Resources Conservation in Microalgae Biodiesel Production: Kelsey Price, Ihab H. Farag
Resources Conservation in Microalgae Biodiesel Production: Kelsey Price, Ihab H. Farag
Resources Conservation in Microalgae Biodiesel Production: Kelsey Price, Ihab H. Farag
Abstract— The aim of this investigation was to reduce cost
and production time of biodiesel from microalgae by conserving Table I. Heating Value and CO2 emissions per gallon of
water, energy and chemical resources. This was done by Petroleum diesel and Biodiesel
investigating two goals. The first is conservation of fresh water
by microalgae growth in treated wastewater using reduced
chemical nutrients while taking advantage of the nitrates and Property Petroleum Biodiesel
phosphates in wastewater. The second is conservation of energy, diesel
while reducing production time and eliminating the use of Heating value, Btu per gallon. 130,500 128,000
hazardous hexane solvent by integrating the two steps of CO2 emissions, lb per gallon. 26.55 5.85
microalgae oil extraction and transesterification of extracted oil
into a one-step in-situ process. To accomplish the first goal, the
project included growth, monitoring, and harvesting Chlorella Biodiesel produces virtually the same heating value as
vulgaris microalgae in various reduced chemical nutrient petroleum diesel. However, biodiesel produces about a fifth
solutions and analyzing the algae growth and the lipid of the carbon emissions per gallon
production results. The added chemical nitrate and phosphate Figure 1 shows the process diagram of producing biodiesel
nutrients to the growth medium were reduced from the from algae. The steps include algae growth, harvesting,
“standard” chemical recipe by a 25% step. The second goal was
accomplished using a sonicator to disrupt algae cells. The in-situ dewatering, oil extraction, and transesterification of algae oil
process was used to obtain the biodiesel production from the dry to fatty acid methyl ester (FAME) or biodiesel. However,
harvested algae. The results were promising, a reduced nutrient there are still technical challenges that have limited the
supply, specifically the trials of 50% phosphates and 50% commercial production of microalgae biodiesel. These
nitrates resulted in a higher lipid production than the include 1- Water Requirements, which range from 300 to
“standard” full nutrients requirement.
2000 gallons of water per gallon of biodiesel [1 - 10]; 2- Lipid
extraction is energy intensive [11], uses hazardous hexane
Index Terms— Biodiesel, Chlorella vulgaris, municipal
wastewater, in-situ process, alternative renewable fuel, Fresh solvent and is the most costly step in the algae growth process;
water conservation, Chemical use reduction. 3- The high cost of algae nutrient chemicals [12].
I. INTRODUCTION
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Resources Conservation in Microalgae Biodiesel Production
division slows due to the lack of resources necessary for bubbled into the algae medium to provide the CO2 needed
growth; 5- Death/ Lysis: Cells begin to die due to lack of for photosynthesis and to mix the solution.
resources. Figure 2 shows these five phases. 4- pH: Most algae species grow well in a pH range
between 7 and 9. The optimum pH range is usually between
8.2-8.7.
6- Temperature: Most microalgae species grow in the
temperature range of 16 C (61 F) and 27 C (81 F). Below
16 C (61 F), the growth will slow down growth.
Temperatures higher than 35 C (95 F) may harm the algae.
The experimental work in the present investigation was
done at room temperature of about 21 C (70 F).
