Analytical tools for evaluating algal biodiesel production collin beal - may 2010

Analytical Tools for Evaluating Algal Biodiesel Production

Beal C.M., Webber M.E., Ruoff R.S., Hebner R.E., Romanovicz D., and Connelly R.

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Colin M BealASME 2010

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Algal biodiesel production can be represented in three steps

Beal C.M. et al., “A Framework to Report the Production of Biodiesel from Algae,” BioEnergy Research, In Review.


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Algal biodiesel production can be represented in three steps

Three commonly studied pathways are:1) Transesterification of Algal Lipids2) Thermochemical Conversion of Algal Biomass3) Excretion of Oils by Genetically Modified Organisms


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Transesterification of Algal Lipids

Beal et al., “A Framework to Report the Production of Biodiesel from Algae,” BioEnergyResearch, In Review.


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The Center for Electromechanics at UT-Austin has a multi-disciplinary team

S. BriggsE. ChenR. ConnellyJ. ChoiM. FountainR. Hebner

J. JonesL. KatzK. Kinney M. MontoyaB. MorrisonR. Pearsall

M. PoenieM. RandazzoD. SchmidF. SeibertJ. Van ReetM. Werst

I would like to acknowledge:

I would like to thank OpenAlgae for funding:


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Algae are grown in flasks, reactors, tanks, and ponds

UT-CEM, 2009-2010


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Then, the algae are concentrated by ~100XMultiple harvesting methods are

being evaluated:1) Centrifugation2) pH induced floc/defloc3) Dissolved Air Flotation4) Electro-flocculation5) Filtration6) Resin collection7) Electro-wicking

UT-CEM, 2009-2010


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• Electromechanical pulsing is a cost effective technology developed at UT-CEM

After harvesting, the cells are compromised with electromechanical pulsing

UT-CEM, 2010UT-CEM, 2010


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Before After

• Electromechanical pulsing is a cost effective technology developed at UT-CEM

After harvesting, the cells are compromised with electromechanical pulsing

Beal C.M., and Romanovicz D., 2008UT-CEM, 2010


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An enhanced coalescence membrane is used to extract the algal lipids

UT-CEM 2009, 2010


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The lipids are then upgraded to fuel

Algal lipids are converted to fatty acid methyl esters, the primary component in biodiesel, by transesterification.


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Evaluating the intermediate products requires many analytical tools


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Evaluating the intermediate products requires many analytical tools


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Generally, microscopy provides structural information.

1) Light Microscopy


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1) Light Microscopy

Fluorescence microscopy using Nile Red dyeBeal C.M., 2008


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1) Light Microscopy2) Scanning Electron Microscopy

- Useful for evaluating electromechanical lysing

Beal C.M. and Romanovicz D., 2008, 2010

Before


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1) Light Microscopy2) Scanning Electron Microscopy

- Useful for evaluating electromechanical lysing

Beal C.M. and Romanovicz D., 2008, 2010

After


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1) Light Microscopy2) Scanning Electron Microscopy3) Transmission Electron Microscopy

Beal C.M. and Romanovicz D., 2008

Electrically PulsedControl

chloroplasts

LB

C

G

LB

CW

CW

Control: Compact, well-ordered structures.

Pulsed: Disordered, initiating apoptosis

Chloroplasts (C), Golgi (G), Cell walls (CW) , Lipid Bodies (LB)


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Usually, spectroscopy provides composition analysis

1) UV-Vis Spectrophotometry- Measure of chlorophyll absorption (a.u.)- Absorption corresponds to algal density (g/L)- Algal productivity calculated from changes in algal density

Bidwell 1979

Grown Mass Productivity


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1) UV-Vis Spectrophotometry2) Raman Spectroscopy

Huang et al., “Micro-Raman Spectroscopy of Algae: Composition Analysis and Fluorescence Background Behavior,” Biotechnology and Bioengineering, 2010.

Algal Spectra(Nitrogen Starved Chlorella and Neochloris)


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1) UV-Vis Spectrophotometry2) Raman Spectroscopy

Huang et al., “Micro-Raman Spectroscopy of Algae: Composition Analysis and Fluorescence Background Behavior,” Biotechnology and Bioengineering, 2010.


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1) UV-Vis Spectrophotometry2) Raman Spectroscopy3) Infrared Spectroscopy (IR)

IR has been used to:1) Analyze nutrient status in algae 2) Identify algal species

- Some groups are using it for lipid analysis, but it requires dry samples.


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1) UV-Vis Spectrophotometry2) Raman Spectroscopy3) Infrared Spectroscopy (IR)4) Nuclear Magnetic Resonance Spectroscopy (NMR)

Beal et al., “Lipid Analysis of Neochloris oleoabundansby Liquid State NMR,” Biotechnology and Bioengineering, 2010.


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Chromatography also provides composition analysis, but requires extraction

1) High Performance (Pressure) Liquid Chromatography (HPLC)

Signal (a.u.)

Time(min)

Signal (a.u.)

Time(min)

Healthy Starved

Beal C.M., 2009Acquired by Poenie M.


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1) High Performance (Pressure) Liquid Chromatography (HPLC)2) Thin Layer Chromatography (TLC)

Connelly R., 2009

Polarity

Lipid StandardsProcessing Samples


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1) High Performance (Pressure) Liquid Chromatography (HPLC)2) Thin Layer Chromatography (TLC)3) Gas Chromatography – Mass Spectrometry (GC-MS)

- Provides molecular information for volatile samples.

- Generally limited to biodiesel, or FAME derived from algae

MS fragmentation patter for FAME

CB062309b #673-677 RT: 6.85-6.87 AV:5 NL: 4.02E6T: {0,0} + c EI det=450.00 Full ms [ 50.00-400.00]

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400m/z

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90

95

100

Rel

ativ

e A

bund

ance

55.167.0

74.0

83.187.0

97.1

98.1

110.1

123.1

264.2125.1

137.1265.2222.2

180.2138.1141.0

152.1

166.1221.1207.0

223.2 296.3193.1 266.3246.2 281.0 297.3 341.0 355.0327.0 383.0 397.0

MS fragmentation patter for FAME

RelativeAbundance(a.u.)

m/z


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Summary: Tricks (or tools) of the trade


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Questions?

Colin M Beal, MSGraduate Student

Thermal/Fluids Systems DivisionMechanical Engineering

The University of Texas at [email protected]

Webber Energy GroupRuoff Nanoscience and Technology Group

andThe Center for Electromechanics

Analytical tools for evaluating algal biodiesel production collin beal - may 2010

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