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Thiobarbituric acid reactive substances in flavored phytosterol-enriched drinking yogurts during storage: formation and matrix interferences


The aim of this study was to evaluate the level of thiobarbituric acid reactive substances (TBARs) in flavored phytosterol-enriched drinking yogurts during storage, with particular attention to matrix interference. Two phytosterol-enriched drinking yogurts (with white vanilla-WV and blood orange-BO) were kept under controlled conditions (16 h light at 25 °C/8 h dark at 15 °C) for 64 h. During the TBARs assay, a yellow complex was formed. The UV–Vis spectra showed two absorption bands around 450 and 530 nm, respectively. In both flavored phytosterol-enriched drinking yogurts, the more intense absorption was around 450 nm. In conclusion, the development of TBA adducts leads to a higher overestimation of TBARs at 450 nm. The study of TBA reaction with some of the matrix compounds shows that propanal, pentanal, hexanal, p-anisaldehyde, and vanillin favor TBARs450 formation. Instead, acetaldehyde, nonanal, lactose, decanal, t-cinnamaldehyde, octanal, limonene, and lactic acid favor TBARs530 formation. The interference of volatile compounds in the TBARs assay is much higher in BO.

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  1. 1.

    Aubourg SP (1993) Interaction of malondialdehyde with biological molecules—new trends about reactivity and significance. Int J Food Sci Technol 28:323–335

    Article  CAS  Google Scholar 

  2. 2.

    De las Heras A, Schoch A, Gibis M, Fischer A (2003) Comparison of methods for determining malondialdehyde in dry sausage by HPLC and the classic TBA test. Eur Food Res Technol 217:180–184

    Article  CAS  Google Scholar 

  3. 3.

    King RL (1962) Oxidation of milk fat globule membrane material. I. Thiobarbituric acid reaction as measure of oxidized flavour milk and model systems. J Dairy Sci 45:1165–1171

    Article  CAS  Google Scholar 

  4. 4.

    Kosugi H, Kikugawa K (1985) Thiobarbituric acid reaction of aldehydes and oxidized lipids in glacial acetic acid. Lipids 20:915–921

    Article  CAS  Google Scholar 

  5. 5.

    Guillén-Sans R, Guzmán-Chozas M (1998) The thiobarbituric acid (TBA) reaction in foods: a review. Crit Rev Food Sci Nutr 38:315–330

    Article  Google Scholar 

  6. 6.

    Fenaille F, Mottier P, Turesky RJ, Ali S, Guy PA (2001) Comparison of analytical techniques to quantify malondialdehyde in milk powders. J Chromatogr A 921:237–245

    Article  CAS  Google Scholar 

  7. 7.

    Salih AM, Smith DM, Price JF, Dawson LE (1987) Modified extraction 2-thiobarbituric acid method for measuring lipid oxidation in poultry. Poult Sci 66:1483–1488

    Article  CAS  Google Scholar 

  8. 8.

    Ott A, Fay LB, Chaintreau A (1997) Determination and origin of the aroma impact compounds of yogurt flavor. J Agric Food Chem 45:850–858

    Article  CAS  Google Scholar 

  9. 9.

    Ott A, Germond JE, Baumgartner M, Chaintreau A (1999) Aroma comparisons of traditional and mild yogurts: headspace gas chromatography quantification of volatiles and origin of α-diketones. J Agric Food Chem 47:2379–2385

    Article  CAS  Google Scholar 

  10. 10.

    Alonso L, Fraga MJ (2001) Simple and rapid analysis for quantitation of the most important volatile flavor compounds in yogurt by headspace gas chromatography-mass spectrometry. J Chromatogr Sci 39:297–300

    Article  CAS  Google Scholar 

  11. 11.

    Frederiksen CS, Haugaard VK, Poll L, Miquel Becker E (2003) Light-induced quality changes in plain yoghurt packed in polylactate and polystyrene. Eur Food Res Technol 217:61–69

    Article  CAS  Google Scholar 

  12. 12.

    Carrillo-Carrión C, Cárdenas S, Valćarcel M (2007) Vanguard/rearguard strategy for the evaluation of the degradation of yoghurt samples based on the direct analysis of the volatiles profile through headspace-gas chromatography-mass spectrometry. J Chromatogr A 1141:98–105

    Article  Google Scholar 

  13. 13.

    Tarladgis BG, Pearson AM, Dugan LR Jr (1964) Chemistry of the 2-thiobarbituric acid test for determination of oxidative rancidity in foods. II. Formation of the TBA malonaldehyde complex without acid heat treatment. J Sci Food Agric 15:602–607

    Article  Google Scholar 

  14. 14.

