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Application No.C137
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The LCMS-8050 system was used with a phospholipid For the 33 possible fatty acid compositions, the peak
st
MRM library 1 method (MRM of 422 phospholipid areas obtained from MRM monitoring of polar head
constituents) to profile a lipid extract obtained from groups are shown in the vertical axis in Fig. 3. For PC
mouse brain. As a result, the peaks were detected for (38:4), the MRM chromatogram obtained from the 2 nd
130 constituents, and the peak heights of 102 method is shown on the bottom left of Fig. 3. The
constituents were 10,000 or above. MRM chromatogram obtained when monitoring for
nd
MRM Event Link Editor was used to create a 2 the polar head group (choline), which is shown above
nd
method, which is needed to determine the fatty acid the 2 method chromatogram, detected a main peak
st
composition of the peaks detected by the 1 method, at 15.5 minutes. The MRM chromatogram below is for
and analysis was performed. Taking PC (38:4) as an fatty acid product ions with compositions of 18:0
example, the MRM Event Link Editor software creates (stearic acid) and 20:4 (arachidonic acid). The MRM
a method from the MRM transitions expected for all chromatograms obtained for other fatty acid
possible combinations of fatty acids (18:0/20:4, combinations did not detect a peak at 15.5 minutes,
18:1/20:3, 18:2/20:2, 18:3/20:1) in ESI negative mode showing that the main fatty acid combination is
(ESI (-)), and takes those transitions from the 867 18:0/20:4.
nd
MRM transitions in the MRM library. The 2 method This article shows the LC/MS/MS MRM library
st
would also include the MRM transitions used in the 1 (phospholipids) can be used for easy phospholipid
method to monitor for eliminated polar head groups. profiling and fatty acid composition determination.

1.E+08

1.E+07

1.E+06

1.E+05
PC (30:1-14:0/16:0) PC (32:0-16:0/16:0) PC (32:1-16:0/16:1) PC (34:0-16:0/18:0) PC (34:1-16:0/18:1) PC (34:2-16:0/18:2) PC (36:0-16:0/20:0) PC (36:1-18:0/18:1) PC (36:2-18:1/18:1) PC (36:3-18:0/20:3) PC (36:4-18:0/20:4) PC (38:1-18:0/20:1) PC (38:2-18:1/20:1) PC (38:3-18:0/20:3) PC (38:4-18:0/20:4) PC (38:5-18:1/20:4) PC (38:6-16:0/22:6) PC (40:1-18:1/22:0) PC (40:5-20:1/20:4) PC (40:6-18:0/22:6) PC (40:7-18:1/22:6) PE (36:2-

5.0E+6 810.6>184 (+) 283 (–)
2.5E+6 O O
184 (+)
O O
0 O O O O
854.6>283 (–) nd
1.0E+5 2 Method + O P O O
854.6>303 (–) N – – O O
O
0.5E+5
303 (–) PC (18:0/20:4)
0
14.0 15.0 16.0 17.0 min
Fig. 3 Profiling results for PC in mouse brain lipid extract (top image). The MRM chromatogram used to determine the fatty acid
composition of PC (38:4) (bottom left) showed the fatty acid composition was 18:0/20:4 (bottom right).

First Edition: Oct. 2016

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