Second and Third Generation Biofuels Krish Jayachandran Professor & Graduate Director Dept of Earth and Environment Florida International University Miami, Florida 8/20/2012 1
Why Biofuels?? Improve energy security. High oil price. Mitigate climate change. 8/20/2012 2
What is Biodiesel?
Biodiesel From Transesterification
Transesterification Transesterification is the stepwise interchangeable organic reaction of a triglyceride with an alcohol to create glycerol and esters. (R is a fatty acid, R is the length of the acyl acceptor, and R is the rest of the triglyercide molecule) The reason for transesterification is that raw oils have high viscosity and low volatility, which cause problem for engines. Through this chemical process the oil is broken down into its derivatives that have more similar properties to that of conventional diesel fuel.
Individual Energy Consumption
U.S. Energy Distribution and Projections for Next 25 years History 2009 Projections U.S. primary energy consumption quadrillion Btu per year 7% Renewables (excluding liquid biofuels) 10% 21% 21% Coal 25% 1% 37% Natural gas Liquid biofuels Oil and other liquid fuels 24% 3% 33% 9% Nuclear 8% 7 Source: EIA, Annual Energy Outlook 2011
U.S. Liquid Fuel Consumption Per Day History U.S. liquid fuels consumption million barrels per day 2009 Projections 4% Biofuels including imports 10% 12% Natural gas plant liquids 13% 2% 34% Liquids from coal Petroleum supply 31% 52% Net petroleum imports 42% 8 Source: EIA, Annual Energy Outlook 2011
U.S. Production of Biodiesel 1,690 Liters = 446.5Gallons 2007 Energy Independence and Security Act mandates: 500 Million Gallons in 2009 1 Billion Gallons in 2012 In 2007 63 Billion gallons of diesel were sold.
Sources of Production
Edible Seed Oils
Characteristics of Edible Seed Oils and Non Edible Seed Oils
U.S. Pricing of Conventional Diesel Compared to Biodiesel Brent Crude Price 4/1/11= 125.46USD/bbl
Research Questions How efficient is the production process of Simarouba biodiesel? What are the processes involved biodiesel production system? How efficient is each process of the production system? Can the production system serve as a model for other second generation biofuels?
Production Efficiency Production Process Decortication: Separating seed kernel from seed husk. Drying: Removing some of the moisture from seed kernels. Expelling: Removing oil from seed kernels using screw press. Transesterification: Chemical process of separating glycerides and esters.
Decortication
Expeller
Expeller
Chemical Extraction
Transesterification Transeserification Glycerin Percent Esters Batch Oil (ml) Methanol + Catalyst (ml) Biodiesel (ml) Unreacted Material (ml) Time (min) Recovered from Oil 1 1000 250 901 305 60 90.1 2 1000 250 898 300 60 89.8 3 1000 250 900 298 60 90 4 1000 250 910 295 60 91 5 1000 250 911 290 60 91.1 6 1000 250 905 302 75 90.5 7 1000 250 895 310 75 89.5 8 1000 250 901 300 75 90.1 9 1000 250 901 312 75 90.1 AVG 1000 250 902.4444444 301.3333333 90.24444444 AVG 60MIN 1000 250 904 297.6 90.4 AVG 75MIN 1000 250 900.5 306 90.05
Research Questions How does Simarouba oil compare to conventional fuels? What are the fuel properties of Simarouba oil and its Biodiesel? Acid Value, Viscosity, Calorification, Saponification, Density, Cloud Point, Pour Point, Iodine Number, Flash Point, and Ash Content. How does the overall fuel efficiency of Simarouba biodiesel perform compared to conventional diesel in electric power generator when mixed with conventional diesel at varying percentages?