    Witte VC, Krause GF, Bailey ME (1970) A new extraction method for determining 2-thiobarbituric acid values of pork and beef during storage. J Food Sci 35:582–585

    Article  CAS  Google Scholar 

  15. 15.

    Robards K, Kerr AF, Patsalides E (1988) Rancidity and its measurement in edible oils and snack foods. A review. Analyst 113:213–224

    Article  CAS  Google Scholar 

  16. 16.

    Vichi S, Pizzale L, Conte LS, Buxaderas S, López-Tamames E (2003) Solid-phase microextraction in the analysis of virgin olive oil volatile fraction: modifications induced by oxidation and suitable markers of oxidative status. J Agric Food Chem 51:6564–6571

    Article  CAS  Google Scholar 

  17. 17.

    Cardoso DR, Libardi SH, Skibsted LH (2012) Riboflavin as a photosensitizer. Effects on human health and food quality. Food Funct 3:487–502

    Article  CAS  Google Scholar 

  18. 18.

    Ng TB, Liu F, Wang ZT (2000) Antioxidative activity of natural products from plants. Life Sci 66:709–723

    Article  CAS  Google Scholar 

  19. 19.

    Londoño-Londoño J, Rodriguez de Lima V, Jaramillo C, Crecsynski Pasa T (2010) Hesperidin and hesperetin membrane interaction: understanding the role of 7-O-glycoside moiety in flavonoids. Arch Biochem Biophys 499:6–16

    Article  Google Scholar 

  20. 20.

    Guzmán-Chozas M, Vicario IM, Guillén-Sans R (1997) Spectrophotometric profiles of off-flavor aldehydes by using their reactions with 2-thiobarbituric acid. J Agric Food Chem 45:2452–2457

    Article  Google Scholar 

  21. 21.

    Almandós ME, Giannini DH, Ciarlo AS, Boeri RL (1986) Formaldehyde as an interference of the 2-thiobarbituric acid test. J Sci Food Agric 37:54–58

    Article  Google Scholar 

  22. 22.

    Lindmark Månsson H (2008) Fatty acids in bovine milk fat. Food Nutr Res 52:1–3

    Google Scholar 

  23. 23.

    Stapelfeldt H, Nielsen BR, Skibsted LH (1997) Effect of heat treatment, water activity and storage temperature on the oxidative stability of whole milk powder. Int Dairy J 7:331–339

    Article  CAS  Google Scholar 

  24. 24.

    Hedegaard RV, Kristensen D, Nielson JH, FrøstMB Østdal H, Hermansen JE, Kröger-Ohlsen M, Skibsted LH (2006) Comparison of descriptive sensory analysis and chemical analysis for oxidative changes in milk. J Dairy Sci 89:495–504

    Article  CAS  Google Scholar 

  25. 25.

    Patton S, Kurtz GW (1955) A note on the thiobarbituric acid test for milk oxidation. J Dairy Sci 38:901

    Article  CAS  Google Scholar 

  26. 26.

    Jennings WG, Dunkley WL, Reiber HG (1955) Studies of certain red pigments formed from 2-thiobarbituric acid. J Food Sci 20:13–22

    Article  CAS  Google Scholar 

  27. 27.

    Cheng H (2010) Volatile flavor compounds in yogurt: a review. Crit Rev Food Sci Nutr 50:938–950

    Article  CAS  Google Scholar 

  28. 28.

    Routray W, Mishra HN (2011) Scientific and technical aspects of yogurt aroma and taste: a review. Compr Rev Food Sci Food 10:208–220

    Article  CAS  Google Scholar 

  29. 29.

    Kaminarides S, Stamou P, Massouras T (2007) Comparison of the characteristics of set type yoghurt made from ovine milk of different fat content. Int J Food Sci Technol 42:1019–1028

    Article  CAS  Google Scholar 

  30. 30.

    Laye I, Karleskind D, Morr CV (1993) Chemical, microbiological and sensory properties of plain nonfat yogurt. J Food Sci 58:991–995

    Article  CAS  Google Scholar 

  31. 31.

    Hruškar M, Vahčić N, Ritz M (1995) Aroma profiles and sensory evaluation of yogurt during storage. Mljekarstvo 45:175–190

    Google Scholar 

  32. 32.

