analytical method (either sample preparation or GC instrumentation). Results outside of upper or lower control limits,
or 3Â standard deviation (99% confidence limits), indicate loss
of control, and results of the run should be discarded. Two
consecutive analyses falling on one side of the mean between
the warning limits and the control limits also indicate loss of
control.
Acknowledgments
This work was supported by the US Department of Energy under
Contract No. DE-AC36-08GO28308 with the National Renewable Energy Laboratory. Funding provided by the US DOE Office
of Energy Efficiency and Renewable Energy Bioenergy Technologies Office. The US Government retains, and the publisher, by
accepting the article for publication, acknowledges that the US
Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this
work, or allow others to do so, for US Government purposes.
References
1. Laurens LML, Nagle NJ, Davis R et al (2015)
Acid-catalyzed algal biomass pretreatment for
integrated lipid and carbohydrate-based biofuels
production. Green Chem 17:1145–1158.
https://doi.org/10.1039/c4gc01612b
2. Dong T, Knoshaug EP, Davis R et al (2015)
Combined algal processing: a novel integrated
biorefinery process to produce algal biofuels
and bioproducts. Algal Res 19:216–323.
https://doi.org/10.1016/j.algal.2015.12.
021
3. Christie WW (2005) Lipid analysis: isolation,
separation, identification and structural analysis
of lipids, 3rd edn. American Oil Chemists Society, Champaign, IL
4. Iverson SJ, Lang SLC, Cooper MH (2001) Comparison of the Bligh and dyer and Folch methods
for total lipid determination in a broad range of
marine tissue. Lipids 36:1283–1287
5. Bligh EG, Dyer WJ (1959) A rapid method of
total lipid extraction and purification. Can J
Biochem Physiol 37:911–917
6. Harwood JL (1998) Membrane lipids in algae.
In: Siegenthaler P-A, Murata N (eds) Lipids
photosynth. Structure, function and genetics.
Springer, Dordrecht, The Netherlands, pp
53–64
7. Rainuzzo JR, Reitan KI, Olsen Y (1994) Effect
of short and long term lipid enrichment on
total lipids, lipid class and fatty acid composition in rotifiers. Aqauculture Int 2:19–32
8. Guschina IA, Harwood JL (2006) Lipids and
lipid metabolism in eukaryotic algae. Prog
Lipid Res 45:160–186. https://doi.org/10.
1016/j.plipres.2006.01.001
9. Laurens LML, Van Wychen S, McAllister JP
et al (2014) Strain, biochemistry, and
cultivation-dependent measurement variability
of algal biomass composition. Anal Biochem
452:86–95. https://doi.org/10.1016/j.ab.
2014.02.009
10. Lepage G, Roy CC (1986) Direct transesterification of all classes of lipids in a one-step reaction. J Lipid Res 27:114–120
11. Haas MJ, Wagner K (2011) Simplifying biodiesel production: the direct or in situ transesterification of algal biomass. Eur J Lipid Sci
Technol 113:1219–1229
12. Griffiths MJ, van Hille RP, Harrison STL
(2010) Selection of direct transesterification
Total Fatty Acid Content Determination of Whole Microalgal Biomass Using In. . .
213
or 3Â standard deviation (99% confidence limits), indicate loss
of control, and results of the run should be discarded. Two
consecutive analyses falling on one side of the mean between
the warning limits and the control limits also indicate loss of
control.
Acknowledgments
This work was supported by the US Department of Energy under
Contract No. DE-AC36-08GO28308 with the National Renewable Energy Laboratory. Funding provided by the US DOE Office
of Energy Efficiency and Renewable Energy Bioenergy Technologies Office. The US Government retains, and the publisher, by
accepting the article for publication, acknowledges that the US
Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this
work, or allow others to do so, for US Government purposes.
References
1. Laurens LML, Nagle NJ, Davis R et al (2015)
Acid-catalyzed algal biomass pretreatment for
integrated lipid and carbohydrate-based biofuels
production. Green Chem 17:1145–1158.
https://doi.org/10.1039/c4gc01612b
2. Dong T, Knoshaug EP, Davis R et al (2015)
Combined algal processing: a novel integrated
biorefinery process to produce algal biofuels
and bioproducts. Algal Res 19:216–323.
https://doi.org/10.1016/j.algal.2015.12.
021
3. Christie WW (2005) Lipid analysis: isolation,
separation, identification and structural analysis
of lipids, 3rd edn. American Oil Chemists Society, Champaign, IL
4. Iverson SJ, Lang SLC, Cooper MH (2001) Comparison of the Bligh and dyer and Folch methods
for total lipid determination in a broad range of
marine tissue. Lipids 36:1283–1287
5. Bligh EG, Dyer WJ (1959) A rapid method of
total lipid extraction and purification. Can J
Biochem Physiol 37:911–917
6. Harwood JL (1998) Membrane lipids in algae.
In: Siegenthaler P-A, Murata N (eds) Lipids
photosynth. Structure, function and genetics.
Springer, Dordrecht, The Netherlands, pp
53–64
7. Rainuzzo JR, Reitan KI, Olsen Y (1994) Effect
of short and long term lipid enrichment on
total lipids, lipid class and fatty acid composition in rotifiers. Aqauculture Int 2:19–32
8. Guschina IA, Harwood JL (2006) Lipids and
lipid metabolism in eukaryotic algae. Prog
Lipid Res 45:160–186. https://doi.org/10.
1016/j.plipres.2006.01.001
9. Laurens LML, Van Wychen S, McAllister JP
et al (2014) Strain, biochemistry, and
cultivation-dependent measurement variability
of algal biomass composition. Anal Biochem
452:86–95. https://doi.org/10.1016/j.ab.
2014.02.009
10. Lepage G, Roy CC (1986) Direct transesterification of all classes of lipids in a one-step reaction. J Lipid Res 27:114–120
11. Haas MJ, Wagner K (2011) Simplifying biodiesel production: the direct or in situ transesterification of algal biomass. Eur J Lipid Sci
Technol 113:1219–1229
12. Griffiths MJ, van Hille RP, Harrison STL
(2010) Selection of direct transesterification
Total Fatty Acid Content Determination of Whole Microalgal Biomass Using In. . .
213