Sensory active substances causing off-odour in liquid whey during storage


  • Mariana Hanková University of Chemistry and Technology, Prague, Faculty of Food and Biochemical Technology, Department of Food Preservation, Technická 3, 166 28, Prague, Czech Republic, Tel.: +420 220 443 064
  • Helena Čížková University of Chemistry and Technology, Prague, Faculty of Food and Biochemical Technology, Department of Food Preservation, Technická 3, 166 28, Prague, Czech Republic, Tel.: +420 220 443 064



whey, SPME-GC-MS/O, off-odor, analysis


Liquid whey is a nutritious product with high water activity and neutral pH. Therefore, it is very susceptible to microbiological spoilage that results in undesirable off-odors. Additionally, minimally processed foods are the recent trend so setting an appropriate shelf life is essential. The commonly used microbiological methods are lengthy and time-demanding, so a quick and early identification of microbial degradation would be a significant benefit. Here we tested a solid-phase microextraction, gas chromatography with mass spectrometry coupled with olfactometry analysis (SPME-GC-MS/O) on samples of sweet unpasteurized liquid whey stored at 6 °C, 12 °C and 25 °C for a week. We compared the common methods – plate methods, measurement of pH, and dry matter determination with our proposed SPME-GC-MS/O. We have identified seven sensory active compounds while octanoic acid and a compound not reliably identified by the MS detector (with main m/z observed 133 (100), 151 (65), and 135 (26)) being the most prominent. Microbiological methods proved irreplaceable for proper setting of storage conditions (with the growth of coliforms being significant (p <0.001) at 25 °C). However, SPME-GC-MS/O was able to identify volatile substances responsible for off-odors and can be used as a powerful tool to detect the cause of undesirable chemical and microbial changes in whey beverages.


Download data is not yet available.


Anand, S., Khanal, S. N., Chenchaiah, M. 2013. Whey and Whey Products. In Park, Y. W., Haenlein, G. F. W. Milk and Dairy Products in Human Nutrition. New Jersey, USA : John Wiley & Sons, p. 477-497

Brattoli, M., Cisternino, E., Rosario Dambruoso, P., de Gennaro, G., Giungato, P., Mazzone, A., Palmisani, J., Tutino, M. 2013. Gas chromatography analysis with olfactometric detection (GC-O) as a useful methodology for chemical characterization of odorous compounds. Sensors, vol. 13, no. 12, p. 16759-16800.

Campbell, R. E., Miracle, R. E., Gerard, P. D., Drake, M. A. 2011. Effects of Starter Culture and Storage on the Flavor of Liquid Whey. Journal of Food Science, vol. 76, no. 5, p. 354-361.

Carunchia Whetstine, M. E., Parker, J. D., Drake, M. A., Larick, D. K. 2003. Determining flavor and flavor variability in commercially produced liquid cheddar whey. Journal of Dairy Science, vol. 86, no. 2, p. 439-448.

Cometto-Muñiz, J. E., Abraham, M. H. 2010. Structure–activity relationships on the odor detectability of homologous carboxylic acids by humans. Experimental Brain Research, vol. 207, no. 1-2, p. 75-84.

Croissant, A. E., Kang, E. J., Campbell, R. E., Bastian, E., Drake, M. A. 2009. The effect of bleaching agent on the flavor of liquid whey and whey protein concentrate. Journal of Dairy Science, vol. 92, no. 12, p. 5917-5927.

Dragone, G., Mussatto, S. I., Oliveira, J. M., Teixeira, J. A. 2009. Characterisation of volatile compounds in an alcoholic beverage produced by whey fermentation. Food Chemistry, vol. 112, no. 4, p. 929-935.

Evans, J., Zulewska, J., Newbold, M., Drake, M. A., Barbano, D. M. 2009. Comparison of composition, sensory, and volatile components of thirty-four percent whey protein and milk serum protein concentrates. Journal of Dairy Science, vol. 92, no. 10, p. 4773-4791.