Fuel Properties Properties SG B100 SG Oil Jatropha B100 Castor Rubber Seed ASTM D 6751-02 EN 14214 Acid Value (mg KOH/g) - 0.24905 2.2465 0.4 0.118 <0.8 <0.50 Viscosity at 40C (mm2/s)- 12.609 60.535 4.8 5.81 1.9-6.0 3.5-5.0 Calorification (MJ/kg) - 32,143 39,230 39,500 36,500 Saponification- 179.561 185.9317 Density- 867 860-900 Cloud Point (Celsius)- 18 Room Temperature -12 4 Report Pour Point (Celsius)- 15 2-32 Report Iodine Number- 56.03 54.28 Flash Point (Celsius)- 160 135 260 130 >130 >120 Ash Content (%)- 0.00485 0.012 0.02 <0.02 <0.02 Properties Palm Soybean Diesel Acid Value (mg KOH/g) - 0.08 Viscosity at 40C (mm2/s)- 4.42 4.08 2.6 Calorification (MJ/kg) - 39,760 Saponification- Density- Cloud Point (Celsius)- 15 Pour Point (Celsius)- 15-20 Iodine Number- Flash Point (Celsius)- 182 69 68 Ash Content (%)- 0.02 0.01
Fuel Properties for Blends
Data for Engine Trials
Third Generation Biofuels Algal fuel or Oilgae. 30 100 times more oil per acre than corn and soybean. No sulfur, non toxic. Grown in marginal land. Biodegradable. Less water consumption. Carbon sequestration. Tolerate brackish and saline waters. 8/20/2012 25
Source: http://www.dailymarkets.com/stocks/2008/07/02/investing-inalgae-biofuel/ 8/20/2012 26
Disadvantages: Not economically feasible. Major issues with harvesting and labor costs. Contamination issues. 8/20/2012 27
Objectives: To screen various strains of native green algal strains from the Florida Everglades to identify those with potential for biodiesel production. Quantify the lipid content in algal cells using gravimetric method. Assess the effect of environmental conditions on accumulation of cell lipid. 8/20/2012 28
Algae Algae are eukaryotes having chlorophyll and other pigments for carrying out oxygen producing photosynthesis. Lipid of interest: Neutral lipids ( in the form of Triacyl Glycerol) best substrate for producing biodiesel. 8/20/2012 29
Materials and Methods Organisms: 31 algal strains from the FIU culture collection (Dr. M. Gantar) are being studied. Botyrococcus braunii reference strain Genus Strain Chlamydomonas EV 29 Chlorella EV 2-4,71-4 Selanstrum EV 2-7,34-4 Scenedesmus EV 3-11, 66-1, 79-1, 80-15, 81-5, 103-4 Chlorococcum EV 5-1, 45-3, 55-2, 55-5 Coelenstrum EV 46-4, 108-5 Coccoid Green EV 56-5, 56-4, 81-7, 103-6, 64-12 Stirgeoclonium EV 64-8 Dactylococcus EV 64-10 Pediastrum EV 81-6, 104-6, 108-4 Prochloro EV 104-1a 8/20/2012 30 Kirchneriella EV 104-7
Culture conditions: Algal biomass will be produced by growing algal strains in 3 l flasks in BG11 medium (Rippka et al. 1979) under cool white light (30µ E m 2 sec 1 ) at 27 C with aeration with sterile air. 8/20/2012 31
Screening of lipid: Nile Red Fluorescence technique (Greenspan et al.,1985). a lipophilic dye. Spectroflurometer analysis at excitation and emission. wavelengths of 530 nm and 575 nm. Calibration Curve Lipid standard Triolein. 8/20/2012 32
Quantification of Lipid (Johnson & Wen, 2009): Freeze dried algal biomass (1g) Solvent Chloroform methanol water system. Solvent evaporated using nitrogen gas Mass of lipid estimated gravimetrically (Bligh &Dyer, 1959). 8/20/2012 33
