Food Analytical Methods

, Volume 12, Issue 11, pp 2582–2590 | Cite as

A Simple HS-SPME/GC-MS Method for Determination of Acrolein from Sourdough to Bread

  • Saša DrakulaEmail author
  • Dubravka Novotni
  • Nikolina Čukelj Mustač
  • Bojana Voučko
  • Marina Krpan
  • Mirjana Hruškar
  • Duška Ćurić


Acrolein is a toxic compound present in food. It can be formed by bacterial species isolated from sourdough or applied as sourdough starter cultures, such as Lactobacillus reuteri. To the best of our knowledge, the content of acrolein in sourdough bread has not yet been determined, neither has a method for its quantification in such a sample been published. The aim of this study was to develop and validate a simple method for determining acrolein in sourdough, batter, and bread by headspace solid-phase microextraction combined with gas chromatography–mass spectrometry. The selected conditions for acrolein extraction were temperature 40 °C and time 20 min. A significant effect of the matrix on acrolein signal was observed. Linearity and precision were achieved within the examined range for all evaluated matrices. The obtained limits of detection and quantification were 1.21 μg/kg of sample and 4.05 μg/kg of sample, respectively. The acrolein solution with the addition of 0.2% hydroquinone was stable for 48 h. The developed method was applied for the quantification of acrolein in gluten-free sourdough (with L. reuteri DSM 20016 or L. brevis DSM 20054 as a starter culture), and batter and bread with or without added sourdough. Acrolein was not detected in the samples prepared without L. reuteri DSM 20016. Acrolein content in bread prepared with L. reuteri DSM 20016 sourdough was 161 μg/kg, which is below the WHO provisional tolerable concentration, but its consumption would significantly contribute to the overall acrolein exposure.


Acrolein Lactobacillus reuteri Sourdough bread SPME GC-MS Validation 



The authors wish to thank The Dow Chemical Company for supplying Methocel™ K4M and Wellence™ Gluten Free 47129, Palco Ltd. for supplying emulsifier MONO 40, and Lidija Drobac for her technical assistance.


This study was funded by Croatian Science Foundation (projects VH/VT 09/01/279 and GbP-FFood IP-06-2016 3789).

Compliance with Ethical Standards

Conflict of Interest

Saša Drakula declares that there is no conflict of interest. Dubravka Novotni declares that there is no conflict of interest. Nikolina Čukelj Mustač declares that there is no conflict of interest. Bojana Voučko declares that there is no conflict of interest. Marina Krpan declares that there is no conflict of interest. Mirjana Hruškar declares that there is no conflict of interest. Duška Ćurić declares that there is no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Not applicable.


