . . . . . . . .
4.
Effects of the magnetic harvesting process . . . . . . . . . . . . . . .
4.1.
Algal species . . . . . . . . . . . . . . . . . . . . . . . . .
4.2.
Growth stage of microalgae . . . . . . . . . . . . . . . . . . .
4.3.
Magnetic particle dosage . . . . . . . . . . . . . . . . . . . .
4.4.
pH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
4.5.
Ions in the medium . . . . . . . . . . . . . . . . . . . . . .
4.6.
Temperature . . . . . . . . . . . . . . . . . . . . . . . . .
4.7.
Gradient of the magnetic eld . . . . . . . . . . . . . . . . . .
5.
Magnetic separator for algal harvesting . . . . . . . . . . . . . . . . .
6.
Detachment of aggregates and the reusability of magnetic particles . . . . .
7.
The biocompatibility of magnetic particles and the reuse of culture medium
8.
Conclusions, challenges, and further directions . . . . . . . . . . . . .
Acknowledgment . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1. Introduction
Over the past few decades, microalgae have received intensive basic
research and applied research to sustain biofuel production [13].
Corresponding author.
E-mail address: czliu@ipe.ac.cn (C.-Z. Liu).

http://dx.doi.org/10.1016/j.algal.2015.03.005
2211-9264/ 2015 Elsevier B.V. All rights reserved.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.
.

178
179
179
179
179
180
180
180
180
180
181
181
182
182
182
183
184
184
184
184

Microalgae are a potential feedstock for the production of transportation fuels due to several advantages, such as the ability to be cultivated
on barren land, a high growth rate, and higher lipid content compared
to other feedstocks [4,5]. In addition, microalgae have the potential to
accumulate high-value substances such as omega-3 fatty acids, vitamins, and antibiotics, as well as antioxidants for food supplements, animal feed, and pharmaceuticals [68]. However, the commercialization

S.-K. Wang et al. / Algal Research 9 (2015) 178185

of microalgae-based products is still limited, especially for biofuel production. This is mainly due to technological and economic limitations
in the production process [911]. The key stages in the production
process are cultivation, biomass harvesting, extraction of the desired
components, and production of the desired substances [12,13]. The harvesting step, in particular, is determined to be energy intensive and the
sustainability of harvesting techniques is sometimes limited by the energy requirement [14]. In addition, the cost of harvesting step is usually
high and contributes to 2030% of the total cost of the process [1517].
Harvesting microalgae at low cost and with a positive energy balance is
signicant for the production of algal biofuels [14].
Overviews of microalgae harvesting methods have been included in
multiple reviews [1822]. Algal cells can be harvested by various
methods, including centrifugation, sedimentation, occulation, ltration, otation, or by a combination of these methods. However, there
is no harvesting method that is considered superior, no one that is suited to all algal species. Current harvesting methods have various disadvantages which include high cost, high energy consumption, or the
requirement for a time-consuming process [19,22]. Therefore, it is
necessary to develop a reliable and cost-effective approach for the
industrial-scale production of algal based products.
Magnetic separation is a simple separation process. For the removal
of magnetic contaminants, it simply requires a magnetic separator
while for the recovery of a desired product, selective magnetic adsorbents are necessary. The separation is achieved based on the intrinsic
paramagnetic movement of the magnetic particle tagged products in
the response to the magnetic eld [23,24]. Due to its advantages,
which include simple operation, low energy consumption, and low
cost, it has been widely applied in diverse industries [23]. Magnetic separation has been demonstrated to be effective and reliable in applications such as kaolin decolorization, wastewater treatment in steel
factories and power plants, enrichment of ores-mineral beneciation,
the removal of specic elements in the food industry, and the removal
of arsenic and metals in water treatment [23,25,26]. In addition to
industrial applications, magnetic solutions have been used in many
biochemical processes, such as protein and DNA purication, drug
targeting and delivery, biocatalysis, and diagnostics [2729]. Furthermore, by tagging non-magnetic target cells with magnetic beads, the
target cells can be rapidly isolated from the medium using very gentle
conditions [24]. Due to this principle, magnetic separation can be utilized for the harvesting of microalgal cells.
Recently, different types of magnetic particles have been synthesized and have shown potential when utilized for the separation of
microalgae. In this review, the research progress of the magnetic separation of microalgae is reviewed, and the challenges and further directions are discussed.
2. Characteristics of algal broth
The harvesting of microalgae faces three major challenges. First is
the dilute nature of the algal broth, which is typically less than 0.5 g/L
in commercial production systems [18]. Therefore, large volumes of
broth need to be handled to recover the algal biomass [18]. Second,
algal cells are small; they typically range from 2 m to 20 m, which
makes the harvesting difcult via some general techniques, such as
ltration [30]. Third, these small cells generally have an electronegative
surface charge at a wide pH range [19]. The aggregative of algal cells is
difcult due to the electrostatic repulsion effect between algal cells
and the cells are generally stably suspended in the broth, which further
increases the difculty in harvesting [31]. In addition, the variety in size,
shape, and motility among different algal species makes it difcult to
develop a single technique that is suitable for the recovery of all species
[19].
The magnetic separation process is based on the interaction between the algal cells and particles. Therefore, the surface characteristics
of algal cells are important for the magnetic recovery efciency (RE). It

179

has been reported in multiple studies that the majority of algal species
have a negative surface charge over a wide pH range [3236]. This is
mainly due to the functional groups present in the proteins, lipids, and
sugars on the surface of algal cells [37]. Fourier transform infrared
(FTIR) spectra analysis indicated that there are abundant COOH
and OH groups on the surface of Chlorella sp. [38,39]. Furthermore,
atomic force microscopy (AFM) analysis revealed that the surfaces of
algal cells are not smooth, rather they contain groove-like indentations,
and striated and sphere-like mound structures; in Chlorella ellipsoidea,
these mound heights ranged from 60 to 60 nm [38,40].
3. Magnetic particles for the separation of microalgae
Magnetic particles have been used for algal separation for almost
forty years. They were initially used for the removal of harmful algae
from lakes [41,42]. Due to their advantages and potential, in recent
years, magnetic particles have received much attention and stimulated
research efforts for microalgae separation. Various types of magnetic
particles have been synthesized and studied for the recovery of algal
cells.
3.1. Naked magnetic particles
It is well-known that naked magnetite is effective for the removal of
microalgal biomass [41]. Recently, these naked magnetic particles have
been synthesized using various methods and successfully applied for
the harvesting of both freshwater and marine microalgae. For example,
Fe3O4 particles synthesized by chemical co-precipitation with an
average diameter of approximately 10 nm and an isoelectric point of
approximately 7 (suspended in deionized water), were efcient in
harvesting the freshwater algal species Botryococcus braunii and
C. ellipsoidea, and the marine species Nannochloropsis maritima [34,43].
In addition, a new agent with a broad size range of 0.1520 m and an
isoelectric point at pH 6.2 was prepared using ferrous sulfate as a precursor using assisted microwave treatment. High RE was achieved
using naked iron oxide magnetic microparticles (IOMMs). The synthetic
process was much simpler and cheaper than more commonly used approaches [33]. It has been conrmed that naked magnetite has ion exchange characteristics and the separation is primarily based on the
electrostatic interactions between the magnetite and the algal cells
[33,34,43]. In addition, Fe ions released from the IOMMs surface may
act as occulating agents and benet the harvesting process [33]. However, the released ions may increase the metal content in the harvested
algal cells and this can inuence the downstream rening of algal biomass. For example, the Fe can poison the catalysts for desulfurization
and decrease the gasoline yields [44]. Therefore, the choice of magnetic
adsorbent should consider the potential inuence on the downstream
process.
3.2. Surface functionalized magnetic particles
Due to the negative surface charge of algal cells, a positive charge on
the surface of the particles improves separation. As previously mentioned,
the surface charge of naked magnetite is pH-based with an isoelectric
point around neutral conditions [45]. Therefore, functionalizing the surface of the naked particles with cationic groups is an effective method
to enhance the RE. The tagging of a polyelectrolyte is commonly achieved
using one of two strategies: either the attached-to or the immobilizedon strategy [46]. The attached-to strategy is based on rst coating the
cells with a polymer binder and then attaching the magnetic particles.
In the immobilized-on approach, the naked magnetic particles are rst
surface functionalized with a polyelectrolyte and then bound to the
algal cells [39]. However, naked particles usually have poor dispersibility
and aggregate into large particle clusters due to magnetostatic and Van
der Waals functions. A lower RE was achieved with an equal dosage of
particles in the attached-to approach compared with that of the

