A validated liquid chromatography method for the simultaneous determination of vitamins A and E in human plasma

Danuta Siluk; Regina V. Oliveira; Maria Esther-Rodriguez-Rosas; Shari Ling; Angelo Bos; Luigi Ferrucci; Irving W. Wainer

doi:10.1016/j.jpba.2007.03.033

A validated liquid chromatography method for the simultaneous determination of vitamins A and E in human plasma

Danuta Siluk, Regina V. Oliveira, Maria Esther-Rodriguez-Rosas, Shari Ling, Angelo Bos, Luigi Ferrucci, Irving W. Wainer

Research output: Contribution to journal › Article › peer-review

55 Scopus citations

Abstract

A fast (15 min) and simple HPLC method for determination of all-trans-retinol and α-, γ- and δ-tocopherols in human plasma has been developed. The assay utilized 200 μl of plasma to which 20 μl of internal standard solution (retinol acetate) was added followed by 200 μl of water, 400 μl of ethanol and 800 μl of hexane. The hexane layer was collected, evaporated, the residue dissolved in 200 μl of methanol and analyzed on a Zorbax Eclipse XDB-C18 column using a step gradient with a polar organic mobile phase composed of acetonitrile and methanol and variable wavelength fluorescence detection. The quantification limits for all-trans-retinol and γ-tocopherol were 20 ng/ml and for α- and δ-tocopherols 500 ng/ml and 10 ng/ml, respectively. The procedure was validated and applied to the analysis of plasma samples from the Baltimore Longitudinal Study of Aging.

Original language	English (US)
Pages (from-to)	1001-1007
Number of pages	7
Journal	Journal of Pharmaceutical and Biomedical Analysis
Volume	44
Issue number	4
DOIs	https://doi.org/10.1016/j.jpba.2007.03.033
State	Published - Aug 15 2007
Externally published	Yes

Keywords

All-trans-retinol
Fluorescence detection
HPLC
Vitamins
α-Tocopherol
γ-Tocopherol
δ-Tocopherol

ASJC Scopus subject areas

Analytical Chemistry
Pharmaceutical Science
Drug Discovery
Spectroscopy
Clinical Biochemistry

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10.1016/j.jpba.2007.03.033

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title = "A validated liquid chromatography method for the simultaneous determination of vitamins A and E in human plasma",

abstract = "A fast (15 min) and simple HPLC method for determination of all-trans-retinol and α-, γ- and δ-tocopherols in human plasma has been developed. The assay utilized 200 μl of plasma to which 20 μl of internal standard solution (retinol acetate) was added followed by 200 μl of water, 400 μl of ethanol and 800 μl of hexane. The hexane layer was collected, evaporated, the residue dissolved in 200 μl of methanol and analyzed on a Zorbax Eclipse XDB-C18 column using a step gradient with a polar organic mobile phase composed of acetonitrile and methanol and variable wavelength fluorescence detection. The quantification limits for all-trans-retinol and γ-tocopherol were 20 ng/ml and for α- and δ-tocopherols 500 ng/ml and 10 ng/ml, respectively. The procedure was validated and applied to the analysis of plasma samples from the Baltimore Longitudinal Study of Aging.",

keywords = "All-trans-retinol, Fluorescence detection, HPLC, Vitamins, α-Tocopherol, γ-Tocopherol, δ-Tocopherol",

author = "Danuta Siluk and Oliveira, {Regina V.} and Maria Esther-Rodriguez-Rosas and Shari Ling and Angelo Bos and Luigi Ferrucci and Wainer, {Irving W.}",

note = "Funding Information: This study was supported by the Intramural Research Program, National Institutes of Health, National Institute on Aging. ",

year = "2007",

month = aug,

day = "15",

doi = "10.1016/j.jpba.2007.03.033",

language = "English (US)",

volume = "44",

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T1 - A validated liquid chromatography method for the simultaneous determination of vitamins A and E in human plasma

AU - Siluk, Danuta

AU - Oliveira, Regina V.

AU - Esther-Rodriguez-Rosas, Maria

AU - Ling, Shari

AU - Bos, Angelo

AU - Ferrucci, Luigi

AU - Wainer, Irving W.

N1 - Funding Information: This study was supported by the Intramural Research Program, National Institutes of Health, National Institute on Aging.

PY - 2007/8/15

Y1 - 2007/8/15

N2 - A fast (15 min) and simple HPLC method for determination of all-trans-retinol and α-, γ- and δ-tocopherols in human plasma has been developed. The assay utilized 200 μl of plasma to which 20 μl of internal standard solution (retinol acetate) was added followed by 200 μl of water, 400 μl of ethanol and 800 μl of hexane. The hexane layer was collected, evaporated, the residue dissolved in 200 μl of methanol and analyzed on a Zorbax Eclipse XDB-C18 column using a step gradient with a polar organic mobile phase composed of acetonitrile and methanol and variable wavelength fluorescence detection. The quantification limits for all-trans-retinol and γ-tocopherol were 20 ng/ml and for α- and δ-tocopherols 500 ng/ml and 10 ng/ml, respectively. The procedure was validated and applied to the analysis of plasma samples from the Baltimore Longitudinal Study of Aging.

AB - A fast (15 min) and simple HPLC method for determination of all-trans-retinol and α-, γ- and δ-tocopherols in human plasma has been developed. The assay utilized 200 μl of plasma to which 20 μl of internal standard solution (retinol acetate) was added followed by 200 μl of water, 400 μl of ethanol and 800 μl of hexane. The hexane layer was collected, evaporated, the residue dissolved in 200 μl of methanol and analyzed on a Zorbax Eclipse XDB-C18 column using a step gradient with a polar organic mobile phase composed of acetonitrile and methanol and variable wavelength fluorescence detection. The quantification limits for all-trans-retinol and γ-tocopherol were 20 ng/ml and for α- and δ-tocopherols 500 ng/ml and 10 ng/ml, respectively. The procedure was validated and applied to the analysis of plasma samples from the Baltimore Longitudinal Study of Aging.

KW - All-trans-retinol

KW - Fluorescence detection

KW - HPLC

KW - Vitamins

KW - α-Tocopherol

KW - γ-Tocopherol

KW - δ-Tocopherol

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ER -

A validated liquid chromatography method for the simultaneous determination of vitamins A and E in human plasma

Abstract

Keywords

ASJC Scopus subject areas

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