Analytical and Bioanalytical Chemistry

, Volume 403, Issue 10, pp 2983–2995 | Cite as

Rapid identification, by use of the LTQ Orbitrap hybrid FT mass spectrometer, of antifungal compounds produced by lactic acid bacteria

  • Brid Brosnan
  • Aidan Coffey
  • Elke K. Arendt
  • Ambrose FureyEmail author
Original Paper


Fungal contamination of food causes health and economic concerns. Several species of lactic acid bacteria (LAB) have antifungal activity which may inhibit food spoilage fungi. LAB have GRAS (generally recognised as safe) status, allowing them to be safely integrated into food systems as natural food preservatives. A method is described herein that enables rapid screening of LAB cultures for 25 known antifungal compounds associated with LAB. This is the first chromatographic method developed which enables the rapid identification of a wide range of antifungal compounds by a single method with a short analysis time (23 min). Chromatographic separation was achieved on a Phenomenex Gemini C18 100A column (150 mm × 2.0 mm; 5 μm) by use of a mobile-phase gradient prepared from (A) water containing acetic acid (0.1%) and (B) acetonitrile containing acetic acid (0.1%), at a flow rate of 0.3 µL min−1. The gradient involved a progressive ramp from 10–95% acetonitrile over 13 min. The LC was coupled to a hybrid LTQ Orbitrap XL fourier-transform mass spectrometer (FTMS) operated in negative ionisation mode. High mass accuracy data (<3 ppm) obtained by use of high resolution (30,000 K) enabled unequivocal identification of the target compounds. This method allows comprehensive profiling and comparison of different LAB strains and is also capable of the identification of additional compounds produced by these bacteria.


LAB Antifungal compounds LTQ Orbitrap XL High mass accuracy data LC–FTMS 



We gratefully acknowledge funding from the Food Institutional Research Measure (FIRM) Department of Agriculture, Fisheries and Food Ireland (project reference 08RDC607). The Council of Directors, Technological Sector Research-Strand III 2006 Grant Scheme, awarded to Dr A. Furey is also acknowledged for funding the formation of the “Team Elucidate” research group. The Higher Education Authority (Programme for Research in Third-Level Institutions, Cycle 4 (PRTLI IV) National Collaboration Programme on Environment and Climate Changes: Impacts and Responses is also acknowledged. We also thank Dr Mary Lehane and Dr Sharon Hutchinson for their critical reading of the manuscript.

Supplementary material

216_2012_5955_MOESM1_ESM.pdf (138 kb)
ESM 1 (PDF 138 KB)


