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Unravelling the matrix effect of fresh sampled cells for in vivo unbiased FTIR determination of the absolute concentration of total lipid content of microalgae

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Abstract

Over the past years, the substitution of the classical biochemical quantification techniques by Fourier transform infrared (FTIR) spectroscopy has been widely studied on microalgae because of its tremendous application potential for bioprocess monitoring. In the present work, mandatory aspects that have never been approached by FTIR end-users working onto fresh biomass were assessed. We demonstrated first that fresh cells’ FTIR spectra main characteristics could be severely and unspecifically altered when the properties of the sampled biomass were not monitored. Microscopy indicated that important cell reorganization could occur when diminishing the cells density of the sample. Molecular probing approach suggested that such a modification could provoke an alteration of the hydrogen-bonding network of the sample. The sample heterogeneity was found to impact also the shape and intensity of the recorded FTIR bands, participating then to a matrix effect uncharacterized until now. In the second part of our study, we selected FTIR spectra not influenced by this matrix effect and the corresponding accurate calibration data obtained by the whole cell analytical procedure to elaborate an optimized total lipid quantification PLS-R model. Results demonstrated that our strategy could provide a small volume sampling (1 mL of fresh culture), rapid (within minutes), robust (physiological condition independent), and accurate (as accurate as the reference method could be) FTIR absolute quantification method to determine the fresh microalgae intracellular total lipid content. To validate our unbiased FTIR approach, a photobioprocess monitoring pipeline was developed and allowed assessing the effect of light attenuation on total lipid production by the marine microalga Nannochloropsis oculata.

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Abbreviations

FA:

Fatty acids

FAME:

Fatty acid methyl ester

FTIR:

Fourier transform infrared spectroscopy

GC-FID:

Gas chromatography coupled to flam ionization detector

HTSXT:

High throughput screening eXTension

PBR:

Photobioreactor

PLS-R:

Partial least square regression

TL:

Total lipid

TL-FA:

Total lipid fatty acids

RMSE:

Root mean square error

RMSECV:

Root mean square error of cross validation

RMSEP:

Root mean square error of prediction for the external validation

RSD:

Relative standard deviation

WCA:

Whole cell analytic

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Acknowledgments

Part of this work has been funded by the French National Research Agency project DIESALG (ANR-12-BIME-0001) and the CAER project (alternative fuel for aeronautics).

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Authors and Affiliations

  1. Université de Nantes, GEPEA UMR CNRS 6144, Bât. CRTT, 37 bd de l’Université, BP 406, 44602, Saint-Nazaire Cedex, France

    Rémy Coat, Valeria Montalescot, Esteban Serrano León, Delphine Kucma, Candice Perrier, Sébastien Jubeau, Gérald Thouand, Jack Legrand, Jérémy Pruvost & Olivier Gonçalves

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  1. Rémy Coat
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  2. Valeria Montalescot
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  3. Esteban Serrano León
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  9. Jérémy Pruvost
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  10. Olivier Gonçalves
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Correspondence to Olivier Gonçalves.

Additional information

R. Coat and V. Montalescot contributed equally to this work.

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Coat, R., Montalescot, V., León, E.S. et al. Unravelling the matrix effect of fresh sampled cells for in vivo unbiased FTIR determination of the absolute concentration of total lipid content of microalgae. Bioprocess Biosyst Eng 37, 2175–2187 (2014). https://doi.org/10.1007/s00449-014-1194-5

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  • DOI: https://doi.org/10.1007/s00449-014-1194-5

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