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 journalArticle

19 Citations (Scopus)

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

Fingerprint

Carotenoids
Vitamin A
Tissue
Plasmas
Saponification
Surgery
Colon
Breast
Fats
Biological Phenomena
Liquid chromatography
Retinoids
Tissue Distribution
Liquid Chromatography
Liver
Organic solvents
Calibration
Muscle
Neoplasms
Triglycerides

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Method for the simultaneous determination of retinol and β-carotene concentrations in human tissues and plasma. / Lunetta, Jennine M.; Zulim, Rebecca A.; Dueker, Stephen R.; Lin, Yumei; Flaig, Vicky; Schneider, Philip D; Wolfe, Bruce M.; Clifford, Andrew J.

In: Analytical Biochemistry, Vol. 304, No. 1, 01.05.2002, p. 100-109.

Research output: Contribution to journalArticle

Lunetta, Jennine M. ; Zulim, Rebecca A. ; Dueker, Stephen R. ; Lin, Yumei ; Flaig, Vicky ; Schneider, Philip D ; Wolfe, Bruce M. ; Clifford, Andrew J. / Method for the simultaneous determination of retinol and β-carotene concentrations in human tissues and plasma. In: Analytical Biochemistry. 2002 ; Vol. 304, No. 1. pp. 100-109.
@article{1e68afcf404a4b6d842ec894d1b886d9,
title = "Method for the simultaneous determination of retinol and β-carotene concentrations in human tissues and plasma",
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.",
keywords = "β-carotene, Analysis, Human, RP-HPLC, Tissue concentrations, Vitamin A",
author = "Lunetta, {Jennine M.} and Zulim, {Rebecca A.} and Dueker, {Stephen R.} and Yumei Lin and Vicky Flaig and Schneider, {Philip D} and Wolfe, {Bruce M.} and Clifford, {Andrew J.}",
year = "2002",
month = "5",
day = "1",
doi = "10.1006/abio.2001.5556",
language = "English (US)",
volume = "304",
pages = "100--109",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Method for the simultaneous determination of retinol and β-carotene concentrations in human tissues and plasma

AU - Lunetta, Jennine M.

AU - Zulim, Rebecca A.

AU - Dueker, Stephen R.

AU - Lin, Yumei

AU - Flaig, Vicky

AU - Schneider, Philip D

AU - Wolfe, Bruce M.

AU - Clifford, Andrew J.

PY - 2002/5/1

Y1 - 2002/5/1

N2 - 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.

AB - 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.

KW - β-carotene

KW - Analysis

KW - Human

KW - RP-HPLC

KW - Tissue concentrations

KW - Vitamin A

UR - http://www.scopus.com/inward/record.url?scp=0036570545&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036570545&partnerID=8YFLogxK

U2 - 10.1006/abio.2001.5556

DO - 10.1006/abio.2001.5556

M3 - Article

C2 - 11969193

AN - SCOPUS:0036570545

VL - 304

SP - 100

EP - 109

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -