Bioresource Technology 216 (2016) 824–829

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Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Microalgae harvesting by pH adjusted coagulation-flocculation,

recycling of the coagulant and the growth media
Probir Das ⇑, Mahmoud Ibrahim Thaher, Mohammed Abdul Quadir Mohd Abdul Hakim,
Hareb Mohammed S.J. Al-Jabri, Ghamza Saed H.S. Alghasal
Algal Technologies Program, Center for Sustainable Development, College of Arts and Sciences, Qatar University, P.O. 2713, Doha, Qatar

h i g h l i g h t s

Almost 90% iron in the harvested biomass was recovered using acidic solution.

Iron extracted solution can be reused in the subsequent biomass harvesting.

Microalgae can grow in the supernatant and utilize the residual iron in it.

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

Article history: Coagulation-flocculation can be considered as one of the least energy intensive microalgae biomass har-
Received 28 April 2016 vesting processes. However, cost of the coagulant and biomass contamination are two critical issues that
Received in revised form 2 June 2016 need to be considered. In this study, ferric chloride (72–96 mg/L) was used to effectively harvest
Accepted 3 June 2016
Scenedesmus sp. (530 mg/L) – grown in BG-11 media and wastewater. Reducing the culture pH below
Available online 11 June 2016
6.5, greatly improved the harvesting efficiency. Acidic solution (pH 1.0) was very effective to recover
(almost 90%) the associated iron from the harvested biomass. Scenedesmus sp. was able to grow in the
Keywords:
supernatant and utilize the residual iron in it. Iron extracted solution, with a supplementation of
Harvesting
Microalgae
9.8 mg/L ferric chloride, was able to achieve similar harvesting efficiency. The potential recovery of iron
Ferric chloride from the harvested biomass and its reuse in the harvesting can improve the biomass quality for subse-
Recycling culture medium quent downstream processing while reducing the cost.
Recovery of coagulant Ó 2016 Elsevier Ltd. All rights reserved.

1. Introduction microalgal biofuel (Milledge and Heaven, 2012; Uduman et al.,

Potential application of large scale microalgae cultivation
encompasses carbon sequestration, biofuel application, wastewa-
ter treatment etc. (Christenson and Sims, 2011; Lam et al., 2012;
Slade and Bauen, 2013; Williams and Laurens, 2010). At the end
of the cultivation process, microalgae biomass needs to be sepa-
rated from the culture. Microalgae are tiny little cells usually in
the range of 2–30 lm (Fon Sing et al., 2011). Production of high
density microalgae biomass is often observed in small scale culti-
vation, especially in photo-bioreactors with short optical depth.
However, in large scale cultivation microalgal biomass density
doesn’t exceed 0.5 g/L (Borowitzka, 1999). Because of lower bio-
mass density and very small cell size, harvesting of biomass is con-
sidered as one of the major bottlenecks for commercializing
⇑ Corresponding author.
E-mail address: probir.das@qu.edu.qa (P. Das).
2010).
Dewatering of the microalgae biomass can be divided into two
steps: in the first step, microalgae biomass is separated from the
bulk of the culture to a slurry of 1–2% solid content; in the second
step, a biomass paste of 10–30% solid content can be achieved
based on the requirement of the downstream processing. Floccula-
tion of microalgae cells can be a potential preliminary microalgae
biomass harvesting option, especially for low-value products
(Vandamme et al., 2013). Although, auto-flocculation (Sukenik
and Shelef, 1984) and bio-flocculation (Salim et al., 2010;
Talukder et al., 2014) are very promising, these processes are strain
specific and need to be demonstrated in large scale (González-Fer
nández and Ballesteros, 2012). On the contrary, chemical coagu-
lants based microalgae biomass harvesting are relatively consis-
tent, inexpensive and effective (Vandamme et al., 2013).
In many previous studies, ferric chloride was used effectively to
harvest different microalgae biomass (Rakesh et al., 2013; Seo
et al., 2015; Wyatt et al., 2012). However, a downside effect of