Table II. “Standard” Nutrients used in the Present Work II. PROJECT PURPOSE AND GOALS
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International Journal of Engineering and Technical Research (IJETR)
ISSN: 2321-0869, Volume-1, Issue-8, October 2013
III. METHODOLOGY displays different peaks. These peaks are used to identify the
content and amount of the main lipids. The GC injection is
A. Experiment Layout done using a needle to gather 3 microliters of (lipid +
The experimental work to study the effect of reducing chloroform) solution, this is inserted into the machine. The
nitrates and phosphates chemical nutrients on algae growth GC then runs for roughly 20 minutes and analyzes the
and lipid production (Goal 1) was done in 4 phases. The aim solution. Because each needle’s cylinder is very tiny, they can
of phases 1 and 2 was to establish baseline data on the be easily clogged. When this were to happen, the following
performance/growth of microalgae in fresh water and in procedure was conducted; about 10 mL of solvent (methanol
wastewater. Phases 3 and 4 were to decrease the or chloroform) is added to 200-300 mL of fresh (RO) water in
concentrations of nitrates and phosphates in wastewater. a beaker (approximate, no exact numbers). Then, the syringe
Algae growth was done in 2L flasks supplied with air and is separated into parts and placed in the solution. The solution
illuminated with fluorescent lighting, as shown in Figure 3. is sonicated for 10 minutes. Then all of the pieces are dried
using paper towel. It was important to be careful and allow
time before opening the sonicator because the chloroform
odor could be strong. The GC was first tested by running pure
biodiesel (B100) sample. The B100 GC results were
comparable to those of [21]. Table III summarizes the
methodology used in this investigation.
Phase Description
1- Obtain The trials of growth were done in 2L flasks
baseline supplied with air and illuminated with
data fluorescent lighting for consistency. There
were six flasks per baseline run of varying
Figure 3. Algae growth in 2 L flasks supplied with air and conditions. These 6 flasks included
illuminated with fluorescent lighting. Two Flasks with RO (fresh) water and
1- An initial nutrient and algae supply,
In-situ (Goal 2) approach combines the two-step procedure 2- No initial nutrient or algae supply
of lipid extraction from algae and transesterification of Four Flasks with wastewater and
extracted algae lipid/oil to FAME into one-step. The 3- an initial nutrient and algae supply
experimental work starts by dissolving 1g of dry algae in 40 4- just an initial algae supply
mL of KOH in methanol. This solution is then placed in a cell 5- just an initial nutrient supply
disrupter/ Sonicator, shown in Figure 4. 6- no initial nutrient or algae supply
2- Repeat Repeat Phase 1 runs to ensure accuracy.
3- Reduced Six flasks per experimental run of varying
nutrients conditions. The added nitrate and phosphate
experiments levels were adjusted in increments of 25%
in less nutrients from the recipe of Table II,
wastewater i.e.,
0.4mM phosphates varying nitrates
1- 75% nitrates (3.9 mM)
2- 50% nitrates (2.6 mM),
3- 25% nitrates (1.3 mM)
5.2 nM nitrates varying phosphates
4- 75% phosphate (0.3 mM)
5- 50% phosphate (0.2 mM)
6- 25% phosphate (0.1 mM)
4- Repeat Repeat Phase 3 runs to ensure accuracy.
Figure 4. Model W-375 Sonicator (cell disrupter) 5- Algae Dissolve dry algae in a solution of KOH in
Sonication methanol. Place the solution in a sonicator
The sonicator runs for 10 minutes and breaks the algae to disrupt the algae cells and release the oil.
cells. It was set to a power of 7-8 on the output dial and the The methanol transesterifies the released
algae oil into FAME/biodiesel. Remove
power meter less than 75% in continuous mode.
algae by filtration. Evaporate methanol.
The disrupted algae are filtered from the (methanol+ algae
6- Gas Dissolve the FAMEs in chloroform. Inject
oil) solution using a vacuum flask. The methanol
Chromato- the dissolved solution into the GC. Analyze
transesterifies the algae oil into FAME/biodiesel. The excess
graph (GC) the peaks. Use the areas under the peaks to
methanol is evaporated using a hot water bath. The remaining
Analysis determine the FAME production.
algae oil is then dissolved in chloroform to be injected into the
gas chromatograph (GC) to analyze the lipid content. The GC
is equipped with an integrator, which plots the data and
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Resources Conservation in Microalgae Biodiesel Production
B. Analytical procedures: pH and Nitrates Table IV. Measured Variables and Metrics Calculated.
It is important to measure the pH, nitrite and nitrate levels
to maintain a consistent growth environment. These readings Measurement Purpose/ metrics
were taken every other day. The procedure was to collect 5 Daily Algae Solution Algae growth, from
mL of each algae solution in a test tube then use Mardel 5 test Absorbance (using absorbance/turbidity.
