Part 3. Dienoic Fatty Acids

Mass Spectrometry of 3-Pyridylcarbinol Esters

 

Here, the electron-impact mass spectra of 3-pyridylcarbinol esters of those dienoic acids that are most likely to be encountered in nature, together with some useful model compounds, are described. The widely used term 'picolinyl ester' is inaccurate and should now be avoided (see our web page on the saturated derivatives). There is a separate document dealing with Mass spectra of allenic fatty acids.

 

Methylene-Interrupted Dienoic Fatty Acids

The mass spectra of 3-pyridylcarbinol esters of dienoic fatty acids, like those of monoenes, tend to be distinctive and permit location of the double bond. As an example, the mass spectrum of 3-pyridylcarbinyl 9,12-octadecadienoate (linoleate or 18:2(n-6)), the most important dienoic fatty acid in nature, is illustrated below (Harvey, 1982) -

Mass spectrum of picolinyl 9,12-octadecadienoate

As with 3-pyridylcarbinol esters of saturated and monoenoic fatty acids, interpretation is easiest if we start with the molecular ion and work down the chain. Ions corresponding to cleavage on either side of the terminal double bond are seen at m/z = 300 and 274 (26 amu apart), there is then a gap of 14 amu to m/z = 260, then one of 26 amu for the internal double bond to m/z = 234. When examining the spectra of unknowns, however, it is often easier to locate gaps of 40 amu for the double bond and the associated methylene group on the carboxyl side, i.e. in this instance between m/z = 300 and 260, and between 260 and 220. Regular series of ions 14 amu apart on either side of the double bond confirm that there are no further functional groups in these regions of the molecule.

Similar series of ions are seen in spectra from 3-pyridylcarbinol esters of most methylene-interrupted dienes, but it is advantageous to have access to spectra of authentic standards when the double bonds are close to either end of the molecule when confusion is most likely to arise. Details of the spectra of the complete series of isomeric methylene-interrupted octadecadienoates have been published (Christie, Brechany and Holman, 1987a), but only a few of the spectra were depicted in the paper for practical reasons. All are now illustrated below, but please consult the original reference for tabulated data. Some of these may not occur in nature, but a surprising number do. Spectra of other homologous fatty acid isomers displaying the same diagnostic features may have been described elsewhere in some instances. Spectra for the complete C18 series follow.

3-Pyridylcarbinyl 2,5-octadecadienoate (2,5-18:2) (Christie et al., 1987) -

Mass spectrum of 3-pyridylcarbinyl 2,5-octadecadienoate

The molecular ion is the base peak, presumably because of the double bond in conjugation with the carboxyl group. Ions that identify the position of the double bond are not apparent, but the doublet of ions at m/z = 214 and 230 are a useful indicator (see the web pages on 3-pyridylcarbinol esters of monoenes). The regular series of ions 14 amu apart between m/z = 214 and the molecular ion confirm that there are no functional groups in this part of the alkyl chain.

3-Pyridylcarbinyl 3,6-octadecadienoate (3,6-18:2) -

Mass spectrum of 3-pyridylcarbinyl 3,6-octadecadienoate

In this spectrum, the gaps of 26 amu between m/z = 151 and 177, and 190 and 216 serve to locate the double bonds in positions 3 and 6, respectively.

3-Pyridylcarbinyl 4,7-octadecadienoate (4,7-18:2). Some of the marked ions that locate the double bonds would be difficult to identify from first principles, so a model spectrum is essential.

Mass spectrum of 3-pyridylcarbinyl 4,7-octadecadienoate

3-Pyridylcarbinyl 5,8-octadecadienoate (5,8-18:2) – this fatty acid is a major component of human sebum lipids. As the distance of double bonds from the carboxyl group increases, the expected fragmentation patterns for the double bonds emerge as marked.

Mass spectrum of 3-pyridylcarbinyl 5,8-octadecadienoate

3-Pyridylcarbinyl 6,9-octadecadienoate (6,9-18:2). Here, and with many of the subsequent spectra in this series, the double bonds are easily located.

Mass spectrum of 3-pyridylcarbinyl 6,9-octadecadienoate

3-Pyridylcarbinyl 7,10-octadecadienoate (7,10-18:2).

Mass spectrum of 3-pyridylcarbinyl 7,10-octadecadienoate

3-Pyridylcarbinyl 8,11-octadecadienoate (8,11-18:2) -

Mass spectrum of 3-pyridylcarbinyl 8,11-octadecadienoate

3-Pyridylcarbinyl 9,12-octadecadienoate - see the start of this web page.

