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An LC-MS/MS method for stable isotope dilution studies of γ-carotene bioefficacy and vitamin A status in humans

Published online by Cambridge University Press:  30 August 2013

Anthony Oxley
Affiliation:
Human Nutrition Research Centre, School of Agriculture, Food and Rural Development, Newcastle University, UK
Philip Berry
Affiliation:
Northern Institute for Cancer Research, Newcastle University, UK
Joseph Cowell
Affiliation:
School of Chemistry, Newcastle University, UK
Michael Hall
Affiliation:
School of Chemistry, Newcastle University, UK
John Hesketh
Affiliation:
Institute for Cell and Molecular Biosciences, Newcastle University, UK
Georg Lietz
Affiliation:
Human Nutrition Research Centre, School of Agriculture, Food and Rural Development, Newcastle University, UK
Alan V. Boddy
Affiliation:
Northern Institute for Cancer Research, Newcastle University, UK
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Abstract

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Abstract
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Copyright © The Authors 2013 

Isotope dilution is currently the most accurate technique in humans to determine vitamin A status and bioavailability/bioconversion of provitamin A carotenoids( Reference Furr, Green and Haskell 1 ). However, limits of MS detection, coupled with extensive isolation procedures, have hindered investigations of physiologically-relevant doses of stable isotopes( Reference van Lieshout, West and van Breemen 2 ). We developed a sensitive liquid chromatography-tandem-mass spectrometry (LC-MS/MS) analytical method to study the plasma response from co-administered oral doses of 2 mg [13C10]-β-carotene and 1 mg [13C10]-retinyl acetate in human subjects. 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 (RBP).

Fig. 1. Quantitative LC-MS/MS analysis of mean plasma responses from 45 human subjects (+/- sem) over the first 48 h (A, C) and whole 14 day study period (B). Administered [13C10]-β-carotene (βC) and resulting [13C5] cleavage products (RET, retinol; RL, retinyl linoleate; RPO, retinyl palmitate+oleate; RS, retinyl stearate) are shown in A, B, and C.

Co-administration of [13C10]-retinyl acetate with [13C10]-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. In summary, this new analytical method enables the detection of physiological concentrations of provitamin A carotenoids, their cleavage products as well as preformed vitamin A for a period of at least 2 weeks post dose administration and allows high throughput analysis due to its simplicity and short run times.

This work was supported by the BBSRC.

References

1. Furr, HC, Green, MH, Haskell, M et al. (2005) Public Health Nutrition 8(6), 596607.CrossRefGoogle Scholar
2. van Lieshout, M, West, CE and van Breemen, RB (2003) Am J Clin Nutr 77(1), 1228.CrossRefGoogle Scholar

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An LC-MS/MS method for stable isotope dilution studies of γ-carotene bioefficacy and vitamin A status in humans
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