Effects of Algal Hydrophobicity and Bubble

Size on Flotation Separation of Microalgae

from Aqueous Medium
1

Sourabh Garg1, Liguang Wang2 and Peer M. Schenk1

Algae Biotechnology Laboratory, School of Agriculture and Food Sciences, The University of
Queensland, Brisbane, QLD 4072
Email: s.garg@uq.edu.au, p.schenk@uq.edu.au
2
School of Chemical Engineering, The University of Queensland, Brisbane, QLD 4072
Email: l.wang2@uq.edu.au

Abstract - Microalgae have great potential to be a

feedstock for producing biofuel. One of the major
impediments
towards
the
industrial-scale
manufacturing of biofuel from microalgae is the
ineffective separation of microalgae from water. In this
communication, we first carried out froth flotation tests
to harvest freshwater microalgae (Chlorella sp. BR2)
and marine microalgae (Tetraselmis sp. M8) from
dilute culture with and without a collector, tetradecyl
trimethylammonium bromide (C14TAB). The surface
hydrophobicity of microalgae was measured by using a
modified adherence-to-hydrocarbon method. If no
collector was added, BR2 showed high hydrophobicity,
and its flotation tests in a mechanically agitated cell
produced an algal concentrate with an enrichment
ratio of 13.5 and 90.3% algae recovery. The natural
hydrophobicity of M8 was low, so was its flotation
recovery (6.4%). Addition of C14TAB improved M8
recovery to 81.7% with a low enrichment ratio of 2.0.
Overall, the flotation performance correlated well with
algal hydrophobicity. In search of more effective
collectors for marine algae flotation, we measured the
hydrophobicity of M8 in aqueous solutions of
dodecylammonium hydrochloride (DAH) at different
pHs and concentrations. It was found that addition of
dodecylammonium hydrochloride (DAH) at 25 ppm
and pH 6 significantly enhanced the hydrophobicity of
M8. Subsequent mechanical flotation results confirmed
that at this chemical condition, M8 enrichment ratio
was increased to 5.8 with 86.8% algae recovery.
Further improvement was achieved by using a
Jameson cell with relatively small air bubbles. The
Jameson flotation for M8 gave an enrichment ratio of
11.4 with 97.4% algae recovery.
Key words: Marine microalgae; froth flotation; surface
hydrophobicity; cationic surfactant
I.

INTRODUCTION

Microalgae have great potential to be a feedstock for

producing biofuel as well as are capable of capturing
carbon dioxide. The overall production of biofuel from

microalgae can be divided into three major steps;

production, harvesting and processing [1]. Among these
steps, the major bottleneck for commercial scale
production of biofuels is the downstream processing,
where algal biomass has to be concentrated and separated
from water for further processing [2, 3]. This step can
constitute 20 to 30 % of total biofuel production cost [2].
Mass production of microalgal biodiesel requires efficient
harvesting of the biomass from cultivation media [4].
Various methods such as flocculation, sedimentation,
filtration, flotation, centrifugation and membrane
separation process have been proposed for separating algae
from the aqueous media [5].However, each method has its
own limitation; for example, they are either of low
efficiency or high capital cost with excessive energy
consumption.
Froth flotation is a promising method for harvesting of
microalgae. It utilises the ability of microalgaes natural
characteristics of relatively low density and self-float [5].
Froth flotation is a highly versatile method for physically
separating the particles with small footprint [6, 7]. It
consists of three phases: water, solid particles and air
bubbles. In dispersed air flotation, fine bubbles with the
diameter of about 1mm are generated either by agitation
combined with the air injection or bubbling air through
porous media. The air bubbles selectively pick up
hydrophobic particles and rise up to the top zone of the
flotation cell to achieve the separation of hydrophobic
particles from hydrophilic particles [8-10].
Microalgae are tiny particles whose size is typically in
the range of 1 to 20 micron. Algal hydrophobicity has been
recognised as a critical factor determining microalgae
flotation, and a missing link between flotation performance
and algal surface hydrophobicity was identified by Garg et
al [7]. Addition of surfactants is commonly used to render
algae surface hydrophobic, making it possible to use
flotation to separate algae from water [6, 7, 11, 12].
Surfactant consists of a charged hydrophilic head and a
hydrophobic carbon tail. The hydrophilic head is adsorbed
on the microalgae surface and the hydrophobic tail creates
a hydrophobic layer on microalgae surface, which

