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Liquid chromatography–mass spectrometry for metabolic footprinting of co-cultures of lactic and propionic acid bacteria

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Abstract

Co-cultures of specific lactic and propionic acid bacteria have been shown to have an antagonistic effect against yeast and moulds in dairy systems. In studies of these co-cultures by bioassay-guided fractionation and analysis, numerous compounds have been reported to inhibit yeast and moulds. Although active, the compounds do not account for the full effect observed. Instead, the inhibitory action in the co-culture is believed to be a result of synergy between known exo-metabolites, depletion of nutrients, and/or compounds not yet identified. Untargeted metabolomics or metabolic footprinting could be a potent approach to elucidation of the mechanism. The purpose of this review is to discuss the two pre-requisites for such a study—the compound classes expected in the co-cultures, and on the basis of these, the most suitable analytical technique(s). Ultrahigh-performance liquid chromatography (UPLC) coupled to high-resolution mass spectrometry (MS) via electrospray ionisation (ESI) operated in both positive and negative modes is regarded as the optimum instrumental technique. The applicability of a range of liquid chromatographic techniques ranging from ion-pair (IPC) and hydrophilic interaction (HILIC) to reversed-phase chromatography (RPC) is discussed in terms of the expected metabolome. Use of both HILIC and RPC is suggested, on account of the complementarity of these modes. The most promising strategy uses a combination of the two electrospray polarities and two modes of LC. The strategy recommended in this study does not include all compound classes, and suggestions for supplementary methods are listed.

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Abbreviations

ADP:

Adenosine diphosphate

ATP:

Adenosine triphosphate

APCI:

Atmospheric pressure chemical ionisation

CoA:

Coenzyme A

ESI:

Electrospray ionisation

FT-ICR:

Fourier-transform ion cyclotron resonance

FWHM:

Full width half maximum

HILIC:

Hydrophilic interaction liquid chromatography

HPLC:

High-performance liquid chromatography

GC–MS:

Gas chromatography–mass spectrometry

IPC:

Ion-pairing chromatography

IT:

Ion trap

LAB:

Lactic acid bacteria

LIT:

Linear ion trap

LC–MS:

Liquid chromatography–mass spectrometry

m/z :

Mass-to-charge ratio

MALDI:

Matrix-assisted laser-desorption ionisation

NAD:

Nicotinamide adenine dinucleotide

NADH:

Reduced nicotinamide adenine dinucleotide

OH-PLA:

Hydroxyphenyllactic acid

PAB:

Propionic acid bacteria

PFP:

Pentafluorophenyl

PLA:

Phenyllactic acid

Q:

Quadrupole

QTOF:

Quadrupole time-of-flight

RPC:

Reversed-phase chromatography

Rt:

Retention time

TOF:

Time-of-flight

UHPLC/UPLC:

Ultrahigh/ultra-performance liquid chromatography

ZIC:

Zwitterionic

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Acknowledgements

This work was partially financed by The Danish Ministry of Science, Innovation and Higher Education.

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Authors and Affiliations

  1. DuPont Nutrition Biosciences Aps, Advanced Analysis, Edwin Rahrs Vej 38, 8220, Brabrand, Denmark

    Anders H. Honoré & Michael Thorsen

  2. Department of Food Science, Quality and Technology, University of Copenhagen, Faculty of Science, Rolighedsvej 30, 1958, Frederiksberg C, Denmark

    Anders H. Honoré & Thomas Skov

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  1. Anders H. Honoré
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  2. Michael Thorsen
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Correspondence to Anders H. Honoré.

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Honoré, A.H., Thorsen, M. & Skov, T. Liquid chromatography–mass spectrometry for metabolic footprinting of co-cultures of lactic and propionic acid bacteria. Anal Bioanal Chem 405, 8151–8170 (2013). https://doi.org/10.1007/s00216-013-7269-3

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