Assessment of the effect of environmental factors: Biomass and Lipid accumulation: 1. Determined over a 45 day period. 2. Biomass concentration (Dere et al., 1997) Ca = 15.65A 666 7.340 A 653 (Methanol) 3. Lipid accumulation: Nile red method Nitrogen Depletion and Lipid accumulation (Widjaja et al., 2008). 1. Cells washed thoroughly with N free medium before transferred to fresh media. 2. Concentrations:0%,50% and 100%. Phosphorous Depletion and Lipid accumulation (Rodolfi et al., 2008). 1. Cells washed thoroughly with P free medium before transferred to fresh media. 2. Concentrations: 0%,50% and 100%. 3. Lipid accumulation: Nile red. 8/20/2012 34
Results for Screening: 8/20/2012 35
Standard curve : Triolein Y= 47.206e 0.1148x Y represents fluorescence intensity. Unknown x is the lipid concentration in the algal cells. 8/20/2012 36
Strain Lipid concentration on13th day (ug) Lipid concentration on 45 th day (ug) 81-6 Pediastrum 10.4374154819987 15.2742197328845 108-4 Pediastrum 11.5478990293633 14.7902409291716 108-5 Coelastrum 10.8782524317431 16.8931598545643 2-4 Chlorella 10.3231310670479 10.7881125153986 EV 29 Chlamydomonas 10.5579447163199 11.8504042799696 56-5 Coccoid green 9.89767174415345 14.214319533937 34-4 Selanstrum 10.4375505490238 14.2959393185065 55-2 Chlorococcum 9.79642908926664 12.1779334529304 55-5 Chlorococcum 10.1043498095117 16.1912263161184 64-10 Dactylococcus 9.66323740781956 20.7521647457002 46-4 Coelastrum 10.2788334785514 32.7072335595228 103-4 Scenedesmus 9.92390643824308 13.5432801267255 45-3 Chlorococcum 10.2058177595784 15.2267692289687 8/20/2012 37
64-12 Coccoid Green 66-1 Scenedesmus 10.3235521636551 28.8980659619241 10.7890128247306 11.9264850756913 5-1 Chlorococcum 11.1657055972044 16.0240790059667 80-15 Pediastrum 9.71383311489515 11.5957867692921 71-4 Chlorella 9.622160922159 12.1040128421204 81-5 Scenedesmus 9.72632286620263 11.1127898854546 104-1a Prochloro 9.77726455585745 11.4201424820779 81-7 Coccoid Green 9.88243685284034 12.8819237441846 79-1 Scenedesmus 9.74505749316385 16.0508912015053 64-8 Stigeoclonium 10.2839646211756 18.5071770989367 56-4 Coccoid Green 10.1848625273624 11.3956883053848 103-6 Coccoid green 11.0580814551602 12.8410195017856 104-7 Kircherniella 9.97844779925473 12.4077787987834 Botryococcus braunii (Control) 24.033823691502 16.5273695160237 8/20/2012 38
Results: 46 4 Coelastrum,64 12 Coccoid green, 64 8 Stirgeoclonium, 64 10 Dactylococcus and 108 5 Coelastrum showed higher amount of lipid accumulation on the 45 th day. Due to depletion of nutrients like phosphorous and nitrogen. Botryococcus showed a decrease in lipid content. Due to decrease in intensity of light that can alter the lipid composition when high cell densities are reached. 8/20/2012 39
Neutral lipid extraction by gravimetric technique: Neutral lipid extraction using gravimetric technique 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% Series1 Neutral lipid (% dry weight) 64-12 Coccoid green 46-4 Coelastrum 108-5 Coelastrum 64-8 Stigeoclonium 64-10 Dactylococcus Botyrococcus braunii Strain 8/20/2012 40
Assessment of environmental factors on lipid accumulation : 8/20/2012 41
Biomass and Cellular accumulation of neutral lipid over a 45 day period. Fluorescence intensity normalized with chlorophyll Biomass and cellular accumulation of neutral lipid in 64-12 coccoid green over a 45 day cultivation period 3500 3000 2500 2000 1500 1000 500 0 Control 0 5 10 15 20 25 30 35 40 45 Cultivation time (Days) 1.8 1.6 1.4 1.2 1 0.8 0.6 0.4 0.2 0 Biomass (ug/ml chlorophyl) 64-12 Fluorscence intensity 64-12 Biomass Biomass and cellular accumulation of neutral lipid in 64-10 Dactylococcus over a 45 day cultivation period