180

S.-K. Wang et al. / Algal Research 9 (2015) 178185

immobilized-on approach due to the enhanced distribution and colloidal stability of the immobilized-on particles [39,46]. Therefore, the
immobilized-on strategy was more effective and was widely applied
in the majority of studies.
Recently, magnetic particles coated with different cationic groups
using the immobilized-on approach have been synthesized. After
they were surface functionalized, the electrophoretic mobility of the
particles generally increased and the isoelectric point also increased
due to the additional functional groups present on the surface of the
coated particles. For example, the isoelectric points of iron oxide
magnetic particles (IONPs), nanoscale zero-valent iron (nZVI), naked
Fe3O4 nanoparticles, and BrFe12O19 particles were obviously increased
after being surface functionalized by poly(diallyldimethylammonium
chloride) (PDDA), aminoclay, polyethylenimine (PEI), and (3aminopropyl)triethoxysilane (APTES), respectively [3638,47]. In
addition, the average size or hydrodynamic diameter also increased
after surface functionalization [35,37,38,46]. In most cases, the saturation
magnetization was constant after surface functionalization, which indicates that the coated particles can maintain their superparamagnetic
behavior, which is benecial for the magnetic separation process [35,
38]. However, an approximate 3-fold reduction of the saturation
magnetization value occurred after particles were coated with nanoscale
zero-valent iron (nZVI) aminoclay. This was mainly because of the
organofunctional pendants in aminoclay [47]. Furthermore, measurements of the contact angles (CAs) and calculations of the total surface
tensions (TOT) of two magnetic beads carrying diethylaminoethyl
(DEAE) and PEI, respectively, indicated that the bead surfaces have
strong hydrophilic characteristics and can provide strong polar interactions [32]. As claried in many studies, the main mechanism in these
separation processes was electrostatic interactions between the negatively charged surface of the algal cells and the coated particles with cationic properties [3638,4749]. In addition, in other types of interactions,
such as the nanoscale interactions of the nanoparticles (Fe3O4-PEI nanocomposites), the released ions act as the occulating agent (from the
surface of IOMMs), and bridging of the polyelectrolytes (CPAM), also occurred in the magnetic separation process [32,35,38].
In addition to cationic polyelectrolytes, other materials have also
been applied in the surface functionalization of magnetic particles and
used in algal separation. Hydrophilic silica-coated magnetic particles
were synthesized and applied in the separation of both freshwater
and marine microalgae. Meanwhile, the adsorption mechanism was
not electrostatic attraction and it was speculated that the interaction
was dependent on the ion-exchange characteristics of the particles. Localized electrostatic attraction may be induced by the NH+
3 groups on
the cell surface and O produced by the deprotonation of OH on
the surface of Fe3O4 nanoparticles [50]. In addition, a magnetic coagulant prepared by acid-modied y ash compounded with Fe3O4 magnetite was used in the removal of algal blooms from freshwater. Based on
the physical adsorption and chemical coagulation effects of the modied y ash, a high algal RE in conjunction with a decrease in chemical
oxygen demand (COD), total nitrogen, and total phosphorus was
achieved [51].

factors including the characteristics of magnetic beads and algal cells,

as well as the separation conditions. These factors are described below.
4.1. Algal species
As reported by Xu et al. [43] and Hu et al. [34], different adsorption capacities were achieved for different algal species using naked Fe3O4 nanoparticles. As shown in Table 1, the adsorption capacity in the harvesting of
B. braunii was nearly 10-times higher than that of C. ellipsoidea. This was
mainly due to the size distinction. For C. ellipsoidea, the unicellular cells
are smaller and have a much higher specic surface area than the colonial
microalga, B. braunii. Therefore, more magnetic particles were necessary
in order to achieve the same RE with B. braunii, resulting in a lower
adsorption capacity [43]. Similar results were obtained using the surface
functionalized particles, CPAM-Fe3O4 [35]. However, the adsorption
capacity of naked Fe3O4 nanoparticles was distinct for the freshwater microalgae C. ellipsoidea compared to the marine microalgae
N. maritima even though the cell sizes of the two species are similar [34,43]. This may be due to the differences in the separation conditions. The culture medium of N. maritima is relatively salty and the ionic
strength was also higher than that for C. ellipsoidea, which may greatly inuence the harvesting efciency. In addition, although high recovery efciencies were achieved in harvesting the freshwater microalgae B. braunii
and C. ellipsoidea and the marine microalgae N. maritima, the naked Fe3O4
nanoparticles were inefcient for the harvesting of Isochrysis galbana (unpublished data). The reason for this is still unclear and further research is
necessary to clarify the differences in the adsorption capacities among different algal species.
4.2. Growth stage of microalgae
As the algal biomass increases with culture time, it is important to
determine the optimal growth stage for harvesting. Hu et al. demonstrated that the maximum recovery capacity was obtained at a growth
stage when the biomass reached the peak value [34,38]. As the algal biomass increases, the opportunity for collisions between the cells and particles also increases. In addition, the surface functional groups of algal
cells decrease over time, which absorbs less particles and enhances
the adsorption capacity of the magnetic particles [52]. After the peak
biomass stage, the dissolved organic matter released by algal cells and
produced by cell autolysis will accumulate and the dissolved organic
matter will compete with algal cells for the surface available active
sites of particles. As a result, the recovery capacity clearly decreases
after the exponential phase [34,38,52,53]. In addition, the surface charge
of algal cells also uctuates during the cultivation process [46,49]. As the
separation process is primarily based on electrostatic interactions, the
uctuation of the electrophoretic mobility of the cells is signicant for
the RE. As reported by Lim et al., the maximum electronegativity was
achieved on the tenth day in a fourteen day cultivation of Chlorella sp.
This was most likely due to the increases in the kinetics of microalgae
growth and cell mobility, which further induced a net electronegative
zeta shield around the algal cells. Therefore, the tenth day was chosen
as the optimal harvesting time [46].

