Detection of 430 Fatty Acid Methyl Esters from a Transesterified Butter Sample

Original Paper

Abstract

Milk fat is known to contain one of the highest number of fatty acids of all edible oils. Some of these fatty acids are known to be valuable (e.g. conjugated linoleic acids, furan fatty acid) and other as undesirable (e.g. saturated and some trans-fatty acids) food ingredients. However, a comprehensive picture on the presence of many trace fatty acids has not been achieved. For this reason we have developed an analysis scheme based on the conversion of the fatty acids into methyl esters. The fatty acid methyl esters were then fractionated by urea complexation. Both the filtrate of the urea complexation (~4 % of the sample weight) and the original sample were fractionated by high-speed counter-current chromatography (HSCCC). The resulting fractions were analyzed by GC/MS analysis. With this method 430 fatty acids were detected in one single butter sample. More than 230 fatty acids had two or more double bonds. In addition to the widely known spectrum of fatty acids we also detected a range of cyclohexyl fatty acids (five homologues) and methyl-branched fatty acids (including short chain and even-numbered anteiso-fatty acids), conjugated tetradecadienoic acids along with the novel ω-oxo-fatty acids (seven homologues). The reported relative retention time on the polar column may serve as a data base for the screening of other samples for this profusion of fatty acids.

Keywords

Butter Fatty acid analysis Urea complexation High-speed counter-current chromatography Mass spectrometry ω-Oxo-fatty acids Cyclohexyl fatty acids 

Abbreviations

9D5/F-VII

9-(3,4-Dimethyl-5-pentylfuran-2-yl)-nonanoic acid

9M5/F-III

9-(3-Methyl-5-pentylfuran-2-yl)-nonanoic acid

11D5/F-VIII

11-(3,4-Dimethyl-5-pentylfuran-2-yl)-undecanoic acid

11M5/F-IV

11-(3-Methyl-5-pentylfuran-2-yl)-undecanoic acid

a11:0

8-Methyl-decanoic acid

a15:0

12-Methyl-tetradecanoic acid

a17:0

14-Methyl-hexadecanoic acid

CLA

Conjugated linoleic acid

CTA

Conjugated tetradecadienoic acid

FAME

Fatty acid methyl ester(s)

GC

Gas chromatography

GC/MS

Gas chromatography with mass spectrometry

HSCCC

High-speed counter-current chromatography

i15:0

13-Methyl-tetradecanoic acid

i17:0

15-Methyl-hexadecanoic acid

KU/L

Partitioning coefficient (upper to lower phase)

MBFA

Methyl branched fatty acid(s)

PUFA

Polyunsaturated fatty acid(s)

SIM

Selected ion monitoring mode

Supplementary material

11746_2013_2218_MOESM1_ESM.doc (1.9 mb)
Supplementary material 1 (DOC 1950 kb)

