Simultaneous quantification of various retinoids by high performance liquid chromatography: Its relevance to alcohol research

Hirokazu Yokoyama, Michinaga Matsumoto, Haruko Shiraishi, Hiromasa Ishii

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: We established a high performance liquid chromatography system that allowed simultaneous quantification of various retinoids. Methods: We applied the retinoids to a high performance liquid chromatography system with a silica gel absorption column. Samples were separated by the system with a binary multistep gradient with two kinds of solvent that contained n-Hexan, 2-propanol, and glacial acetic acid in different ratios. Each retinoid was detected at a wavelength of 350 nm. Results: This condition allowed separation of 13-cis-retinoic acid, 9-cis-retinoic acid, all-trans- retinoic acid, 13-cis-retinol, all-trans-retinol, all-trans-4-oxo-retinoic acid, and 13-cis-4-oxo-retinoic acid as distinct single peaks. Each retinoid was also analyzed separately and its retention time determined. To ascertain the reliability of this system for retinoid quantification, retinoids at various concentrations were applied to the system. We observed the linearities between the concentration and area under the curve of the peak for each retinoid by linear least-squares regression analysis up to 2.5 ng/ml for all retinoic acids and up to 5 ng/ml for all retinols. There was no significant scattering in tests of within-day reproducibility or day-to-day reproducibility. Using this system, we examined effects of light exposure, on isomerization of retinoids. When retinoids were exposed to room light for 2 hr, the amounts of all but 13-cis-retinol changed significantly. In particular, the amounts of all-trans-retinoic acid and 9-cis-retinoic acid were reduced by 40% and 60%, respectively. Conclusion: The HPLC system established in this study should be useful for studying the oxidation pathway of retinol to retinoic acid. A light-shielded condition is required when particular retinoic acids are analyzed.

Original languageEnglish
JournalAlcoholism: Clinical and Experimental Research
Volume24
Issue number4 SUPPL.
Publication statusPublished - 2000 Apr

Fingerprint

Retinoids
High performance liquid chromatography
High Pressure Liquid Chromatography
Alcohols
Tretinoin
Vitamin A
Research
Light
Isotretinoin
2-Propanol
Silica Gel
Isomerization
Least-Squares Analysis
Regression analysis
Acetic Acid
Area Under Curve
Regression Analysis
Scattering
Wavelength
Oxidation

Keywords

  • Photoisomerization
  • Retinoic Acid
  • Retinol

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Toxicology

Cite this

Simultaneous quantification of various retinoids by high performance liquid chromatography : Its relevance to alcohol research. / Yokoyama, Hirokazu; Matsumoto, Michinaga; Shiraishi, Haruko; Ishii, Hiromasa.

In: Alcoholism: Clinical and Experimental Research, Vol. 24, No. 4 SUPPL., 04.2000.

Research output: Contribution to journalArticle

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abstract = "Background: We established a high performance liquid chromatography system that allowed simultaneous quantification of various retinoids. Methods: We applied the retinoids to a high performance liquid chromatography system with a silica gel absorption column. Samples were separated by the system with a binary multistep gradient with two kinds of solvent that contained n-Hexan, 2-propanol, and glacial acetic acid in different ratios. Each retinoid was detected at a wavelength of 350 nm. Results: This condition allowed separation of 13-cis-retinoic acid, 9-cis-retinoic acid, all-trans- retinoic acid, 13-cis-retinol, all-trans-retinol, all-trans-4-oxo-retinoic acid, and 13-cis-4-oxo-retinoic acid as distinct single peaks. Each retinoid was also analyzed separately and its retention time determined. To ascertain the reliability of this system for retinoid quantification, retinoids at various concentrations were applied to the system. We observed the linearities between the concentration and area under the curve of the peak for each retinoid by linear least-squares regression analysis up to 2.5 ng/ml for all retinoic acids and up to 5 ng/ml for all retinols. There was no significant scattering in tests of within-day reproducibility or day-to-day reproducibility. Using this system, we examined effects of light exposure, on isomerization of retinoids. When retinoids were exposed to room light for 2 hr, the amounts of all but 13-cis-retinol changed significantly. In particular, the amounts of all-trans-retinoic acid and 9-cis-retinoic acid were reduced by 40{\%} and 60{\%}, respectively. Conclusion: The HPLC system established in this study should be useful for studying the oxidation pathway of retinol to retinoic acid. A light-shielded condition is required when particular retinoic acids are analyzed.",
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N2 - Background: We established a high performance liquid chromatography system that allowed simultaneous quantification of various retinoids. Methods: We applied the retinoids to a high performance liquid chromatography system with a silica gel absorption column. Samples were separated by the system with a binary multistep gradient with two kinds of solvent that contained n-Hexan, 2-propanol, and glacial acetic acid in different ratios. Each retinoid was detected at a wavelength of 350 nm. Results: This condition allowed separation of 13-cis-retinoic acid, 9-cis-retinoic acid, all-trans- retinoic acid, 13-cis-retinol, all-trans-retinol, all-trans-4-oxo-retinoic acid, and 13-cis-4-oxo-retinoic acid as distinct single peaks. Each retinoid was also analyzed separately and its retention time determined. To ascertain the reliability of this system for retinoid quantification, retinoids at various concentrations were applied to the system. We observed the linearities between the concentration and area under the curve of the peak for each retinoid by linear least-squares regression analysis up to 2.5 ng/ml for all retinoic acids and up to 5 ng/ml for all retinols. There was no significant scattering in tests of within-day reproducibility or day-to-day reproducibility. Using this system, we examined effects of light exposure, on isomerization of retinoids. When retinoids were exposed to room light for 2 hr, the amounts of all but 13-cis-retinol changed significantly. In particular, the amounts of all-trans-retinoic acid and 9-cis-retinoic acid were reduced by 40% and 60%, respectively. Conclusion: The HPLC system established in this study should be useful for studying the oxidation pathway of retinol to retinoic acid. A light-shielded condition is required when particular retinoic acids are analyzed.

AB - Background: We established a high performance liquid chromatography system that allowed simultaneous quantification of various retinoids. Methods: We applied the retinoids to a high performance liquid chromatography system with a silica gel absorption column. Samples were separated by the system with a binary multistep gradient with two kinds of solvent that contained n-Hexan, 2-propanol, and glacial acetic acid in different ratios. Each retinoid was detected at a wavelength of 350 nm. Results: This condition allowed separation of 13-cis-retinoic acid, 9-cis-retinoic acid, all-trans- retinoic acid, 13-cis-retinol, all-trans-retinol, all-trans-4-oxo-retinoic acid, and 13-cis-4-oxo-retinoic acid as distinct single peaks. Each retinoid was also analyzed separately and its retention time determined. To ascertain the reliability of this system for retinoid quantification, retinoids at various concentrations were applied to the system. We observed the linearities between the concentration and area under the curve of the peak for each retinoid by linear least-squares regression analysis up to 2.5 ng/ml for all retinoic acids and up to 5 ng/ml for all retinols. There was no significant scattering in tests of within-day reproducibility or day-to-day reproducibility. Using this system, we examined effects of light exposure, on isomerization of retinoids. When retinoids were exposed to room light for 2 hr, the amounts of all but 13-cis-retinol changed significantly. In particular, the amounts of all-trans-retinoic acid and 9-cis-retinoic acid were reduced by 40% and 60%, respectively. Conclusion: The HPLC system established in this study should be useful for studying the oxidation pathway of retinol to retinoic acid. A light-shielded condition is required when particular retinoic acids are analyzed.

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