    Güler Z, Taşdelen A, Şenol H, Kerimoğlu N, Temel U (2009) The determination of volatile compounds in set-type yogurt using static headspace gas chromatographic method. Gida 34:137–142

    Google Scholar 

  33. 33.

    Güler Z, Gürsoy-Balci AC (2011) Evaluation of volatile compounds and free fatty acids in set types yogurts made of ewes’, goats’ milk and their mixture using two different commercial starter cultures during refrigerated storage. Food Chem 127:1065–1071

    Article  Google Scholar 

  34. 34.

    Erkaya T, Şengül M (2011) Comparison of volatile compounds in yoghurts made from cows’, buffaloes’, ewes’ and goats’ milks. Int J Dairy Technol 64:240–246

    Article  CAS  Google Scholar 

  35. 35.

    Şenel E (2011) Some carbonyl compounds and free fatty acid composition of Afyon Kaymagı (clotted cream) and their effects on aroma and flavor. Grasas Aceites 62:418–427

    Article  Google Scholar 

  36. 36.

    Saint-Eve A, Lévy C, Le Moigne M, Ducruet V, Souchon I (2008) Quality changes in yogurt during storage in different packaging materials. Food Chem 110:285–293

    Article  CAS  Google Scholar 

  37. 37.

    Kora EP, Souchon I, Latrille E, Martin N, Marin M (2004) Composition rather than viscosity modifies the aroma compound retention of flavored stirred yogurt. J Agric Food Chem 52:3048–3056

    Article  CAS  Google Scholar 

  38. 38.

    Gocmen D, Gurbuz O, Rouseff RL, Smoot JM, Dagdelen AF (2004) Gas chromatographic-olfactometric characterization of aroma active compounds in sun-dried and vacuum-dried tarhana. Eur Food Res Technol 218:573–578

    Article  CAS  Google Scholar 

  39. 39.

    Matthäus B, Rubner M (2010) In: Decker EA, Elias RJ, McClements DJ (eds) Oxidation in food and beverages and antioxidant applications, vol 2: management in different industry sectors. Woodhead Publishing, Cambridge

  40. 40.

    Beshkova D, Simova E, Frengova G, Simov Z (1998) Production of flavor compounds by yogurt starter cultures. J Ind Microbiol Biotechnol 20:180–186

    Article  CAS  Google Scholar 

  41. 41.

    Harasawa N, Tateba H, Ishizuka N, Wakayama T, Kishino K, Ono M (1998) Flavor deterioration in yogurt. Adv Exp Med Biol 434:285–296

    Article  CAS  Google Scholar 

  42. 42.

    Adhicari K, Grün IU, Mustapha A, Fernando LN (2002) Changes in the profile of organic acids in plain set and stirred yogurts during manufacture and refrigerated storage. J Food Qual 25:435–451

    Article  Google Scholar 

  43. 43.

    Seckin AK, Ozkilinc AY (2011) Effect of some prebiotics usage on quality properties of concentrated yogurt. J Anim Vet Adv 10:1117–1123

    Article  CAS  Google Scholar 

  44. 44.

    De Jager LS, Perfetti GA, Diachenko GW (2008) Comparison of headspace-SPME-GC-MS and LC-MS for the detection and quantification of coumarin, vanillin, and ethyl vanillin in vanilla extract products. Food Chem 107:1701–1709

    Article  Google Scholar 

  45. 45.

    McSweeney PLH, Sousa MJ (2000) Biochemical pathways for the production of flavor compounds in cheeses during ripening: a review. Lait 80:293–324

    Article  CAS  Google Scholar 

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This study was partly supported by the POSDRU/89/1.5/S/62371 project from Romania, as a postdoctoral training fellowship. The authors thank Dr. Lorenzo Barbanti for lending the room with control environmental conditions and Dr. Andrea Borsari of Granarolo s.p.a. (Bologna) for providing samples of flavored phytosterol-enriched drinking yogurt.

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Correspondence to Maria Teresa Rodriguez-Estrada or Sevastiţa Muste.

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The authors declare that they have no conflict of interest.

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This study does not contain any experiment involving human or animal subjects.

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Semeniuc, C.A., Mandrioli, M., Rodriguez-Estrada, M.T. et al. Thiobarbituric acid reactive substances in flavored phytosterol-enriched drinking yogurts during storage: formation and matrix interferences. Eur Food Res Technol 242, 431–439 (2016).

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  • TBARs
  • Flavored phytosterol-enriched drinking yogurt
  • Aldehydes
  • Lipid oxidation
  • Storage