  1. Abraham K, Andres S, Palavinskas R, Berg K, Appel KE, Lampen A (2011) Toxicology and risk assessment of acrolein in food. Mol Nutr Food Res 55(9):1277–1290. CrossRefPubMedGoogle Scholar
  2. ATSDR (2007) Toxicological Profile for Acrolein. U.S. Department of Health and Human Services, Public Health Service, Agency for Toxic Substances and Disease Registry AtlantaGoogle Scholar
  3. AOAC Peer Verified Methods Advisory Committee (1998) AOAC peer verified methods program, manual on policies and procedures. AOAC International, GaithersburgGoogle Scholar
  4. Bauer R, Cowan DA, Crouch A (2010a) Acrolein in Wine: Importance of 3-hydroxypropionaldehyde and derivatives in production and detection. J Agric Food Chem 58:3243–3250. CrossRefPubMedGoogle Scholar
  5. Bauer R, du Toit M, Kossmann J (2010b) Influence of environmental parameters on production of the acrolein precursor 3-hydroxypropionaldehyde by Lactobacillus reuteri DSMZ 20016 and its accumulation by wine lactobacilli. Int J Food Microbiol 137(1):28–31. CrossRefPubMedGoogle Scholar
  6. Bianchi F, Careri M, Chiavaro E, Musci M, Vittadini E (2008) Gas chromatographic–mass spectrometric characterisation of the Italian protected designation of origin “Altamura” bread volatile profile. Food Chem 110(3):787–793. CrossRefGoogle Scholar
  7. Bianchi F, Careri M, Mangia A, Musci M (2007) Retention indices in the analysis of food aroma volatile compounds in temperature-programmed gas chromatography: database creation and evaluation of precision and robustness. J Sep Sci 30(4):563–572. CrossRefPubMedGoogle Scholar
  8. Corsetti A, Settanni L (2007) Lactobacilli in sourdough fermentation. Food Res Int 40(5):539–558. CrossRefGoogle Scholar
  9. EFSA BIOHAZ Panel (2017) Scientific Opinion on the update of the list of QPS-recommended biological agents intentionally added to food or feed as notified to EFSA. EFSA J.
  10. Engels C, Schwab C, Zhang J, Stevens MJ, Bieri C, Ebert M-O, McNeill K, Sturla SJ, Lacroix C (2016) Acrolein contributes strongly to antimicrobial and heterocyclic amine transformation activities of reuterin. Sci Rep.
  11. Faroon O, Roney N, Taylor J, Ashizawa A, Lumpkin MH, Plewak DJ (2008) Acrolein health effects. Toxicol Ind Health 24(7):447–490. CrossRefPubMedGoogle Scholar
  12. Galle S, Schwab C, Dal Bello F, Coffey A, Gänzle MG, Arendt EK (2012) Influence of in-situ synthesized exopolysaccharides on the quality of gluten-free sorghum sourdough bread. Int J Food Microbiol 155(3):105–112. CrossRefPubMedGoogle Scholar
  13. Gobbetti M, Gänzle MG (eds) (2013) Handbook on sourdough biotechnology. Springer, BostonGoogle Scholar
  14. Gomes R, Meek ME, Eggleton M (2002) Concise International Chemical Assessment Document 43: Acrolein. World Health Organization, GenevaGoogle Scholar
  15. Green JM (1996) Peer reviewed: a practical guide to analytical method validation. Anal Chem 68(9):305A–309A. CrossRefGoogle Scholar
  16. ICH (2005) ICH harmonised tripartite guideline - validation of analytical procedures: text and methodology Q2(R1). In: International Conference on Harmonisation, GenevaGoogle Scholar
  17. Kächele M, Monakhova YB, Kuballa T, Lachenmeier DW (2014) NMR investigation of acrolein stability in hydroalcoholic solution as a foundation for the valid HS-SPME/GC-MS quantification of the unsaturated aldehyde in beverages. Anal Chim Acta 820:112–118. CrossRefPubMedGoogle Scholar
  18. Langa S, Martín-Cabrejas I, Montiel R, Peirotén Á, Arqués JL, Medina M (2018) Protective effect of reuterin-producing Lactobacillus reuteri against Listeria monocytogenes and Escherichia coli O157: H7 in semi-hard cheese. Food Control 84:284–289. CrossRefGoogle Scholar
  19. Lim H-H, Shin H-S (2012) Simple determination of acrolein in surface and drinking water by headspace SPME GC–MS. Chromatographia 75:943–948. CrossRefGoogle Scholar
  20. Mansur AR, Nam TG, Jang HW, Cho YS, Yoo M, Seo D, Ha J (2017) Determination of 2-propenal using headspace solid-phase microextraction coupled to gas chromatography–time-of-flight mass spectrometry as a marker for authentication of unrefined sesame oil. J Chemother 2017:1–12. CrossRefGoogle Scholar