References

  1. 1.
    Hu FB, Manson JE, Willett WC (2001) Types of dietary fat and risk of coronary heart disease: a critical review. J Am Coll Nutr 20:5–19Google Scholar
  2. 2.
    Shorland FB, Weenink RO, Johns AT, McDonalds IRC (1957) The effect of sheep-rumen contents on unsaturated fatty acids. Biochem J 67:328–333Google Scholar
  3. 3.
    Polan CE, McNeill JJ, Tove SB (1964) Biohydrogenation of unsaturated fatty acids by rumen bacteria. J Bacteriol 88:1056–1064Google Scholar
  4. 4.
    Chilliard Y, Ferlay A, Mansbridge RM, Doreau M (2000) Ruminant milk fat plasticity: nutritional control of saturated, polyunsaturated, trans and conjugated fatty acids. Ann Zootech 49:181–205CrossRefGoogle Scholar
  5. 5.
    Lindmark Månsson H (2008) Fatty acids in bovine milk fat. Food Nutr Res 52. doi:10.3402/fnr.v52i0.1821
  6. 6.
    Jensen RG (2002) The composition of bovine milk lipids: January 1995 to December 2000. J Dairy Sci 85:295–350CrossRefGoogle Scholar
  7. 7.
    Willett WC, Stampfer MJ, Manson JE, Colditz GA, Speizer FE, Rosner BA, Sampson LA, Hennekens CH (1993) Intake of trans fatty acids and risk of coronary heart disease among women. Lancet 341:581–585CrossRefGoogle Scholar
  8. 8.
    Wahle KWJ, Heys SD, Rotondo D (2004) Conjugated linoleic acids: are they beneficial or detrimental to health? Prog Lipid Res 43:553–587CrossRefGoogle Scholar
  9. 9.
    Parodi PW (2009) Milk fat nutrition. In: Tamime AY (ed) Dairy fats and related products. Blackwell, Oxford (ISBN: 978-1-405-15090-3)Google Scholar
  10. 10.
    Spitteler G (2005) Furan fatty acids: occurrence, synthesis, and reactions. Are furan fatty acids responsible for the cardioprotective effects of a fish diet? Lipids 40:711–755Google Scholar
  11. 11.
    Bengen MF (1940) Deutsche Patentanmeldung O.Z. 12438Google Scholar
  12. 12.
    Bengen F, Schlenk W (1949) Über neuartige Additionsverbindungen des Harnstoffs. Experientia 5:200CrossRefGoogle Scholar
  13. 13.
    Täufel K, Müller G, Franzke CL (1958) Über die Adduktbildung langkettiger Fettsäuren mit Harnstoff. Nahrung 2:255–267CrossRefGoogle Scholar
  14. 14.
    Ito Y (2005) Golden rules and pitfalls in selecting optimum conditions for high-speed counter-current chromatography. J Chromatogr A 1065:145–168CrossRefGoogle Scholar
  15. 15.
    Degenhardt A, Engelhardt UH, Lakenbrink C, Winterhalter P (2000) Preparative separation of polyphenols from tea by high-speed countercurrent chromatography. J Agric Food Chem 48:3425–3430CrossRefGoogle Scholar
  16. 16.
    Montilla EC, Hillebrand S, Butschbach D, Baldermann S, Watanabe N, Winterhalter P (2010) Preparative isolation of anthocyanins from Japanese purple sweet potato (Ipomoea batatas L.) varieties by high-speed countercurrent chromatography. J Agric Food Chem 58:9899–9904CrossRefGoogle Scholar
  17. 17.
    Koehler N, Wray V, Winterhalter P (2008) Preparative isolation of procyanidins from grape seed extracts by high-speed counter-current chromatography. J Chromatogr A 1177:114–125CrossRefGoogle Scholar
  18. 18.
    Zhou Y, Chen F, Li Z (2002) Preparative separation of beta-sitosterol by high speed countercurrent chromatography. J Liq Chromatogr Relat Technol 25:1693–1701CrossRefGoogle Scholar
  19. 19.
    Schröder M, Vetter W (2011) High-speed counter-current chromatographic separation of phytosterols. Anal Bioanal Chem 400:3615–3623CrossRefGoogle Scholar
  20. 20.
    Schröder M, Vetter W (2012) Investigation of unsaponifiable matter of plant oils and isolation of eight phytosterols by means of high-speed counter-current chromatography. J Chromatogr A 1237:96–105CrossRefGoogle Scholar
  21. 21.
    Huang L, Cao X, Xu H, Chen G (2011) Separation and purification of ergosterol and stigmasterol in Anoectochilus roxburghii (wall) lindl by high-speed counter-current chromatography. J Sep Sci 34:385–392CrossRefGoogle Scholar
  22. 22.
    Cao X, Ito Y (2003) Supercritical fluid extraction of grape seed oil and subsequent separation of free fatty acids by high-speed countercurrent chromatography. J Chromatogr A 1021:117–124CrossRefGoogle Scholar
  23. 23.
    Bousquet O, Le Goffi F (1995) Counter-current chromatographic separation of polyunsaturated fatty acids. J Chromatogr A 704:211–216CrossRefGoogle Scholar
  24. 24.
