R W Browne1, D Armstrong. 1. Department of Clinical Laboratory Science, State University of New York at Buffalo, 14214, USA. rwbrowne@acsu.buffalo.edu
Abstract
BACKGROUND: Lipid peroxidation is a prominent manifestation of free radical activity and oxidative stress in biological systems. Diverse methodologies have been developed that measure a variety of lipid peroxidation products used as markers of lipid peroxidation processes. METHODS: Hydroxy and hydroperoxy polyunsaturated fatty acid (PUFA) peroxidation products were analyzed in human blood plasma by reversed-phase HPLC after liquid-liquid extraction of total lipids and alkaline hydrolysis of lipid esters to liberate free PUFAs. An isocratic mobile phase containing 1 g/L acetic acid-acetonitrile-tetrahydrofuran (52:30:18, by volume) over 60 min duration, with ultraviolet absorbance detection at 236 nm by photodiode array, enabled the resolution and quantification of 13 regioisomeric hydroxy and hydroperoxy PUFAs. RESULTS: As little as 250 microL of human plasma was utilized with an analytical range of 0.033-1.6 micromol/L for each compound. Intra- and interassay CVs for all compounds detected in normal or oxidatively modified human plasma were 3.2-11% and 4.7-12%, respectively. Analytical recoveries were 87-103%. Analysis of human plasma exposed to artificial oxidation with Cu(2+) ion and hydrogen peroxide, a free radical-generating reaction, showed marked increases in hydroxy and hydroperoxy PUFA concentrations. CONCLUSION: Lipid-derived hydroxy and hydroperoxy PUFAs may be useful as clinical markers of lipid peroxidation and oxidative stress in the peripheral circulation.
BACKGROUND:Lipid peroxidation is a prominent manifestation of free radical activity and oxidative stress in biological systems. Diverse methodologies have been developed that measure a variety of lipid peroxidation products used as markers of lipid peroxidation processes. METHODS:Hydroxy and hydroperoxy polyunsaturated fatty acid (PUFA) peroxidation products were analyzed in human blood plasma by reversed-phase HPLC after liquid-liquid extraction of total lipids and alkaline hydrolysis of lipid esters to liberate free PUFAs. An isocratic mobile phase containing 1 g/L acetic acid-acetonitrile-tetrahydrofuran (52:30:18, by volume) over 60 min duration, with ultraviolet absorbance detection at 236 nm by photodiode array, enabled the resolution and quantification of 13 regioisomeric hydroxy and hydroperoxy PUFAs. RESULTS: As little as 250 microL of human plasma was utilized with an analytical range of 0.033-1.6 micromol/L for each compound. Intra- and interassay CVs for all compounds detected in normal or oxidatively modified human plasma were 3.2-11% and 4.7-12%, respectively. Analytical recoveries were 87-103%. Analysis of human plasma exposed to artificial oxidation with Cu(2+) ion and hydrogen peroxide, a free radical-generating reaction, showed marked increases in hydroxy and hydroperoxy PUFA concentrations. CONCLUSION:Lipid-derived hydroxy and hydroperoxy PUFAs may be useful as clinical markers of lipid peroxidation and oxidative stress in the peripheral circulation.
Authors: Anna Z Pollack; Enrique F Schisterman; Lynn R Goldman; Sunni L Mumford; Neil J Perkins; Michael S Bloom; Carole B Rudra; Richard W Browne; Jean Wactawski-Wende Journal: Am J Epidemiol Date: 2012-02-02 Impact factor: 4.897
Authors: Richard W Browne; Michael S Bloom; Enrique F Schisterman; Kathy Hovey; Maurizio Trevisan; Chengqing Wu; Aiyi Liu; Jean Wactawski-Wende Journal: Biomarkers Date: 2008-03 Impact factor: 2.658
Authors: R Colas; A Sassolas; M Guichardant; C Cugnet-Anceau; M Moret; P Moulin; M Lagarde; C Calzada Journal: Diabetologia Date: 2011-08-17 Impact factor: 10.122
Authors: Zhi-Xin Yuan; Stanley I Rapoport; Steven J Soldin; Alan T Remaley; Ameer Y Taha; Matthew Kellom; Jianghong Gu; Maureen Sampson; Christopher E Ramsden Journal: Biomed Chromatogr Date: 2012-10-05 Impact factor: 1.902
Authors: Anna Z Pollack; Lindsey Sjaarda; Katherine A Ahrens; Sunni L Mumford; Richard W Browne; Jean Wactawski-Wende; Enrique F Schisterman Journal: PLoS One Date: 2014-03-28 Impact factor: 3.240