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Suitability of Soxhlet Extraction to Quantify Microalgal Fatty Acids as Determined by Comparison with In Situ Transesterification

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Lipids

Abstract

To assess Soxhlet extraction as a method for quantifying fatty acids (FA) of microalgae, crude lipid, FA content from Soxhlet extracts and FA content from in situ transesterification (ISTE) were compared. In most cases, gravimetric lipid content was considerably greater (up to sevenfold) than the FA content of the crude lipid extract. FA content from Soxhlet lipid extraction and ISTE were similar in 12/18 samples, whereas in 6/18 samples, total FA content from Soxhlet extraction was less than the ISTE procedure. Re-extraction of residual biomass from Soxhlet extraction with ISTE liberated a quantity of FA equivalent to this discrepancy. Employing acid hydrolysis before Soxhlet extraction yielded FA content roughly equivalent to ISTE, indicating that acidic conditions of ISTE are responsible for this observed greater recovery of FA. While crude lipid derived from Soxhlet extraction was not a useful proxy for FA content for the species tested, it is effective in most strains at extracting total saponifiable lipid. Lipid class analysis showed the source of FA was primarily polar lipids in most samples (12/18 lipid extracts contained <5% TAG), even in cases where total FA content was high (>15%). This investigation confirms the usefulness of ISTE, reveals limitations of gravimetric methods for projecting biodiesel potential of microalgae, and reinforces the need for intelligent screening using both FA and lipid class analysis.

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Abbreviations

FA:

Fatty acid

FAME:

Fatty acid methyl ester

TE:

Transesterification

ISTE:

In-situ transesterification

R-ISTE:

Residual ISTE

TE-AH:

TE preceded by acid hydrolysis

PUFA:

Polyunsaturated fatty acid

GC:

Gas chromatography

TAG:

Triacylglycerol

HPLC-CAD:

High performance liquid chromatography-charged aerosol detection

LC-MS:

Liquid chromatography/mass spectrometry

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Acknowledgments

Many people contributed to this project, either by providing biomass or assisting with lipid analysis. We would like to thank (in alphabetical order): Mather Carscallen, Katie Dickinson, Laura Garrison, Jenny MacPherson, Scott MacQuarrie, Ron Melanson, Stephen O’Leary, Kyoung Park and Crystal Whitney. This work was funded by the National Bioproducts Program in Microalgal Biofuels. This is NRC Publication No. 54055.

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Author notes

  1. Jesse McNichol

    Present address: MIT/Woods Hole Oceanographic Institution Joint Program in Biological Oceanography, 266 Woods Hole Rd, MS# 52, Woods Hole, MA, 02543-1050, USA

Authors and Affiliations

  1. Institute for Marine Biosciences, National Research Council of Canada, Halifax, NS, B3H 3Z1, Canada

    Jesse McNichol, Karen M. MacDougall, Jeremy E. Melanson & Patrick J. McGinn

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  1. Jesse McNichol
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  2. Karen M. MacDougall
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Correspondence to Jesse McNichol.

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McNichol, J., MacDougall, K.M., Melanson, J.E. et al. Suitability of Soxhlet Extraction to Quantify Microalgal Fatty Acids as Determined by Comparison with In Situ Transesterification. Lipids 47, 195–207 (2012). https://doi.org/10.1007/s11745-011-3624-3

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