AJAC  Vol.4 No.10 B , October 2013
Molecular Probes in Tandem Electrospray Ionization Mass Spectrometry: Application to Tracing Chemical Changes of Specific Phospholipid Molecular Species
Abstract: New ionization and detection techniques in mass spectrometry have been successfully applied for efficient analyses of complex biological systems. It is, however, still difficult to trace structural changes of a specific molecular species in such systems. In the present study, a molecular probe strategy in combination with tandem electrospray ionization mass spectrometry has been examined using synthetic deuterium-labeled phosphatidylcholine hydroperoxide (PC-OOH/D3) and ethyl-labeled phosphatidylcholine having docosahexaenoic acid side chain (DHA-PC/Et). Administration of a mixture of PC-OOH/D3 and DHA-PC/Et to human blood and human skin surface, followed by extraction and analysis with collision-induced tandem electrospray ionization mass spectrometry demonstrated that metabolites of both molecular probes can be detected simultaneously with strict selectivity. The present method is also found to be useful in tracing chemical changes of the unstable docosahexaenoyl group on the surface of processed fish. The activity of phospholipase A2 can also be assessed using a phospholipid molecular probe with a linoleoyl and a deuteriomethyl group via selective detection of the lyso-phospholipid product by mass spectrometry. The advantage of the present method is that no chromatographic separation is required and analysis can be performed under strictly the same condition for different molecular probes, affording multiple data by one experiment. The present strategy may be useful for tracing time-dependent phenomena in dynamic phospholipid biochemistry, and can be widely used for any biological and food systems.
Cite this paper: H. Tominaga, T. Ishihara, A. Shah, R. Shimizu, A. Onyango, H. Ito, T. Suzuki, Y. Kondo, H. Koaze, K. Takahashi and N. Baba, "Molecular Probes in Tandem Electrospray Ionization Mass Spectrometry: Application to Tracing Chemical Changes of Specific Phospholipid Molecular Species," American Journal of Analytical Chemistry, Vol. 4 No. 10, 2013, pp. 16-26. doi: 10.4236/ajac.2013.410A2003.

[1]   A. Shevchenko and K. Simons, “Lipidomics: Coming to Grips with Lipid Diversity,” Nature Reviews Molecular Cell Biology, Vol. 11, No. 8, 2010, pp. 593-598.

[2]   E. Fahy, D. Cotter, M. Sud and S. Subramaniam, “Lipid Classification, Structures and Tools,” Biochimica et Biophysica Acta (BBA)—Molecular and Cell Biology of Lipids, Vol. 1811, No. 11, 2011, pp. 637-647.

[3]   A. Lamaziere, D. Richard, U. Barbe, K. Kefi, P. Bausero, C. Wolf and F. Visiol, “Differential Distribution of DHA-Phospholipids in Rat Brain after Feeding: A Lipidomic Approach,” Prostaglandins Leukot Essent Fatty Acid, Vol. 84, No. 1, 2011, pp. 7-11.

[4]   P. Fagone and S. Jackowski, “Membrane Phospholipid Synthesis and Endoplasmic Reticulum Function” The Journal of Lipid Research, Vol. 50, 2009, pp. S311-S316.

[5]   K. Halder, A. F. de Amorim and A. M. Cross, “Transport of Fluorescent Phospholipid Analogues from the Erythrocyte Membrane to the Parasite in Plasmodium falciparum-Infected Cells,” The Journal of Cell Biology, Vol. 108, No. 6, 1989, pp. 2183-2102.

[6]   D. Marsh, “Electron Spin Resonance in Membrane Research: Protein-Lipid Interactions from Challenging Beginnings to State of The Art,” European Biophysics Journal, Vol. 39, No. 4, 2010, pp. 513-525.

[7]   M. Pulfer and R. C. Murphym, “Electrospray Mass Spectrometry of Phospholipids,” Mass Spectrometry Reviews, Vol. 22, No. 5, 2003, pp. 332-364.

[8]   V. E. Kagan, G. G. Borisenko, Y. Y. Tyurina, T. A. Tyurin, J. Jiang, A. I. Potapovich, V. Kini, A. A. Amoscato and Y. Fuji, “Oxidative Lipidomics of Apoptosis: Redox Catalytic Interactions of Cytochrome c with Cardiolipin and Phosphatidylserine,” Free Radical Biology & Medicine, Vol. 37, No. 12, 2004, pp.1963-1985.

[9]   Katja Dettmer, P. A. Aronov and B. D. Hammock, “Mass Spectrometry-Based Metabolomics,” Mass Spectrometry Reviews, Vol. 26, No. 1, 2007, pp. 51-78.

[10]   M. Rosario, M. Domingues, A. Reis and P. Domingues, “Mass Spectrometry Analysis of Oxidized Phospholipids,” Chemistry and Physics of Lipids, Vol. 156, No. 1-2, 2008, pp. 1-12.

[11]   A. D. Postle and A. N. Hunt, “Dynamic Lipidomics with Stable Isotope Labeling,” Journal of Chromatography B, Vol. 877, No. 26, 2009, pp. 2716-2721.

