An isocratic high-performance liquid chromatography method for the simultaneous analysis of plasma retinol, α-tocopherol, and various carotenoids

Kenneth W. Miller, Chung Yang

Research output: Contribution to journalArticle

154 Citations (Scopus)

Abstract

An isocratic high-performance liquid chromatography method for the simultaneous determination of various fat-soluble vitamins and carotenoids is reported. The method utilizes a Radial-Pak C-18, 5-μm column and an elution solvent composed of methanol:acetonitrile:chloroform (25:60:15). Only 100 μl of plasma sample is required for one determination. Retinol, α-tocopherol, α-carotene, β-carotene, lycopene, zeaxanthin, and two other unidentified carotenoids can be clearly separated and quantified in one HPLC run using α-tocopheryl acetate or tocol as the internal standard. The eluted peaks are quantified by either a photodiodearray detector at preprogrammed wavelengths at the absorption maxima of the compounds or a dual-wavelength detector at 280 and 436 nm. The total run time is 16 min. With an automatic injector and a programmable detector, the system allows unattended operation. The within-run and day-to-day coefficients of variation range from 1 to 8%. The lower limits of determination are 2, 40, and 2 ng for retinol, α-tocopherol, and carotenes, respectively. In addition, the system monitors the absorption spectra of the eluant during the HPLC run; this allows the spectral identification of various compounds separated in the same run.

Original languageEnglish (US)
Pages (from-to)21-26
Number of pages6
JournalAnalytical Biochemistry
Volume145
Issue number1
DOIs
StatePublished - Feb 15 1985

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Tocopherols
High performance liquid chromatography
Carotenoids
Vitamin A
High Pressure Liquid Chromatography
Plasmas
Detectors
Wavelength
alpha-Tocopherol
Chloroform
Vitamins
Methanol
Absorption spectra
Fats

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

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title = "An isocratic high-performance liquid chromatography method for the simultaneous analysis of plasma retinol, α-tocopherol, and various carotenoids",
abstract = "An isocratic high-performance liquid chromatography method for the simultaneous determination of various fat-soluble vitamins and carotenoids is reported. The method utilizes a Radial-Pak C-18, 5-μm column and an elution solvent composed of methanol:acetonitrile:chloroform (25:60:15). Only 100 μl of plasma sample is required for one determination. Retinol, α-tocopherol, α-carotene, β-carotene, lycopene, zeaxanthin, and two other unidentified carotenoids can be clearly separated and quantified in one HPLC run using α-tocopheryl acetate or tocol as the internal standard. The eluted peaks are quantified by either a photodiodearray detector at preprogrammed wavelengths at the absorption maxima of the compounds or a dual-wavelength detector at 280 and 436 nm. The total run time is 16 min. With an automatic injector and a programmable detector, the system allows unattended operation. The within-run and day-to-day coefficients of variation range from 1 to 8{\%}. The lower limits of determination are 2, 40, and 2 ng for retinol, α-tocopherol, and carotenes, respectively. In addition, the system monitors the absorption spectra of the eluant during the HPLC run; this allows the spectral identification of various compounds separated in the same run.",
author = "Miller, {Kenneth W.} and Chung Yang",
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N2 - An isocratic high-performance liquid chromatography method for the simultaneous determination of various fat-soluble vitamins and carotenoids is reported. The method utilizes a Radial-Pak C-18, 5-μm column and an elution solvent composed of methanol:acetonitrile:chloroform (25:60:15). Only 100 μl of plasma sample is required for one determination. Retinol, α-tocopherol, α-carotene, β-carotene, lycopene, zeaxanthin, and two other unidentified carotenoids can be clearly separated and quantified in one HPLC run using α-tocopheryl acetate or tocol as the internal standard. The eluted peaks are quantified by either a photodiodearray detector at preprogrammed wavelengths at the absorption maxima of the compounds or a dual-wavelength detector at 280 and 436 nm. The total run time is 16 min. With an automatic injector and a programmable detector, the system allows unattended operation. The within-run and day-to-day coefficients of variation range from 1 to 8%. The lower limits of determination are 2, 40, and 2 ng for retinol, α-tocopherol, and carotenes, respectively. In addition, the system monitors the absorption spectra of the eluant during the HPLC run; this allows the spectral identification of various compounds separated in the same run.

AB - An isocratic high-performance liquid chromatography method for the simultaneous determination of various fat-soluble vitamins and carotenoids is reported. The method utilizes a Radial-Pak C-18, 5-μm column and an elution solvent composed of methanol:acetonitrile:chloroform (25:60:15). Only 100 μl of plasma sample is required for one determination. Retinol, α-tocopherol, α-carotene, β-carotene, lycopene, zeaxanthin, and two other unidentified carotenoids can be clearly separated and quantified in one HPLC run using α-tocopheryl acetate or tocol as the internal standard. The eluted peaks are quantified by either a photodiodearray detector at preprogrammed wavelengths at the absorption maxima of the compounds or a dual-wavelength detector at 280 and 436 nm. The total run time is 16 min. With an automatic injector and a programmable detector, the system allows unattended operation. The within-run and day-to-day coefficients of variation range from 1 to 8%. The lower limits of determination are 2, 40, and 2 ng for retinol, α-tocopherol, and carotenes, respectively. In addition, the system monitors the absorption spectra of the eluant during the HPLC run; this allows the spectral identification of various compounds separated in the same run.

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