Dufour Claire, Loonis Michèle
Institut National de la Recherche Agronomique, UMR A408, Safety and Quality of Plant Products, Agroparc, 84914 Avignon Cedex 9, France.
Chem Phys Lipids. 2005 Dec;138(1-2):60-8. doi: 10.1016/j.chemphyslip.2005.08.003. Epub 2005 Sep 12.
An efficient RP-HPLC method was developed for the detection of the oxidation products derived from the AAPH-initiated peroxidation of linoleic acid bound to human serum albumin. Diode array UV-detection allowed the quantification at 234 nm of four regioisomeric hydroperoxyoctadecadienoic acids (HPODE) and four hydroxyoctadecadienoic acids (HODE) while at 280 nm four oxooctadecadienoic acid isomers (KODE) were detected. Full identification of the different underivatized HODE, HPODE and KODE isomers was achieved by negative ESI-mass spectrometry outlining common fragmentation pathways for 9- and 13-regioisomers. Chemical synthesis of 9-(E,Z)-, 9-(E,E)-, 13-(Z,E)- and 13-(E,E)-KODE helped to their structural characterization by 1H NMR. Lipid peroxidation in the presence of albumin proved to be regioselective with a larger accumulation of 13-HPODE and 9-KODE isomers. Thermodynamically more stable E,E-stereoisomers were also favored by albumin for both HPODE and KODE.
开发了一种高效的反相高效液相色谱法，用于检测与人类血清白蛋白结合的亚油酸在2,2'-偶氮二异丁腈引发的过氧化反应中产生的氧化产物。二极管阵列紫外检测可在234 nm处对四种区域异构体氢过氧十八碳二烯酸（HPODE）和四种羟基十八碳二烯酸（HODE）进行定量，而在280 nm处可检测到四种氧代十八碳二烯酸异构体（KODE）。通过负离子电喷雾质谱法对不同的未衍生化HODE、HPODE和KODE异构体进行了全面鉴定，概述了9-和13-区域异构体的常见裂解途径。9-(E,Z)-、9-(E,E)-、13-(Z,E)-和13-(E,E)-KODE的化学合成有助于通过1H NMR对其结构进行表征。在白蛋白存在下的脂质过氧化反应具有区域选择性，13-HPODE和9-KODE异构体的积累量更大。白蛋白对HPODE和KODE的热力学更稳定的E,E-立体异构体也更有利。
