TitleNovel liquid chromatography-mass spectrometry method shows that vitamin E deficiency depletes arachidonic and docosahexaenoic acids in zebrafish (Danio rerio) embryos.
Publication TypeJournal Article
Year of Publication2013
AuthorsLebold, KM, Kirkwood, JS, Taylor, AW, Choi, J, Barton, CL, Miller, GW, La Du, J, Jump, DB, Stevens, JFrederik, Tanguay, RL, Traber, MG
JournalRedox Biol
Volume2
Pagination105-13
Date Published2013
ISSN2213-2317
Keywordsalpha-Tocopherol, Animals, Arachidonic Acid, Cholesterol, Chromatography, High Pressure Liquid, Docosahexaenoic Acids, Embryo, Nonmammalian, Fatty Acids, Unsaturated, Lipid Peroxidation, Mass Spectrometry, Solid Phase Extraction, Vitamin E, Vitamin E Deficiency, Zebrafish
Abstract

To test the hypothesis that embryogenesis depends upon α-tocopherol (E) to protect embryo polyunsaturated fatty acids (PUFAs) from lipid peroxidation, new methodologies were applied to measure α-tocopherol and fatty acids in extracts from saponified zebrafish embryos. A solid phase extraction method was developed to separate the analyte classes, using a mixed mode cartridge (reverse phase, π-π bonding, strong anion exchange), then α-tocopherol and cholesterol were measured using standard techniques, while the fatty acids were quantitated using a novel, reverse phase liquid chromatography-mass spectrometry (LC-MS) approach. We also determined if α-tocopherol status alters embryonic lipid peroxidation products by analyzing 24 different oxidized products of arachidonic or docosahexaenoic (DHA) acids in embryos using LC with hybrid quadrupole-time of flight MS. Adult zebrafish were fed E- or E+ diets for 4 months, and then were spawned to obtain E- and E+ embryos. Between 24 and 72 hours post-fertilization (hpf), arachidonic acid decreased 3-times faster in E- (21 pg/h) compared with E+ embryos (7 pg/h, P<0.0001), while both α-tocopherol and DHA concentrations decreased only in E- embryos. At 36 hpf, E- embryos contained double the 5-hydroxy-eicosatetraenoic acids and 7-hydroxy-DHA concentrations, while other hydroxy-lipids remained unchanged. Vitamin E deficiency during embryogenesis depleted DHA and arachidonic acid, and increased hydroxy-fatty acids derived from these PUFA, suggesting that α-tocopherol is necessary to protect these critical fatty acids.

DOI10.1016/j.redox.2013.12.007
Alternate JournalRedox Biol
PubMed ID24416717
PubMed Central IDPMC3887274
Grant ListR01 DK094600 / DK / NIDDK NIH HHS / United States
HL081721 / HL / NHLBI NIH HHS / United States
HD062109 / HD / NICHD NIH HHS / United States
DK094600 / DK / NIDDK NIH HHS / United States
R01 HD062109 / HD / NICHD NIH HHS / United States
R01 HL081721 / HL / NHLBI NIH HHS / United States
S10 RR027878 / RR / NCRR NIH HHS / United States
NIEHS ES000210. / ES / NIEHS NIH HHS / United States
P30 ES000210 / ES / NIEHS NIH HHS / United States