Fluorscence intensity mormalized with chlorophyll 1600 1400 1200 1000 800 600 400 200 0 ControlD0 D5 D10D15D20D25D30D35D40D45 Cultivation time (days) 1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 Biomass (ug/ml chlorophyll) 64-10 Fluorscence intensity 64-10 Amount of Biomass Biomass and cellular accumulation of neutral lipid in 64-8 stirgeoclonium over a 45 day cultivation period Biomass and cellular accumulation of neutral lipid of 46-4 Ceolastrum over a 45 day cultivation period Fluorscence intensity normalized with chlorophyll 1400 1200 1000 800 600 400 200 0 Control0 5 10 15 20 25 30 35 40 45 Cultivation time(days) 1.4 1.2 1 0.8 0.6 0.4 0.2 0 Biomass ( ug/ml chlororphyll) 64-8 Fluorscence intensity 64-8 Biomass Fluorscent intensity normalized with chlorophyll 8/20/2012 42 1400 1200 1000 800 600 400 200 0 Control 0 5 10 15 20 25 30 35 40 45 Cultivation time (Days) 3.5 3 2.5 2 1.5 1 0.5 0 Biomass (ug/ml chlorophyll) 46-4 Fluorescence intensity 46-4 Biomass
Results continued.. Biomass and cellular accumualtion of neutral lipid for 108-5 Coelastrum over a 45 day period Biomass and cellular accumulation of neutral lipid of B.braunii over a 45 day period cultivation period Fluorscent intensity norm alized with chlorophyll 1600 1400 1200 1000 800 600 400 200 0 Control0 5 10 15 20 25 30 35 40 45 1.6 1.4 1.2 1 0.8 0.6 0.4 0.2 0 Biom ass (ug/ m l chlorophyll) 108-5 Fluorscence intensity 108-5 Biomass Fluorscence intensity norm alized with chlorophyll 3500 3000 2500 2000 1500 1000 500 0 Control0 5 10 15 20 25 30 35 40 45 2 1.8 1.6 1.4 1.2 1 0.8 0.6 0.4 0.2 0 Biom ass (ug/m l chlorophyll) B.braunii Fluor intensity B.b Biomass Cultivation time(days) Cultivation time(days) 8/20/2012 43
Results As the biomass concentration decreases the amount of lipid concentration increases. 64 12 coccoid green has the maximum amount of lipid. Botryococcus braunii showed a decrease in lipid content as the biomass concentration increases. 8/20/2012 44
Effect of nitrogen depletion on neutral lipid accumulation 8/20/2012 45
Results contd 0%N 50%N 100%N 64-8 Stirgeoclonium responded to depletion of nitrogen by showing the maximum concentration of lipid. 64-10 Dactylococcus and Botyrococcus braunii did not seem to respond to Nitrogen depleted conditions. 8/20/2012 46
Effect of Phosphorous depletion 8/20/2012 47
108-5 Coelastrum Results continued: 0%P 50%P 100%P Influence of phosphorous deprivation on lipid accumulation of 46-4 Coelastrum Influence of phosphorous deprivation on lipid accumulation of B.braunii Fluorescence intensity 2000 1500 1000 500 0 Control D0 D5 D10 Time(Days) 0%P 50%P 100%P Fluorscence intensity 2000 1500 1000 500 0 Control D0 D5 D10 Time(Days) 0%P 50%P 100%P 64-12 Coccid green showed a maximum fluorescence intensity over a 10 day period. 64-10 Dactylococcus showed no response. 8/20/2012 48
Limitations: Lag in algal taxonomic definition. Lipid contents and biomass and lipid productivities, not the only characteristics to ensure a cost effective and feasible biodiesel production. Contamination, tolerance to conditions like light, temperature, ionic strength, nutrient requirements, ease of harvesting and downstream processing impacts the success of 8/20/2012 large scale production. 49
Acknowledgements Ms. Priyanka Narendar Dr. Miroslav Gantar Dr. Kateel Shetty Mr. Andrew Jungman USDA NIFA Hispanic Serving Institutions Grant 2010 38422 21261 Thank you 8/20/2012 50