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1.
J Pharm Biomed Anal ; 127: 156-69, 2016 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-26964480

RESUMO

Tocopherols and tocotrienols, widely described as vitamin E derivatives, have been proven to take part in a number of important biological functions. Among them, antioxidant properties had been investigated and documented in the literature. Since tocochromanols have revealed their plausible beneficial impact on several pathological processes, such as cancerogenesis or cognitive impairment diseases, there is a growing interest in quantitative determination of these compounds in biological fluids, tissues and plant organs. However, due to vitamin E chemical features, such as lipophilic and non-polar characteristics, quantitative determination of the compounds seems to be problematic. In this paper we present current analytical approaches in tocopherols and tocotrienols determination in biological and food matrices with the use of chromatographic techniques, especially gas chromatography (GC) and high performance liquid chromatography (HPLC) coupled with mass spectrometry. Derivatization techniques applied for GC-MS analysis in the case of tocol derivatives, especially silylation and acylation, are described. Significant attention is paid to ionization process of tocopherols and tocotrienols.


Assuntos
Cromatografia Líquida/métodos , Produtos Agrícolas/química , Análise de Alimentos/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Tocoferóis/análise , Tocotrienóis/análise , Animais , Cromatografia Líquida/instrumentação , Fezes/química , Análise de Alimentos/instrumentação , Cromatografia Gasosa-Espectrometria de Massas/instrumentação , Humanos , Fígado/química , Tocoferóis/sangue , Tocoferóis/urina , Tocotrienóis/sangue , Tocotrienóis/urina
2.
Arch Phys Med Rehabil ; 93(12): 2223-8.e2, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22772083

RESUMO

OBJECTIVE: To determine the plasma and urine levels of antioxidants and oxidative stress biomarkers in subjects with spinal cord injury (SCI) the first year after injury. DESIGN: Descriptive 1-year follow-up study. SETTING: Rehabilitation and research center. PARTICIPANTS: SCI subjects (n=37; age range, 18-70 y) consecutively enrolled within the first month after injury. A healthy, able-bodied control group (n=346) was also included. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Blood and urine levels of antioxidants and oxidative stress biomarkers were measured at inclusion and after 3 and 12 months postinjury. RESULTS: One month after injury, the plasma antioxidants (total and oxidized glutathione and 6 different carotenoids and α-tocopherol) were reduced by 19% to 71% among the SCI subjects compared with the controls. The redox potential was reduced by 7% among the SCI subjects. The oxidative stress biomarker urinary 8-epi prostagladin F2α (PGF2α) increased to 161% in the SCI subjects compared with the controls. After 3 and 12 months, most of the antioxidant biomarkers were still significantly reduced compared with the controls, while urinary 8-epi PGF2α had increased to 208% compared with the controls. CONCLUSIONS: The levels of antioxidants were significantly lower, while the marker of oxidative stress was higher in the SCI subjects compared with the controls. This observation demonstrates that SCI patients experience increased oxidative stress and reduced antioxidant defense the first year after injury. Our findings warrant intervention studies where SCI patients receive dietary antioxidant support as part of their rehabilitation.


Assuntos
Antioxidantes/análise , Adolescente , Adulto , Idoso , Antioxidantes/metabolismo , Biomarcadores , Carotenoides/sangue , Carotenoides/urina , Feminino , Glutationa/sangue , Glutationa/urina , Humanos , Masculino , Pessoa de Meia-Idade , Oxirredução , Estresse Oxidativo , Centros de Reabilitação , Traumatismos da Medula Espinal , Fatores de Tempo , Tocoferóis/sangue , Tocoferóis/urina , Adulto Jovem
3.
J Biol Chem ; 287(31): 26077-86, 2012 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-22665481

