Part 5. Tetra- to Hexaenoic Fatty Acids

Mass Spectrometry of DMOX Derivatives

Methylene-Interrupted Tetraenoic Fatty Acids

As with trienes, the mass spectra of DMOX derivatives of tetraenoic fatty acids permit location of the double bonds, but less easily than those of monoenes and dienes. The diagnostic ions often occur at the centres of clusters, and do not stand out well. However, different isomers tend to have very different spectra so that characterization is possible when spectra of authentic fatty acids can be compared. Few model compounds are available, so most of the following spectra have been obtained from analyses of natural products as part of our research programme, and fatty acids with a variety of chain-lengths are described. As with other nitrogen-containing derivatives, the molecular ion is always clearly delineated, in contrast to the mass spectra of methyl esters, for example. References are listed when we are aware of prior formal publication of spectra in the scientific literature.

For example, the mass spectrum of the DMOX derivative of 6,9,12,15-octadecatetraenoate (stearidonate or 18:4(n-3)), a minor component of fish and some seed oils, is illustrated first -

Mass spectrum of the DMOX derivative of 6,9,12,15-octadecatetraenoate

The double bond in position 6 is recognized by the characteristic fingerprint of ions at m/z = 152, 167 and 180 (Mass spectra of DMOX derivatives. Part 2. Monoenoic fatty acids). The remainder are located by the gaps of 12 amu between m/z = 194 and 206, 234 and 246, and 274 and 286, i.e. for positions 9, 12 and 15, respectively, as can be deduced by extrapolation from our document on monoenes (Yu et al., 1989; Sayanova et al., 1997). Also, the gaps of 40 amu between m/z = 194, 234 and 274, and between m/z = 206, 246 and 286, are useful diagnostic guides for the three terminal double bonds. Indeed, these can sometimes be easier to detect than the gaps of 12 amu.

The DMOX derivative of 8,11,14,17-octadecatetraenoate or 18:4(n-1) - a minor component of fish oils -

Mass spectrum of the DMOX derivative of 8,11,14,17-octadecatetraenoate

The double bonds in positions 8, 11 and 14 can be recognized by the appropriate gaps of 12 amu, as indicated. That in position 17 can only be inferred, although the gaps of 40 amu (m/z = 168 to 208 to 248 to 288 to 329) provide some additional evidence. However, terminal double bonds are not easily recognized with pyrrolidides and 3-pyridinylcarbinol ('picolinyl') esters either.

The DMOX derivative of 5,8,11,14-eicosatetraenoate (20:4(n-6) or arachidonate) - the important essential fatty acid.

Mass spectrum of the DMOX derivative of 5,8,11,14-eicosatetraenoate

The double bond in position 5 is confirmed by the fingerprint ion at m/z = 153, together with the relative abundance of the ion at m/z = 113, while the remainder are located by the gaps of 12 amu as indicated (Zhang et al., 1988; Yu et al., 1989; Wolff et al., 1999). The same diagnostic ions are present in the spectrum of the DMOX derivative of 5,8,11,14-18:4 or 18:4(n-4).

DMOX derivative of 8,11,14,17-eicosatetraenoate (20:4(n-3)) (Yu et al., 1989; Luthria and Sprecher, 1993). The diagnostic ions are marked.

Mass spectrum of the DMOX derivative of 8,11,14,17-eicosatetraenoate

We also have the mass spectra of the DMOX derivatives of several more methylene-interrupted tetraenes, including C22 isomers, on file in the Archive Section of the web pages, but without interpretation.

 

Bis- and Polymethylene-Interrupted Tetraenoic Fatty Acids

We have a few relevant spectra on file garnered from natural sources. For example, the DMOX derivative of 5,11,14,17-eicosatetraenoate (5,11,14,17-20:4) from the seed oil of a gymnosperm (Berdeaux and Wolff, 1996) .

Mass spectrum of the DMOX derivative of 5,11,14,17-eicosatetraenoate

Again, the double bond in position 5 can be deduced from the characteristic fingerprint ion at m/z = 153, together with the relative abundance of the ion at m/z =113, while the remaining double bonds are located by the appropriate gaps of 12 amu, as indicated on the spectrum.

DMOX derivative of 5,9,19,23-tricosatetraenoate (5,9,19,23-30:4) -

Mass spectrum of the DMOX derivative of 5,9,19,23-tricosatetraenoate

This unusual fatty acid from a sponge has two bis-methylene-interrupted double bond systems, recognized most easily by the characteristic fragmentations at the centre of each, i.e. at m/z = 180 and 372, for the 5,9- and 19,23-double bond systems, respectively.

We also have the mass spectrum of the DMOX derivative of 5,9,21,25-32:4 on file in the Archive Section of the web pages, but without interpretation.

 

Pentaenoic Fatty Acids

Only a few natural fatty acids of this type exist, but they are all of great biological importance. Interpretation of these spectra is as for DMOX derivatives of other polyunsaturated fatty acids.