http://dx.doi.org/10.1016/j.biortech.2016.06.014
0960-8524/Ó 2016 Elsevier Ltd. All rights reserved.
 P. Das et al. / Bioresource Technology 216 (2016) 824–829
ferric chloride (or other coagulant) based microalgae harvesting is
that a major fraction of the coagulant is expected to be associated
with the microalgae biomass. Any downstream processing with
this harvested biomass will reduce the overall efficiencies and it
may also impair the final products (Knuckey et al., 2006). There-
fore, before applying any downstream processing, it might be very
useful to separate the coagulant. It is known that acidic solution
can leach elemental metals from microalgae biomass (Darnall
et al., 1986; Mehta and Gaur, 2005). Therefore, the associated iron
in the harvested biomass could be recovered using appropriate
acidic solution. This approach will not only remove the iron from
the biomass, it may also allow reusing the iron for next
coagulation-flocculation process. Addition of ferric chloride
reduces the pH of the culture and forms positively charged hydrox-
ide iron species which can bind negatively charged microalgae
cells onto it. pH of the culture determines the net surface charge
(positive, negative or even neutral) on the microalgae cells. There-
fore, initial culture pH can influence the coagulant dose require-
ment for optimum harvesting (Chen et al., 2013; Seo et al.,
2015). Iron recovery solution will be acidic, even after the extrac-
tion process. If this solution is reused in a fresh culture, it can serve
two purposes: (1) it will reduce the pH of the culture, and (2) it will
supply iron in the culture which later may help in microalgal floc
formation. However, for optimum harvesting efficiencies addi-
tional iron chloride may be needed – based on the iron extraction
efficiency.
Scenedesmus species are widely available freshwater microal-
gae. Because of the nutritional benefits, these species are used in
aquaculture (Ovie and Egborge, 2002). Scenedesmus species are also
know to accumulate copious amount of lutein (Sanchez et al.,
2008). Furthermore, Scenedesmus species are also studied as feed-
stock for biofuel, due to their high growth rate, and desired intrin-
sic metabolites (Miranda et al., 2012). Scenedesmus was also found
effective in treating municipal waste water and other industrial
wastewater (Mandal and Mallick, 2011). First objective of this
study was to evaluate the improvement potential of ferric chloride
based harvesting efficiency by adjusting the pH of Scenedesmus sp.
culture-grown in BG-11 media and municipal wastewater (or,
MWW). The second objective of this study was to evaluate the
recycling potential of the iron from the harvested biomass slurry
and reuse of the growth medium.
2. Methods
2.1. Strains and cultivation
A local Scenedesmus sp. (GenBank: KM985413) was used in this
study (Saadaoui et al., 2016). To determinate the biomass density
of the Scenedesmus sp. culture at any point of time, optical density
measurement at 750 nm was used. Previously, a calibration curve
between biomass density and optical density at 750 nm was made
for this strain. All the indoor cultures of Scenedesmus sp. were
grown in a temperature controlled room (at 25 °C) and using arti-
ficial blue fluorescent light (200 l mol E/m2/s; 12 h of photoperiod
per day) while mixing the cultures with air at 0.5 v/v/m rate. Apart
from sodium nitrate and sodium phosphate monobasic, all other
nutrients were added as BG-11 media concentrations for indoor
and outdoor cultivation of Scenedesmus sp; urea (78 mg/L) and
sodium phosphate dibasic (15 mg/L) were used instead of sodium
nitrate and sodium phosphate monobasic. No nutrients were
added in the municipal wastewater (characteristics given in Sup-
plementary file) for cultivating Scenedesmus sp. 40 L of indoor
grown Scenedesmus sp. culture (OD @ 750 nm = 0.5) was mixed
with 960 L BG-11 growth media in an outdoor raceway tank. Same
amount inoculum was used for the MWW culture. Light intensity
825
and air temperature during these outside growth experiments
are included in the Supplementary file. Bleach was added in the
outdoor tanks, before inoculation of Scenedesmus sp. culture. In
every two days, samples from outdoor cultures were collected
and checked under the microscope for possible contamination by
other microalgae; although no microalgal contamination was
found in the outdoor tanks. Outdoor cultures received 99.99% pure