promotes the attachment of microalgae to air bubble

surface and the subsequent transport of this algae-bubble
aggregate through the froth [13, 14]. Flotation
performance can be affected by chemical and
hydrodynamic factors. Among the variables that affect the
chemical condition of froth flotation, pH and surfactant
type and concentration play critically important role in
affecting the electrical charge and hydrophobicity of
particle surface [15]. Different types of flotation machines
which provide different hydrodynamic conditions may also
affect
flotation
separation
efficiency.
Different
hydrodynamics can be generated by using different type of
flotation machines. The Jameson cell is one of the
advanced flotation machines which uses a plunging jet to
generate smaller air bubbles than the mechanical flotation
cell. The Jameson Cell technology was applied by Yan and
Jameson [16] to treat wastewater, with algal removal
efficiencies (defined on the basis of the difference in algal
concentration between feed and tail) being over 98%. In
the present work, a comparative study was undertaken to
understand the effects of different surfactants and
machines on microalgae flotation efficiency.
II.

EXPERIMENTALS

A. Algal culture and characterization

Marine microalga Tetraselmis sp. M8 was isolated
from the Sunshine Coast, Queensland, Australia
(2639'39"S, 1536'18"E; Genbank accession number
JQ423158) and freshwater microalga Chlorella sp. BR2
was isolated from the Brisbane River, Tennyson,
Queensland, Australia (2731'21.36"S, 1530'32.87"E;
Genbank accession number JQ423156).Microalgae stocks
are maintained in the Algae Biotechnology Laboratory at
The University of Queensland (UQ). They were cultivated
in silicate free f/2 medium, on an orbital shaker (100 rpm)
under 120 mol photon m-2 s-1 with 12-hour light/dark
cycles, at 26C 1C. Under the same conditions, they
were scaled up in two polyethylene bags with continuous
air and nutrients supplies. When the microalgae reached
the exponential growth phase, they were nutrient-starved
for two days for efficient lipid induction [17]. Then the
microalgae cultures were collected for flotation
experiments.
B. Froth flotation
The majority of the flotation experiments in the present
work were carried out using a 1.5-litre bottom-driven
mechanically agitated (Agitair) cell. Prior to the flotation
process, the microalgae cultures were stirred vigorously
for 2 min. Then each culture was subdivided into aliquots
of 1.3 litres, weighed and transferred into the flotation cell.
The pH of the flotation pulp was adjusted by adding NaOH
or HCl before adding the collector, tetradecyl trimethyl
ammonium bromide (C14TAB) or dodecyl ammonium
hydrochloride (DAH). The microalgae suspension was
first conditioned by stirring at 800 rpm for 5 mins. In the
flotation tests, the agitation rate was 600 rpm when
C14TAB was used and 800 RPM when DAH was used.
The air flow rate was 5litre/min. The mechanical flotation
lasted for 6 minutes.
At an optimal reagent scheme determined by the
above-mentioned mechanical flotation tests, we also
carried out Jameson cell flotation tests to understand the

effect of bubble size (or flotation hydrodynamics) on

microalgae flotation. The diameter size of Jameson Cell
was 150 mm with an orifice diameter of 3.83 mm. A 35litre slurry was fed into the Jameson Cell flotation at
pressure of 150 kPa. The air flow rate was 10 L/min. The
Jameson cell flotation time was around 15 minutes, during
which the tailing was continuously recycled to the feed
sump and pumped back to the Jameson cell. The Jameson
Cell flotation was initially developed by Mount Isa Mines
and Professor G.J. Jameson of The University of
Newcastle, Australia [18]. An excellent review of Jameson
cell flotation was given elsewhere [19].
The concentrates collected in trays and the tailings left
in the flotation machine underwent weighing and
microalgae cell counting. The cell count for each sample
was taken in three duplicates by loading 10 L of sample
on a haemocytometer (Brightline, USA), and the averaged
value was reported. The microalgae recovery (Y) was
determined using the following equation:
= 1

