Abstract
High performance liquid chromatography/electron spin resonance/mass spectrometry (HPLC/ EPR/MS) analyses of radicals is performed for the reaction mixture of 13-hydroperoxy-(9Z,11E)-octadeca-9,11-dienoic acid (13ZE-OOH) with ferrous ions under an aerobic condition, or an anaerobic condition. Radicals are identified from 13ZE-OOH by using high performance liquid chromatography/electron spin resonance spectrometry (HPLC/EPR) and HPLC/EPR/MS. The pentyl radical and isomers of epoxylinoleic acid radicals from 13ZE-OOH are identified under an anaerobic condition and the 7-carboxyheptyl radical and pentyl radical from 13ZE-OOH under an aerobic condition. These results suggest that the formation of the respective radical species depends to a great extent on oxygen concentration in the reaction mixtures.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Li, A.S.W., Cummings, K.B., Roethling, H.P., Buettner, G.R., and Chignell, C.F. (1988) A spin-trapping database implemented on the IBM PC/AT. J. Magn. Reson., 79, 140–142.
Rokushika, S., Taniguchi, H., and Hatano, H. (1975) Flow ESR detector for liquid chromatography of radicals. Anal. Lett., 8, 205–213.
Makino, K., and Hatano, H. (1979) Separation and characterization of short-lived radicals in DL-methionine aqueous solution by high speed liquid chromatograph equipped with ESR spectrometer. Chem. Lett., 119ñ122.
Iwahashi, H., Negoro, Y., Ikeda, A., Morishita, H., and Kido, R. (1986) Inhibition by chlorogenic acid of haematin-catalysed retinoic acid 5,6-epoxidation. Biochem. J., 239, 641–646.
Iwahashi, H., Parker, E.C., Mason, R.P., and Tomer, K.B. (1992) Combined liquid chromatography/electron paramagnetic resonance spectrometry/electrospray ionization mass spectrometry for radical identification. Anal. Chem., 64, 2244–2252.
Iwahashi, H., Hirai, T., and Kumamoto, K. (2006) High performance liquid chromatography/electron spin resonance/mass spectrometry analyses of radicals formed in anaerobic reaction of 9- (or 13-) hydroperoxide octadecadienoic acids with ferrous ions. J. Chromotogr. A, 1132, 67–75.
Minakata, K., Okuno, E., Nakamura, M., and Iwahashi, H. (2007) Identification of radicals formed in the reaction mixtures of rat liver microsomes with ADP, Fe31 and NADPH Using HPLC-EPR and HPLC-EPR-MS. J. Biochem., 142, 73–78.
Iimura, S., and Iwahashi, H. (2006) Enhancement by cigarette smoke extract of the radical formation in a reaction mixture of 13-hydroperoxide octadecadienoic acid and ferric ions. J. Biochem., 139, 671–676.
Kumamoto, K., Hirai, T., Kishioka, S., and Iwahashi, H. (2007) Identification of a radical formed in the reaction mixture of rat brain homogenate with a ferrous ion/ascorbic acid system using HPLC-EPR and HPLC-EPR-MS. Free Radic. Res., 41, 650–654.
Iwahashi, H., Deterding, L.J., Parker, C.E., Mason, R.P., and Tomer, K.B. (1996) Identification of radical adducts formed in the reactions of unsaturated fatty acids with soybean lipoxygenase uing continuous flow fast atom bombardment with tandem mass spectrometry. Free Radic. Res., 25, 255–274.
Spiteller, G. (1996) Enzymic lipid peroxidation ñ consequence of cell injury? Free Radic. Biol. Med., 25, 1003–1009.
Jira, W., Spiteller, G., Carson, W., and Schramm, A. (1998) Strong increase in hydroxy fatty acids derived from linoleic acid in human low density lipoproteins of atheroscleorotic patients. Chem. Phys. Lipids., 91, 1–11.
Jira, W., Richter, A., and Spiteller, G. (1997) Increased levels of lipid oxidation products in low density lipoproteins patients suffering from rheumatoid arthritis. Chem. Phys. Lipids., 87, 81–89.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Iwahashi, H. (2008). High Performance Liquid Chromatography/Electron Spin Resonance/Mass Spectrometry Analyses of Lipid-Derived Radicals. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress I. Methods In Molecular Biology, vol 477. Humana Press. https://doi.org/10.1007/978-1-60327-517-0_6
Download citation
DOI: https://doi.org/10.1007/978-1-60327-517-0_6
Publisher Name: Humana Press
Print ISBN: 978-1-60327-218-6
Online ISBN: 978-1-60327-517-0
eBook Packages: Springer Protocols