[12]   R. J. Mishur and S. L. Rea, “Applications of Mass Spectrometry to Metabolomics and Metabonomics: Detection of Biomarkers of Againg and of Age-Related Diseases,” Mass Spectrometry Reviews, Vol. 31, No. 1, 2012, pp. 70-95.

[13]   A. Thomas, J. Déglon, S. Lenglet, F. Mach, P. Mangin, J. Wolfender, S. Steffens and C. Staub, “High-Throughput Phospholipidic Fingerprinting by Online Desorption of Dried Spots And Quadrupole-Linear Ion Trap Mass Spectrometry: Evaluation of Atherosclerosis Biomarkers In Mouse Plasma,” Analytical Chemistry, Vol. 82, No. 15, 2010, pp. 6687-6694.

[14]   V. N. Bochkov and N. Leitinger, “Anti-Inflammatory Properties of Lipid Oxidation Products,” Journal of Molecular Medicine, Vol. 81, No. 10, 2003, pp. 613-626.

[15]   G. K. Marathe, A. Harrison, R. C. Murphy, S. M. Prescott, G. A. Zimmerman and T. M. McIntyre, “Bioactive Phospholipid Oxidation Products,” Free Radical Biology & Medicine, Vol. 28, No. 12, 2006, pp. 1762-1770.

[16]   C. Erridge, S. Kennedy, C. M. Spickett and D. J. Webb, “Oxidized Phospholipid Inhibition of Toll-Like Receptor (TLR) Signaling Is Restricted to TLR2 and TLR4, “The Journal of Biological Chemistry,” Vol. 283, 2008, pp. 24748-24759.

[17]   R. A. Siddiqui, K. Harvey and W. Stillwell, “Anticancer Properties of Oxidation Products of Docosahexaenoic Acid,” Chemistry and Physics of Lipids, Vol. 153, No. 1, 2008, pp. 47-56.

[18]   G. K. Marathe, K. A. Harrison, R. C. Murphy, S. M. Prescott, G. A. Zimmerman and T. M. ScIntyre, “Bioactive Phospholipid Oxidation Products”, Free Radical Biology & Medicine, Vol. 28, No. 12, 2000, pp. 1762-1770.

[19]   T. Miyazawa, T. Suzuki, K. Fujimoto and M. Kinoshita, “Age-Related Change of Phosphatidylcholine Hydroperoxide and Phosphatidylethanolamine Hydroperoxide Levels in Normal Human Red Blood Cells,” Mechanisms of Ageing and Development, Vol. 86, No. 3, 1996, pp. 145-150.

[20]   V. Frisardi, F. Panza, D. Seripa, T. Farooqui and A. A. Farooqui, “Glycerophospholipids and Glycerophospholipid-Derived Lipid Mediators: A Complex Meshwork in Alzheimer’s Disease Pathology,” Progress in Lipid Research, Vol. 50, No. 4, 2011, pp. 313-330.

[21]   R. Shimizu, A. Nagai, H. Tominaga, H. Tominaga, M. Imura, A. N. Onyango, M. Izumi, S. Nakajima, S. Tahara, T. Kaneko and N. Baba, “A Combination of Molecular Probe-Tandem Electrospray Ionization Mass Spectrometry: A Technique for Tracing Structural Changes in Phospholipid Hydroperoxides,” Bioscience, Biotechnology, and Biochemistry, Vol. 73, No. 3, 2009, pp. 781-784.

[22]   N. Baba, H. Daido, T. Kosugi, M. Miyake and S. Nakajima, “Analysis of Glycerophospholipid Hydroperoxides by Ion Spray Mass Spectrometry,” Bioscience, Biotechnology, and Biochemistry, Vol. 62, No. 1, 1998, pp. 160-163.

[23]   A. N. Onyango, N. Kumura, H.Tominaga and N. Baba, “Dihydroperoxidation Facilitates the Conversion of Lipids to Aldehydic Products via Alkoxyl Radicals,” Food Research International, Vol. 43, No. 3, 2010, pp. 925-929.

[24]   H. Kern, T. Volk, S. Knaueru-Schiefer, T. Mieth, B. Bustow, W. J. Kox and M. Schlame, “Stimulation of Monocytes and Platelets by Short-Chain Phosphatidylcholines with and without Terminal Carboxyl Group,” Biochimica et Biophysica Acta (BBA)—Molecular and Cell Biology of Lipids, Vol. 1394, No. 1, 1998, pp. 33-42.

[25]   S. Hong and Y. Lu, “Omega-3 Fatty Acid-Derived Resolvins and Protectins in Inflammation Resolution and Leukocyte Functions: Targeting Novel Lipid Mediator Pathways in Mitigation of Acute Kidney Injury,” Frontiers in Immunology, Vol. 4, 2013, p. 13.

[26]   C. D. Luca and G. Valacchi, “Surface Lipids as Multifunctional Mediators of Skin Responses to Environmental Stimuli,” Mediators of Inflammation, Vol. 2010, 2010, pp. 1-11.

[27]   A. K. M. Azad Shah, T. Ishihara, M. Ogasawara, H. Kurihara, N. Baba and K. Takahashi, “Mechanism Involved in the Formation of Characteristic Taste and Flavor During the Production of Dried Herring (Clupea pallasii) Fillet,” Food Science and Technology Research, Vol. 16, No. 3, 2010, pp. 201-208.