Quantitative analyses of ß-carotene and retinol in serum and feces in support of clinical bioavalailability studies

D. Zhu, Y. Wang, Y. Pang, A. Liu, Jian Guo, C.A. Bouwman, C.E. West, R.B. van Breemen

Research output: Contribution to journalArticleAcademicpeer-review

10 Citations (Scopus)

Abstract

Among more than 50 provitamin carotenoids, beta-carotene is the most metabolically active source of retinol. Despite diets rich in fruits and vegetables containing beta-carotene, vitamin A deficiency is the leading cause of blindness and childhood mortality in developing countries. In addition, the uncertainty of beta-carotene bioconversion into vitamin A suggests that new data are needed to update the nutritional guidelines in developed countries. Previously, we reported the development of a carotene/retinol plateau isotopic enrichment method (CarRet PIE) for the determination of beta-carotene bioavailability and bioconversion into retinol, which utilizes positive ion atmospheric pressure chemical ionization (APCI) liquid chromatography/mass spectrometry (LC/MS). While seeking to validate the CarRet PIE using a mass balance approach requiring fecal measurements of beta-carotene and retinol, interference was encountered that required substantial modifications of the LC/MS assay. Here we report a new LC/MS assay that is based on the detection of molecular anions of beta-carotene using negative ion APCI with a reversed-phase C-30 column for HPLC separation. Sample preparation required saponification to eliminate interfering triglycerides. The limit of detection (LOD) of beta-carotene was 0.25 pmol calculated on the basis of an injection of 20 mu L of 0.0125 mu M beta-carotene, and the limit of quantitation (LOQ) was 1.0 pmol based on the injection of 20 mu L of 0.050 mu M beta-carotene. The linear range was 1.1 to 2179 pmol on-column. The wide linear range and low LOD and LOQ of this assay facilitated the sensitive and selective quantitative analysis of beta-carotene in both serum and fecal samples in support of an on-going clinical investigation of beta-carotene bioavailability and bioconversion into vitamin A. Copyright (c) 2006 John Wiley & Sons, Ltd.
Original languageEnglish
Pages (from-to)2427-2432
JournalRapid Communications in Mass Spectrometry
Volume20
Issue number16
DOIs
Publication statusPublished - 2006

Keywords

  • chromatography-mass-spectrometry
  • bioefficacy
  • children
  • humans

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