(1)

Ff

where T is the mass of tailing (or sink), F is the mass of

feed, t is the microalgae concentration in the tailing, and f
is the microalgae concentration in the feed.
The enrichment ratio (ER) is defined as the ratio of the
concentration of algae in the concentrate to the
concentration of algae in the feed. It was calculated using
the following formula:
=

(2)

where WRR represents the water rejection rate being equal

to T/F.
C. Hydrophobicity test
The hydrophobicity of microalgae was measured by
using the modified adherence-to-hydrocarbon method [20].
The test assesses essentially the distribution ratio of cells
between water and an organic phase. A total of 4 mL of
the algae sample was placed in a test tube to which 1 mL
of 98% pure n-hexane was added and shaken vigorously
by hand for 1 min; the emulsion was allowed to settle for
20 seconds. Then, 1 mL liquid from the bottom layer was
carefully obtained from the bottom aqueous layer of the
test tube and its absorbance was read at 620 nm using a
spectrophotometer (Hitachi, Model U-2800) to represent
the concentration of microalgae. The extractability (H) of
the hexane layer on organic substances in the algal
suspension was calculated using the following expression:
100%

(3)

where Ao is the initial absorbance of the microalgae

suspension and Aw is the absorbance of the aqueous phase
after being settled for 20 seconds.
III.

RESULTS AND DISCUSSION

Tab.I shows algal surface hydrophobicity and the

overall recoveries of freshwater microalgae BR2 and
marine microalgae M8 after 6 minutes of flotation in the
absence of any collector. BR2 had a much higher level of

TABLE
I.
COMPARISON
FOR
ALGAL
SURFACE
HYDROPHOBICITY, ALGAL RECOVERY, AND ENRICHMENT
RATIO BETWEEN FRESHWATER MICROALGAE (BR2) AND
MARINE MICROALGAE (M8) AT pH 9.5 IN THE ABSENCE OF
ANY COLLECTOR

Algal
type

Hydrophobicity
(%)

Recovery
(%)

Enrichment
ratio

BR2

30.1

90.3

13.5

M8

1.2

6.4

0.6

The flotation response of the marine microalgae in the

absence of any collector was poor. To improve it, one can
use appropriate collectors which can render microalgae
surface hydrophobic. According to the literature, most
microalgae are negatively charged at natural pHs, so
cationic surfactants are commonly used as algae flotation
collectors [6, 21]. In the present work, as a first attempt,
we performed a series of flotation tests using C14TAB. By
varying the dosage of this collector, we systematically
examined its effect on algal surface hydrophobicity and
flotation performance, and the results are shown in Fig. 1.
As shown, addition of C14TAB considerably improved
M8s surface hydrophobicity and its flotation response of
M8. When C14TAB dosage was increased from 0 to 80
ppm, the algal hydrophobicity was increased from 1.2% to
25.0%, the flotation recovery was increased from 6.4% to
81.7%, and the enrichment ratio reached a peak at 3.1 at a
low concentration before decreasing at higher
concentrations.

60

100

50

Fig. 2b shows that increasing DAH concentration also

increased the flotation recovery. It was also found that pH
had a large effect on M8 flotation. At a given
concentration, the flotation recovery was higher at pH 6
than at pHs 4 and 9.5. Fig. 2c shows that increasing DAH
first increased the enrichment ratio, reaching a peak before
decreasing at higher concentrations. The decrease in
enrichment ratio at concentrations above 10 ppm was
related to the decrease in water rejection rate. Specifically,
at pH 6, when DAH concentration was increased from 10
ppm to 50 ppm, the water rejection ratio was significantly
reduced from 93.0% to 68.7%. It appears that better
flotation performance was achieved at pH 6 than at pHs 4
and 9.5.