RESUMO

Vitamin E is a family of naturally occurring and structurally related lipophilic antioxidants, one of which, α-tocopherol (α-TOH), selectively accumulates in vertebrate tissues. The ω-hydroxylase cytochrome P450-4F2 (CYP4F2) is the only human enzyme shown to metabolize vitamin E. Using cDNA cloning, cell culture expression, and activity assays, we identified Cyp4f14 as a functional murine ortholog of CYP4F2. We then investigated the effect of Cyp4f14 deletion on vitamin E metabolism and status in vivo. Cyp4f14-null mice exhibited substrate-specific reductions in liver microsomal vitamin E-ω-hydroxylase activity ranging from 93% (γ-TOH) to 48% (γ-tocotrienol). In vivo data obtained from metabolic cage studies showed whole-body reductions in metabolism of γ-TOH of 90% and of 68% for δ- and α-TOH. This metabolic deficit in Cyp4f14(-/-) mice was partially offset by increased fecal excretion of nonmetabolized tocopherols and of novel ω-1- and ω-2-hydroxytocopherols. 12'-OH-γ-TOH represented 41% of whole-body production of γ-TOH metabolites in Cyp4f14(-/-) mice fed a soybean oil diet. Despite these counterbalancing mechanisms, Cyp4f14-null mice fed this diet for 6 weeks hyper-accumulated γ-TOH (2-fold increase over wild-type littermates) in all tissues and appeared normal. We conclude that CYP4F14 is the major but not the only vitamin E-ω-hydroxylase in mice. Its disruption significantly impairs whole-body vitamin E metabolism and alters the widely conserved phenotype of preferential tissue deposition of α-TOH. This model animal and its derivatives will be valuable in determining the biological actions of specific tocopherols and tocotrienols in vivo.


Assuntos
Sistema Enzimático do Citocromo P-450/genética , Oxigenases de Função Mista/genética , Tocoferóis/metabolismo , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Família 4 do Citocromo P450 , Fezes/química , Feminino , Expressão Gênica , Técnicas de Inativação de Genes , Recombinação Homóloga , Hidroxilação , Fígado/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microssomos Hepáticos/enzimologia , Tocoferóis/química , Tocoferóis/urina
4.
Clin Chem ; 52(7): 1339-45, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16728468

RESUMO

BACKGROUND: Vitamin E supplementation has been recommended for persons with familial hypobetalipoproteinemia (FHBL), a rare disorder of lipoprotein metabolism that leads to low serum alpha-tocopherol and decreased LDL-cholesterol and apolipoprotein (apo) B. We examined the effect of truncated apoB variants on vitamin E metabolism and oxidative stress in persons with FHBL. METHODS: We studied 9 individuals with heterozygous FHBL [mean (SE) age, 40 (5) years; body mass index (BMI), 27 (10) kg/m2] and 7 normolipidemic controls [age, 41 (5) years; BMI, 25 (2) kg/m2]. We also studied 3 children-2 with homozygous FHBL (apoB-30.9) and 1 with abetalipoproteinemia-who were receiving alpha-tocopherol supplementation. We used HPLC with electrochemical detection to measure alpha- and gamma-tocopherol in serum, erythrocytes, and platelets, and gas chromatography-mass spectrometry to measure F2-isoprostanes and tocopherol metabolites in urine as markers of oxidative stress and tocopherol intake, respectively. RESULTS: Compared with controls, persons with FHBL had significantly lower fasting plasma concentrations of total cholesterol [2.4 (0.2) vs 4.7 (0.2) mmol/L], triglycerides [0.5 (0.1) vs 0.9 (0.1) mmol/L], LDL-cholesterol [0.7 (0.1) vs 2.8 (0.3) mmol/L], apoB [0.23 (0.02) vs 0.84 (0.08) g/L], alpha-tocopherol [13.6 (1.0) vs 28.7 (1.4) micromol/L], and gamma-tocopherol [1.0 (0.1) vs 1.8 (0.3) micromol/L] (all P < 0.03). Erythrocyte alpha-tocopherol was decreased [5.0 (0.2) vs 6.0 (0.3) micromol/L; P < 0.005], but we observed no differences in lipid-adjusted serum tocopherols, erythrocyte gamma-tocopherol, platelet alpha- or gamma-tocopherol, urinary F2-isoprostanes, or tocopherol metabolites. CONCLUSION: Taken together, our findings do not support the recommendation that persons with heterozygous FHBL receive vitamin E supplementation.