3.3. Summary
4.3. Magnetic particle dosage
The performance of various magnetic particles including naked and
coated magnetic reagents on microalgae recovery is summarized in
Table 1. As shown in Table 1, these reagents were effective for the harvesting of algal cells. In addition, the separation process generally required less than 10 min. These particles show potential and provide a
basis for a rapid and efcient microalgae harvesting method.
4. Effects of the magnetic harvesting process
Based on the interaction between magnetic particles and algal cells,
the magnetic separation process is signicantly inuenced by many

In most cases, the RE increases as the dosage of magnetic particles

increases until it reaches a peak RE value. At that point, it generally remains constant even if the particle dosage is increased [3235,38,43,
47]. This relationship is mainly due to the ratio of the algal cells and
the magnetic particles. At a low particle dosage, the magnetic particles
cannot completely adsorb the algal cells or the algal cells cannot adsorb
a sufcient amount of magnetic particles and cannot be completely harvested by the external magnetic eld due to the insufcient magnetic
force. At the optimal dosage, the quantity of the particles is sufcient
to interact with the algal cells and can provide sufcient magnetic

S.-K. Wang et al. / Algal Research 9 (2015) 178185

181

Table 1
Performance of various magnetic particles on microalgae recovery.
Recovery Adsorption
Process Cost of particle
efciency capacity
time
(US$/kg-particles) c
(g/g-particles) (min)

Separation cost Reference

(US$/kg-algal
biomass) d

~98%
pH 7
95%
pH 7
N98%
pH 7
N90%
pH 912
99%

Magnetic particle

Algal species Algal

concentration

Dosage Working
(mg/L) medium

Naked Fe3O4 particles

(chemical precipitation)
CPAM-Fe3O4 magnetic
occulant
Naked Fe3O4 particles
(chemical precipitation)
Silica-coated magnetic
particles
PDDA-rodlike Fe3O4
nanoparticles
Naked Fe3O4 particles
(microwave
synthesized)
CPAM-Fe3O4 magnetic
occulant
Fe3O4-PEI nanocomposites
PEI magnetic beads

B. braunii

1.8 g/L

75

CM b

B. braunii

1.8 g/L

25

CM

C. ellipsoidea 0.8 g/L

300

CM

C. vulgaris

1.3 g/L

1300

CM

Chlorella. sp.

200

CM

C. vulgaris

0.3 g/L

C. ellipsoidea 0.7 g/L

120

C. ellipsoidea 0.75 g/L

C. vulgaris
0.2 g/L

20

DEAE magnetic beads

C. vulgaris

0.2 g/L

Chitosan/magnetic
nanoparticle
Aminoclay-nZVI composite

Chlorella sp.
KR-1
Chlorella sp.
KR-1
Chlorella sp.

1 g/L

1400

~100%
Mineral
medium
lacking KH2PO4
CM
96%
pH 7
CM
97%
10 mM KCl
90%
pH 4
10 mM KCl
90%
pH 4
CM
99%

19130

CM

3 10 cells/mL

300

N. maritima

2.12.9 g/L

N. maritima

1.02 g/L

Chitosan-iron oxide
particles
Silica-coated magnetic
particles
Naked Fe3O4 particles
(chemical precipitation)
a
b
c
d

1.5 g/L
7

23.52

6.86

0.29

[42]

68.4

10

7.28

0.10

[34]

2.61

6.86

2.61

[42]

615

[48]

N15

[44]

0.33

1112

[32]

21.4

10

7.28

1.24

[34]

32.61
10

2
40

35.34

0.95

[37]
[31]

40

[31]

0.71

[46]

~100%

0.078

[45]

CM

97.19%

0.63

[38]

1680

CM

1120

[48]

120

CM

20%40%
pH 8
97.5%
pH 8

10.05

6.86

0.82

[33]

: not mentioned.
CM: Culture medium.
The calculation was based on Wang et al. (2014c).
This only considers the cost of magnetic particles regardless of the operation cost.

force to completely collect algal cells by an external magnetic eld and

the maximum RE is achieved. At this point, the spare particles will be
separated by the magnetic eld and have no inuence on the RE even
with further increases in the particle dosage. However, in algal cell
removal using a magnetic coagulant prepared by acid-modied y ash
compounded with Fe3O4 magnetite, the RE clearly decreased when
the coagulant dosage was increased after the optimum threshold
dosage was reached [51]. Similar results were also found in the occulation of microalgae using cationic starch [54]. This was primarily due to a
different mechanism in that separation process compared with others;
in that study, the separation was mainly based on coagulation which
formed thick ocs by bridging. Improved colloidal cohesion was
achieved as the dosage increased and a higher turbidity removal
efciency was obtained. However, adsorption saturation and overlay
formation on the colloids resulted in a second stable phenomenon led
by the overdose of coagulant, which decreased the turbidity removal
efciency [51].
4.4. pH
The pH value of the solution inuences the ionization degree and the
surface charge of both algal cells and magnetic particles as is indicated
by the zeta potential variation at different pH values [32,35,38,43]. As
a result, the RE is dependent on the pH value in the environment. In addition, these magnetic particles have different pH optimum values for
the harvesting of different algal species. In the harvesting of Chlorella
vulgaris using DEAD-beads and PEI-beads in a model environment
(10 mM KCl solution), lower pH values achieved higher REs [32]. Similar
results were obtained in the separation of B. braunii using naked Fe3O4
magnetic nanoparticles, and C. ellipsoidea using Fe3O4-PEI nanocomposite processes in culture medium [38,43]. However, in harvesting the

freshwater algae C. vulgaris and Chlamydomonas reinhardtii, and the marine algae Phaeodactylum tricornutum and Nannochloropsis salina in different media, higher REs were obtained at higher pH values [50]. In
contrast, neutral conditions beneted the harvesting of C. ellipsoidea
using naked Fe3O4 magnetic nanoparticles in the culture medium,
while negatively impacting the REs in the separation of N. salina using
naked Fe3O4 magnetic nanoparticles and the harvesting of B. braunii
and C. ellipsoidea using CPAM surface functionalized Fe3O4 (CPAMFe3O4) magnetic occulants in culture medium [34,35,43]. The differences were primarily due to the different surface ionization characteristics of the particles. In addition, other functions were also inuenced by
variations in the pH in the separation process. As reported by Hu et al., at
elevated pH values, the diameter of cell aggregates was observed to increase, which enhanced the RE [34]. This was mainly due to the algal
occulation induced by the pH increase [55]. Furthermore, in acidic or
alkali conditions, large chain deformation of CPAM can occur, which
can improve the interaction between the occulant and the algal cells.
This resulted in a higher RE in the separation of B. braunii and
C. ellipsoidea using CPAM-Fe3O4 magnetic occulant [35,56]. However,
the effect of pH on the RE was not signicant in the separation of Chlorella sp. using chitosan/magnetic nanoparticle composites in a pH range
from 2 to 12 [47]. This may be due to the strong cationic character of
chitosan.
4.5. Ions in the medium
The RE is also impacted by the ions present in the medium. In the
separation of freshwater microalgae using silica-coated magnetic
beads, a higher RE was obtained in IGV-medium compared to that in
modied TAP medium due to its high concentration of Mg2 + ions
[50]. Once Mg2 + ions are hydrolyzed at high pH values and form

