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SPME-GC×GC-TOF MS fingerprint of virally-infected cell culture: Sample preparation optimization and data processing evaluation.

Analytica chimica acta (2018-06-06)
Giorgia Purcaro, Pierre-Hugues Stefanuto, Flavio A Franchina, Marco Beccaria, Wendy F Wieland-Alter, Peter F Wright, Jane E Hill
ABSTRACT

Untargeted metabolomics study of volatile organic compounds produced by different cell cultures is a field that has gained increasing attention over the years. Solid-phase microextraction has been the sampling technique of choice for most of the applications mainly due to its simplicity to implement. However, a careful optimization of the analytical conditions is necessary to obtain the best performances, which are highly matrix-dependent. In this work, five different solid-phase microextraction fibers were compared for the analysis of the volatiles produced by cell culture infected with the human respiratory syncytial virus. A central composite design was applied to determine the best time-temperature combination to maximize the extraction efficiency and the salting-out effect was evaluated as well. The linearity of the optimized method, along with limits of detection and quantification and repeatability was assessed. Finally, the effect of i) different normalization techniques (i.e. z-score and probabilistic quotient normalization), ii) data transformation (i.e. in logarithmic scale), and iii) different feature selection algorithms (i.e. Fisher ratio and random forest) on the capability of discriminating between infected and not-infected cell culture was evaluated.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly(dimethylsiloxane), viscosity 1.0 cSt (25 °C)
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SPME Portable Field Sampler, coating Polydimethylsiloxane (PDMS)
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SPME fiber assortment kit 3
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SPME/HPLC Interface, rheodyne valve version
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SPME FFA PTFE Sealing Cups, size 23ga Beveled (No line)
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SPME fiber assembly Carboxen/Polydimethylsiloxane (CAR/PDMS), df 75 μm, for use with manual holder, needle size 24 ga
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SPME fiber assembly Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS), needle size 24 ga, for use with manual holder
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SPME fiber assembly polyacrylate (PA), df 85 μm, for use with manual holder, needle size 24 ga
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SPME Fiber Assembly Polydimethylsiloxane/Divinylbenzene (PDMS/DVB), df 60 μm, for use with autosampler/HPLC, needle size 24 ga, StableFlex fiber
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SPME fiber assortment kit 2, for use with manual holder, needle size 24 ga
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SPME fiber assortment kit 4, for use with manual holder, size 24 ga
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SPME fiber assortment kit 5, needle size 23 ga, for use with autosampler
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SPME-OC Fiber Assembly, Polydimethylsiloxane/Divinylbenzene (PDMS/DVB), df 75 μm (Coating thickness includes 65 μm coating + 10 μm OC (overcoating)), needle size 23 ga, pkg of 3 ea, fiber L 1 cm
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SPME FFA PTFE Sealing Cups, 24ga BLUNT, size 24ga blunt (2 Lines)
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SPME fiber assembly Carboxen/Polydimethylsiloxane (CAR/PDMS), df 75 μm, needle size 23 ga, for use with manual holder
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SPME fiber assembly Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS), needle size 24 ga, StableFlex, for use with manual holder/autosampler
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SPME fiber assembly polyacrylate (PA), df 85 μm, for use with autosampler, needle size 24 ga
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SPME fiber assembly Polydimethylsiloxane (PDMS), df 7 μm, needle size 24 ga, for use with autosampler
Supelco
SPME Fiber Assembly Polydimethylsiloxane/Divinylbenzene (PDMS/DVB), df 65 μm, needle size 24 ga, for use with manual holder

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