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Novel liquid chromatography–mass spectrometry method shows that vitamin E deficiency depletes arachidonic and docosahexaenoic acids in zebrafish (Danio rerio) embryos

ScholarsArchive at Oregon State University

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Title Novel liquid chromatography–mass spectrometry method shows that vitamin E deficiency depletes arachidonic and docosahexaenoic acids in zebrafish (Danio rerio) embryos
Names Lebold, Katie M. (creator)
Kirkwood, Jays S. (creator)
Taylor, Alan W. (creator)
Choi, Jaewoo (creator)
Barton, Carrie L. (creator)
Miller, Galen W. (creator)
La Du, Jane (creator)
Jump, Donald B. (creator)
Stevens, Jan Frederik (creator)
Tanguay, Robert L. (creator)
Traber, Maret G. (creator)
Date Issued 2014 (iso8601)
Note This is the publisher’s final pdf. The published article is copyrighted by the author(s) and published by Elsevier. The published article can be found at: http://www.journals.elsevier.com/redox-biology.
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.
Genre Article
Access Condition http://creativecommons.org/licenses/by-nc-sa/3.0/us/
Topic Vitamin E
Identifier Lebold, K. M., Kirkwood, J. S., Taylor, A. W., Choi, J., Barton, C. L., Miller, G. W., ... & Traber, M. G. (2014). Novel liquid chromatography–mass spectrometry method shows that vitamin E deficiency depletes arachidonic and docosahexaenoic acids in zebrafish (Danio rerio) embryos. Redox Biology, 2, 105-113. doi:10.1016/j.redox.2013.12.007

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