Mass spectra of alkyl esters

W.W. Christie lipidlibrary.aocs.org

1

MASS SPECTRA OF ALKYL ESTERS



Alkyl esters occur can occur naturally in animal and plant tissues, and wax esters, for example,
constitute an important natural lipid class. In addition, there may be good practical reasons to
prepare alkyl esters other than methyl for chromatographic and mass spectrometric analysis (see
our web page on this topic). Many of the spectra that follow may not have been published (or at
least illustrated) formally elsewhere.


Fatty Acid Ethyl Esters

Fatty acid ethyl esters can be found naturally in animal tissues at very low levels in certain
circumstances, and in particular they can be a useful marker for excessive alcohol consumption in
humans. I have encountered them as artefacts when methylating reagents have been accidentally
contaminated with ethanol, and I am aware that this has happened to others. In addition, purified
fatty acid preparations intended for pharmaceutical or nutritional applications are often converted to
ethyl esters prior to administration or encapsulation. The mass spectrum of ethyl palmitate
(Ryhage and Stenhagen, 1959) is

The spectrum resembles that of the methyl ester, except that the McLafferty ion (the base peak
see our webpage on methyl esters of saturated fatty acids) is at m/z = 88 instead of 74. In the
high mass range, following the molecular ion, there is an ion at m/z = 255 for loss of the ethyl
group, then one at m/z = 241 from a rearrangement reaction involving expulsion of a C
3
fragment
(C2 to C4). An ion at m/z = 239 adjacent to this represents loss of the ethoxide ion ([M-45]
+
).

Similar features are seen in the spectra that follow as further examples. As with other simple
esters, the McLafferty ion becomes less abundant with an increasing degree of unsaturation.

60 80 100 120 140 160 180 200 220 240 260 280
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
70
88
101
115
129
143
157
185
199 213
227
241
255
273
284
%
M
+
CH
3
CH
2
OOC
239

Mass spectra of alkyl esters
W.W. Christie lipidlibrary.aocs.org

2
Mass spectrum
of ethyl oleate













In this instance, the ion for loss of the ethyl group (m/z = 282) is much less abundant than that for
the loss of the ethoxide moiety (m/z = 264).

Mass spectrum of
ethyl linoleate -













Mass spectrum of ethyl 5,8,11,14,17-eicosapentaenoate (20:5(n-3) or EPA) -

















Note that the ion at m/z = 108, typical of fatty acids of the (n-3) family in the spectra of methyl
esters, is independent of the nature of the alcohol moiety so is also prominent in this spectrum, but
the position of the -ion at m/z = 194 is dependent on the nature of the alcohol moiety (see our
web page on methyl esters of trienes).
60 80 100 120 140 160 180 200 220 240 260 280 300
10
20
30
40
50
60
70
80
90
m/z
Abundance 55
69
83
88
111
123
137 152
180
194 207
222
235
246
264
310
%
M
+
CH
3
CH
2
OOC

60 80 100 120 140 160 180 200 220 240 260 280 300
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
67
81
95
109
123
135
150
178
192
206
220
234
253
263/4
279
293
308
%
M
+
CH
3
CH
2
OOC

60 80 100 120 140 160 180 200 220 240 260 280 300
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
67
79
88
91
119
105
133
147
175
194
194
201
215
234
248
261
287
301
330
%
108
108
M
+
CH
3
CH
2
OOC



Mass spectra of alkyl esters
W.W. Christie lipidlibrary.aocs.org

3

Mass spectra of many more ethyl esters are available in our Archive pages, but without
interpretation.


n-Propyl-, i-Propyl and n-Butyl Esters

Occasionally, it may be necessary to prepare esters other than methyl for specific purposes, and
some examples follow. Propan-2-ol esters may have a slight advantage over methyl esters in mass
spectrometric and chromatographic terms as regards separation of geometric isomers and of
highly unsaturated fatty acids (see our webpage on alternatives to methyl esters).

Mass spectrum of n-propyl hexadecanoate (16:0)















The ion at m/z = 239 ([M-59]
+
) reflects the loss of a propyloxy ion, while that at m/z = 257 ([M-41]
+

or RCO
2
H
2
+
) is presumably due to the loss of a propenyl moiety. The latter is often the most
abundant ion in spectra of saturated long-chain esters. The McLafferty ion is at m/z = 102.