182

S.-K. Wang et al. / Algal Research 9 (2015) 178185

magnesium hydroxide precipitate, the ions can effectively coagulate

algal cells by occulation and charge neutralization [55]. Similar results
were observed in the harvesting of marine algae in ASW-medium compared to modied MannMyers-medium; the higher concentration of
Mg2 +- and Ca2 +-ions in the ASW-medium effectively improved the
occulation efciency [50,57]. Furthermore, Prochazkova et al., found
that the presence and/or absence of certain ions can affect the surface
charge of magnetic beads [33]. The surface charge of the microwave
synthesized magnetic particles changed from negative to positive after
KH2PO4 was excluded from the medium, and it was conrmed that
this was due to the negatively charged phosphate ions. In addition,
phosphate ions can block the functional binding sites of the particles
by electrostatic attraction and have a negative effect on the RE [33]. In
addition, the separation mechanism was also signicantly inuenced
by the ions in the medium. Using Extended DerjaguinLandau
VeweyOverbeek (XDLVO) analysis, Toh et al. found that the main interaction mechanism between Chlorella sp. and bared IONPs was electrostatic (ES) interaction in freshwater. However, the Lewis acidbase
(AB) and van der Waals functions (vdW) played a key role in the separation of marine Nannochloropsis sp. using surface functionalized IONPs
in marine water [58]. Therefore, it is crucial for an effective harvesting
process that the competing ions are absent. Generally, in the microalgae
cultivation process, these ions are usually exhausted at the later stages
of the cultivation which is benecial to achieve a high RE. However,
the dissolved organic matter would accumulate at the later culture
stages, which is adverse for the efcient harvesting of algal cells as previously stated. These factors should be considered when determining
the harvesting time.
4.6. Temperature
In outdoor cultivation, the temperature may uctuate greatly during
the day; because the temperature impacts the RE, this inuences the optimal harvesting time point. For example, the adsorption capacity of
naked Fe3O4 nanoparticles and Fe3O4-PEI nanocomposites for the separation of N. maritima and C. ellipsoidea, respectively, increased as the
temperature increased [34,38]. This was mainly due to two reasons.
First, a higher temperature can reduce the solution viscosity of the medium, which can enhance the distribution of nanoparticles [59]. Second,
the elevated temperature can promote the mobility of the particles and
provide sufcient energy for the particles to adsorb the microalgal cells.
These results indicated that for these two separation processes, a higher
temperature is superior for efcient microalgae harvesting, and that it is
better to conduct the harvesting at a higher temperature point during
the day for outdoor cultivations [34,38]. However, the inuence of temperature in other separation processes should be further investigated.
This can direct the choice of the harvesting time for the outdoor
microalgal cultures due to the temperature uctuation in outdoor
conditions.

magnet arrays in LGMS. Therefore, the LGMS system was more costeffective due to its low energy consumption, and a system can be easily
designed [37]. Although HGMS has been widely used in manufacturing,
LGMS is more widely used in biotechnology processes [25,6062]. Furthermore, the LGMS has been used in magnetic harvesting of microalgae
and shows good potential [46,49]. The magnetic eld in LGMS can be
generated without extra power and the separation time is even less
than 3 min using nanorod particles, and it may be concluded that the
LGMS technique is more energy and time-efcient and is a better option
for microalgae harvesting compared with HGMS technology [46].
5. Magnetic separator for algal harvesting
For the large scale separation of microalgae using magnetic particles,
an efcient magnetic separation system is necessary. The high-gradient
magnetic separation (HGMS) system has been widely used in industrial
processes for several decades [63,64]. Currently, the development of
high gradient magnetic elds as high as 104 T/m provides forces large
enough to capture weakly magnetic particles from solutions, which allows magnetic separation to be widely used in various industrial processes [6567]. Magnetic separators of many different shapes and
scales have been applied for the recovery of magnetic material from
non-magnetic matter in industrial minerals, for the separation of ball
segments from discharge, the separation of low susceptibility minerals,
and water purication in the steel industry [23,66]. However, these
magnetic separators are not suitable for use in the magnetic harvesting
of microalgal cells due to the specic characteristics of the algal broth
that differ from that of industrial processes. A magnetic separator
consisting of a chamber, a magnet drum, a scraper blade, an inlet, and
two outlet ports, has been developed for efcient microalgae separation
by Hu et al. [68]. This magnetic separator can efciently harvest
microalgal cells using two modes. As shown in Fig. 2-procedure I, in
the batch separation mode, a mixture of magnetic particles and algal
broth was added into the separation chamber. After adsorption by the

Particle-cell aggregates
Procedure II

Procedure I

Dissolve particles using acid

Extract desired products

Residual
aggregates

Desired
products

Filtration

Dissolve particles using acid

4.7. Gradient of the magnetic eld

High-gradient magnetic separation (HGMS) with B N 1000 T/m
[37] has been successfully used for the separation of microalgae from
lakes for more than thirty years [42]. Recently, a lab-scale HGMS was applied for the magnetic separation of both freshwater algae and marine
microalgae, and demonstrated the potential of HGMS for efcient
microalgae harvesting using magnetic particles [50]. Toh et al., investigated the performance of both HGMS and low gradient magnetic separation (LGMS, B b 80 T/m) with varying dosages of magnetic particles.
At a low particle dosage, the HGMS resulted in a higher RE compared
with that of LGMS due to the high magnetophoresis kinetics under the
high eld gradient. However, the LGMS achieved an equal RE with
that of HGMS when a high particle dosage was tested [37]. In HGMS,
the high power consumption is necessary for magnetic power generation while the magnetic eld can be simply generated by permanent

Filtration

Microalgal
cells

Filtrate

Filtrate

Residual
cells

Regenerate particles using alkali

Magnetic particle
Fig. 1. A ow chart for the detachment of aggregates and the reusability of magnetic
particles.

S.-K. Wang et al. / Algal Research 9 (2015) 178185

Algal broth

and P. tricornutum within 5 min using silica-coated magnetic particles

at pH values ranging from 8.0 to 8.4. In the harvesting of C. reinhardtii,
the initial biomass concentration, particles-to-cells ratio, and medium
were 12.1 g/L, 0.74 g/g, and TAP-medium, respectively. Meanwhile, in
the separation of P. tricornutum, the initial biomass concentration,
particles-to-cells ratio, and medium were 3.5 g/L, 1.1 g/g, and mod.
MannMyers-medium, respectively. However, the separation capacity
was low and extra energy is necessary for the generation of the magnetic eld using electromagnet devices [50].

