Harvesting marine algae for biodiesel feedstock

Dinh Trinh Thanh Xuan

Department of Environmental Engineering, National University of Singapore

Singapore 117576

ABSTRACT
Harvesting is one of the key processes that determine the feasibility of algal biodiesel
production. This paper proposes a cost-effective system to harvest Nannochloropsis sp., using
ferric chloride for coagulation and air sparging for flocculation. For an influent concentration of
0.5g algal dry biomass per liter, the system can achieve up to 90% removal efficiency. Optimum
conditions are ferric chloride dosage of 0.18 mg/L and air sparging at 2 L/min for 2 min,
followed by unagitated settling for 10 min.

1. Introduction
The compromise between harvesting efficiency and cost is a critical problem in algal
biodiesel production. Evidently, poor harvesting process not only is a waste of manufacturing
material, but also poses a threat to the environment as high algal concentration in effluent may
cause eutrophication. However, due to the small size of micro-algae (2-30μm) and its dilute
concentration in race-way ponds (approximately 0.5-1.0g dry biomass per liter), effective
harvesting methods can be very costly. The process is estimated to contribute up to 20-30% of
the total cost, and thus, harvesting optimization has been emphasized as one of the key factors
determining the feasibility of algal biodiesel development in the future (Sheehan et al, 1988).
There are a number of possible methods for harvesting algae, including centrifugation,
filtration, electro-flocculation and coagulation. Centrifugation seems to be the most efficient yet
is too costly and therefore not suitable for mass biomass production. Similarly, filtration is not a
practical solution because of the formation of a filter-cake, which substantially increases head
loss and requires frequent maintenance.
Electro-flocculation, on the other hand, has been proven to effectively remove up to 95%
of algae in fresh water (Poleman et al, 1997). Nevertheless, the efficiency in harvesting marine
algae has not been tested. One reason is the high normality of seawater, which would compete
for positive charges from an electrode source, thus increase the current level required to
destabilize algae to form flocs. Furthermore, there is a possibility of cell oxidation that leads to
undesirable changes in lipid profile and final product quality.
In comparison with all the above methods, coagulation has many advantages. Unlike
centrifugation and filtration, coagulation is a practical technique that has been widely used for
algal removal in wastewater treatment and some industrial mass manufacture (Moraine et al,
1980). The process is relatively fast, with a reasonable cost and high efficiency. Besides, since
coagulation uses additional cations such as Fe3+ and Al3+ to destabilize algae for floc formation,
the problem of cell oxidation is minor. Therefore, with regard to algal biodiesel production,
coagulation should be the most appropriate method for harvesting.
This paper reviews our experience in using coagulation to harvest algae from laboratory
race-way ponds. The work focuses on three main targets: dosage optimization, types of
mechanical mixing and the effects of time period on flocculation and sedimentation.

2. Materials and Methods

Algal growth conditions
The marine algae selected for study was Nannochloropsis sp. isolated from Singapore
seawater. Firstly, algae were cultured in 10 liter bottles with Guillard F/2-Si medium for 15
days. After that, they were diluted and transferred into an indoor race-way pond where glycerol
was added as fixed carbon source. On photoperiod, continuous illumination was provided by
fluorescent light tubes at 6000 lux. After 1 week growing in the race-way pond, the culture
reached stationary phase and was collected for experimentation.
Calibration curve of cell density vs optical density
Optical density (OD) was used to represent the algal concentration in solution before and
after the experiments. Hence, the very first step was to construct a calibration curve of cell
density vs OD. Cell numbers were counted using a hemecytometer and OD was measured at
wavelength 680 nm by a Hitachi 2900 UV-visible spectrophotometer.
Flocculation experiments
There were two types of mechanical mixing studied: paddle-driven flocculation and air
sparging. The conventional paddle flocculation was provided by a lab-scale Stuart Scientific
SW1 Flocculator, with the blades immersed to 4 cm from the bottom of 500 ml jars. The
procedures followed Sukenik’s tests4: after coagulant addition, solutions were stirred at 80 rpm
for 2 min, then at 20 rpm for 30 min, and then left unagitated for 30 min for settling.
Air sparging was produced by letting the air supplied from a HP air pump go through an
air stone (diameter 3cm and height 2cm) to form micro bubbles. Between the pump and the air
stone connected a TSI 4000 flow meter to monitor air flows. There were various tests with
different agitating and settling times conducted to specify air sparging.
Coagulation experiments
Dosage optimization was tested using two inorganic coagulants, i.e. ferric chloride and
aluminum sulfate. Concentrations of ferric chloride and aluminum sulfate used in stock solution
were 0.0600 mg/ml and 0.0565 mg/ml, respectively. Different amounts of chemicals were added
in after ionic strength and pH had been adjusted to 0.7 and 5.0, respectively. After a settling
period, samples were taken specifically at the 200 ml position for OD measurement.