strips. Spectrophotometer)
pH, nitrite and nitrate Nutrient concentration
C. Analytical procedures: Algae Growth Monitoring levels with a Mardel 5 in 1 and if there is nutrient
Algae growth was monitored daily by measuring the test strip. depletion or starvation
absorptivity at 680 nm using a spectrophotometer. These
readings were important in determining when the algae Algae mass after Algae production
reached its stationary phase and needed to be harvested. harvesting and drying (g dry algae/L)
The algae growth runs varied in duration, ranging roughly (using a balance)
from nine days to two weeks. Once the algae was harvested, Mass of algae Oil after Algae oil yield (g
they were dewatered, centrifuged, and freeze-dried. The final extraction (using a gas oil/100 g dry algae)
chromatographer)
algae powder mass of each run was measured using a digital
Algae biodiesel production Algae FAME/biodiesel
scale. This would allow the determination of the algae mass
from the one-step in-situ production
production per initial volume of algae solution for each run.
process (g FAME/L)
D. Algae Harvesting, Dewatering and Drying
Algae harvesting is the process of separating algae from
its growth medium. The challenge is that the algae are in a IV. RESULTS/DISCUSSION/ACCOMPLISHMENTS
dilute solution. The high water content of algae must be
removed to enable harvesting. Several processes have been A. Algae Growth and Monitoring Results
developed to harvest microalgae. These include using Figure 5 shows the absorbance/turbidity of the algae
microscreens, centrifugation, flocculation, and broth growth solution measured over the growth period. The data
filtration. High-speed centrifugation (5000 rpm for 10 covered several “growth solution” cases to establish the
minutes) was used in the present work [5], [7], [12] and [21] confidence in the data. These cases include fresh (RO) water,
because it did not require the addition of flocculants, and for RO water with algae and nutrients, wastewater with algae,
the ease of separating the solid from the liquid solution. Algae wastewater with nutrient, and wastewater with algae and
drying was accomplished by freeze-drying for approximately nutrients. For fresh (RO) water with no nutrient and no algae,
48 hours. This produced dry algae flakes, which were crushed the absorbance remains at zero, as expected.
into powder using a mortar and pestle.
E. Calculation of Oil yield from GC Measurements
The GC Integrator generated graphs (chromatograms) of
each mixture injected into it. The graphs contained peaks that
corresponded to each algae biodiesel fatty acid in the injected
mixture. A series of standard FAME cocktails were prepared,
injected into the GC and the results were analyzed [21]. The
GC calculations of the in-situ biodiesel determined the ratios
(area of produced peaks to specified standard) [21]. This was
done after matching the in-situ biodiesel peaks to the
appropriate FAME standard. The results the composition,
concentration, and mg of FAME per gram of algae.
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International Journal of Engineering and Technical Research (IJETR)
ISSN: 2321-0869, Volume-1, Issue-8, October 2013
Figure 8 indicates that when the algae grows the algae
growth media reaches pH of 8.4. This is within the
optimum algae growth pH range of 8.2-8.7.
C. Algae Mass Production
Figure 9 displays the algae mass production in grams per
liter of growth medium under different conditions. A key is
provided below the figure to correspond the number labels
of the x-axis to its growth conditions (WW abbreviating
wastewater and RO abbreviating reverse osmosis fresh
water). The green bars are for growth solutions with 100%
of chemical nutrients nitrates and phosphates. The blue
bars are for growth solutions with reduced chemical
nutrients (25%, 50% and 75%) nitrates and phosphates.
The average algae production of the six blue lines is 0.751
Fig. 6. Nitrates concentration (ppm) variation with culture g/L. The algae production with reduced nutrients are within
age (days) in different growth media. 25% of the average algae production rate of all runs with
reduced chemical nutrients.
Similarly, Figures 7 and 8 show the variation in nitrites
concentration (ppm) and pH of the nutrient medium in
different growth medium.
