Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS)
Abstract
:1. Introduction
2. Results
2.1. Extraction of PIPs
2.2. Mass Spectrometric Characterization of DAGs and PIPs
2.3. Identification and Quantification of DAG, PIP and PIP2 in Human T Cells
Species | DAG | PIP | PIP2 | |||
---|---|---|---|---|---|---|
m/z | mol% | m/z | mol% | m/z | mol% | |
28:1 | 528.5 | 2.1 ± 0.6 | - | - | - | - |
28:0 | 530.5 | 0.5 ± 0.2 | - | - | - | - |
30:2 | 554.5 | 1.0 ± 0.1 | - | - | - | - |
30:1 | 556.5 | 3.1 ± 0.8 | - | - | - | - |
30:0 | 558.5 | 1.6 ± 0.6 | - | - | - | - |
32:2 | 582.6 | 1.9 ± 0.5 | - | - | - | - |
32:1 | 584.6 | 5.4 ± 0.9 | - | - | - | - |
32:0 | 586.6 | 3.9 ± 0.6 | - | - | - | - |
34:2 | 610.6 | 11.8 ± 2.5 | 932.5 | 0.5 ± 0.2 | - | - |
34:1 | 612.6 | 12.6 ± 1.8 | 934.5 | 1.1 ± 0.4 | 1014.5 | 3.6 ± 2.8 |
34:0 | 614.6 | 3.2 ± 0.8 | - | - | - | - |
36:4 | 634.6 | 6.4 ± 1.9 | 956.5 | 3.6 ± 1.5 | 1036.5 | 5.5 ± 3.1 |
36:3 | 636.6 | 6.0 ± 4.1 | 958.5 | 0.8 ± 0.1 | 1038.5 | 2.2 ± 2.2 |
36:2 | 638.6 | 5.5 ± 0.8 | 960.5 | 2.4 ± 0.5 | 1040.5 | 4.2 ± 0.3 |
36:1 | 640.6 | 11.2 ± 1.1 | 962.5 | 0.9 ± 0.2 | 1042.5 | 3.3 ± 2.3 |
36:0 | 642.6 | 2.4 ± 0.4 | - | - | - | - |
38:5 | 660.6 | 3.3 ± 0.6 | 982.5 | 5.0 ± 0.6 | 1062.5 | 5.5 ± 0.9 |
38:4 | 662.6 | 10.8 ± 2.2 | 984.5 | 82.2 ± 1.5 | 1064.5 | 70.0 ± 11.1 |
38:3 | 664.6 | 2.1 ± 0.9 | - | - | - | - |
40:6 | - | - | 1008.5 | 0.6 ± 0.3 | 1088.5 | 1.4 ± 0.8 |
40:5 | 688.6 | 2.7 ± 0.4 | 1010.5 | 1.6 ± 0.3 | 1090.5 | 2.1 ± 1.8 |
40:4 | 690.6 | 2.3 ± 0.3 | 1012.5 | 1.5 ± 0.2 | 1092.5 | 2.2 ± 1.5 |
2.4. Effect of TCR Stimulation on DAG, PIP and PIP2 in Human T Cells
3. Discussion
4. Experimental Section
QStar® Elite | QTrap® 5500 | |
---|---|---|
Curtain Gas | 10 | 10 |
CAD Gas | 2 | 5 |
Operating pressure (torr) | 2.5 × 10−5 | 1.6 × 10−5 |
Interface heater temperature IHT (°C) | 40 | 40 |
Declustering potential (DP) | 40 | 100 |
Focusing potential (FP) | 200 | - |
Declustering potential 2 (DP2) | 10 | - |
Entrance potential (EP) | - | 7 |
Collision cell exit potential (CXP) | - | 19 |
Detector (CEM) | 2,500 | 2,100 |
Quadrupole resolution | Unit (Q1) | Unit (Q1 and Q3) |
MAG-H2O | 14:1 | 14:0 | 16:2 | 16:1 | 16:0 | 17:0 | 18:3 | 18:2 | 18:1 |
m/z | 283.2 | 285.2 | 309.2 | 311.3 | 313.3 | 327.3 | 335.3 | 337.3 | 339.3 |
MAG-H2O | 18:0 | 20:5 | 20:4 | 20:3 | 20:2 | 20:1 | 20:0 | 22:6 | 22:5 |
m/z | 341.3 | 359.3 | 361.3 | 363.3 | 365.3 | 367.3 | 369.3 | 385.3 | 387.3 |
5. Conclusions
Supplementary Materials
Acknowledgments
Conflict of Interest
References
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Haag, M.; Schmidt, A.; Sachsenheimer, T.; Brügger, B. Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS). Metabolites 2012, 2, 57-76. https://doi.org/10.3390/metabo2010057
Haag M, Schmidt A, Sachsenheimer T, Brügger B. Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS). Metabolites. 2012; 2(1):57-76. https://doi.org/10.3390/metabo2010057
Chicago/Turabian StyleHaag, Mathias, Angelika Schmidt, Timo Sachsenheimer, and Britta Brügger. 2012. "Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS)" Metabolites 2, no. 1: 57-76. https://doi.org/10.3390/metabo2010057