3-Pyridylcarbinyl 10,13-octadecadienoate (10,13-18:2) -

Mass spectrum of 3-pyridylcarbinyl 10,13-octadecadienoate

3-Pyridylcarbinyl 11,14-octadecadienoate (11,14-18:2) (Christie et al., 1987a) -

Mass spectrum of 3-pyridylcarbinyl 11,14-octadecadienoate

3-Pyridylcarbinyl 12,15-octadecadienoate (12,15-18:2) -

Mass spectrum of 3-pyridylcarbinyl 12,15-octadecadienoate

3-Pyridylcarbinyl 13,16-octadecadienoate (13,16-18:2) -

Mass spectrum of 3-pyridylcarbinyl 13,16-octadecadienoate

3-Pyridylcarbinyl 14,17-octadecadienoate (14,17-18:2). It is possible even with this last of the isomers to locate the double bonds by the specific fragments marked, especially if we look for the gaps of 40 amu as well as those of 26 amu.

Mass spectrum of 3-pyridylcarbinyl 14,17-octadecadienoate

Spectra of 3-pyridylcarbinol esters of many more dienoic fatty acids of this type of differing chain lengths are illustrated in our Archive section, but without interpretation. Many of these have not been formally published elsewhere.

 

Conjugated Dienoic Fatty Acids

3-Pyridylcarbinol esters may not be the best derivatives for fatty acids with conjugated double bond systems, and for example, DMOX derivatives may be better in this instance (see Mass spectra of DMOX derivatives - dienoic fatty acids), or MTAD adducts of methyl esters are uniquely valuable, especially for the complex mixtures found in commercial ‘conjugated linoleic acid’ (CLA) and some natural samples. However, 3-pyridylcarbinol esters can be useful if authentic standards are available for comparison, especially when isomers are well resolved on the GC column.

The mass spectrum of 3-pyridylcarbinyl 9-cis,11-trans-octadecadienoate, the most abundant natural isomer (e.g. in cows' milk fat) follows -

Mass spectrum of 3-pyridylcarbinyl 9-cis,11-trans-octadecadienoate

There is a gap of 52 amu between m/z = 234 and 286 at either end of the conjugated double bond system, with intervals of 14 amu on either side for cleavage at successive methylene groups. The molecular ion at m/z = 371 is the base peak.

3-Pyridylcarbinyl 10-trans,12-cis-octadecadienoate -

Mass spectrum of 3-pyridylcarbinyl 10-trans,12-cis-octadecadienoate

In this instance, the diagnostic ions are shifted up by 14 amu from the previous spectrum, but not very convincingly if a definitive identification were to be required. These spectra do not appear to have been published formally elsewhere.

 

Bis-Methylene-Interrupted Dienoic Fatty Acids

It is becoming apparent that bis- and polymethylene-interrupted dienoic fatty acids are more common in nature than may have been supposed. In particular, fatty acids with a 5,9-double bond system and their chain elongation products are common in seed oils from Gymnosperms or in certain marine invertebrates such as sponges. The spectrum of 5,9-18:2 and of some synthetic fatty acids with the bis-methylene-interrupted double bonds (useful for confirming fragmentation mechanisms) follow next (Christie et al., 1987b).

3-Pyridylcarbinyl 5,9-octadecadienoate (5,9-18:2) (Hierro et al., 1996) -

Mass spectrum of 3-pyridylcarbinyl 5,9-octadecadienoate

The truly distinctive feature is the ion at m/z = 219 (unusual in being odd-numbered) representing cleavage at the centre of the bis-methylene-interrupted double bond system. In addition, gaps of 26 amu, between m/z = 178 and 206 and m/z = 232 and 258 help to locate the double bonds in positions 5 and 9, respectively.

3-Pyridylcarbinyl 2,6-octadecadienoate (2,6-18:2) -

Mass spectrum of 3-pyridylcarbinyl 2,6-octadecadienoate

In this spectrum (unpublished), the ion at m/z = 177 representing cleavage at the centre of the bis-methylene-interrupted double bonds is the base peak in fact. The expected ion at m/z = 164 is missing, but the ions at m/z = 190 and 216 locate the double bond in position 6.

3-Pyridylcarbinyl 6,10-octadecadienoate (6,10-18:2) (Christie et al., 1987b) -

Mass spectrum of 3-pyridylcarbinyl 6,10-octadecadienoate

All the ions that locate the double bonds (marked) are easily located, and the same is true for the 7,11-isomer (not illustrated). It appears that ions diagnostic for the positions of the double bonds become easier to locate as they move further from the carboxyl group.

3-Pyridylcarbinyl 8,12-octadecadienoate (8,12-18:2) (Christie et al., 1987b) -

Mass spectrum of 3-pyridylcarbinyl 8,12-octadecadienoate

Note that the ion representing cleavage at the centre of the bis-methylene-interrupted double bonds is now even numbered (m/z = 260). The same is true of the 9,13-isomer as in 3-pyridylcarbinyl 9,13-eicosadienoate (here..).

Spectra of 3-pyridylcarbinol esters of many more bis-methylene-interrupted dienoic fatty acids are illustrated in our Archive section, but without interpretation. Many of these have not been formally published elsewhere.

 

Poly-Methylene-Interrupted Dienoic Acids

Fatty acids with more than two methylene groups between double bonds are perhaps less common than bis-methylene-interrupted isomers in nature, but some examples of mass spectra of 3-pyridylcarbinol esters of synthetic and natural fatty acids of this type are illustrated below. Spectra from two synthetic fatty acids are available (Christie et al., 1987b)), and the first of these has been found naturally in a sponge (Carballeira and Maldona, 1989).