CO2 from a rack of 16 cylinders. Flow velocity of the cultures in
these tanks were approximately 20 cm/s. Depth of the cultures in
these raceway tanks were maintained at 20 cm. Evaporation of
water was balanced everyday by adding same amount of
freshwater.
2.2. Harvesting experiment
The most common and conventional harvesting experiment is
conducted by using a jar test. However, in this experiment a differ-
ent approach was investigated. Initially, pH of the culture was
adjusted to desired values by adding different volume of H2SO4.
Next, 1 L cultures were kept in Duran glass bottles. 10 g FeCl3
(anhydrous) was mixed in 100 mL deionized water and different
volume of this coagulant solution was added in each bottle. For
each batch of harvesting experiment, coagulant solution was pre-
pared separately and used as fresh. Next, the cultures were mixed
with the help of air-infusion. A fixed 0.5 v/v/m airflow rate was
used for 2 min to mix the cultures. Once the mixing was stopped,
the biomass was allowed to settle for 20 min. Next, supernatant
sample was collected from the 200 mL mark of the Duran bottle
and OD of the supernatant was determined at 750 nm. pH of the
supernatant was also recorded using a handheld pH meter. Har-
vesting efficiency of any treatment was calculated by the following
formula.
ODi  ODf
Harvesting efficiency ¼  100;
ODi
where ODi = initial OD of the culture and ODf = final OD after the
settlement.
2.3. Comparison with low and high pH induced coagulation-
flocculation
pH adjusted flocculation of Scenedesmus sp. was also studied at
low and high pHs. pH of the original 1 L culture was adjusted to
either 3.75 or, 11.5 by adding appropriate amount of 1 M H2SO4
and 1 M NaOH solution, respectively. The volume of H2SO4 and
NaOH solutions were recorded for comparing with optimum ferric
chloride dose. After one hour, the sample from supernatant was
collected for determining the harvesting efficiency.
2.4. Recovery of iron
15 L culture of Scenedesmus sp. was harvested using optimum
conditions found in Section 2.2. The supernatant was decanted
from the top and collected in a container. Later the separated bio-
mass was concentrated further using a benchtop Thermo-Scientific
SL 16R centrifuge (5000 rpm for 10 min) and a biomass paste was
obtained. The liquid, obtained after the centrifuge, was then mixed
with the initial supernatant volume. The paste was then divided
equally into 15 parts and each part was placed in a 50 mL cen-
trifuge tube. Later, 10 mL volume of each iron extraction solution
was added in these centrifuge tubes. All these tubes were then vor-
texed for 1 min and placed horizontally in an orbital shaker. Mixing
inside the orbital shaker was maintained at 120 rpm for 1 h. Next,
these tubes were centrifuged (5000 rpm for 10 min) to separate the
liquid solution. Iron concentration in the liquid was then measured
826 P. Das et al. / Bioresource Technology 216 (2016) 824–829
colorimetrically using HACH method. Similarly, iron concentration
in the mixed supernatant was also determined.
2.5. Extract and reuse of iron as coagulant
Initially, biomass from one liter culture was harvested using
optimized coagulant dose. The settled biomass slurry was then
separated by decanting the supernatant. Later, the slurry was cen-
trifuged to obtain a biomass paste. 10 mL of acidic extracting water
was added to the paste and mixed for 1 h. This mixture was then
centrifuged and the supernatant was collected. The concentration
of extracted iron in the supernatant was calculated. Next extra fer-
ric chloride was added in this supernatant to match the optimum
coagulant requirement. Finally, this iron containing solution was
used for harvesting 1 L culture in the second batch.
2.6. Determination of iron in liquid
The concentration of iron in the liquid was determined by Hach
Method 8008 using HACH reagent kit. In brief, the powder of a
FERROVERÒ reagent kit was emptied in 10 mL sample containing
cuvette. After a gentle mixing, the cuvette was left for 10 min to
form pink color. Later the cuvette was placed in the HACH DR
3900 spectrophotometer for direct measurement of iron.
2.7. Reuse of the culture medium
After coagulation-flocculation of the biomass, 90% of the super-
natant was collected. pH of the collected water was adjusted to the
7.1 by adding appropriate amount of NaOH. Later 10% inoculum
was added to subculture Scenedesmus sp. Nutrients and growth
conditions for Scenedesmus sp. were the same as described in Sec-
