Method for the simultaneous determination of retinol and β-carotene concentrations in human tissues and plasma

Jennine M. Lunetta, Rebecca A. Zulim, Stephen R. Dueker, Yumei Lin, Vicky Flaig, Philip D. Schneider, Bruce M. Wolfe, Andrew J. Clifford

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

To understand differential tissue distribution of retinoids and carotenoids, as it might influence biological processes in humans, we developed and demonstrated a method for measuring them in selected human tissues. The method includes internal standards and a secondary reference standard to eliminate the need for external standard calibration and to minimize sample-handling errors. Tissues were digested (saponified) in ethanolic KOH. Retinol and β-carotene were extracted with organic solvent containing internal standards. Analytes were separated using isocratic liquid chromatography and quantified at 325 nm for retinol and 450 nm for β-carotene. Plasma was analyzed in a similar way but without saponification. Retinal-O-ethyloxime and β-apo-12′-carotenal-O-t-butyloxime served as internal standards. Plasma, breast, and fat from breast surgery patients and colon, liver, muscle, and fat from colon surgery patients were analyzed. Within-day relative standard deviations (RSDs) for plasma were <0.04 for β-carotene and <0.03 for retinol, between-day RSDs were <0.05 for β-carotene and <0.04 for retinol. Saponification ensured complete extraction of retinol and β-carotene and removal of triglycerides that "foul" chromatographic columns. It seems retinol and β-carotene concentrations in tissues and blood of cancer patients are the same or higher than those in corresponding tissues of patients without these cancers.

Original languageEnglish (US)
Pages (from-to)100-109
Number of pages10
JournalAnalytical Biochemistry
Volume304
Issue number1
DOIs
StatePublished - May 1 2002

Keywords

  • Analysis
  • Human
  • RP-HPLC
  • Tissue concentrations
  • Vitamin A
  • β-carotene

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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