    Kapp T, Vetter W (2009) Offline coupling of high-speed counter-current chromatography and gas chromatography/mass spectrometry generates a two-dimensional plot of toxaphene components. J Chromatogr A 1216:8391–9397CrossRefGoogle Scholar
  25. 25.
    Vetter W, Kirres J, Bendig P (2011) Bromination of 2-methoxydiphenyl ether to an average of tetrabrominated 2-methoxydiphenyl ethers. Chemosphere 84:1117–1124CrossRefGoogle Scholar
  26. 26.
    Du Q, Shu A, Ito Y (1996) Purification of fish oil ethyl esters by high-speed countercurrent chromatography using non-aqueous solvent systems. J Liq Chromatogr Relat Technol 19:1451–1457CrossRefGoogle Scholar
  27. 27.
    Murayama W, Kosuge Y, Nakaya N, Nunogaki Y, Nunogaki K, Cazes J, Nunogaki H (1988) Preparative separation of unsaturated fatty acid esters by centrifugal partition chromatography (CPC). J Liq Chromatogr 11:283–300CrossRefGoogle Scholar
  28. 28.
    Hauff S, Vetter W (2010) Exploring the fatty acids of vernix caseosa in form of their methyl esters by off-line coupling of non-aqueous reversed phase high performance liquid chromatography and gas chromatography coupled to mass spectrometry. J Chromatogr A 1217:8270–8278CrossRefGoogle Scholar
  29. 29.
    Vetter W, Schröder M (2010) Concentrations of phytanic acid and pristanic acid are higher in organic than in conventional dairy products from the German market. Food Chem 119:746–752CrossRefGoogle Scholar
  30. 30.
    Thurnhofer S, Vetter W (2005) A gas chromatography/electron ionization-mass spectrometry-selected ion monitoring method for determining the fatty acid pattern in food after formation of fatty acid methyl esters. J Agric Food Chem 53:8896–8903CrossRefGoogle Scholar
  31. 31.
    Thurnhofer S, Vetter W (2006) Application of ethyl esters and d 3-methyl esters as internal standards for the gas chromatographic quantification of transesterified fatty acid methyl esters in food. J Agric Food Chem 54:3209–3214CrossRefGoogle Scholar
  32. 32.
    Li D, Schröder M, Vetter W (2012) Isolation of 6,9,12,15-hexadecatetraenoic fatty acid (16:4n–1) methyl ester from transesterified fish oil by HSCCC. Chromatographia 75:1–6CrossRefGoogle Scholar
  33. 33.
    Crexi VT, Mote ML, Monte ML, Pinto LAA (2012) Polyunsaturated fatty acid concentrates of carp oil: chemical hydrolysis and urea complexation. J Am Oil Chem Soc 89:32–334CrossRefGoogle Scholar
  34. 34.
    Gámez-Meza N, Noriega-Rodríguez JA, Medina-Juárez LA, Ortega-García J, Monroy-Rivera J, Toro-Vázquez FJ, García HS, Angulo-Guerrero O (2003) Concentration of eicosapentaenoic acid and docosahexaenoic acid from fish oil by urea complexation. Food Res Int 36:721–727CrossRefGoogle Scholar
  35. 35.
    Liu S, Zhang C, Hong P, Ji H (2006) Concentration of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) of tuna oil by urea complexation: optimization of progress parameters. J Food Eng 73:203–209CrossRefGoogle Scholar
  36. 36.
    Chakraborty K, Vijayagopal P, Chakraborty RD, Vijayan KK (2010) Preparation of eicosapentaenoic acid concentrates from sardine oil by Bacillus circulans lipase. Food Chem 120:433–442CrossRefGoogle Scholar
  37. 37.
    Ackman RG, Hooper SN (1968) Examination of isoprenoid fatty acids as distinguishing characteristics of specific marine oils with particular reference to whale oil. Comp Biochem Physiol 24:549–565CrossRefGoogle Scholar
  38. 38.
    Hayes DG, Bengtsson YC, van Alstine JM, Setterwall F (1998) Urea complexation for the rapid, ecologically responsible fractionation of fatty acids from seed oil. J Am Oil Chem Soc 75:1403–1409CrossRefGoogle Scholar
  39. 39.
    June Brown P, Mei G, Gibberd FB, Burston D, Mayne PD, McClinchy JE, Sidey M (1993) Diet and Refsum’s disease. The determination of phytanic acid and phytol in certain foods and the application of this knowledge to the choice of suitable convenience foods for patients with Refsum’s disease. J Human Nutr Diet 6:295–305CrossRefGoogle Scholar
  40. 40.
    Josten H, Fieg G, Gutsche B, Johannisbauer W (2004) Harnstoffällung als konkurrenzfähiges Trennverfahren für schwierige Stofftrennungen in der Oleochemie. Chem Ing Tech 76:1700–1703CrossRefGoogle Scholar
  41. 41.
    Schröder M, Vetter W (2011) GC/EI-MS determination of the diastereomer distribution of phytanic acid in food samples. J Am Oil Chem Soc 88:341–349CrossRefGoogle Scholar
  42. 42.
    Jensen RG (1973) Composition of bovine milk lipids. J Am Oil Chem Soc 50:186–192CrossRefGoogle Scholar