tion 2.1. However, no iron was supplemented during the inocula-
tion. Growth of Scenedesmus sp. in the recycled medium was
monitored daily by recording the OD at 750 nm. These procedures
were also repeated for the second time growth medium recycling.
2.8. Microalgae biomass harvesting in large scale
100 L culture of Scenedesmus sp. was transferred into a con-
tainer (1.2 m deep) using a pump. Later optimum amount of ferric
chloride was added in the culture, and immediately the culture
was aerated with the help of an air pump for 5 min. After 30 min
of settling time, the biomass was recovered from the bottom. The
harvested biomass was centrifuged and iron concentration in the
separated water was determined. Acidic extraction solution was
used to extract the iron from the harvested biomass. Concentration
of iron in the extracted solution was also quantified. The difference
between optimum iron requirement and iron concentration in the
extract solution was calculated and this amount of iron was added
(as ferric chloride) in the extracted solution for a second batch of
harvesting.
2.9. Statistical analysis
All the culture samples, either in BG-11 media, or MWW, were
collected from outdoor 1000 L raceway tanks. Each coagulation-
flocculation test was performed in triplicate experiments. To deter-
mine whether initial culture pH and iron chloride dose affect
microalgae harvesting efficiency, analysis of variance was used
with p < 0.05. For the medium recycling experiment, three individ-
ual cultures were grown, harvested and re-inoculated separately.
Iron extraction efficiency was also reported by the average of three
individual samples. Standard errors were included in all the
graphs.
3. Results and Discussions
3.1. Biomass density and pH of outdoor cultures
For Scenedesmus sp., the calibration curve of biomass concentra-
tion (Y, g/L) and optical density at 750 nm (X) was determined as
Y = 0.7269X (r2 = 0.9813). Although Scenedesmus sp. was grown in
BG-11 media and MWW in two different times, culture samples
from both tanks were collected after the biomass density reached
0.53 g/L. During the cultivation of Scenedesmus sp. in the BG-11
media, pH of the culture was maintained in the range of 7.0–7.5,
whereas in MWW culture it was in the range of 7.2–7.5. As the bio-
mass concentrations in both these cultures were reaching in the
stationery phase, CO2 supplementation was stopped. Therefore,
the pH values of these cultures continued to increase. During the
harvesting experiment, pH values of the Scenedesmus sp. cultures
in BG-11 media and MWW were 8.0 and 7.91 respectively.
3.2. Effect of pH adjustment on the harvesting efficiency of
Scenedesmus sp
3.2.1. BG-11 media culture
Harvesting efficiencies of Scenedesmus sp. cultures for different
initial pH and different ferric chloride doses are shown in Fig. 1a.
There was significant effect of initial pH and coagulant dose on bio-
mass harvesting efficiency (p < 0.05); within the ferric chloride
dose range 24–72 mg/L, higher harvesting efficiencies were always
observed at lower initial pH, for the same dose. For 72 mg/L ferric
chloride dose, harvesting efficiency in the original culture was
72.9%; however, for the same dose, harvesting efficiency in the
pH 6.5 adjusted culture was 97.2%. Seo et al. (2015) also found that
harvesting efficiencies were higher at low pH adjusted cultures of
Chlorella sp. Growth medium and cultivation period of different
microalgae species will vary. Additionally, surface groups on the
microalgae cells will vary among different strains. Therefore, dif-
ferent amount of coagulant would be required for harvesting dif-
ferent microalgae cultures (Table 1). Additionally, if the initial
culture pH values were higher, higher doses of ferric chloride were
required for optimum harvesting efficiency.
Harvesting efficiency in the pH 6.5 adjusted culture was
decreasing slightly with the increase in ferric chloride dose. When
72 mg/L ferric chloride was added in the culture, the resulting pH
of the supernatant was 3.88 and it continued to decrease for adding
more coagulant. It was, therefore, also possible that Scenedesmus
sp. cells could have gained positive surface charge at pH below
3.88 and it reduced the harvesting efficiencies (Wyatt et al.,
Fig. 1a. Effect of pH on the harvesting efficiencies of Scenedesmus sp. cultures,
grown in BG-11 media.
 P. Das et al. / Bioresource Technology 216 (2016) 824–829 827

