Second and Third Generation Biofuels
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1 Second and Third Generation Biofuels Krish Jayachandran Professor & Graduate Director Dept of Earth and Environment Florida International University Miami, Florida 8/20/2012 1
2 Why Biofuels?? Improve energy security. High oil price. Mitigate climate change. 8/20/2012 2
3 What is Biodiesel?
4 Biodiesel From Transesterification
5 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.
6 Individual Energy Consumption
7 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
8 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
9 U.S. Production of Biodiesel 1,690 Liters = 446.5Gallons 2007 Energy Independence and Security Act mandates: 500 Million Gallons in Billion Gallons in 2012 In Billion gallons of diesel were sold.
10 Sources of Production
11 Edible Seed Oils
12 Characteristics of Edible Seed Oils and Non Edible Seed Oils
13 U.S. Pricing of Conventional Diesel Compared to Biodiesel Brent Crude Price 4/1/11= USD/bbl
14 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?
15 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.
16 Decortication
17 Expeller
18 Expeller
19 Chemical Extraction
20 Transesterification Transeserification Glycerin Percent Esters Batch Oil (ml) Methanol + Catalyst (ml) Biodiesel (ml) Unreacted Material (ml) Time (min) Recovered from Oil AVG AVG 60MIN AVG 75MIN
21 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?
22 Fuel Properties Properties SG B100 SG Oil Jatropha B100 Castor Rubber Seed ASTM D EN Acid Value (mg KOH/g) <0.8 <0.50 Viscosity at 40C (mm2/s) Calorification (MJ/kg) - 32,143 39,230 39,500 36,500 Saponification Density Cloud Point (Celsius)- 18 Room Temperature Report Pour Point (Celsius) Report Iodine Number Flash Point (Celsius) >130 >120 Ash Content (%) <0.02 <0.02 Properties Palm Soybean Diesel Acid Value (mg KOH/g) Viscosity at 40C (mm2/s) Calorification (MJ/kg) - 39,760 Saponification- Density- Cloud Point (Celsius)- 15 Pour Point (Celsius) Iodine Number- Flash Point (Celsius) Ash Content (%)
23 Fuel Properties for Blends
24 Data for Engine Trials
25 Third Generation Biofuels Algal fuel or Oilgae 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/
26 Source: 8/20/
27 Disadvantages: Not economically feasible. Major issues with harvesting and labor costs. Contamination issues. 8/20/
28 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/
29 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/
30 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, Chlorococcum EV 5-1, 45-3, 55-2, 55-5 Coelenstrum EV 46-4, Coccoid Green EV 56-5, 56-4, 81-7, 103-6, Stirgeoclonium EV 64-8 Dactylococcus EV Pediastrum EV 81-6, 104-6, Prochloro EV 104-1a 8/20/ Kirchneriella EV 104-7
31 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/
32 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/
33 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/
34 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 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/
35 Results for Screening: 8/20/
36 Standard curve : Triolein Y= e x Y represents fluorescence intensity. Unknown x is the lipid concentration in the algal cells. 8/20/
37 Strain Lipid concentration on13th day (ug) Lipid concentration on 45 th day (ug) 81-6 Pediastrum Pediastrum Coelastrum Chlorella EV 29 Chlamydomonas Coccoid green Selanstrum Chlorococcum Chlorococcum Dactylococcus Coelastrum Scenedesmus Chlorococcum /20/
38 64-12 Coccoid Green 66-1 Scenedesmus Chlorococcum Pediastrum Chlorella Scenedesmus a Prochloro Coccoid Green Scenedesmus Stigeoclonium Coccoid Green Coccoid green Kircherniella Botryococcus braunii (Control) /20/
39 Results: 46 4 Coelastrum,64 12 Coccoid green, 64 8 Stirgeoclonium, Dactylococcus and 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/
40 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) Coccoid green 46-4 Coelastrum Coelastrum 64-8 Stigeoclonium Dactylococcus Botyrococcus braunii Strain 8/20/
41 Assessment of environmental factors on lipid accumulation : 8/20/
42 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 coccoid green over a 45 day cultivation period Control Cultivation time (Days) Biomass (ug/ml chlorophyl) Fluorscence intensity Biomass Biomass and cellular accumulation of neutral lipid in Dactylococcus over a 45 day cultivation period Fluorscence intensity mormalized with chlorophyll ControlD0 D5 D10D15D20D25D30D35D40D45 Cultivation time (days) Biomass (ug/ml chlorophyll) Fluorscence intensity 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 Control Cultivation time(days) Biomass ( ug/ml chlororphyll) 64-8 Fluorscence intensity 64-8 Biomass Fluorscent intensity normalized with chlorophyll 8/20/ Control Cultivation time (Days) Biomass (ug/ml chlorophyll) 46-4 Fluorescence intensity 46-4 Biomass
43 Results continued.. Biomass and cellular accumualtion of neutral lipid for 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 Control Biom ass (ug/ m l chlorophyll) Fluorscence intensity Biomass Fluorscence intensity norm alized with chlorophyll Control Biom ass (ug/m l chlorophyll) B.braunii Fluor intensity B.b Biomass Cultivation time(days) Cultivation time(days) 8/20/
44 Results As the biomass concentration decreases the amount of lipid concentration increases coccoid green has the maximum amount of lipid. Botryococcus braunii showed a decrease in lipid content as the biomass concentration increases. 8/20/
45 Effect of nitrogen depletion on neutral lipid accumulation 8/20/
46 Results contd 0%N 50%N 100%N 64-8 Stirgeoclonium responded to depletion of nitrogen by showing the maximum concentration of lipid Dactylococcus and Botyrococcus braunii did not seem to respond to Nitrogen depleted conditions. 8/20/
47 Effect of Phosphorous depletion 8/20/
48 108-5 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 Control D0 D5 D10 Time(Days) 0%P 50%P 100%P Fluorscence intensity Control D0 D5 D10 Time(Days) 0%P 50%P 100%P Coccid green showed a maximum fluorescence intensity over a 10 day period Dactylococcus showed no response. 8/20/
49 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
50 Acknowledgements Ms. Priyanka Narendar Dr. Miroslav Gantar Dr. Kateel Shetty Mr. Andrew Jungman USDA NIFA Hispanic Serving Institutions Grant Thank you 8/20/
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