Novel liquid chromatography-mass spectrometry method shows that vitamin E deficiency depletes arachidonic and docosahexaenoic acids in zebrafish (Danio rerio) embryos.
Redox Biol
; 2: 105-13, 2013.
Article
en En
| MEDLINE
| ID: mdl-24416717
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.
Palabras clave
ARA, 20:4 ω-6, arachidonic acid; Arachidonic acid; CNS, central nervous system; DHA, 22:6 ω-3, docosahexaenoic acid; Embryogenesis; HDHA, hydroxy-DHA; HETEs, hydroxy-eicosatetraenoic acids; Hybrid quadrupole-time of flight MS; LTA4, leukotriene A4; Neurogenesis; Vitamin E; dn, deuterium-labeled; hpf, hours post-fertilization; δT3, delta-tocotrienol
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Espectrometría de Masas
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Pez Cebra
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Ácidos Docosahexaenoicos
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Cromatografía Líquida de Alta Presión
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Ácido Araquidónico
Límite:
Animals
Idioma:
En
Revista:
Redox Biol
Año:
2013
Tipo del documento:
Article
País de afiliación:
Estados Unidos