Title | Novel liquid chromatography-mass spectrometry method shows that vitamin E deficiency depletes arachidonic and docosahexaenoic acids in zebrafish (Danio rerio) embryos. |
Publication Type | Journal Article |
Year of Publication | 2013 |
Authors | Lebold KM, Kirkwood JS, Taylor AW, Choi J, Barton CL, Miller GW, La Du J, Jump DB, Stevens JFrederik, Tanguay RL, Traber MG |
Journal | Redox Biol |
Volume | 2 |
Pagination | 105-13 |
Date Published | 2013 |
ISSN | 2213-2317 |
Keywords | alpha-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. |
DOI | 10.1016/j.redox.2013.12.007 |
Alternate Journal | Redox Biol |
PubMed ID | 24416717 |
PubMed Central ID | PMC3887274 |
Grant List | R01 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 |