Separation and characterization of the aldehydic products of lipid peroxidation stimulated by ADP-Fe2+ in rat liver microsomes

Abstract

1. Methods using t.l.c. and high-pressure liquid chromatography (h.p.l.c.) have been used to separate the complex variety of substances possessing a carbonyl function that are produced during lipid peroxidation. 2. The major type of lipid peroxidation studied was the ADP-Fe2+-stimulated peroxidation of rat liver microsomal phospholipids. Preliminary separation of the polar and non-polar products was achieved by t.l.c.: further separation and identification of individual components was performed by h.p.l.c. Estimations were performed on microsomal pellets and the supernatant mixture after incubation of microsomes for 30 min at 37 degrees C. 3. The polar fraction was larger than the non-polar fraction when expressed as nmol of carbonyl groups/g of liver. In the non-polar supernatant fraction the major contributors were n-alkanals (31% of the total), alpha-dicarbonyl compounds (22%) and 4-hydroxyalkenals (37%) with the extraction method used. 4. Major individual contributors to the non-polar fraction were found to be propanal, 4-hydroxynonenal, hexanal and oct-2-enal. Other components identified include butanal, pent-2-enal, hex-2-enal, hept-2-enal, 4-hydroxyoctenal and 4-hydroxyundecenal. The polar carbonyl fraction was less complex than the non-polar fraction, although the identities of the individual components have not yet been established. 5. Since these carbonyl compounds do not react significantly in the thiobarbituric acid reaction, which largely demonstrates the presence of malonaldehyde, it is concluded that considerable amounts of biologically reactive carbonyl derivatives are released in lipid peroxidation and yet may not be picked up by the thiobarbituric acid reaction.

Effect of supplementation of drumstick (Moringa oleifera) and amaranth (Amaranthus tricolor) leaves powder on antioxidant profile and oxidative status among postmenopausal women

“…Biochemical analysis Blood samples were analysed before and after supplementation for serum retinol (Bessey et al 1946), plasma ascorbic acid (Raghuramulu et al 2003) and malondialdehyde using thiobarbituric acid (Esterbauer et al 1982). Glutathione peroxidase and superoxide dismutase were analysed by kit provided by RANDOX Laboratories Limited, United Kingdom.…”

Increased levels of 4‐hydroxynonenal in human monocytes fed with malarial pigment hemozoin A possible clue for hemozoin toxicity

“…HNE analysis was performed as described [17][18][19][20] after derivatization of HNE with dinitrophenylhydrazine and separation of dinitrophenylhydrazones by thinlayer chromatography and subsequently by HPLC. Peak identification was performed by comparison of retention times of peaks of biological extracts and of standard extracts (pure HNE was a kind gift from Professor H. Esterbauer, Graz, Austria), by co-elution of biological extracts with the reference compounds and by comparison of spectra [19,20].…”

Chemistry and analysis of HNE and other prominent carbonyl-containing lipid oxidation compounds

“…Formed via 15-hydroperoxyeicosatetraenoic acid 15-HpETE (from arachidonic acid) and from 13-hydroperoxy-octadecadienoic acid 13-HpODE (from linoleic acid). [37,144] 4-hydroperoxy-2-nonenal (HpNE) C9…”

“…[ 49,143,144] 4-hydroxy-2-octenal (HOE) C8 Derived from peroxidation products of ω-6 polyunsaturated fatty acids.…”