3. Results and discussion

Calibration curve of cell density vs optical density

Cell density
(million/ml)

Optical density

Fig 1: Calibration curve of cell density vs optical density

 The calibration curve shows a strong linear relationship between cell density and optical
density (R2 = 0.9932). Hence, the efficiency of coagulation tests can be directly calculated from
the OD data as follows.
finalOD
Efficiency = 1 - Eq 1
initialOD
It should be noted that the calibration curve is only valid for OD between 0.075 and
1.675. For other values beyond this range, cell density may not be directly proportional to OD,
thus equation 1 will not be applicable.
The calibration curve may change from time to time during the algal lifetime. In
different growth phases, variation in algal biological activity may lead to changes in chlorophyll
concentration inside the cells. Therefore, even though cell numbers may be relatively constant,
there would be dissimilar OD results because the OD measurement was based on absorption of
chlorophyll by visible light. In this study, the calibration curve was constructed on the same day
as coagulation experiments in order to minimize any errors when estimating cell density from
OD data.
Effect of coagulant on harvesting efficiency
The study focuses on two coagulants: ferric chloride and aluminum sulfate (alum),
which are most common in use for coagulation in water treatment. After different amounts of
coagulants were added, all samples were agitated using the same air sparge rate of 2 ml/min for
2 min. The results are shown in Fig 2 and Fig 3 below.

Harvesting
efficiency
(%)

Time (minutes)

Fig 2: Effect of ferric chloride concentration on harvesting efficiency

 Harvesting
efficiency
(%)

Time (minutes)

Fig 3: Effect of Alum concentration on harvesting efficiency

Comparing the two figures, it is obvious that ferric chloride is more efficient for
harvesting Nannochloropsis sp. The optimum dosage of ferric c hloride, 3ml/L (equivalent to
0.18 mg/L) gives 90% efficiency after 10 min settling, while the optimum dosage of alum 2ml/L
only gives 55% efficiency in the same conditions. Therefore, in all experiments following on
effects of air sparging duration and types of mechanical mixing on harvesting efficiency, only
ferric chloride was used.
The two figures also show that after 2 min settling, the change in harvesting efficiency
with respect to time was relatively small where the difference between the harvesting efficiency
at 2 min and 10 min is only 1% for ferric chloride, and 5% for alum. Hence, if ferric chloride is
used in practical algal biodiesel production, the settling duration can be reduced to 2 min to
maximize harvesting capacity.
Effect of air sparging duration on harvesting efficiency
To study the effect of agitating duration on harvesting efficiency, experiments were
conducted with various air sparge times of 1 min, 2 min, 3 min and 4 min. All samples were
pretreated with the same amount of coagulant, i.e. 3ml ferric chloride per liter solution. After
mixing, samples were left to settle for 5 min before being collected for OD measurement. The
harvesting efficiency obtained for each sparge time is demonstrated in Fig 4 below.
 Harvesting
efficiency
(%)

Sparing time (minutes)

Fig 4: Effect of air sparging time on harvesting efficiency

Figure 4 shows that the optimum air sparging duration for harvesting Nannochloropsis
sp. by ferric chloride coagulation is 2 min, with an efficiency of 89%. Lower than 2 min will be
too short for algae to form flocs. However, a longer mixing time up to 3-4 min may destabilize
the flocs or result in too much micro bubble exposure which tends to bring the flocs up to the
surface instead of settling; hence, decreasing the overall harvesting efficiency.
Effect of air sparging flow rate on harvesting efficiency

Harvesting
efficiency
(%)

Time (minutes)

Fig 5: Effect of air sparge flow rate on harvesting efficiency

The effect of air sparge flow rate on harvesting efficiency is demonstrated in Fig 5,
where the highest efficiency, approximately 90%, was obtained for an air flow rate of between
1.5 L/min to 2.5 L/min after 10 min settling. For air flow rates less than 1 L/min, the mixing was
too weak to provide effective chemical dispersion or suitable velocity gradients for floc
formation. In contrast, too high sparging flow rates leads to overexposure to bubbles that tend to
float algal flocs, thus negatively impacting on the consequent settling process. As shown in the
figure, using a 2.5 L/min flow rate required up to 6 min of settling to achieve 89% efficiency,
while it only took approximately 2 min using 1.5 L/min and 2 L/min flow rates to achieve the
same result. Furthermore, shorter settling time implies a higher harvesting capacity for biodiesel
production, and lower air sparging flow rates are also beneficial in terms of operational costs.
Effect of mechanical mixing types on harvesting efficiency
The study compared two types of mechanical mixing: conventional paddle-driven
flocculation and air sparge flocculation. Samples were pretreated with the same amount of
coagulation, i.e. 3 ml ferric chloride per liter solution. For paddle-driven flocculation, the
procedure followed Sukenik’s experiment as described in Materials and Methods section. Air
sparge flocculation was based on flow rate of 2 L/min for 2 min agitating, then a settling period
of 10 min before OD measurement.

From fig 6, it can be noted that the harvesting efficiencies obtained from paddle-driven
and air sparge flocculation were 85% and 89%, respectively. Although the difference in
effectiveness of the two mixing types is relatively small, the total retention time (agitating time
+ settling time) for paddle-driven flocculation was 62 min compared to 12 min for air sparging.
Hence, with regard to harvesting capacity, air sparging flocculation is more favorable.

Harvesting
efficiency
(%)

Mechanical types of mixing

Fig 6: Effect of mixing types on harvesting efficiency

 4. Conclusion
We have developed a cost-effective procedure to harvest Nannochloropsis sp. for algal
biodiesel production. The selected coagulant is ferric chloride with an optimum dosage of 18
mg/L algal solution. With an air sparge flow rate of 2 L/min flow rate for 2 min agitating and 10
min settling, the process achieved up to 90% efficiency with a total retention time of 12 min.

5. Acknowledgement
I would like to thank Dr Jeff Obbard for the opportunity to participate in UROP and Probir Das
for his guidance.

6. References
[1] Sheehan, J., Dunahay, T., Benemann, J. & Roessler, P (1988). A look back at the US
Department of Energy’s Aquatic Species Program – Biodiesel from Algae.
Hyperlink: http://www.nrel.gov/docs/legosti/fy98/24190.pdf
Retrieved on 8th November, 2008.
[2] Poleman, E., De Pauw, N. & Jeurissen, B. 1996. Potential of electrolytic flocculation for
recovery of micro-algae. Resources, Conservation and Recycling 19 (1997) 1-10.
[3] Moraine, R., Shelef, G., Sandbank, E., Bar-Moshe, Z. & Shvartzbard, L. Recovery of
sewage borne algae: flocculation and centrifugation techniques. In: Algae biomass, G. Shelef
and C. J. Soeder (eds). Elsevier/North Holland, Amsterdam, 1980, pp. 531-46.
[4] Sukenik, A., Bilanovic, D. & Shelef G. 1988. Flocculation of microalgae in brackish and sea
waters. Biomass 15, pp. 187-199.