α tocopherol

Reagents were obtained from the following sources: CBD from THC Pharm GmbH (Frankfurt, Germany); 8-iso prostaglandin F_2α-d4 (8-isoPGF_2α–d4), anandamide-d8 (AEA-d8), 2-arachidonylglycerol-d8 (2-AG-d8), oleoylethanolamide-d4 (OEA-d4), cannabidol-d9 (CBD-d9), leukotriene B4-d4 (LTB4-d4), 5(S)-hydroxy-5Z,8Z,11Z,13E-eicosatetraenoic-5,6,8,9,11,12,14,15-d8 acid (15-HETE-d8) and prostaglandin D2-d4 (PGD2-d4) from Cayman Chemical Company (Ann Arbor, MI, USA); α-tocopherol, retinol, L-glutathione reduced (GSH), glutathione peroxidase (GSH-Px), glutathione reductase (GSSG-R), catalase (CAT), thioredoxin (Trx), β-nicotinamide adenine dinucleotide phosphate hydrate (NADP), β-nicotinamide adenine dinucleotide phosphate, reduced tetra (cyclohexylammonium) salt (NADPH), hexane, butylated hydroxytoluene (BHT), ethanol, O-(2,3,4,5,6-pentafluoro-benzyl) hydroxylamine hydrochloride (PFBHA-HCl), 4-hydroxynonenal (4-HNE) and benzaldehyde-d6 from Sigma-Aldrich (Saint Louis, MO, USA)

Isoprenoids (carotenoids and tocopherols) were extracted from triplicate aliquots of freeze dried tomato powder (0.1 g) as reported by Sadler et al. [41 (link)] modified by Perkins-Veazie et al. [42 (link)]. The extracts were assayed as described by Fraser et al. [43 (link)] using an 1100 Series HPLC system (Agilent Technologies, California, USA) equipped with a YMC Reversed-Phase HPLC C30 Columns (Wilmington, NC, USA). The chromatographic separation was performed using the following solvents: methanol (A), water/methanol (20:80 v/v) containing 0.2% ammonium acetate (B), and tert-methyl butyl ether (C). Samples were initially eluted with 95% A and 5% B; 0 to 12 min, 80% A, 5% B, and 15% C; 12 to 42 min, 30% A, 5% B, and 65% C; 42 to 60 min, 30% A, 5% B, and 65% C; 60 to 62 min, 95% A, and 5% B. Ten microliter aliquots of the samples were injected in the HPLC-DAD system. The flow rate was 1.0 mL min⁻¹, and the column temperature was maintained at 25 °C. Diode array detection was performed at 475 nm for carotenoids and 290 nm for tocopherols. Peaks were identified by comparing their retention times and UV-vis spectra to those of authentic standards. α-tocopherol, ß-tocopherol and γ-tocopherol were purchased from Cayman chemicals (Ann Arbor, MI, USA). Lutein, α-Carotene, ß-Carotene and lycopene were purchased from CaroteNature (Lupsingen, Base-land, Switzerland).