Mixing

Add magnetic
particles

Pump
Magnet
drum

Scraper
blade

Procedure I
Pumped into
the separator
for one time

Medium

6. Detachment of aggregates and the reusability of magnetic particles

Procedure II

Outlet I

Outlet II

183

Continuously
pumped into
the separator

Medium

Fig. 2. Schematic diagrams of the magnetic separation process using a magnetic separator
[66].

magnet drum, the particle-algal cell aggregates can be collected by the

scraper blade and the medium drained from Outlet I. A RE of 95% can
be obtained within 40 s in the harvesting of C. ellipsoidea using naked
Fe3O4 nanoparticles [68]. Using the continuous separation mode
(Fig. 2-Procedure II), the mixture was continuously pumped into the
separation chamber while the aggregates were continuously collected
by the scraper blade by the rotation of the magnet drum. At ow rates
less than 100 mL/min, recovery efciencies of more than 95% were
achieved when the naked Fe3O4 nanoparticle dosage, biomass concentration, and algal broth pH were 250 mg/L, 0.75 g/L, and around 7.0, respectively [68]. In addition, an in situ magnetic separation system was
constructed by Wang et al. [69]. At the end of cultivation, 120 mg/L of
CPAM-Fe3O4 particles was directly added into the B. braunii broth
with a biomass concentration of 1.23 g/L and the pH was around 7.0.
After being mixed with the algal cells by aeration, the mixture was
pumped into the magnetic separator operated in continuous mode. A
RE of 90% was obtained at a ow rate of 100 mL/min [69]. The energy
consumption in this process is low, which only needs 4.50 kW h for
the harvesting 1000 L algal broth, and the particle cost for the harvesting
of 1 kg of dry B. braunii cells is only US$0.73. The capital investment of
magnetic separator is the highest part, which accounts for 40.61% of
the total harvesting cost. The total harvesting cost is only US$2.07 to
harvest 1 kg dry biomass. Overall, the separation process using the magnetic separator was simple and economic [69], and it shows great potential for the harvesting of microalgae. In addition, a high gradient
magnetic ltration (HGMF) system was constructed by Cerff et al.
[50]. Using an electromagnet and a lter chamber, separation efciencies of more than 90% were obtained in the harvesting of C. reinhardtii

After algal cells have been separated from the medium using magnetic particles, it is necessary to determine how to utilize the algal
cells in the collected cell-particle aggregates and determine the reusability of the magnetic particles. As initially reported by Xu et al. [43],
the cell-particle aggregates can be treated by one of two procedures
(Fig. 1). In procedure I, the demagnetized algal cells can be collected
by micro-ltration after the magnetic particles have been dissolved
using HCl. The cells can be used to produce the desired products while
the magnetic particles can be regenerated by adding alkali. In procedure
II, the aggregates can be rst used for the extraction of the desired products and then the residual aggregates disposed of as in the rst procedure. The method of choice is dependent on the characteristics of the
algal species and the desired products, such as cell wall structure, the location and stability of the products, the application of the products etc.
For example, it is more effective to treat C. ellipsoidea cell-particle aggregates using the rst procedure due to the tough cell wall of C. ellipsoidea
and the desired products may be contaminated by the magnetic particles if directly extracted from the cell-particle aggregates. However,
the second procedure is better for the treatment of B. braunii cellparticle aggregates, because the extraction and quality of hydrocarbon
(that is the main product) was not inuenced by the existence of magnetic particles [43]. In addition, the regenerated magnetic particles
show equivalent efciency following ve reuse cycles compared with
newly synthesized particles [43]. Furthermore, an efcient recovery of
demagnetized-cells based on treating the cell-particle aggregates with
10% (v/v) H2SO4 under constant agitation in an ultrasonic bath and
heating at 40 C was developed by Prochavkova et al. [33]. Using this
procedure, a recovery efciency of nearly 100% of the magnetized
algal cells was obtained within 60 min [33]. The feasibility of this acid
dissolving procedure was also conrmed by Lee et al. and integrated
with the recovery of Fe3+ using polyphenols [47].
However, the above mentioned method based on the dissolution
under acidic conditions is not desirable for all algal cell compounds or
downstream processing methods [32]. In procedure I, the desired compounds may be destroyed in the acidic conditions while the existence of
the magnetic particle may inuence the extraction of the desired products in procedure II. In this case, mild conditions are superior. Based on
the pH-dependent characteristics of the surface charge of the particles, a
detachment procedure based on pH adjustment was developed. A detachment efciency of 90% was obtained in model environments
(10 mM KCl) at pH 12 within 90 min of agitation for DEAE magnetic
beads. However, the detachment efciency of PEI magnetic beads was
low due to the strong interactions between PEI and the functional
groups on the surface of algal cells which may form covalent bonds
[32]. In addition, Seo et al. found that APTES surface functionalized
BaFe12O19 magnetic particles with larger particle size were more efciently detached at pH 12 aqueous solution while separated at a neutral
medium. This is mainly because with a larger surface area, the smaller
magnetic particles provide more contact sites for algal cells and form
stronger electrostatic bonds compared to larger magnetic particles. In
addition, the saturation magnetization of magnetic particles has no effect on the detachment from the microalgae although it can inuence
the harvesting rate [36]. Furthermore, a repeated detachment of bare
Fe3O4 magnetic particles from algal slurry by increasing pH in dH2O

184

S.-K. Wang et al. / Algal Research 9 (2015) 178185

assisted by vortexing (30 s) or sonication (1 min) was developed by Lee

et al. [70]. Cell harvesting efciencies of 94%-99% and particle recovery
efciencies of 90%-97% were obtained for 10 reuse cycles [70]. These detachment procedures based on the electrostatic repulsion above the isoelectric points showed efciency [36,70]. However, it has been reported
that the magnetic particles, including CuO nanoparticles, naked-IONPs,
and surface functionalized-IONPs may enter the algal cells by collision
and interactions between particles and algal cells [39,71], in this case,
it would be impossible to completely detach the magnetic particles
from the algal biomass, and this may impact the production of certain
compounds. Currently, there are no superior techniques for the detachment of particles and cells from the aggregates and for the reuse of magnetic particles. Further investigation of the interaction of cells and
particles is necessary for the development of detachment techniques
and the industrial application of magnetic particles. In addition, the associated costs for the detachment should also be considered.
7. The biocompatibility of magnetic particles and the reuse of
culture medium
Ample studies have conrmed the toxicity of both naked magnetic
particles and coated particles on animal cells [72,73]. In addition, the
commonly found nanoparticles are potentially toxic to algal cells due
to various effects, such as agglomeration and physical interactions, oxidative stress, and sequestration of nutrient elements [39]. Therefore, the
biocompatible magnetic particles are necessary for some cases. Lee et al.
[48] reported that the harvested microalgae-particles ocs obtained in
the harvesting of Chlorella sp. KR-1 using a chitosan/Fe3O4 nanoparticle
composite could survive and grow well on an agar plate. In addition, the
microalgae-attached particle composite showed a typical cell growth
pattern in a 1 L bubble-column photobioreactor when used as an inoculum. The results indicated that the chitosan/Fe3O4 composite was biocompatible with Chlorella sp. A reduction of cell growth was found in
the cultivation of Chlorella sp. adding both naked-IONPs and surface
functionalized IONPs at a dosage of 50 mg/L into the medium compared
with the culture without particles. However, similar nal cell densities
were achieved. This inhibition effect was mainly due to light shading
rather than severe damage onto the cell membrane and DNA [39].
Microalgae cultivation is a water-consuming process and improving
the water-use efciency signicantly impacts the economics of
microalgae production. The reusability of the culture medium has
been investigated by many studies. Hu et al. conrmed that the recycled
culture medium resulting from the separation using naked Fe3O4 nanoparticles had no negative effects on the cultivation of N. maritima compared with that from centrifugation and fresh medium [34]. In addition,
the medium recycled after separation using surface functionalized
magnetic particles, including Fe3O4-PEI nanocomposites and chitosan/
magnetic nanoparticles, showed similar cell growth to that of the
recycled medium resulting from centrifugation and fresh medium [38,
48]. In contrast, microalgal cell growth was inhibited in the reused medium after harvesting microalgae using traditional inorganic Alum occulant at different dosages [48]. This indicates that the use of the nakedor coated-magnetic particles has less inhibitory effect on the medium
for the growth of microalgae. In addition, the reusability of medium
after pH adjustment in the harvesting or detachment process was also
investigated. The results indicated that the use of bared Fe3O4 magnetic
particles and pH adjustment showed no signicant adverse effect on the
growth of Chlorella sp. KR-1 along with the co-existing bacterial species
[70]. They also found that some dissolved organic matter can also be
separated by the magnetic particles from the medium. However, in
the large-scale microalgae production process, the application of acid
or alkali may increase the cost. This may be solved by displacing the addition of acid or alkali by controlling the CO2 supply using exhaust ue
gas and the pH values can be adjusted in the range of pH 4 to 8 by
using exhaust ue gas [70]. The reused medium following magnetic
separation can be effectively used for further cultivation of microalgae,