Table 1
Use of ferric chloride as a coagulant to harvest different freshwater microalgae species.

Strain name Biomass density (mg/L) pH of the culture Dosage of ferric chloride (mg/L) Harvesting efficiency Reference
Chlorella sp. 1700 3.0 200 90 Seo et al. (2015)
Scenedesmus sp. 540 10.3 150 97.3 Chen et al. (2013)
Chlorella zofingiensis 500 10.6 200 >90 Wyatt et al. (2012)
Chlorella sp. 710a >6.0 122 93 Udom et al. (2013)
Chlorella vulgaris 500 7.5 125 99a Chatsungnoen and Chisti (2016)
Choricystis minor 500 7.5 100 98a
Scenedesmus sp. 530 6.5 72 97.2 This study
a
Estimated from graph.

2012). Ferric chloride, being acidic in nature, reduced the final pH

of the cultures (Fig. 1b). Lower the pH of the supernatant, higher
would be the base (e.g., NaOH) requirement to neutralize it for
recycling as growth medium or even for discharge.

3.2.2. MWW culture

Ferric chloride is commonly used in wastewater treatment pro-
cess as it is relatively inexpensive and very effective in removing
suspended and colloidal particles, or even microalgae (Tatsi et al.,
2003; Udom et al., 2013). The effects of different iron dose and ini-
tial culture pH on the harvesting efficiencies of MWW grown Sce-
nedesmus sp. are shown in Fig. 2a. Maximum harvesting efficiency
was 98.87% when the culture pH was adjusted to 7.0, before adding
144 mg/L ferric chloride; however, when the pH was adjusted to
6.0 harvesting efficiency of 93.2% and 95.6% were achieved for
48 mg/L and 72 mg/L ferric chloride dose. Although the initial pH
of the MWW grown culture was lesser than BG-11 media grown
Fig. 2a. Effect of pH on the harvesting efficiencies of MWW-grown Scenedesmus sp.
culture, MWW grown culture required either lower initial pH
(<6.0) or higher coagulant dose (96 mg/L ferric chloride for pH
6.5 adjusted MMW culture) for similar harvesting efficiency. Ionic
strength in the MWW grown culture would have been higher com-
pared to freshwater grown culture and this could be the reason for
higher coagulant requirement for the MWW grown culture. For the
same reason, the final pH value of the supernatant was slightly
higher for same ferric chloride dose and initial culture pH (Fig. 2b).

3.3. Comparison with high and low pH induced coagulation-

flocculation

As the pH of Scenedesmus sp. culture was reduced to 3.75 by

adding 0.157 g/L H2SO4, there was spontaneous sedimentation of
the biomass and corresponding harvesting efficiency was 93.7%
(see Fig. 3, Supplementary S5). However, it took more than one

Fig. 2b. Effect of iron dosage on the final pH on the MWW-grown Scenedesmus sp.
cultures.

hour for the biomass settlement. Furthermore, as the pH of the

Fig. 1b. Effect of iron dosage on the final pH of Scenedesmus sp. cultures, grown in
BG-11 media.
original culture was adjusted to 3.0 by adding 0.192 g/L of H2SO4,
the harvesting efficiency was 99.5% and it took less than 30 min
for complete settlement. On the contrary, when the culture pH
was increased to 11.5 by adding 0.28 g/L NaOH, harvesting effi-
ciency was 70.6%. For a ferric chloride dose of 0.096 g/L, the har-
vesting efficiency was 91.4% and biomass settlement took place
under 10 min. Furthermore, after separating the biomass the
supernatant had a pH of 6.37 which was closer to the pH value
of the freshwater. Similarly, when 0.058 g/L H2SO4 and 0.072 g/L
of ferric chloride were added in the same culture, the biomass har-
vesting efficiency was 97.2% and the pH of the supernatant was
3.83. It was also demonstrated that microalgae biomass formed
spontaneous flocs when the culture pH was reduced to 4 by adding
828 P. Das et al. / Bioresource Technology 216 (2016) 824–829

was also shown in s 3.2.1 and 3.2.2 that lowering the pH of the cul-
ture reduced the coagulant requirement. As the extracted solution
was acidic, no reduction of culture pH was necessary for the subse-
quent biomass harvesting. As the total chemical requirement
would be lower, cost of producing microalgae biomass would also
be lower.