60

80

ER

60
30

Y (%)

10

40
20

ER
)
m
p
p
(
B
A
T
4
C1

pH 6

pH 9.5

40
20

0
100
80
60
40
20
0
15

20

10
0

pH 4

M8

40

H (%)

15

In search of more effective collectors for marine

microalgae flotation, we measured the hydrophobicity of
M8 in aqueous solutions of dodecylammonium
hydrochloride (DAH) at different pHs and concentrations.
It was found that DAH is more capable of rendering M8
hydrophobic, and the hydrophobicity values were shown in
Fig. 2a. Increasing DAH concentration from 0 to 50 ppm
steadily increased the algal hydrophobicity. At a given
DAH concentration, pH 6 gave higher algal
hydrophobicity than pHs 4 and 9.5. Algal hydrophobicity
above 50% was achieved at 50 ppm at pHs 4 9.5 or 25
ppm at pH 6.

Y (%)

Hydrophobicity
Recovery
Enrichment ratio

flotation concentration stream, thus the water rejection rate

was decreased, leading to the decrease in enrichment ratio
at higher collector concentrations. This phenomenon can
be accounted for by the slower liquid drainage in a saline
water film confined between air bubbles at higher
concentrations of flotation reagents [22]. It was observed
that, in our experiment, the flotation froth became overly
stable when increasing C14TAB concentration to high
levels. Ideally, a metastable froth is wanted for achieving a
high microalgae recovery and a high enrichment ratio.

H (%)

natural surface hydrophobicity than M8. The flotation

recovery of BR2 reached more than 90.3% with
enrichment ratio being 13.5, whereas under the same
process conditions only 6.4% recovery with enrichment
ratio of 0.6 was attainable for M8. Note that the
enrichment ratio of M8 flotation is less than 1, which
might be accounted for by the (downward) gravitational
sedimentation, which prevailed over the (upward) flotation
of microalgae.

10
5

10 20 30 40 50 60 70 80 90 100

10

20

30

40

50

DAH (ppm)
Figure 1. Algal surface hydrophobicity (H), flotation recovery (Y), and
enrichment ratio (ER) for M8 at different concentrations of C14TAB at pH
9.5. The lines were drawn to guide the eye.

While increasing C14TAB dosage is beneficial for

improving the flotation recovery of the marine microalgae,
it brought about increased amount of water reported to the

Figure 2. Hydrophobicity (H), recovery (Y) and Enrichment ratio (ER) of

M8 at different concentrations of DAH at pHs 4, 6 & 9.5. The lines were
drawn to guide the eye.

Over the tested DAH concentration range, we could not

concurrently achieve the highest flotation recovery and the
largest enrichment ratio. Therefore, the flotation condition
of 25 ppm DAH and pH 6 was selected as a compromise,
at which the flotation recovery was 85.0% with an
enrichment ratio of 5.6. This test was then repeated twice,
and the results were compared with the best outcomes of
C14TAB. Fig. 3 shows that DAH was capable of giving
higher flotation recovery and larger enrichment ratio than
C14TAB. Specifically, with DAH, the enrichment ratio
could be increased to 5.8 with 86.8% algae recovery; in
comparison, at 50 ppm C14TAB the enrichment ratio was
3.4 with a flotation recovery of 71.1% and at 80 ppm
C14TAB the enrichment ratio was 2.0 with a flotation
recovery of 81.7%.

recovery of M8, take 86% for instance, the Jameson cell

gave a remarkably high enrichment ratio (that is, 25.9),
which was 4.5 times of the one obtained by using the
mechanical cell.
That the Jameson cell was superior to the mechanical
cell in M8 flotation was mainly attributed to the smaller
bubbles generated by the former. However, other features
of Jameson cell may also play a role. The Jameson cell has
intense mixing with small bubbles, tailings recycling, no
mechanical agitation and no external air supply. Further
research is needed to understand the flotation of marine
microalgae using the Jameson cell technology.
100
C14TAB
80

10
DAH (25 ppm) at pH 6
C14TAB (50 and 80 ppm) at pH 9.5

40
20

Y (%)

ER

pH 9.5

60

0
100
DAH
pH4
pH6
pH9.5

80

60
40

2
20

0
70

75

80

85

90

95

100

10

20

30

40

50

60

H (%)

Y (%)

Figure 3. Comparison of flotation performance of M8 between DAH and

C14TAB.