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Resources Conservation in Microalgae Biodiesel Production
A B C D E F G H I
1 RO Yes 100 100 1.59 21.82 34.8 40.5
2 RO No No No 0 0 0 0
3 WW Yes 100 100 0.76 23.0 17.5 20.3
4 WW No 100 100 1.35 21.97 29.6 34.4
5 WW Yes No No 0.21 7.065 1.48 1.72
6 WW No No No 0.19 7.528 1.45 1.68
7 WW Yes 25 100 0.79 21.31 16.9 19.6
8 WW Yes 50 100 0.58 30.46 17.6 20.5
9 WW Yes 75 100 0.77 12.46 9.53 11.1
10 WW Yes 100 25 0.75 29.63 22.2 25.8 1- RO, nutrients, algae, 2- RO, 3- WW, nutrients, algae,
11 WW Yes 100 50 0.89 42.59 37.9 44.0 4- WW, nutrients, 5-WW, algae, 6- WW, 7- 25% nitrates,
12 WW Yes 100 75 0.73 27.15 19.9 23.2
8- 50% nitrates, 9- 75% nitrates, 10- 25% phosphates,
11- 50% phosphates, 12- 75% phosphates.
Column Headings:
A Trial Number
Fig. 11. Algae Oil production (microL of FAME/L of
B Water Source (RO or WW)
growth medium) of the base run trials (green bars) compared
C Algae present (Yes or No)
to the experimental trials (blue bars).
D Nitrates, % of “standard” nitrates feed
E Phosphates, % of “standard” phosphates feed
Figures 10 and 11 indicate that almost all of the
F Algae Production (g dry algae/L of initial solution)
experimental trials in wastewater with reduced nitrates or
G Algae Oil concentration (mg FAMEs/g algae)
phosphates chemical nutrients (Runs 7 – 12 with blue bars)
H Algae Oil Production (mg FAMEs per L of initial
had higher lipid production per g of algae and per L of initial
medium solution)
growth solution than the base trials of both RO water and
I Algae Oil Production (microL FAMEs per L of initial
wastewater with 100% of the “standard” nitrates and
medium solution), or L FAME/million liters on initial
phosphates chemical nutrients (the green bars in Figure 10
solution.
and 11). In both cases, for the phosphates and nitrates, the
trials with 50% of the original chemical nutrient had the
Figure 10 compares the oil/lipid production (mg of
higher FAME production. The highest FAMES production
FAME/g of algae) for the different growth conditions. It
was obtained with 50% phosphates, Trial 11. This trial show
shows how the different conditions affect lipid/oil production
lipid production of 42.59 (mg FAMEs/g algae), 37.9 (mg
FAMEs per L of initial medium solution), 44 microL FAMEs
per L of initial medium solution). Nonetheless, all of the cases
of reduced nutrient solutions look promising.
E. Production time and Energy use in two-stage vs.
one-stage in-situ Biodiesel/FAMES Production
One of the main advantages of the in-situ process is the
reduction time in biodiesel production. In the two-step
method, the microalgae oil extraction would take 2 hours
alone, plus evaporation and oven requirement time, roughly
another 40 minutes. The extracted oil still needs to be
traansestrified to biodiesel. Each in-situ would only take 10
minutes plus roughly 20 minutes to run through the gas
1- RO, nutrients, algae, 2- RO, 3- WW, nutrients, algae, chromatograph. In addition, the in-situ process avoids the use
4- WW, nutrients, 5-WW, algae, 6- WW, 7- 25% nitrates, of a large amount of water, which is required by the flask
8- 50% nitrates, 9- 75% nitrates, 10- 25% phosphates, condenser during lipid extractions. In addition, the in-situ
11- 50% phosphates, 12- 75% phosphates. process does not require the use of a hot water bath and oven,
which are both energy intensive. Finally, in-situ avoids the use
Fig. 10. Algae Oil production (mg of FAME/g of algae) of of a hazardous solvent, hexane, which is required for lipid
the base run trials (green bars) compared to the experimental extractions from the dry harvested algae.
trials (blue bars).