DMOX derivative of of 5,8,11,14,17-eicosapentaenoate (20:5(n-3) or EPA) (Yu et al., 1989; Wolff et al., 1999)-

Mass spectrum of the DMOX derivative of 5,8,11,14,17-eicosapentaenoate

The double bond in position 5 can be deduced from the characteristic fingerprint ion at m/z = 153, while all the remaining double bonds are located by the appropriate gaps of 12 amu as indicated on the spectrum.

DMOX derivative of 4,7,10,13,16-docosapentaenoate (22:5(n-6)) (Yu et al., 1989) -

Mass spectrum of the DMOX derivative of 4,7,10,13,16-docosapentaenoate

The double bond in position 4 is defined by the fingerprint ion at m/z = 152, but only the later double bonds are clearly delineated otherwise. The relative abundance of the ion at m/z = 113 in comparison to that at m/z = 126 is a further aid to identification.

DMOX derivative of 7,10,13,16,19-docosapentaenoate (22:5(n-3)) (Yu et al., 1989). When the first double bond is a little further from the carboxyl group, it is not difficult to locate each of the double bonds in the spectrum as indicated.

Mass spectrum of the DMOX derivative of 7,10,13,16,19-docosapentaenoate

We also have the mass spectrum of the DMOX derivative of 6,9,12,15,18-heneicosapentaenoate (21:5(n-3)), a minor but ubiquitous component of fish oils, on file in the Archive Section of the web pages, but without interpretation.

 

Hexaenoic Fatty Acids

4,7,10,13,16,19-Docosahexaenoate (22:6(n-3) or 'DHA') is a vital component of animal cell membranes, especially those in brain and nervous tissue, and of fish oils. The mass spectrum of its DMOX derivative (Yu et al., 1989; Zhang et al., 1988) is -

Mass spectrum of the DMOX derivative of 4,7,10,13,16,19-docosahexaenoate

The double bond in position 4 is defined by the fingerprint ion at m/z = 152, and the subsequent double bonds are located by the appropriate gaps of 12 amu as indicated on the spectrum.

6,9,12,15,18,21-Tetracosahexaenoate is a biosynthetic precursor of the previous fatty acid, but is only occasionally encountered in tissues in significant amounts (in this instance from a jelly fish). The mass spectrum of its DMOX derivative (Nichols et al., 2003) is -

Mass spectrum of the DMOX derivative of 6,9,12,15,18,21-tetracosahexaenoate

The double bond in position 6 is recognized by the characteristic fingerprint of ions at m/z = 152, 167 and 180, while the remainder are located by the gaps of 12 amu as indicated. The ion at m/z = 126 is now more abundant than that at m/z = 113.

 

References

  • Berdeaux, O. and Wolff, R.L. Gas-liquid chromatography-mass spectrometry of the 4,4-dimethyloxazoline derivatives of Δ5-unsaturated polymethylene-interrupted fatty acids from conifer seed oils. J. Am. Oil Chem. Soc., 73, 1323-1326 (1996) (DOI: 10.1007/BF02525463).
  • Luthria, D.L. and Sprecher, H. 2-Alkenyl-4,4-dimethyloxazolines as derivatives for the structural elucidation of isomeric unsaturated fatty acids. Lipids, 28, 561-564 (1993) (DOI: 10.1007/BF02536089).
  • Nichols, P.D., Danaher, K.T. and Koslow, J.A. Occurrence of high levels of tetracosahexaenoic acid in the jellyfish Aurelia sp. Lipids, 38, 1207-1210 (2003) (DOI: 10.1007/s11745-003-1180-z).
  • Sayanova, O., Smith, M.A., Lapinskas, P., Stobart, A.K., Dobson, G., Christie, W.W. and Shewry, P.R. Expression of a borage desaturase cDNA containing an N-terminal cytochrome b5 domain results in the accumulation of high levels of Δ6 -desaturated fatty acids in transgenic tobacco. Proc. Natl. Acad. Sci. USA, 94, 4211-4216 (1997).
  • Wolff, R.L., Christie, W.W., Pedrono, F. and Marpeau, A.M. Arachidonic, eicosapentaenoic, and biosynthetically related fatty acids in the seed lipids from a primitive gymnosperm, Agathis robusta. Lipids, 34, 1083-1097 (1999) (DOI: 10.1007/s11745-999-0460-y).
  • Yu, Q.T., Liu, B.N., Zhang, J.Y. and Huang, Z.H. Location of double bonds in fatty acids of fish oil and rat testis lipids. Gas chromatography-mass spectrometry of the oxazoline derivatives. Lipids, 24, 79-83 (1989) (DOI: 10.1007/BF02535269).
  • Zhang, J.Y., Yu, Q.T., Liu, B.N. and Huang, Z.H. Chemical modification in mass spectrometry IV. 2-Alkenyl-4,4-dimethyloxazolines as derivatives for double bond location of long-chain olefinic acids. Biomed. Environ. Mass Spectrom., 15, 33-44 (1988) (DOI: 10.1002/bms.1200150106).

 Updated November 6, 2013

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