Mass spectrum of n-propyl 9,12-octadecadienoate (linoleate)















As in the spectrum of methyl linoleate, hydrocarbon ions predominate. The ion representing the
loss of the propyloxy ion (m/z = 263) is prominent, but in contrast to the previous spectrum that for
loss of a propenyl ion (m/z = 279) is not.

Fatty acid esters of iso-propanol (propan-2-ol) have excellent gas chromatographic properties, and
for example they elute appreciable before most higher aliphatic esters in general and the
analogous ethyl and n-propyl esters in particular. They have been recommended for the separation
60 80 100 120 140 160 180 200 220 240 260 280
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
61
73
97
102
129
143
157
171
185 199
213
227
239
257
298
115
%
M
+
CH
3
(CH
2
)
2
OOC
*
*
*

60 80 100 120 140 160 180 200 220 240 260 280 300
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
67 81
95
109
135
150
164
178
191
220
234
263
279
322
%
M
+
CH
3
(CH
2
)
2
OOC
*
*

Mass spectra of alkyl esters
W.W. Christie lipidlibrary.aocs.org

4
of positional and geometrical isomers of mono- and polyenoic fatty acids (see an article on this
theme in our Topics pages).

Mass spectrum of i-propyl hexadecanoate (16:0)
















The spectrum is somewhat different from that of the corresponding n-propyl derivative, though
mainly in the relative abundances of ions. The base peak is at m/z = 256 for loss of a propenyl
moiety (m/z = 256 ([M-42]
+
rather than ([M-41]
+
).

Similarly, other than the more abundant ion for loss of a propenyl moiety, the mass spectrum of i-
propyl 9,12-octadecadienoate (linoleate) differs in minor ways only from that of the n-propyl ester
above
















On the other hand, the ion for loss of a propyl unit (now at [M-43]
+
or (m/z = 327)) is apparent even
in the spectrum of i-propyl docosahexaenoate, so that the molecular weight can at least be
determined. The -ion at m/z = 108 is apparent, but the -ion is barely detectable, possibly
because this can undergo elimination of propyl and other moieties too readily.
60 80 100 120 140 160 180 200 220 240 260 280
10
20
30
40
50
60
70
80
90
m/z
Abundance
60
71
97
102
129
143
157
171 185
199
213
239
256
298
%
M
+
CHOOC
CH
3
CH
3
*
*
*

60 80 100 120 140 160 180 200 220 240 260 280 300
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
67
81
95
109
123
151
137
163
181 195
209 223
243
263
279
322
M
+
%
CHOOC
CH
3
CH
3

Mass spectra of alkyl esters
W.W. Christie lipidlibrary.aocs.org

5

In comparison, it is often difficult to determine the molecular weight of methyl esters of
polyunsaturated fatty acids such as this. As isopropyl esters have excellent GC properties, it might
be of value to study them further.

Butyl esters are sometimes favoured for the analysis of short-chain fatty acids such as those in
milk fat. The mass spectrum of n-butyl hexadecanoate (16:0) is

By analogy with the previous spectra, the ion at m/z = 239 ([M-73]
+
) reflects the loss of a butyloxy
ion, while that at m/z = 257 ([M-55]
+
) is presumably due to the loss of a butenyl moiety. The
McLafferty ion is at m/z = 116. The butyl ion at m/z = 56 is the base ion.

Similarly, the mass spectrum of n-butyl 9,12-octadecadienoate (linoleate) has comparable
features to that of the n-propyl ester above
60 80 100 120 140 160 180 200 220 240 260 280 300
10
20
30
40
50
60
70
80
90
m/z
Abundance 56
73
97
116
129
143
157
185
199
213
227
239
257
269
283
312
%
M
+
CH
3
(CH
2
)
3
OOC
*
*
*

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340
10
20
30
40
50
60
70
80
90
m/z
55
67
79
91
105
108
108
119
145
159
194
194
215 241
259
285 311
327
370
M
+
CHOOC
CH
3
CH
3
[M-43]
+
Abundance
%

Mass spectra of alkyl esters
W.W. Christie lipidlibrary.aocs.org

6
















In this instance, the first significant ion in the high mass region at m/z = 279 ([M-57]
+
) represents
loss of a butyl rather than a butenyl moiety. Mass spectra from many more n-propyl, i-propyl and n-
butyl esters are available in our Archive pages but without interpretation.