  43. 43.
    Thurnhofer S, Hottinger G, Vetter W (2007) Enantioselective determination of food-relevant anteiso fatty acids. Anal Chem 79:4696–4701CrossRefGoogle Scholar
  44. 44.
    Hauff S, Vetter W (2010) Creation and evaluation of a two-dimensional contour plot of fatty acid methyl esters after off-line coupling of reversed-phase HPLC and GC/EI-MS. Anal Bioanal Chem 396:2695–2707CrossRefGoogle Scholar
  45. 45.
    Haahti E, Nikkari T, Salmi A-M, Laaksonen A-L (1961) Fatty acids of vernix caseosa. Scand J Clin Lab Invest 13:70–73CrossRefGoogle Scholar
  46. 46.
    Schogt JCM, Haverkamp Begemann P (1965) Isolation of 11-cyclohexylundecanoic acid from butter. J Lipid Res 6:466–470Google Scholar
  47. 47.
    Ohya H, Komai Y, Yamaguchi M (1986) Zinc tolerance of an isolated bacterium containing ω-cyclohexyl fatty acid. FEMS Microbiol Lett 34:257–260Google Scholar
  48. 48.
    Schlosser S, Vetter W (2011) Fatty acids and polar lipid content of cheese and mould-contaminated cheese. Eur J Lipid Sci Technol 113:469–478CrossRefGoogle Scholar
  49. 49.
    Hauff S, Rilfors L, Hottinger G, Vetter W (2010) Structure and absolute configuration of an unsaturated anteiso fatty acid from Bacillus megaterium. J Chromatogr A 1217:1683–1687CrossRefGoogle Scholar
  50. 50.
    The Lipid Library. http://lipidlibrary.aocs.org/
  51. 51.
    Chance DL, Gerhardt KO, Mawhinney TP (1998) Gas-liquid chromatography-mass spectrometry of hydroxyl fatty acids as their methyl esters tert-butyldimethylsilyl ethers. J Chromatogr A 793:91–98CrossRefGoogle Scholar
  52. 52.
    Jenske R, Vetter W (2008) Gas chromatography electron-capture negative-ion mass spectrometry for the quantitative determination of 2- and 3-hydroxy fatty acids in bovine milk fat. J Agric Food Chem 56:5500–5505CrossRefGoogle Scholar
  53. 53.
    Noble AC, Nawar WW (1971) Mass spectrometry of aldehyde esters. J Agric Food Chem 19:1039–1040CrossRefGoogle Scholar
  54. 54.
    Yayli N, Kiran Z, Seymen H, Genc H (2001) Characterization of lipids and fatty acid methyl ester contents in leaves and roots of Crocus vallicola. Turk J Chem 25:391–395Google Scholar
  55. 55.
    Brechany EY, Christie WW (1992) Identification of the saturated oxo fatty acids in cheese. J Dairy Res 59:57–64CrossRefGoogle Scholar
  56. 56.
    Brechany EY, Christie WW (1994) Identification of the unsaturated oxo fatty acids in cheese. J Dairy Res 62:111–115CrossRefGoogle Scholar
  57. 57.
    Márquez-Ruiz G, Rodríguez-Pino V, de la Fuente MA (2011) Determination of 10-hydroxystearic, 10-ketostearic, 8-hydroxypalmitic, and 8-ketopalmitic acids in milk fat by solid-phase extraction plus gas chromatography-mass spectrometry. J Dairy Sci 94:4810–4819CrossRefGoogle Scholar
  58. 58.
    Welke JE, Manfroi V, Zanus M, Lazarotto M, Alcaraz Zini C (2012) Characterization of the volatile profile of Brazilian Merlot wines through comprehensive two dimensional gas chromatography time-of-flight mass spectrometric detection. J Chromatogr A 1226:124–139CrossRefGoogle Scholar
  59. 59.
    Bourgeois G, vivas N, Glories Y, Vitry C (1995) Identification paer spectrométrie de masse couplée à la chromatographie en phase gazeuse des produits de dégradation des hydorperoxydes de lácide linoléique. Sci Aliments 15:625–630Google Scholar
  60. 60.
    Glass RL, Krick TP, Sand DM, Rahn CH, Schlenk H (1975) Furanoid Fatty acids from fish lipids. Lipids 10:695–702CrossRefGoogle Scholar
  61. 61.
    Guth H, Grosch W (1992) Furan fatty acids in butter and butter oil. Z Lebensm Unters Forsch 194:360–362CrossRefGoogle Scholar
  62. 62.
    Vetter W, Laure S, Wendlinger C, Mattes A, Smith AWT, Knight DW (2012) Determination of furan fatty acids in food samples. J Am Oil Chem Soc 89:1501–1508. doi:10.1007/s11746-012-2038-6 Google Scholar
  63. 63.
    Iverson JL, Eisner J, Firestone D (1965) Detection of trace fatty acids in fats and oils by urea fractionation and gas-liquid chromatography. J Am Oil Chem Soc 42:1063–1068CrossRefGoogle Scholar
  64. 64.
    Precht D, Molkentin J (2003) Overestimation of linoleic acid and trans-C18:2 isomers in milk fats with emphasis on transΔ9, transΔ12-octadecadienoic acid. Milchwiss 58:30–34Google Scholar

Copyright information

© AOCS 2013

Authors and Affiliations

  1. 1.Institute of Food Chemistry (170b)University of HohenheimStuttgartGermany

Personalised recommendations