which has great potential for the large scale economic microalgaebased biorenement.
8. Conclusions, challenges, and further directions
Over the past several years, various aspects of the process of magnetic separation of microalgae have been studied, such as the synthesis of
efcient magnetic reagents, the separation process, the detachment of
the particlecell aggregates, the reuse of the magnetic particles, and
the development of an effective magnetic separator. These magnetic
separation technologies show strong potential for the efcient harvesting of microalgae with advantages such as low energy consumption,
rapid separation, and the reusability of medium and magnetic particles.
The costs of the magnetic particles in selected algal harvesting processes have been calculated and listed in Table 1. In the magnetic removal of microalgae from shpond water, the overall separation cost
was US$0.13/cubic meter of treated pond water [35]. In addition, the
overall cost of harvesting B. braunii using CPAM-Fe3O4 particles was
US$2.07 to harvest 1 kg algal biomass (including the particle cost and operation cost) [37]. Although these systems have potential, it is necessary
to further decrease the cost of the magnetic separation process for the
large-scale industrial production of algal biomass. To achieve an effective,
economic, and reusable magnetic separation process, further development on some key technologies in the near future is needed as follows:
(i) The development of more efcient, universal, biocompatible, and reusable magnetic particles with a minimum environmental impact;
(ii) A more thorough understanding of the particlecell interactions in
the separation process is necessary to aid in the design of magnetic
particles and for the optimization of the separation process;
(iii) Increased research on the design of a magnetic separator for
microalgae harvesting, with a focus on the development of a simple,
efcient, versatile, easy-to-operate, and high throughput magnetic
separation systems. In addition, LGMS technology shows potential
in the design of a magnetic separator for microalgae harvesting;
(iv) Further research on the downstream processes following magnetic
separation including the extraction of desired products and the
reuse of magnetic particles and culture medium. In addition, the design process should be based on the characteristics of the desired
end products.

Acknowledgment
This work was nancially supported by the Major State Basic Research
Development Program (973 Project) of China (Nos. 2011CB200903 &
2011CB200905), the National Natural Science Foundation of China (No.
21476242), the Knowledge Innovation Program of the Chinese Academy
of Sciences (Nos. KSZD-EW-Z-015 & YZ200947), and the Chinese Academy of Sciences Fellowships for Young International Scientists (No.
2011Y1GA01).
References
[1] Y. Chisti, Biodiesel from microalgae, Biotechnol. Adv. 25 (2007) 294306.
[2] R.H. Wijffels, M.J. Barbosa, An outlook on microalgal biofuels, Science 329 (2010)
796799.
[3] Y.G. Li, L. Xu, Y.M. Huang, F. Wang, C. Guo, C.Z. Liu, Microalgal biodiesel in China: opportunities and challenges, Appl. Energy 88 (2011) 34323437.
[4] Y. Chisti, Biodiesel from microalgae beats bioethanol, Trends Biotechnol. 26 (2008)
126131.
[5] T.M. Mata, A.A. Martins, N.S. Caetano, Microalgae for biodiesel production and other
applications: A review, Renew. Sust. Energ. Rev. 14 (2010) 217232.
[6] R. Harun, M. Singh, G.M. Forde, M.K. Danquah, Bioprocess engineering of microalgae
to produce a variety of consumer products, Renew. Sustain. Energy Rev. 14 (2010)
10371047.
[7] E. Specht, S. Miyake-Stoner, S. Mayeld, Micro-algae come of age as a platform for
recombinant protein production, Biotechnol. Lett. 32 (2010) 13731383.