Wax Esters

Wax esters are among the most abundant lipids in nature as part of the wax components that
cover every green leaf. In most instances, the fatty acid and alcohol components are saturated or
monoenoic, although polyunsaturated fatty acids are found also in waxes of marine origin. The
early mechanistic studies of mass spectral fragmentation of was esters were concerned with fully
saturated compounds (Aasen et al., 1971; Ryhage and Stenhagen, 1959), but a more
comprehensive study is now available (Urbanova et al., 2012).

The mass spectrum of hexadecanyl tetradecanoate (14:0acid-16:0alcohol) is -

For an ester of the type RCOOR, the important diagnostic ions are for RCO
2
H
2
+
, i.e. derived from
the acid component and at m/z = 229 in this instance, for [R-1]
+
(m/z = 224), and for ROOC
+
(m/z
= 269). There is no dubiety about the molecular ion (m/z = 452), but all other ions, including the
McLafferty rearrangement ion, are small.

60 80 100 120 140 160 180 200 220 240 260 280 300 320
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
67
81
95
109
135
150
164
220
234
263
279
336
%
M
+
CH
3
(CH
2
)
3
OOC

50 100 150 200 250 300 350 400
10
20
30
40
50
60
70
80
90
m/z
Abundance
57
83
97
111
139
171
196
224
211
229
269
297
452
%
M
+
C
O
O

Mass spectra of alkyl esters
W.W. Christie lipidlibrary.aocs.org

7
For comparison, the mass spectrum of the isomeric tetradecanyl hexadecanoate (16:0acid-
14:0alcohol) is

In this example, the important diagnostic ions are for RCO
2
H
2
+
(at m/z = 257), for [R-1]
+
(m/z =
196), and for ROOC
+
(m/z = 241). The McLafferty rearrangement ion at m/z = 312 is just
discernable.

The mass spectra of unsaturated wax esters are somewhat different in that ions containing the
double bond tend to dominate (Spencer, 1979). That of octadecanyl hexadecenoate (16:1acid-
18:0alcohol), i.e. with a double bond in the fatty acid component is -



















The base peak at m/z = 236 now represents [RCO-1]
+
, while the other diagnostic ions for RCO
2
H
2
+

(at m/z = 255), for [R-1]
+
(m/z = 252), and for ROOC
+
(m/z =297), together with the molecular ion,
are much smaller than in fully saturated wax esters.

For comparison, the mass spectrum of the isomeric octadecenyl hexadecanoate (16:0acid-
18:1alcohol), i.e. with the same molecular weight but with a double bond in the fatty alcohol
component is

50 100 150 200 250 300 350 400
10
20
30
40
50
60
70
80
90
m/z
Abundance
57
83 97
111
139
168
196
213
241
257
312
452
%
M
+
C
O
O

50 100 150 200 250 300 350 400 450
10
20
30
40
50
60
70
80
90
m/z
Abundance
57
69
97
83
125
111
152
194
218
236
255
297
325
379
424
506
%
M
+
C
O
O
Mass spectra of alkyl esters
W.W. Christie lipidlibrary.aocs.org

8


















Now the base peak is the [R-1]
+
ion at m/z = 250, i.e. derived from the alcohol component, while
other useful ions are for RCO
2
H
2
+
(m/z = 257), RCO
+
(m/z = 239), and ROOC
+
(m/z =295).

Mass spectra of more wax esters are available in our Archive pages.


References

o Aasen, A.J., Hofstetter, H.H., Iyengar, B.T.R. and Holman, R.T. Identification and analysis of wax esters
by mass spectrometry. Lipids, 6, 502-507 (1971).
o Ryhage, R. and Stenhagen, E. Mass spectrometric studies II. Saturated normal long-chain esters of
ethanol and higher alcohols. Arkiv Kemi, 14, 483-495 (1959).
o Spencer, G.F. Alkyloxy-acyl combinations in the wax esters from winterised sperm oil by gas
chromatography-mass spectrometry. J. Am. Oil Chem. Soc., 56, 642-646 (1979).
o Urbanova, K., Vrkoslav, V., Valterova, I., Hakova, M. and Cvacka, J. Structural characterization of wax
esters by electron ionization mass spectrometry. J. Lipid Res., 53, 204-213 (2012).


William W. Christie

James Hutton Institute (and Mylnefield Lipid Analysis), Invergowrie, Dundee (DD2
5DA), Scotland

Last updated: January 14
th
, 2013
50 100 150 200 250 300 350 400 450
10
20
30
40
50
60
70
80
90
m/z
Abundance
55
69
82
96
124
110
152
138
194
222
250
257
239
295
506
M
+
C
O
O
%