3.5. Reusability of the growth medium

Recycling of culture media has the potential to reduce the bio-

mass production cost. Growth of Scenedesmus sp. was not affected
up to two times of culture medium recycling (see Fig. 4). No flocks
of Scenedesmus sp. biomass were visible during the medium recy-
cling experiments. Iron was added at the first batch, however as
there was residual iron in the recycled medium, iron was not
required in the two subsequent growth trials. Total iron concentra-
Fig. 3. Comparison of harvesting efficiencies for different flocculation methods tions in 2nd and 3rd batches of Scenedesmus sp. cultures were
applied for Scenedesmus sp. culture, grown in BG-11 media. 107.8 ± 19.2 lg/L and 129.7 ± 13.9 lg/L respectively. Therefore,
Scenedesmus sp. not only grew in the supernatant, but also could
utilize the residual iron in the supernatant.
HNO3, or HCl (Liu et al., 2013; Pezzolesi et al., 2015). However, the
harvesting efficiencies were poor when the biomass concentration
3.6. Large scale biomass harvesting study
was below 1 g/L. Similarly, in some studies culture pHs were raised
above 11 to have higher harvesting efficiency (Beevi et al., 2016;
Initially, 7.2 g ferric chloride (2.48 g as iron) was added in 100 L
Castrillo et al., 2013; Knuckey et al., 2006). On the contrary, no floc-
Scenedesmus sp. culture (0.53 g/L biomass density, pH was adjusted
culation or settling was found for Chlorella zofingiensis cultures in
to 6.5) and the corresponding harvesting efficiency was 93.4%,
the pH range of 1.82–11.97 (Wyatt et al., 2012). Therefore, harvest-
which was slightly lower than optimum values of indoor experi-
ing of microalgae biomass using pH adjustment could be strain
ment. After decanting the supernatant, almost 5 L slurry of floccu-
specific (Rakesh et al., 2013). Additionally, adjustment of culture
lated biomass was collected; therefore, the initial biomass density
pH to extreme level may require excess amount of chemicals as
was concentrated almost 20 times using this single step. This
found in this study; similar amount of chemicals would be neces-
slurry was then centrifuged by a bench top centrifuge machine
sary to neutralize the supernatant. Hence, biomass harvesting by
and 222.6 g (22.2% solid content) biomass paste was obtained.
modulating the culture pH alone could be expensive
200 mL, 1 M H2SO4 extraction solution could extract 2.06 g of iron
(Chatsungnoen and Chisti, 2016).
which was 83% of initial added iron. The weight of the biomass
paste, after removing the iron extraction solution by centrifuga-
tion, was 215.4 g which could be due to the loss of some metabo-
3.4. Extraction of iron from the biomass and reuse as coagulant
lites and the associated iron during the iron recovery process. Later
1.22 g of ferric chloride was added in the extraction solution and
Optimum pH (6.5) and ferric chloride dose (72 mg/L) were used
the mixture was used to harvest another batch of 100 L culture.
for this experiment. The supernatant of this experiment had an
Harvesting efficiency in the second batch was 92.5% which was
iron concentration of 0.95 ± 0.02 mg/L and therefore, 96.18% of
very close to the value – obtained in the first batch. The fraction
the added iron was associated with the settled biomass. The con-
of iron that couldn’t be extracted from the harvested biomass
centration of iron in the supernatant was very close to the iron
and the other fraction remained in the supernatant can be consid-
concentration in BG-11 growth media. It was found that lower
ered as loss in each batch of harvesting; however, it was observed
the pH of the extraction solution, higher was the iron extraction
in the small scale study that Scenedesmus sp. could utilize the
efficiency (see Supplementary Fig. S6). Recovery of iron was 9.2%
or lower for any extraction solution-having a pH value 2.0 or
higher. Maximum amount of iron was recovered (86.4 ± 1.2% of
the added iron) for the pH 1.0 extraction solution; 9.8% of the
added iron couldn’t be extracted which remained with the bio-
mass. By increasing the volume of the extracting solution and ade-
quate mixing, iron recovery could have been increased.
Considering the overall iron loss (as dissolved in supernatant
and attached in the biomass) of 13.6% in the extracted solution,
iron was supplemented in it (as ferric chloride) so that the com-
bined solution could provide 72 mg/L iron concentration in the
fresh culture. Harvesting efficiencies were 94.3 ± 0.1% and
93.7 ± 0.3% for fresh iron solution and combined iron solution,
respectively. Although metal based coagulants are cheaper and
allow less energy intensive harvesting, these are known to contam-
inate the biomass in various applications. Additionally, recovery
and recycling of the coagulants were found to be problematic
(Milledge and Heaven, 2012). In this study, it was shown that a
major fraction of the associated iron in the biomass was extracta-
ble using a simple acid solution. Only 4.9 mg iron remained Fig. 4. Growth comparison of Scenedesmus sp. in fresh BG-11 media and recycled
absorbed in 1 g of Scenedesmus sp. biomass (dry weight basis). It culture media.
 P. Das et al. / Bioresource Technology 216 (2016) 824–829
residual iron in the supernatant. Therefore, the net requirement of
ferric chloride, as coagulant, in the subsequent harvesting would
be very low.
4. Conclusion
Low initial culture pH can reduce the coagulant dose for effi-
cient harvesting. Acidic extraction solution can be used to extract
a major fraction of the iron from the harvested biomass. Being
acidic in nature, iron extracted solution can improve the harvesting
efficiency and reduce the coagulant requirement in subsequent
harvesting. Microalgae can be grown in the supernatant and no
iron is needed as microalgae can utilize the residual iron in the
supernatant. Recovery of the iron from the settled microalgae bio-
mass and reuse it to harvest successive batch of microalgae culture
would improve the sustainability of the microalgae based
products.
Acknowledgements
This work was supported by Qatar Science and Technology Park
(QSTP) of Qatar Foundation (QF), Qatar Airways (QA), and Qatar
University (QU).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.biortech.2016.06.
014.
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