An LC/MS/MS method for stable isotope dilution studies of β-carotene bioavailability, bioconversion, and vitamin A status in humans

Isotope dilution is currently the most accurate technique in humans to determine vitamin A status and bioavailability/bioconversion of provitamin A carotenoids such as β-carotene. However, limits of MS detection, coupled with extensive isolation procedures, have hindered investigations of physiologi...

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Bibliographic Details
Published in:Journal of lipid research Vol. 55; no. 2; pp. 319 - 328
Main Authors: Oxley, Anthony, Berry, Philip, Taylor, Gordon A, Cowell, Joseph, Hall, Michael J, Hesketh, John, Lietz, Georg, Boddy, Alan V
Format: Journal Article
Language:English
Published: United States Elsevier 01.02.2014
Subjects:
Adolescent
Adult
beta Carotene - blood
beta Carotene - metabolism
beta Carotene - pharmacokinetics
Biological Availability
Biotransformation
carotenoid metabolism
Chromatography, Liquid - methods
Female
Humans
Isotopes
Male
Middle Aged
retinol metabolism
retinyl esters
tandem mass spectrometry
Tandem Mass Spectrometry - methods
Time Factors
Vitamin A - blood
Vitamin A - metabolism
Young Adult
β-carotene 15,15′-monooxygenase
tandem mass spectrometry
retinyl esters
carotenoid metabolism
retinol metabolism
β-carotene 15,15′-monooxygenase
ISSN:1539-7262, 0022-2275, 1539-7262
Online Access:Get full text
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Summary:Isotope dilution is currently the most accurate technique in humans to determine vitamin A status and bioavailability/bioconversion of provitamin A carotenoids such as β-carotene. However, limits of MS detection, coupled with extensive isolation procedures, have hindered investigations of physiologically-relevant doses of stable isotopes in large intervention trials. Here, a sensitive liquid chromatography-tandem mass spectrometry (LC/MS/MS) analytical method was developed to study the plasma response from coadministered oral doses of 2 mg [(13)C10]β-carotene and 1 mg [(13)C10]retinyl acetate in human subjects over a 2 week period. A reverse phase C18 column and binary mobile phase solvent system separated β-carotene, retinol, retinyl acetate, retinyl linoleate, retinyl palmitate/retinyl oleate, and retinyl stearate within a 7 min run time. Selected reaction monitoring of analytes was performed under atmospheric pressure chemical ionization in positive mode at m/z 537→321 and m/z 269→93 for respective [(12)C]β-carotene and [(12)C] retinoids; m/z 547→330 and m/z 274→98 for [(13)C10]β-carotene and [(13)C5] cleavage products; and m/z 279→100 for metabolites of [(13)C10]retinyl acetate. A single one-phase solvent extraction, with no saponification or purification steps, left retinyl esters intact for determination of intestinally-derived retinol in chylomicrons versus retinol from the liver bound to retinol binding protein. Coadministration of [(13)C10]retinyl acetate with [(13)C10]β-carotene not only acts as a reference dose for inter-individual variations in absorption and chylomicron clearance rates, but also allows for simultaneous determination of an individual's vitamin A status.
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ISSN:1539-7262
0022-2275
1539-7262
DOI:10.1194/jlr.D040204