54 www.erpublication.org
International Journal of Engineering and Technical Research (IJETR)
ISSN: 2321-0869, Volume-1, Issue-8, October 2013
growth medium. The base data were obtained for both growth [12] J. Ferrentino, “Microalgae oil extraction and in situ
transesterification,” M.S. Thesis, Chemical Engineering Dept.,
medium with standard chemical nutrients of Table II. University of New Hampshire (UNH) (I. Farag, advisor). 2007.
[13] S. Kanes, “The Choice of Next-Generation Biofuels,” Equity Research
This investigation indicates that 50% of the nitrate and Industry Report, Scotia Capital, 2009.
phosphate chemical nutrients in the growth medium are [14] "2.3. Algal Production." FAO Corporate Document Repository.
Fisheries and Aquaculture Department”, Wed. 23 Sept. 2012.,
conserved and the biodiesel production is increased when http://www.fao.org/docrep/003/W3732E/w3732e06.htm
microalgae are grown in wastewater. [15] D. Eltringham, M. Elmoraghy, and I. Farag, “Algae Growth in
Photobioreactors using a Removable Jacket of Colored LEDs,” 16th
The in-situ process was also successful in reducing Annual Energy & Environment Conference & Expo (EUEC 2013), Jan
biodiesel production time, conserving energy and water. It 28 – 30, 2013, Phoenix Convention Center, Phoenix, AZ.
[16] D. Eltringham, and I. Farag "Greener Algae Growth in
also eliminated the use of hexane, a hazardous solvent used in Photobioreactors using LEDs to Maximize Lipid Production,” the 17th
the extraction of oil from dry microalgae. Annual Green Chemistry & Engineering (GC&E) Sustainable
Chemistry & Engineering for the 21st Century, June 18-20, 2013
Washington, DC.
[17] L. Xin, G. Ke,, S. Ying-xue, “Effects of different nitrogen and
phosphorus concentrations on the growth, nutrient uptake, and lipid
ACKNOWLEDGMENT accumulation of a freshwater microalga Scenedesmus sp.”,
Bioresource Technology 101: 5494–5500, 2010.
The authors would like to thanks to Dr. Leland Jahnke, UNH [18] EPA (U.S. Environmental Protection Agency). 2008. Clean
Plant Biology Dept. for algae input; Dr. Nancy Whitehouse, Watersheds Needs Survey 2004.
UNH Dairy Research Center, for help with large-scale [19] K. Price, M. Elmoraghy, and I. Farag, "Energy and Water Resource
Efficient Production of Microalgae Lipids,” 16th Annual Energy &
centrifugation and freeze-drying; John Newell and Marian Environment Conference & Expo (EUEC 2013), Jan 28 – 30, 2013,
Elmoraghy for technical assistance and help in conducting Phoenix Convention Center, Phoenix, AZ. 2013.
experiments and operating the GC. [20] S. Tewfik, S. Hawash, N. Atteya, G. El Diwani, and I. Farag.
“Techno-Economic Appraisal of Biodiesel from Jatropha curcas: An
Egyptian case study,” Journal of Agricultural Science and Technology
(JAST) B 2 (2012) 287-297. 2012.
[21] M. Elmoraghy (2013), “Production of Bio-Jet Fuel from Microalgae,”
REFERENCES M.S. Thesis, Chemical Engineering Dept., University of New
Hampshire (UNH) (I. Farag, Mentor). 2013.
[1] G. Chaput, K. Charmanski,, and I. Farag, "Sustainable Production of
Microalgae Oil Feedstock Using Municipal Wastewater and CO2
Fertilization”, International Journal of Engineering Science and
Technology (IJEST), Vol. 4, No. 7, July 2012, p 3489-3499, ISSN:
0975-5462. 2012.
[2] M. Elmoraghy, and I Farag, “Bio-jet Fuel from Microalgae: Reducing
Kelsey Price Junior working for a Bachelor's of Science in Chemical
Water and Energy Requirements for Algae Growth,” International
Engineering with a minor in Applied Mathematics. She intends to pursue a
Journal of Engineering and Sciences, IJES, ISSN: 2278-4721, Vol. 1,
career with a focus on Chemical Engineering to benefit the environment.
Issue 2 (Sept 2012), PP 22-30. 2012, www.researchinventy.com.
Kelsey has two years of research experience working on the production of
[3] M. Elmoraghy, T. Webster, and I Farag,“ Microalgae Lipid Triggering
bio-fuels from microalgae, with a focus on microalgae biodiesel. She
by Cooling Stressing,” Journal of Energy and Power Engineering,
received 2012 and 2013 Summer Undergraduate Research Fellowships from
Volume 6, Number 12, December 2012 (Serial Number 61),
the University of New Hampshire Hamel Research Center. Her research
pp1918-1924 (ISSN: 1934-8983). 2012.
focused on conserving resources to make the process more energy efficient
[4] B. McConnell, and I. Farag, “Offshore Microalgae Growth in
and environmentally friendly.
Wastewater,” International Review of Chemical Engineering
(I.RE.CH.E), ISSN 2035-1755, Vol. 4, N. 4, pp 423-429, July 2012.
[5] N. Mulumba, “Production of Biodiesel from Microalgae,” M.S. Thesis,
Chemical Engineering Dept., University of New Hampshire (UNH) (I.
Farag, advisor). 2010
[6] N. Mulumba, and I. Farag,” Tubular Photobioreactor for Microalgae
Biodiesel Production,” International Journal of Engineering Science and
Technology (IJEST), Vol. 4 No.02 February 2012, pp 703-709. 2012
(ISSN: 0975-5462). http://www.ijest.info/docs/IJEST12-04-02-135.pdf
[7] T. Webster, “Temperature Stressing (Lipid Triggering) of Salt Water
Algae for Enhanced Lipid Production,” B.S. Honors Thesis, Chemical
Engineering Dept., University of New Hampshire (UNH) (I Farag,
advisor). 2009.
[8] R. Wilson, and I Farag, “Parametric Study of Biodiesel Quality and Yield
Using a Bench-Top Processor,” International Journal of Oil, Gas and
Coal Technology 2012 - Vol. 5, No.1 pp. 92 - 105, 2012
[9] R. Wilson, G. Salama, and I., Farag. "Microalgae Growth in Qatar for
CO2 Capture and Biodiesel Feedstock Production,” Global Journal of
Researches in Engineering (GJRE), Chemical Engineering, Volume 12
Issue 1 Version 1.0 Year 2012, Online ISSN: 2249-4596 & Print ISSN: Ihab H. Farag Professor Emeritus, Chemical Engineering at University
0975-5861. 2012. of New Hampshire. In Sept 1976, he joined the faculty at UNH after
[10] Z. Zuka, B. McConnell, and I. Farag, "Comparison of Freshwater and receiving his doctorate of Science (Sc.D.) from the Massachusetts Institute
Wastewater Medium for Microalgae Growth and Oil Production,” of Technology (MIT) in Chemical Engineering. His research has focused on
Journal of American Science 2012; 8(2):392-398. (ISSN: 1545-1003). biofuels, microalgae biodiesel (renewable alternative to petro-diesel liquid
http://www.jofamericanscience.org/journals/am-sci/am0802/055_815 fuel), bio-jet fuels, cleaner production, energy efficiency, and chemical risk
0am0802_392_398.pdf screening applications. He initiated the Bio-Oil Group, the Biodiesel
[11] B. McConnell, “Kinetics of microalgae oil extraction,” B.S. Honors Research Group and the New Hampshire Pollution Prevention Internship
Thesis, Chemical Engineering Dept., University of New Hampshire Program to involve and mentor interdisciplinary students in research and
(UNH) (I. Farag, advisor). 2013. industrial projects. Invited presentations and briefings were given to the
legislators on Pollution Prevention, Biomass, Biodiesel and Bio-Oil. He has
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