S.-K. Wang et al. / Algal Research 9 (2015) 178185

[8] S.K. Wang, Y.R. Hu, F. Wang, A.R. Stiles, C.Z. Liu, Scale-up cultivation of Chlorella
ellipsoidea from indoor to outdoor in bubble column bioreactors, Bioresour. Technol.
156 (2014) 117122.
[9] L. Xu, P.J. Weathers, X.R. Xing, C.Z. Liu, Microalgal bioreactors: challenges and opportunities, Eng. Life Sci. 9 (2009) 178189.
[10] E. Stephens, I.L. Ross, Z. King, J.H. Mussgnug, O. Kruse, C. Posten, M.A. Borowitzka, B.
Hankamer, An economic and technical evaluation of microalgal biofuels, Nat.
Biotechnol. 28 (2010) 126128.
[11] C.Z. Liu, F. Wang, A.R. Stiles, C. Guo, Ionic liquids for biofuel production: opportunities and challenges, Appl. Energy 92 (2012) 406414.
[12] L. Xu, R. Liu, F. Wang, C.Z. Liu, Development of a draft-tube airlift bioreactor for
Botryococcus braunii with an optimized inner structure using computational uid
dynamics, Bioresour. Technol. 119 (2012) 300305.
[13] C.Z. Liu, S. Zheng, L. Xu, F. Wang, C. Guo, Algal oil extraction from wet biomass of
Botryococcus braunii by 1, 2-dimethoxyethane, Appl. Energy 102 (2013) 971974.
[14] J.E. Coons, D.M. Kalb, T. Dale, B.L. Marrone, Getting to low-cost algal biofuels: a
monograph on conventional and cutting-edge harvesting and extraction technologies, Algal Res. 6 (2014) 250270.
[15] L. Xu, F. Wang, H.Z. Li, Z.M. Hu, C. Guo, C.Z. Liu, Development of an efcient
electroocculation technology integrated with dispersed-air otation for harvesting
microalgae, J. Chem. Technol. Biotechnol. 85 (2010) 15041507.
[16] M. otari, D. Klinar, M. Bricelj, J. Golob, M. Berovi, B. Likozar, Growth, lipid extraction and thermal degradation of the microalga Chlorella vulgaris, New Biotechnol. 29
(2012) 325331.
[17] J. Kim, G. Yoo, H. Lee, J. Lim, K. Kim, C.W. Kim, M.S. Park, J.W. Yang, Methods of
downstream processing for the production of biodiesel from microalgae, Biotechnol.
Adv. 31 (2013) 862876.
[18] E.M. Grima, E.H. Belarbi, F.G.A. Fernndez, A.R. Medina, Y. Chisti, Recovery of
microalgal biomass and metabolites: process options and economics, Biotechnol.
Adv. 20 (2003) 491515.
[19] N. Uduman, Y. Qi, M.K. Danquah, G.M. Forde, A. Hoadley, Dewatering of microalgal
cultures: a major bottleneck to algae-based fuels, J. Renew. Sustain. Energy 2
(2010) 012701.
[20] C.Y. Chen, K.L. Yeh, R. Aisyah, D.J. Lee, J.S. Chang, Cultivation, photobioreactor design
and harvesting of microalgae for biodiesel production: a critical review, Bioresour.
Technol. 102 (2011) 7181.
[21] L. Christenson, R. Sims, Production and harvesting of microalgae for wastewater
treatment, biofuels, and bioproducts, Biotechnol. Adv. 29 (2011) 686702.
[22] J.J. Milledge, S. Heaven, A review of the harvesting of micro-algae for biofuel production, Rev. Environ. Sci. Biotechnol. 12 (2013) 165178.
[23] C.T. Yavuz, A. Prakash, J.T. Mayo, V.L. Colvin, Magnetic separations: from steel plants
to biotechnology, Chem. Eng. Sci. 64 (2009) 25102521.
[24] L. Borlido, A.M. Azevedo, A.C.A. Roque, M.R. Aires-Barros, Magnetic separations in
biotechnology, Biotechnol. Adv. 31 (2013) 13741385.
[25] C.Z. Liu, H. Honda, A. Ohshima, M. Shinkai, T. Kobayashi, Development of chitosanmagnetite aggregates containing Nitrosomonas europaea cells for nitrication enhancement, J. Biosci. Bioeng. 89 (2000) 420425.
[26] C.T. Yavuz, J.T. Mayo, W.W. Yu, A. Prakash, J.C. Falkner, S. Yean, L. Cong, H.J. Shipley,
A. Kan, M. Tomson, D. Natelson, V.L. Colvin, Low-eld magnetic separation of monodisperse Fe3O4 nanocrystals, Science 314 (2006) 964967.
[27] Y.Y. Jiang, C. Guo, H.S. Xia, H.S. Gao, I. Mahmood, C.Z. Liu, H.Z. Liu, Lipase immobilization on ionic liquids modied magnetic nanoparticles: ionic liquids controlled esters hydrolysis at oilwater interface, AIChE J. 58 (2012) 12031211.
[28] F. Wang, C. Guo, L.R. Yang, C.Z. Liu, Magnetic mesoporous silica nanoparticles: fabrication and their laccase immobilization performance, Bioresour. Technol. 101
(2010) 89318935.
[29] F. Wang, W. Huang, C. Guo, C.Z. Liu, Functionalized magnetic mesoporous silica nanoparticles: fabrication, laccase adsorption performance and direct laccase capture from
Trametes versicolor fermentation broth, Bioresour. Technol. 126 (2012) 117122.
[30] S.O. Gultom, B. Hu, Review of microalgae harvesting via co-pelletization with lamentous fungus, Energies 6 (2013) 59215939.
[31] H.M. Amaro, A.C. Guedes, F.X. Malcata, Advances and perspectives in using
microalgae to produce biodiesel, Appl. Energy 88 (2011) 34023410.
[32] G. Prochazkova, N. Podolova, I. Safarik, V. Zachleder, T. Branyik, Physicochemical approach to freshwater microalgae harvesting with magnetic particles, Colloids Surf. B
112 (2013) 213218.
[33] G. Prochazkova, I. Safarik, T. Branyik, Harvesting microalgae with microwave synthesized magnetic microparticles, Bioresour. Technol. 130 (2013) 472477.
[34] Y.R. Hu, F. Wang, S.K. Wang, C.Z. Liu, C. Guo, Efcient harvesting of marine
microalgae Nannochloropsis maritima using magnetic nanoparticles, Bioresour.
Technol. 138 (2013) 387390.
[35] S.K. Wang, F. Wang, Y.R. Hu, A.R. Stiles, C. Guo, C.Z. Liu, Magnetic occulant for high efciency harvesting of microalgal cells, ACS Appl. Mater. Interfaces 6 (2014) 109115.
[36] J.Y. Seo, K. Lee, S.Y. Lee, S.G. Jeon, J.G. Na, Y.K. Oh, S.B. Park, Effect of barium ferrite
particle size on detachment efciency in magnetophoretic harvesting of oleaginous
Chlorella sp, Bioresour. Technol. 152 (2014) 562566.
[37] P.Y. Toh, S.P. Yeap, L.P. Kong, B.W. Ng, D.J.C. Chan, A.L. Ahmad, J.K. Lim,
Magnetophoretic removal of microalgae from shpond water: feasibility of high
gradient and low gradient magnetic separation, Chem. Eng. J. 211 (2012) 2230.
[38] Y.R. Hu, C. Guo, F. Wang, S.K. Wang, F. Pan, C.Z. Liu, Improvement of microalgae harvesting by magnetic nanocomposites coated with polyethylenimine, Chem. Eng. J.
242 (2014) 341347.
[39] P.Y. Toh, B.W. Ng, C.H. Chong, A.L. Ahmad, J.W. Yang, C.J.C. Derek, J.K. Lim,
Magnetophoretic separation of microalgae: the role of nanoparticles and polymer
binder in harvesting biofuel, RSC Adv. 4 (2014) 41144121.