40
35

25 ppm DAH
pH 6

Jameson cell
Mechanical cell

30

ER

25
20
15
10
5
0
70

75

80

85

90

95

100

Y (%)

Figure 4. Enrichment ratio (ER) versus flotation recovery (Y) for M8

using mechanical cell and Jameson cell with 25 ppm DAH at pH 6. The
lines are drawn to guide the eye, and they are expected to end up with the
recovery limitation of 100% at which the enrichment ratio is 1.

Three flotation tests with 25 ppm DAH at pH 6 were

done using the Jameson flotation cell, and the results are
shown in Fig. 4. Also plotted in Fig. 4 are the experimental
results obtained under the same chemical condition but
with the mechanical cell. As shown, the Jameson cell gave
an enrichment ratio of 11.4 with 97.4% M8 recovery. The
change in flotation machine type from the mechanical cell
to the Jameson cell allowed the enrichment ratio to be
almost doubled and the flotation recovery to be increased
by at least 10 percentage points. For the same flotation

Figure 5. Flotation recovery (Y) as a function of algal surface

hydrophobicity (H) for M8 with using C14TAB and DAH.

In Fig. 5, the flotation recoveries were plotted as a

function of algal hydrophobicity. It shows that a strong
correlation exists between flotation recovery and algal
hydrophobicity when C14TAB or DAH was used as the
collector. Although C14TAB could be used to improve the
hydrophobicity of the marine microalgae, DAH was much
more effective for rendering M8 hydrophobic. The DAH
molecules can more readily adsorb on the surface of the
microalgae with their hydrocarbon tails exposed to the
aqueous phase than C14TAB. Since the hydrocarbon tails
are hydrophobic, the collector-coated marine microalgae
surfaces acquire hydrophobicity, which can be attached to
hydrophobic air bubbles via hydrophobic interaction. In
general, the higher the packing density of the hydrocarbon
tails on the particle surface, the stronger the surface
hydrophobicity. A collector which is more prone to be
adsorbed onto microalgae surface may also make possible
the formation of microalgae aggregate, which is conducive
to improving fine particle flotation. The good correlation
between microalgae flotation recovery and hydrophobicity
suggests that one can increase the flotation performance of
microalgae by using more effective collectors which can
render microalgae surface more hydrophobic. And the
hydrophobicity test for microalgae is a simple and
effective method for screening the collectors of microalgae
flotation.
CONCLUSIOINS
Flotation of freshwater microalgae (Chlorella sp. BR2)
and marine microalgae (Tetraselmis sp. M8) with and
without collectors were studied. If no collector was added,

BR2 showed high natural hydrophobicity and excellent

flotation response. The natural hydrophobicity of M8 was
low, so was its flotation response. Addition of cationic
surfactants such as tetradecyl trimethylammonium
bromide (C14TAB) and dodecylammonium hydrochloride
(DAH) improved M8s surface hydrophobicity and
flotation recovery. Overall, the flotation performance
correlated well with algal hydrophobicity. DAH was a
more effective collector than C14TAB for M8 flotation.
The mechanical flotation tests found that DAH worked
best at 25 ppm and pH 6, where M8 enrichment ratio was
5.8 with 86.8% recovery. Further improvement was
achieved by using the Jameson flotation technology, which
gave M8 enrichment ratio of 11.4 with 97.4% recovery.
The results suggest that both chemical reagents and bubble
size (hydrodynamics) are important for the flotation of
marine microalgae.

[8]

[9]

[10]

[11]

[12]

[13]

[14]

ACKNOWLEDGEMENT
Financial support for this study, provided by a CIEF
grant by The University of Queensland, is gratefully
acknowledged.

[15]

[16]

REFERENCES
[1]

[2]

[3]

[4]

[5]

[6]

[7]