Fox, P. F., Guinee, T. P., Cogan, T. M., McSweeney, P. L. H. 2016. Fundamentals of Cheese Science. 2nd ed. New York, USA : Springer publishing media, 799 p. ISBN 978-1-4899-7679-6.

Francis, F. 1999. Whey: Composition, Properties, Processing and Uses. In Wiley Encycl. Food Sci. Technol. 2nd ed. NEW YORK, USA : John Wiley and Sons Inc., 799 p. ISBN 978-1-4899-7679-6.

Franco-Luesma, E., Ferreira, V. 2016. Formation and Release of H2S, Methanethiol, and Dimethylsulfide during the Anoxic Storage of Wines at Room Temperature. Journal of Agricultural and Food Chemistry, vol. 64, no. 32, p. 6317-6326.

Gocmen, D., Elston, A., Williams, T., Parish, M., Rouseff, R. L. 2005. Identification of medicinal off-flavours generated by Alicyclobacillus species in orange juice using GC-olfactometry and GC-MS. Letters in Applied Microbiology, vol. 40, no. 3, p. 172-177.

Högnadóttir, Á., Rouseff, R. L. 2003. Identification of Aroma Active Compounds in Orange Essence Oil Using Gas Chromatography-Olfactometry and Gas Chromatography-Mass Spectrometry. Journal of Chromatography A, vol. 998, no. 1-2, p. 201-211.

Hugenholtz, J., Kleerebezem, M., Starrenburg, M., Delcour, J., De Vos, W., Hols, P. 2000. Lactococcus lactis as a cell factory for high-level diacetyl production. Applied and Environmental Microbiology, vol. 66, no. 9, p. 4112-4114.

ISO 21527-1. 2008. Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration of yeasts and moulds — Part 1: Colony count technique in products with water activity greater than 0,95. International Standard Organisation.

ISO 4832. 2006. Microbiology of food and animal feeding stuffs — Horizontal method for the enumeration of coliforms — Colony-count technique. International Standard Organisation.

ISO 7218. 2007. Microbiology of food and animal feeding stuffs — General requirements and guidance for microbiological examinations. International Standard Organisation.

Karagul-Yuceer, Y., Drake, M. A., Cadwallader, K. R. 2003. Aroma-active components of liquid cheddar whey. Journal of Food Science, vol. 68, no. 4, p. 1215-1219.

Kilara, A. 2015. Whey and Whey Products. In Chandan, R. C., Kilara, A., Shah, N. P. Dairy Processing and Quality Assurance. New Jersey, USA : John Wiley & Sons, Ltd, p. 349-366. ISBN: 9781118810279.

Kilcast, D. 2010. Sensory quality control for taint prevention. Sensory Analysis for Food and Beverage Quality Control. Cambridge, US : Woodhead Publishing Limited, p.156-185. ISBN 978-1-84569-476-0.

Leksrisompong, P. P., Miracle, R. E., Drake, M. 2010. Characterization of flavor of whey protein hydrolysates. Journal of Agricultural and Food Chemistry, vol. 58, no. 10, p. 6318-6327.

Liaw, I. W., Evan Miracle, R., Jervis, S. M., Listiyani, M. A. D., Drake, M. A. 2011. Comparison of the Flavor Chemistry and Flavor Stability of Mozzarella and Cheddar Wheys. Journal of Food Science, vol. 76, no. 8, p. 1188–1194.

Lo, R., Xue, T., Weeks, M., Turner, M. S., Bansal, N. 2016. Inhibition of bacterial growth in sweet cheese whey by carbon dioxide as determined by culture-independent community profiling. International Journal of Food Microbiology, vol. 217, p. 20-28.

