Determination of α-tocopherol in plasma and erythrocytes by high-performance liquid chromatography

González-Corbella, M.J; Lloberas-Blanch, N.; Castellote-Bargalló, A.I; López‐Sabater, M. Carmen; Rivero-Urgell, M.

doi:10.1016/0378-4347(94)00293-2

Cited by 24 publications

(5 citation statements)

References 17 publications

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“…Quantitative HPLC with ultraviolet detection [1,2,3] is currently used for the determination of α-tocopherol in biological samples. In this study, we developed and validated a new simple HPLC method for selective and sensitive determination of α-tocopherol in erythrocyte membranes.…”

Section: Resultsmentioning

confidence: 99%

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Development and validation of HPLC method for the determination of α-tocopherol in human erythrocytes for clinical applications

et al. 2003

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In this work, a simple isocratic reversed-phase HPLC method for determination of alpha-tocopherol in human erythrocytes has been developed and validated. After separation of plasma the erythrocytes were washed three times with 0.9% sodium chloride containing 0.01% butylated hydroxytoluene (BHT) as antioxidant and then were diluted 1:1 (v/v) with the same solution. In the liquid-liquid extraction (LLE) procedure, 2500 microL of n-hexane was added to 500 microL of erythrocytes. After 2 min this mixture was deproteinized by addition of cool ethanol (500 microL, 5 min) denatured with 5% methanol containing alpha-tocopherol acetate (20 micromol L(-1)), as internal standard, and then extracted for 5 min by vortex mixing. After centrifugation (10 min, 1600xg) an aliquot (2000 microL) of the clean extract was separated and evaporated under nitrogen. The residue was dissolved in 400 microL methanol and analysed by reversed-phase HPLC on a 4.6 mmx150 mm, 5 microm Pecosphere C18 column; the mobile phase was 100% methanol, flow rate 1.2 mL min(-1). The volume injected was 100 microL and detection was by diode-array detector at a wavelength of 295 nm. The extraction recovery of alpha-tocopherol from human erythrocytes was 100.0+/-2.0%. The detection limit was 0.1 micromol L(-1) and a linear calibration plot was obtained in the concentration range 0.5-20.0 micromol L(-1). Within determination precision was 5.2% RSD (n=10), between determination precision was 6.1% RSD (n=10). The method was applied successfully in a clinical study of patients with acute pancreatitis and for determination of the reference values in the healthy Czech population.

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Section: Resultsmentioning

confidence: 99%

“…α-Tocopherol is able to prevent cell damage by reaction with free radicals. Concentration of α-tocopherol in serum does not reflect the content of α-tocopherol in membranes whereas erythrocyte α-tocopherol may be good indicator of vitamin E nutritional status [1,2,3].…”

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confidence: 99%

Development and validation of HPLC method for the determination of α-tocopherol in human erythrocytes for clinical applications

et al. 2003

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“…Determination of a-tocopherol and retinol in plasma and erythrocytes was performed by reversed-phase HPLC (Gonza Âlez-Corbella et al, 1994). a-tocopherol acetate and retinyl acetate were used as internal standards.…”

Section: Blood Analysismentioning

confidence: 99%

Administration of low doses of fish oil derived N-3 fatty acids to elderly subjects

et al. 1997

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Objectives: To assess the incorporation of n-3 polyunsaturated fatty acids (PUFA) in plasma and erythrocyte lipids of elderly subjects after ingestion of very low doses of ®sh oil. The effects on a-tocopherol and retinol concentrations were also studied. Setting: Municipal nursing home in Barcelona, Spain. Subjects: Forty-®ve elderly subjects aged 60±92 y. Design and intervention: Subjects received a non-commercialized milk formula containing 1% ®sh oil for 15 months, which provided 0.40 g/d of n-3 PUFA. Fatty acid pro®les and antioxidant concentrations were measured before and after the intervention period. Results: Fish oil ingestion was associated with signi®cant increases in total n-3 PUFA in plasma and erythrocytes by 32% and 18%, respectively. Eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid concentrations were higher after the ingestion period both in plasma and erythrocytes (P`0.05), whereas linoleic and arachidonic acids remained unchanged. The n-6/n-3 ratio decreased by 21% in plasma and by 16% in erythrocytes (P`0.05). Moreover, younger subjects showed a greater incorporation of EPA and DHA than older subjects. Plasma a-tocopherol and retinol concentrations did not vary signi®cantly, whereas erythrocyte atocopherol was signi®cantly higher after the intervention period. Conclusion: This study shows that low doses of n-3 PUFA supplemented with adequate amounts of a-tocopherol can be incorporated into blood lipids in elderly subjects without lowering their antioxidant concentrations. Sponsorship: This study was supported by a research grant from the Spanish CDTI.

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“…When only a-tocopherol is to be separated from other tocopherols and quantified, reversed-phase HPLC is often chosen (Rupe´rez et al, 2001;Nilsson et al, 1978;Julianto et al, 1999). Methods for the analysis of tocopherol contents of erythrocytes with and without prior saponification are well described by Vatassery et al (1993) and Gonzalez-Corbella et al (1994), respectively. It is suggested that quantification of b-, g-, and d-tocopherols and tocopherolquinones could be achieved with satisfactory precision and accuracy only by use of a sensitive detector (Mino et al, 1985;Rupe´rez et al, 2001;Sobczak et al, 1999;Vatassery et al, 1993).…”

Section: Introductionmentioning

confidence: 99%

Spectrofluorometric and high-performance liquid chromatographic determination of all-rac-α-tocopheryl acetate in virgin olive oil

Cho

Rima

Chang

et al. 2007

Journal of Food Composition and Analysis

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Determination of α-tocopherol in plasma and erythrocytes by high-performance liquid chromatography

Cited by 24 publications

References 17 publications

Development and validation of HPLC method for the determination of α-tocopherol in human erythrocytes for clinical applications

Development and validation of HPLC method for the determination of α-tocopherol in human erythrocytes for clinical applications

Administration of low doses of fish oil derived N-3 fatty acids to elderly subjects

Spectrofluorometric and high-performance liquid chromatographic determination of all-rac-α-tocopheryl acetate in virgin olive oil

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