C. Ryan, "Cultivating Clean Energy: The Promise of Algae

Biofuels," Report by the Natural Resources Defense Council
(NRDC) and Terrapin Bright Green, LLC. 2009, Washington,
DC NRDC Publications. 92.
E. Molina Grima, E. H. Belarbi, F. G. Acin Fernndez, A.
Robles Medina and Y. Chisti, "Recovery of microalgal biomass
and metabolites: process options and economics," Biotechnology
Advances. vol. 20 (7-8), 2003: p. 491-515.
L. Christenson and R. Sims, "Production and harvesting of
microalgae for wastewater treatment, biofuels, and bioproducts,"
Biotechnology Advances. vol. 29(6), 2011: p. 686-702.
Y.-L. Cheng, et al., "Dispersed ozone flotation of Chlorella
vulgaris," Bioresource Technology. vol. 101( 23), 2010: p. 90929096.
W. Phoochinda and D. A. White, "Removal of algae using froth
flotation," Environmental Technology. vol. 24(1), 2003: p. 8796.
Y. M. Chen, J. C. Liu and Y.-H. Ju, "Flotation removal of algae
from water," Colloids and Surfaces B: Biointerfaces. vol. 12(1),
1998: p. 49-55.
S. Garg, Y. Li, L. Wang and P. M. Schenk, "Flotation of marine
microalgae: Effect of algal hydrophobicity," Bioresource
Technology. vol. 121, 2012: p. 471-474.

[17]

[18]

[19]

[20]

[21]

[22]

L. Pan, S. Jung and R.-H. Yoon, "A fundamental study on the

role of collector in the kinetics of bubbleparticle interaction,"
International Journal of Mineral Processing. vol. 106109, 2012:
p. 37-41.
R. Perea-Carpio, F. Gonzlez-Caballero and J. M. Bruque, "On
the interactions at interfaces in fluorite flotation," International
Journal of Mineral Processing. vol. 23(34), 1988: p. 229-240.
S. Farrokhpay, "The significance of froth stability in mineral
flotation A review," Advances in Colloid and Interface
Science. vol. 166(12), 2011: p. 1-7.
J. C. Liu, Y. M. Chen and Y.-H. Ju, "Separation of Algal Cells
from Water by Column flotation," Separation Science and
Technology. vol. 34(11), 1999: p. 2259-2272.
N. Uduman, Y. Qi, M. K. Danquah, G. M. Forde and A.
Hoadley, "Dewatering of microalgal cultures: A major
bottleneck to algae-based fuels," Journal of Renewable and
Sustainable Energy. vol.2(1), 2010: p. 012701-15.
S. M. Bulatovic, Handbook of Flotation Reagents: Chemistry,
Theory and Practice: Volume 2: Flotation of Gold, PGM and
Oxide Minerals. 2010: Elsevier Science.
J. Drzymaa and A. Swatek, Mineral Processing: Foundations of
Theory and Practice of Minerallurgy. 2007: University of
Technology.
S. M. Bulatovic, Handbook of flotation reagents : chemistry,
theory and practice : flotation of sulphides ores. Vol. 1. 2007,
Amsterdam: Elsevier.
Y.-D. Yan, G. J. Jameson, Application of the Jameson Cell
technology for algae and phosphorus removal from maturation
ponds, International Journal of Mineral Processing, 73 (2004)
23-28
Hu, Q., M. Sommerfeld, E. Jarvis, M. Ghirardi, M. Posewitz, M.
Seibert, A. Darzins, 2008. Microalgal triacylglycerols as
feedstocks for biofuel production: perspectives and advances,
Plant J. 54, 621-639.
G. J. Jameson, A new concept in flotation column design, in:
SASTRY, K. V. S. (ed.) Column flotation '88 : proceedings of an
International Symposium on Column Flotation: SME Annual
Meeting. Arizona: Phoenix, 1988.
Harbort, G., Bono, S.D., Carr, D., Lawson, V., 2003. Jameson
Cell fundamentals - a revised perspective, Minerals
Engineering, 16 (2003) 1091-1101.
M. Rosenberg, D. Gutnick and E. Rosenberg, "Adherence of
bacteria to hydrocarbons: A simple method for measuring cellsurface hydrophobicity," FEMS Microbiology Letters. 9 vol. 1,
1980: p. 29-33.
W. Phoochinda, D. A. White and B. J. Briscoe, "An Algal
Removal Using a Combination Of Flocculation and Flotation
Processes," Environmental Technology. vol. 25(12), 2004: p.
1385-1395.
L. Wang, and R.-H. Yoon, Hydrophobic forces in thin aqueous
films and their role in film thinning, Colloids And Surfaces Aphysicochemical And Engineering Aspects, 263 (2005) 267-274.