Luo, D., Pang, X., Xu, X., Bi, S., Zhang, W., Wu, J. 2018. Identification of Cooked Off-Flavor Components and Analysis of Their Formation Mechanisms in Melon Juice during Thermal Processing. Journal of Agricultural and Food Chemistry, vol. 66, no. 22, p. 5612-5620.

Muñoz, R., Sivret, E. C., Parcsi, G., Lebrero, R., Wang, X., Suffet, I. H. M., Stuetz, R. M. 2010. Monitoring techniques for odour abatement assessment. Water Research, vol. 44, no. 18, p. 5129-5149.

Nadal, I., Rico, J., Pérez-Martínez, G., Yebra, M. J., Monedero, V. 2009. Diacetyl and acetoin production from whey permeate using engineered Lactobacillus casei. Journal of Industrial Microbiology & Biotechnology, vol. 36, no. 9, p. 1233-1237.

Nishibori, N., Sasaki, K., Okimori, Y., Kanai, M., Isogai, A., Yamada, O., Fujii, T., Goto-Yamamoto, N. 2014. Yeast cell lysis enhances dimethyl trisulfide formation in sake. Journal of Bioscience and Bioengineering, vol. 118, no. 5, p. 526-528.

Ömür-Özbek, P., Dietrich, A. M. 2008. Developing hexanal as an odor reference standard for sensory analysis of drinking water. Water Research, vol. 42, no. 10-11, p. 2598-2604.

Panseri, S., Soncin, S., Chiesa, M., Biondi, P. A. 2011. A headspace solid-phase microextraction gas-chromatographic mass-spectrometric method (HS-SPME-GC/MS) to quantify hexanal in butter during storage as marker of lipid oxidation. Food Chemistry, vol. 127, no. 2, p. 886-889.

Plutowska, B., Wardencki, W. 2008. Application of gas chromatography–olfactometry (GC–O) in analysis and quality assessment of alcoholic beverages – A review. Food Chemistry, vol. 107, no. 1, p. 449-463. https://10.1016/j.foodchem.2007.08.058

Qian, M., Reineccius, G. 2003. Potent aroma compounds in Parmigiano Reggiano cheese studied using a dynamic headspace (purge-trap) method. Flavour and Fragrance Journal, vol. 18, no. 3, p. 252-259.

Sattin, E., Andreani, N. A., Carraro, L., Lucchini, R., Fasolato, L., Telatin, A., Balzan, S., Novelli, E., Simionati, B., Cardazzo, B. 2016. A multi-omics approach to evaluate the quality of milk whey used in ricotta cheese production. Frontiers in Microbiology, vol. 7, p. 1-13.

Semmelroch, P., Grosch, W. 1995. Analysis of roasted coffee powders and brews by gas chromatography-olfactometry of headspace samples. LWT – Food Science and Technology, vol. 28, no. 3, p. 310-313.

Smit, G., Smit, B. A., Engels, W. J. M. 2005. Flavour formation by lactic acid bacteria and biochemical flavour profiling of cheese products. FEMS Microbiology Reviews, vol. 29, no. 3, p. 591-610.

Smithers, G. W. 2008. Whey and whey proteins—From ‘gutter-to-gold. International Dairy Journal, vol. 18, no. 7, p. 695-704.

The Good Scents Company. n.d. Available at:

Zellner, B. A., Dugo, P., Dugo, G., Mondello, L. 2008. Gas chromatography-olfactometry in food flavour analysis. Journal of Chromatography A, vol. 1186, no. 1-2, p. 123-143.

Zepka, L. Q., Garruti, D. S., Sampaio, K. L., Mercadante, A. Z., Da Silva, M. A. A. P. 2014. Aroma compounds derived from the thermal degradation of carotenoids in a cashew apple juice model. Food Research International, vol. 56, p. 108-114.



How to Cite

Hanková, M., & Čížková, H. (2020). Sensory active substances causing off-odour in liquid whey during storage. Potravinarstvo Slovak Journal of Food Sciences, 14, 729–734.