185

[40] M.J. Higgins, J.E. Sader, P. Mulvaney, Probing the surface of living diatoms with
atomic force microscopy: the nanostructure and nanomechanical properties of the
mucilage layer, J. Phycol. 39 (2003) 722734.
[41] G. Bitton, J.L. Fox, H.G. Strickland, Removal of algae from Florida Lakes by magnetic
ltration, Appl. Microbiol. 30 (1975) 905908.
[42] R. Yadida, A. Abeliovich, G. Belfort, Algae removal by high gradient magnetic ltration, Environ. Sci. Technol. 11 (1977) 913916.
[43] L. Xu, C. Guo, F. Wang, S. Zheng, C.Z. Liu, A simple and rapid harvesting method for
microalgae by in situ magnetic separation, Bioresour. Technol. 102 (2011)
1004710051.
[44] B.E. Eboibi, D.M. Lewis, P.J. Ashman, S. Chinnasamy, Effect of operating conditions on
yield and quality of biocrude during hydrothermal liquefaction of halophytic
microalga Tetraselmis sp. Bioresour. Technol. 170 (2014) 2029.
[45] A.E. Regazzoni, G.A. Urrutia, M.A. Blesa, A.J.G. Marato, Some observations on the
composition and morphology of synthetic magnetites obtained by different routes,
J. Inorg. Nucl. Chem. 43 (1981) 14891493.
[46] J.K. Lim, D.C.J. Chieh, S.A. Jalak, P.Y. Toh, N.H.M. Yasin, B.W. Ng, A.L. Ahmad, Rapid
magnetophoretic separation of microalgae, Small 8 (2012) 16831692.
[47] Y.C. Lee, K. Lee, Y. Hwang, H.R. Andersen, B. Kim, S.Y. Lee, M.H. Choi, J.Y. Park, Y.K.
Han, Y.K. Oh, Y.S. Huh, Aminoclay-templated nanoscale zero-valent iron (nZVI) synthesis for efcient harvesting of oleaginous microalga, Chlorella sp KR-1, RSC Adv. 4
(2014) 41224127.
[48] K. Lee, S.Y. Lee, J.G. Na, S.G. Jeon, R. Praveenkumar, D.M. Kim, W.S. Chang, Y.K. Oh,
Magnetophoretic harvesting of oleaginous Chlorella sp. by using biocompatible
chitosan/magnetic nanoparticle composites, Bioresour. Technol. 149 (2013) 575578.
[49] P.Y. Toh, B.W. Ng, A.L. Ahmad, D.J.C. Chan, J.K. Lim, Magnetophoretic separation of
Chlorella sp.: role of cationic polymer binder, Process Saf. Environ. (2014)http://
dx.doi.org/10.1016/j.psep.2014.03.010.
[50] M. Cerff, M. Morweiser, R. Dillschneider, A. Michel, K. Menzel, C. Posten, Harvesting
fresh water and marine algae by magnetic separation: screening of separation parameters and high gradient magnetic ltration, Bioresour. Technol. 118 (2012) 289295.
[51] D. Liu, P. Wang, G.R. Wei, W.B. Dong, F. Hui, Removal of algal blooms from freshwater by
the coagulationmagnetic separation method, Environ. Sci. Pollut. Res. 20 (2013) 6065.
[52] X. Zhang, P. Amendola, J.C. Hewson, M. Sommerfeld, Q. Hu, Inuence of growth
phase on harvesting of Chlorella zongiensis by dissolved air otation, Bioresour.
Technol. 116 (2012) 477484.
[53] D. Vandamme, K. Muylaert, I. Fraeye, I. Foubert, Floc characteristics of Chlorella
vulgaris: inuence of occulation mode and presence of organic matter, Bioresour.
Technol. 151 (2014) 383387.
[54] D. Vandamme, I. Foubert, B. Meesschaert, K. Muylaert, Flocculation of microalgae
using cationic starch, J. Appl. Phycol. 22 (2010) 525530.
[55] Z.C. Wu, Y. Zhu, W.Y. Huang, C.W. Zhang, T. Li, Y.M. Zhang, A.F. Li, Evaluation of occulation induced by pH increase for harvesting microalgae and reuse of occulated
medium, Bioresour. Technol. 110 (2012) 496502.
[56] Z.L. Gui, J.W. Qian, X.K. Li, B.Q. Zheng, Q.F. An, Study on chain deformation of polyacrylamides in solutions and its occulation performance during the occulation
process, J. Appl. Polym. Sci. 117 (2010) 29152922.
[57] A. Sukenik, G. Shelef, Algal autoocculation-verication and proposed mechanism,
Biotechnol. Bioeng. 26 (1984) 142147.
[58] P.Y. Toh, B.W. Ng, A.L. Ahmad, D.J.C. Chan, J.K. Lim, The role of particle-to-cell interactions in dictating nanoparticle aided magnetophoretic separation of microalgal
cell, Nanoscale (2014)http://dx.doi.org/10.1039/C4NR03121K.
[59] N.N. Nassar, Rapid removal and recovery of Pb(II) from wastewater by magnetic
nanoadsorbents, J. Hazard. Mater. 184 (2010) 538546.
[60] C.H. Setchell, Magnetic separations in biotechnologya review, J. Chem. Technol.
Biotechnol. 35 (1985) 175182.
[61] M. Lewin, N. Carlesso, C.H. Tung, X.W. Tang, D. Cory, D.T. Scadden, R. Weissleder, Tat
peptide-derivatized magnetic nanoparticles allow in vivo tracking and recovery of
progenitor cells, Nat. Biotechnol. 18 (2000) 410414.
[62] J.M. Nam, C.S. Thaxton, C.A. Mirkin, Nanoparticle-based bio-bar codes for the ultrasensitive detection of proteins, Science 301 (2003) 18841886.
[63] C. Delatour, Magnetic separation in water-pollution control, IEEE Trans. Magn. 9
(1973) 314316.
[64] J. Oberteuffer, High gradient magnetic separation, IEEE Trans. Magn. 9 (1973) 303306.
[65] H.H. Kolm, Large-scale manipulation of small particles, IEEE Trans. Magn. 11 (1975)
15671569.
[66] T. Ohara, H. Kumakura, H. Wada, Magnetic separation using superconducting magnets, Physica C 357360 (2001) 12721280.
[67] G.D. Moeser, K.A. Roach, W.H. Green, T.A. Hatton, P.E. Laibinis, High-gradient magnetic separation of coated magnetic nanoparticles, AIChE J. 50 (2004) 28352848.
[68] Y.R. Hu, C. Guo, L. Xu, F. Wang, S.K. Wang, Z.M. Hu, C.Z. Liu, A magnetic separator for
efcient microalgae harvesting, Bioresour. Technol. 158 (2014) 388391.
[69] S.K. Wang, F. Wang, A.R. Stiles, C. Guo, C.Z. Liu, Botryococcus braunii cells:
ultrasound-intensied outdoor cultivation integrated with in situ magnetic separation, Bioresour. Technol. 167 (2014) 376382.
[70] K. Lee, S.Y. Lee, R. Praveenkumar, B. Kim, J.Y. Seo, S.G. Jeon, J.G. Na, J.Y. Park, D.M.
Kim, Y.K. Oh, Repeated use of stable magnetic occulant for efcient harvest of oleaginous Chlorella sp. Bioresour. Technol. 167 (2014) 284290.
[71] Z. Wang, J. Li, J. Zhao, B. Xing, Toxicity and internalization of CuO nanoparticles to
prokaryotic alga Microcystis aeruginosa as affected by dissolved organic matter, Environ. Sci. Technol. 45 (2011) 60326040.
[72] S.M. Hussain, K.L. Hess, J.M. Gearhart, K.T. Geiss, J.J. Schlager, In vitro toxicity of
nanoparticles in BRL 3A rat liver cells, Toxicol. in Vitro 19 (2005) 975983.
[73] M. Mahmoudi, A. Simchi, A.S. Milani, P. Stroeve, Cell toxicity of superparamagnetic
iron oxide nanoparticles, J. Colloid Interface Sci. 336 (2009) 510518.