  21. Moghe A, Ghare S, Lamoreau B, Mohammad M, Barve S, McClain C, Joshi-Barve S (2015) Molecular mechanisms of acrolein toxicity: relevance to human disease. Toxicol Sci 143(2):242–255. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Nageswara Rao T (2018) Validation of Analytical Methods. In Stauffer, M (ed) Calibration and validation of analytical methods, a sampling of current approaches. IntechOpen. Google Scholar
  23. National Association of Testing Authorities, Australia (2018) General accreditation guidance – validation and verification of quantitative and qualitative test methods. Accessed 25 Feb 2019
  24. Novotni D, Vrana Špoljarić I, Drakula S, Čukelj N, Voučko B, Ščetar M, Galić K, Ćurić D (2017) Influence of barley sourdough and vacuum cooling on shelf life quality of partially baked bread. Food Technol Biotechnol 55(4):464–474. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Osório VM, de Lourdes Cardeal Z (2011) Determination of acrolein in french fries by solid-phase microextraction gas chromatography and mass spectrometry. J Chromatogr A 1218(21):3332–3336. CrossRefPubMedGoogle Scholar
  26. Osório VM, de Lourdes Cardeal Z (2013) Using SPME-GC/MS to evaluate acrolein production in cassava and pork sausage fried in different vegetable oils. J Am Oil Chem Soc 90(12):1795–1800. CrossRefGoogle Scholar
  27. Papoušek R, Pataj Z, Nováková P, Lemr K, Barták P (2014) Determination of acrylamide and acrolein in smoke from tobacco and e-cigarettes. Chromatographia 77(17-18):1145–1151. CrossRefGoogle Scholar
  28. Pawliszyn J (2012) Handbook of solid phase microextraction. Elsevier, LondonGoogle Scholar
  29. Raffo A, Carcea M, Castagna C, Magrì A (2015) Improvement of a headspace solid phase microextraction-gas chromatography/mass spectrometry method for the analysis of wheat bread volatile compounds. J Chromatogr A 1406:266–278. CrossRefPubMedGoogle Scholar
  30. Rietjens IMCM, Dussort P, Günther H, Hanlon P, Honda H, Mally A, O’Hagan S, Scholz G, Seidel A, Swenberg J, Teeguarden J, Eisenbrand G (2018) Exposure assessment of process-related contaminants in food by biomarker monitoring. Arch Toxicol 92(1):15–40. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Schmidt M, Lynch KM, Zannini E, Arendt EK (2018) Fundamental study on the improvement of the antifungal activity of Lactobacillus reuteri R29 through increased production of phenyllactic acid and reuterin. Food Control 88:139–148. CrossRefGoogle Scholar
  32. Schütz H, Radler F (1984) Anaerobic reduction of glycerol to propanediol-1.3 by Lactobacillus brevis and Lactobacillus buchneri. Syst Appl Microbiol 5(2):169–178. CrossRefGoogle Scholar
  33. Seaman VY, Charles MJ, Cahill TM (2006) A sensitive method for the quantification of acrolein and other volatile carbonyls in ambient air. Anal Chem 78(7):2405–2412. CrossRefPubMedGoogle Scholar
  34. Takamoto S, Sakura N, Yashiki M, Kojima T (2001) Determination of acrolein by headspace solid-phase microextraction gas chromatography and mass spectrometry. J Chromatogr B 758(1):123–128. CrossRefGoogle Scholar
  35. Vollenweider S, Evers S, Zurbriggen K, Lacroix C (2010) Unraveling the hydroxypropionaldehyde (HPA) system: an active antimicrobial agent against human pathogens. J Agric Food Chem 58(19):10315–10322. CrossRefPubMedGoogle Scholar
  36. Vollenweider S, Lacroix C (2004) 3-Hydroxypropionaldehyde: applications and perspectives of biotechnological production. Appl Microbiol Biotechnol 64(1):16–27. CrossRefPubMedGoogle Scholar
  37. Yasuhara A, Tanaka Y, Hengel M, Shibamoto T (2003) Gas chromatographic investigation of acrylamide formation in browning model systems. J Agric Food Chem 51(14):3999–4003. CrossRefPubMedGoogle Scholar
  38. Zhao CJ, Kinner M, Wismer W, Gänzle MG (2015) Effect of glutamate accumulation during sourdough fermentation with Lactobacillus reuteri on the taste of bread and sodium-reduced bread. Cereal Chem 92(2):224–230. CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Faculty of Food Technology and BiotechnologyUniversity of ZagrebZagrebCroatia

Personalised recommendations