Identification of trans,trans-2,4-Decadienal Metabolites in Mouse and Human Cells Using Liquid Chromatography–Mass Spectrometry

Abstract

trans,trans-2,4-Decadienal (tt-DDE), a lipid peroxidation product of linolieic acid, is the most abundant aldehyde identified in cooking oil fumes and is readily detectable in food products as well as in restaurant emissions. Previously, we have reported the toxicological effects of tt-DDE in vitro and in vivo. However, the metabolic pathways of tt-DDE in vivo remain unclear. In our present study, we combined liquid chromatography-mass spectrometry with triple quadrupole and time-of-flight to identify tt-DDE metabolites in the urine of mice orally administered tt-DDE. We identified two tt-DDE metabolites, 2,4-decadienoic acid and cysteine-conjugated 2,4-decadien-1-ol, in the urine of mice gavaged with tt-DDE and in human hepatoma cell cultures. The structure of 2,4-decadienoic acid was confirmed upon comparison of its tandem mass spectrometry (MS/MS) spectrum and retention time with those of synthetic standards. The moieties of cysteine and alcohol on cysteine-conjugated 2,4-decadien-1-ol were validated by treating cell cultures with stable-isotope-labeled cysteine and 4-methylpyrazole, an alcohol dehydrogenase inhibitor. The MS/MS spectra of a cysteine standard and ionized cysteine detached from cysteine-conjugated 2,4-decadien-1-ol were identical. Two metabolic pathways for the biotransformation of tt-DDE in vivo are proposed: (i) the oxidation of tt-DDE to the corresponding carboxylic acid, 2,4-decadienoic acid, in liver cells and (ii) glutathione (GHS) conjugation, GSH breakdown, and aldehyde reduction, which generate cysteine-conjugated 2,4-decadien-1-ol in both liver and lung cells. In conclusion, this platform can be used to identify tt-DDE metabolites, and cysteine-conjugated 2,4-decadien-1-ol can serve as a biomarker for assessing exposure to tt-DDE.

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

“…Trans,trans-2,4-decadienal (tt-DDE), a lipid peroxidation product of linoleic acid, can be oxidized to the two metabolites 2,4-decadienoic acid and cysteine-conjugated 2,4-decadien-1-ol. These have been found in cell culture models and urine of mice gavaged with tt-DDE, using liquid chromatography coupled to tandem mass spectrometry to identify the structures [96]. Carboxylic acid metabolites of HHE (4-hydroxy-2E-hexenoic acid; 4-HHA), HNE (4-hydroxy-2E-nonenoic acid; 4-HNA) and HDDE (4-hydroxy-2E,6Z-dodecadienoic acid; 4-HDDA) were detected in human urine using NICI-MS, and occurred at higher concentrations in aging and diabetes [50].…”

Diunsaturated Aldehyde, trans,trans-2,4-Decadienal in the Intestinal Lumen Suppresses Gastric Emptying through Serotonin Signaling in Rats

“…A recent study demonstrated that orally administered 2,4-decadienal was metabolized to 2,4-decadienoic acid and a cysteine-conjugated compound. 19 Unsaturated aldehydes are generated during lipid peroxidation and play critical roles in oxidative stress. 20,21 The structure activity relationship demonstrated in the present study suggests that serotonin-producing enteroendocrine cells sense luminal unsaturated aldehydes to prevent the excess absorption and ingestion of potent peroxides such as unsaturated aldehydes through delaying gastric emptying followed by suppression of appetite.…”

Effect of (E,E)‐2,4‐decadienal on Side‐Chain Modification, Conformation Change, and Aggregation of Bovine Serum Albumin

“…Because (E,E)‐2,4‐decadienal is highly reactive and exhibits teratogenicity and carcinogenicity, most of the research is focused on DNA damage, [ 13 ] cytotoxic effects on smooth muscle cells, [ 14 ] pulmonary changes, [ 15 ] cytotoxicity to human vascular SMC, [ 14 ] effects on human bronchial epithelial cells, [ 16 ] and mitochondrial dysfunction and oxidative stress [ 17 ] induced by the aldehyde, as well as identification of its metabolites. [ 18 ] However, in the field of food science, most of the research focuses on the monitoring of the formation of (E,E)‐2,4‐decadienal during processing [ 19–22 ] or studying the Maillard reaction of (E,E)‐2,4‐decadienal and amino acids in a heating system. [ 23,24 ] Only a few studies have focused on the reaction of (E,E)‐2,4‐decadienal with proteins; for example, mass spectrometry was used to identify the (E,E)‐2,4‐decadienal‐mediated modification sites in cytochrome c [ 25 ] as well as ribonuclease A and lactoglobulin.…”