Assuntos
Hipobetalipoproteinemias/metabolismo , Estresse Oxidativo , Tocoferóis/metabolismo , Abetalipoproteinemia/tratamento farmacológico , Abetalipoproteinemia/genética , Abetalipoproteinemia/metabolismo , Adulto , Apolipoproteínas B/genética , Biomarcadores/urina , Plaquetas/metabolismo , Pré-Escolar , Eritrócitos/metabolismo , F2-Isoprostanos/urina , Feminino , Heterozigoto , Humanos , Hipobetalipoproteinemias/genética , Lactente , Masculino , Tocoferóis/sangue , Tocoferóis/urina , Vitaminas/administração & dosagem , Vitaminas/sangue , Vitaminas/urina , alfa-Tocoferol/administração & dosagem , alfa-Tocoferol/sangue , alfa-Tocoferol/urina , gama-Tocoferol/sangue
5.
Am J Clin Nutr ; 83(1): 95-102, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16400056

RESUMO

BACKGROUND: Some studies have shown potential benefit of vitamin E on platelet function, but several clinical trials failed to show improved cardiovascular outcome with alpha-tocopherol supplementation. Gamma-tocopherol, a major dietary form of vitamin E, may have protective properties different from those of alpha-tocopherol. OBJECTIVE: We compared the effects of supplementation with alpha-tocopherol (500 mg) and a gamma-tocopherol-rich compound (500 mg, containing 60% gamma-tocopherol) on serum and cellular tocopherol concentrations, urinary tocopherol metabolite excretion, and in vivo platelet activation in subjects with type 2 diabetes. DESIGN: Fifty-eight subjects were randomly assigned to receive either 500 mg alpha-tocopherol/d, 500 mg mixed tocopherols/d, or matching placebo. Serum, erythrocyte, and platelet tocopherol and urinary metabolite concentrations were measured at baseline and after the 6-wk intervention. Soluble CD40 ligand, urinary 11-dehydro-thromboxane B2, serum thromboxane B2, soluble P-selectin, and von Willebrand factor were measured as biomarkers of in vivo platelet activation. RESULTS: Serum alpha-tocopherol increased with both tocopherol treatments. Serum and cellular gamma-tocopherol increased 4-fold (P < 0.001) in the mixed tocopherol group, whereas red blood cell gamma-tocopherol decreased significantly after alpha-tocopherol supplementation. Excretion of alpha-carboxyethyl-hydroxychroman increased significantly after supplementation with alpha-tocopherol and mixed tocopherols. Excretion of gamma-carboxyethyl-hydroxychroman increased significantly after supplementation with mixed tocopherols and after that with alpha-tocopherol, which may reflect the displacement of gamma-tocopherol by alpha-tocopherol due to incorporation of the latter into lipoproteins in the liver. Neither treatment had any significant effect on markers of platelet activation. CONCLUSIONS: Supplementation with alpha-tocopherol decreased red blood cell gamma-tocopherol, whereas mixed tocopherols increased both serum alpha-tocopherol and serum and cellular gamma-tocopherol. Changes in serum tocopherol closely reflect changes in cellular concentrations of tocopherols after supplementation.


Assuntos
Diabetes Mellitus Tipo 2/sangue , Eritrócitos/química , Ativação Plaquetária/efeitos dos fármacos , Tocoferóis/administração & dosagem , alfa-Tocoferol/sangue , gama-Tocoferol/análise , Análise de Variância , Plaquetas/efeitos dos fármacos , Plaquetas/fisiologia , Cromanos/urina , Cromatografia Líquida de Alta Pressão , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/urina , Suplementos Nutricionais , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Isomerismo , Masculino , Pessoa de Meia-Idade , Tocoferóis/sangue , Tocoferóis/urina , alfa-Tocoferol/análise , alfa-Tocoferol/metabolismo , gama-Tocoferol/sangue , gama-Tocoferol/metabolismo
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