3-Pyridylcarbinyl 6,11-octadecadienoate (6,11-18:2) has three methylene groups between the double bonds -

Mass spectrum of 3-pyridylcarbinyl 6,11-octadecadienoate

There are prominent ions for cleavages between the double bonds, i.e. at m/z - 233 and 246 (one odd-numbered), and the double bonds themselves can be located by the usual means as marked.

3-Pyridylcarbinyl 7,12-octadecadienoate (7,12-18:2) -

Mass spectrum of 3-pyridylcarbinyl 7,12-octadecadienoate 

The spectrum is very similar to the previous, except that the key ions are all shifted by 14 amu upwards, as might be expected.

3-Pyridylcarbinyl 5,11-octadecadienoate (5,11-18:2) has four methylene groups between the double bonds and is present in the seeds of Gingko biloba. Its mass spectrum has been published (Wolff et al., 1999) -

Mass spectrum of 3-pyridylcarbinyl 5,11-octadecadienoate

In this instance, the double bonds are located in the same way as for isolated monoenes, with that for position 5 being determined by the gap of 26 amu between m/z = 178 and 204, and for position 11 by that between m/z = 260 and 286. Direct comparison with the mass spectrum of 5-18:1 is invaluable here, for confirmation of the position of the first double bond. 3-Pyridylcarbinyl 5,11-eicosadienoate has a very similar spectrum (here...).

3-Pyridylcarbinyl 6,12-octadecadienoate (6,12-18:2) -

Mass spectrum of 3-pyridylcarbinyl 6,12-octadecadienoate

This spectrum resembles that of the 5,11-isomer except that the diagnostic ions are all 14 amu higher.

My colleagues and I have encountered a few long-chain fatty acids with more than four (six) methylene groups between double bonds and some examples from marine invertebrates follow.

3-Pyridylcarbinyl 5,13-eicosadienoate (Christie et al., 1988)). As with the previous, interpretation of the spectrum is similar to those of isolated monoenes, and this is also true of the next spectrum.

Mass spectrum of 3-pyridylcarbinyl 5,13-eicosadecadienoate

3-Pyridylcarbinyl 9,19-tetracosadienoate (9,19-26:2) (Christie et al., 1992) -

Mass spectrum of 3-pyridylcarbinyl 9,19-tetracosadecadienoate

Spectra of 3-pyridylcarbinol esters of many more dienoic fatty acids, including some branched-chain isomers, are illustrated in our Archive section, but without interpretation. Many of these have not been published formally elsewhere.

 

References

  • Carballeira, N.M. and Maldona, M.E. On the isolation of the new fatty acid 6,11-eicosadienoic (20:2) and related 6,11-dienoic acids from the sponge Euryspongia rosea. Lipids, 24, 665-668 (1989) (DOI: 10.1007/BF02535087).
  • Christie, W.W., Brechany, E.Y. and Holman, R.T. Mass spectra of the picolinyl esters of isomeric mono- and dienoic fatty acids. Lipids, 22, 224-228 (1987a) (DOI: 10.1007/BF02533983).
  • Christie, W.W., Brechany, E.Y., Gunstone, F.D., Lie Ken Jie, M.S.F. and Holman, R.T. Mass spectra of the picolinyl esters of some non-methylene-interrupted octadecadienoic acids. Lipids, 22, 664-666 (1987b) (DOI: 10.1007/BF02533946).
  • Christie, W.W., Brechany ,E.Y. and Stefanov, K. Silver ion high-performance liquid chromatography and gas chromatography-mass spectrometry in the analysis of complex fatty acid mixtures: application to marine invertebrates. Chem. Phys. Lipids, 46, 127-136 (1988) (DOI: 10.1016/0009-3084(88)90123-5).
  • Christie, W.W., Brechany, E.Y., Stefanov, K. and Popov, S. The fatty acids of the sponge Dysidea fragilis from the Black Sea. Lipids, 27, 640-644 (1992) (DOI: 10.1007/BF02536125).
  • Harvey, D.J. Picolinyl esters as derivatives for the structural determination of long chain branched and unsaturated fatty acids. Biomed. Mass Spectrom., 9, 33-38 (1982) (DOI: 10.1002/bms.1200090107).
  • Hierro, M.T.G., Robertson, G., Christie, W.W. and Joh, Y.-G. The fatty acid composition of the seeds of Ginkgo biloba. J. Am. Oil Chem. Soc., 73, 575-579 (1996) (DOI: 10.1007/BF02518110).
  • Wolff, R.L., Christie, W.W. and Marpeau, A.M. Reinvestigation of the polymethylene-interrupted 18:2 and 20:2 acids of Ginkgo biloba seed lipids. J. Am. Oil Chem. Soc., 76, 273-276 (1999) (DOI: 10.1007/s11746-999-0230-0).

Updated August 7, 2013

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