  1. 1.
    Carr FJ, Chill D, Maida N (2002) The lactic acid bacteria: a literature survey. Crit Rev Microbiol 28:281–370CrossRefGoogle Scholar
  2. 2.
    Gobetti M, De Angelis M, Corsetti A, Di Cagno R (2005) Biochemistry and physiology of sourdough lactic acid bacteria. Trends Food Sci Tech 16:57–69CrossRefGoogle Scholar
  3. 3.
    Broberg A, Jacobsoson K, Strom K, Schnürer J (2007) Metabolite profiles of lactic acid bacteria in Grass Silage. Appl Environ Microbiol 73:5547–5552CrossRefGoogle Scholar
  4. 4.
    Niku-Paavola ML, Laitila A, Mattila-Sandholm T, Haikara A (1998) New types of antimicrobial compounds produced by Lactobacillus plantarum. J Appl Microbiol 86:29–35CrossRefGoogle Scholar
  5. 5.
    Schnürer J, Magnusson J (2005) Antifungal lactic acid bacteria as biopreservatives. Trends Food Sci Tech 16:70–78CrossRefGoogle Scholar
  6. 6.
    Corsetti A, Gobbetti M, Rossi J, Damiani P (1998) Antimould activity of sourdough lactic acid bacteria identification of a mixture of organic acids produced by Lactobacillus sanfrancisco CB1. Appl Microbiol Biotechnol 50:255–256CrossRefGoogle Scholar
  7. 7.
    Valerio F, Lavermicocca P, Pascale M, Visconti A (2003) Production of phenyllactic acid by lactic acid bacteria an approach to the selection of strains contributing to food quality and preservation. FEMS Microbiol Lett 233:289–295CrossRefGoogle Scholar
  8. 8.
    Ryan LAM, Dal Bello F, Arendt EK (2008) The use of sourdough fermented by antifungal LAB to reduce the amount of calcium propionate in bread. I J Food Microbiol 152:274–278CrossRefGoogle Scholar
  9. 9.
    Rizzello CG, CassanaA CR, Gobbetti M (2011) Antifungal activity of sourdough fermented wheat germ used as an ingredient for bread making. Food Chem 127:952–959CrossRefGoogle Scholar
  10. 10.
    Sathe SJ, Nawani NN, Dhakephalkar PK, Kapadnis BP (2007) Antifungal lactic acid bacteria with potential to prolong shelf-life of fresh vegetables. J Appl Microbiol 103:2622–2628CrossRefGoogle Scholar
  11. 11.
    Ndagano D, Lamoureox T, Dortu C, Vandermoten S, Thonart P (2011) Antifungal activity of 2 lactic acid bacteria of the Weissella genus isolated from food. J Food Sci. doi: 10.1111/j.1750-3841.2011.02257.x
  12. 12.
    Ström K, Sjögren J, Broberg A, Schnürer J (2002) Lactobacillus plantarum MiLAB 393 produces the antifungal Cyclic Dipeptides Cyclo(L-Phe –L-Pro) and Cyclo(L-Phe_trans-4-OH-L-Pro) and 3-phenyllactic acid. Appl Environ Microbiol 68:4322–4327CrossRefGoogle Scholar
  13. 13.
    Sjögren J, Magnusson J, Broberg A, Schnürer J, Kenne L (2003) Antifungal 3-hydroxy fatty from Lactobacillus plantarum MiLAB 14. Appl Environ Microbial 69:7554–7557CrossRefGoogle Scholar
  14. 14.
    Lind H, Sjӧgren J, Gohil S, Kenne L, Schnürer J, Broberg A (2007) Antifungal compounds from cultures of dairy propionibacteria type strains. FEMS Microbiol Lett 271:310–315CrossRefGoogle Scholar
  15. 15.
    Armaforte E, Carri S, Caboni GF (2006) High-performance liquid chromatography determination of phenyllactic acid in MRS broth. J Chromatogra A 1131:281–284CrossRefGoogle Scholar
  16. 16.
    Ryan LAM, Zannini E, Dal Bello F, Pawlowska A, Koehler P, Arendt EK (2011) Lactobacillus amylovorus DSM19280 as a novel food-grade antifungal agent for bakery products. I J Food Microbiol 146:276–283CrossRefGoogle Scholar
  17. 17.
    Furey A, Lehane M, Gillman M, Puente-Fernandez P, James KJ (2001) n Determination of Domoic acid in shellfish using liquid chromatography electrospray ionisation ion-trap mass spectrometry. J Chromatogr A 938:167–174CrossRefGoogle Scholar
  18. 18.
    Furey A, Braña-Magdalena A, Lehane M, Moroney C, James KJ, Satake M, Yasumoto T (2002) Determination of Azaspiracids in shellfish using liquid chromatography-tandem electrospray mass spectrometry. Rapid Commun Mass Spectrom 16:238–242CrossRefGoogle Scholar
  19. 19.
    Furey A, Crowley J, Lehane M, James KJ (2002) Liquid chromatography with Electrospray Ion-trap mass spectrometry for the determination of Anatoxins in Cyanobacteria and drinking water. Rapid Commun Mass Spectrom 17:583–588CrossRefGoogle Scholar
  20. 20.
    Allis O, Dauphard J, Hamilton B, Ni Shuilleabhain A, Lehane M, James KJ, Furey A (2007) Liquid chromatography-tandem mass spectrometry application, for the determination of extracellular hepatotoxins in Irish lake and drinking waters. Anal Chem 79:3436–3447CrossRefGoogle Scholar
  21. 21.
    López-Rivera A, O’Callaghan K, Moriarty M, O’Driscoll D, Hamilton B, Lehane M, James KJ, Furey A (2010) First Evidence of Azaspiracids (AZAs): a family of Lipophilic polyether marine toxins in scallops (Argopecten purpuratus) and mussels (Mytilus chilensis) collected in two regions of chile. Toxicon 55:692–701CrossRefGoogle Scholar
  22. 22.
    European Parliament and Council (1995) Directive No. 95/2/EC of 20 February 1995 on food additives other than colours and sweetenersGoogle Scholar
  23. 23.
    Wessels S, Axelsson L, Hansen EB, De Vuyst L, Laulund S, Lähteenmäki LS, Mollet B, Salminen S, Von Wright A (2004) The lactic acid bacteria, the food chain, and their regulation. Trends Food Sci Technol 15:498–505CrossRefGoogle Scholar
  24. 24.
    Watson JT, Sparkman OD (2008) Introduction to mass spectrometry: Instrumentation, applications and strategies for data interpretation. 4th Edn. WileyGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Brid Brosnan
    • 1
  • Aidan Coffey
    • 2
  • Elke K. Arendt
    • 3
  • Ambrose Furey
    • 1
    • 4
    Email author
  1. 1.Team Elucidate, Department of ChemistryCork Institute of Technology (CIT)CorkIreland
  2. 2.Department of Biological SciencesCork Institute of TechnologyCorkIreland
  3. 3.Department of Food and Nutritional SciencesUniversity College CorkCorkIreland
  4. 4.Department of ChemistryProteobio, Mass Spectrometry Centre for Proteomics and Biotoxin ResearchCorkIreland

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