Development of a high-performance liquid chromatography-based assay for carotenoids in human red blood cells: Application to clinical studies

Kiyotaka Nakagawa, Takehiro Kiko, Keijiro Hatade, Akira Asai, Fumiko Kimura, Phumon Sookwong, Tsuyoshi Tsuduki, Hiroyuki Arai, Teruo Miyazawa

Research output: Contribution to journalArticlepeer-review

38 Citations (Scopus)

Abstract

Peroxidized phospholipid-mediated cytotoxicity is involved in the pathophysiology of many diseases; for example, there is an abnormal increase of phospholipid hydroperoxides in red blood cells (RBCs) of dementia patients. Dietary carotenoids have gained attention as potent inhibitors of RBC phospholipid hydroperoxidation, thereby making them plausible candidates for preventing disease. However, the occurrence of carotenoids in human RBCs is still unclear. This is in contradistinction to plasma carotenoids, which have been investigated thoroughly for analytical methods as well as biological significance. In this study, we developed a method to analyze RBC carotenoids using high-performance liquid chromatography (HPLC) coupled with ultraviolet (UV) diode array detection (DAD) and atmospheric pressure chemical ionization (APCI) mass spectrometry (MS). Under optimized conditions that included extraction, separation, and detection procedures, six carotenoids (lutein, zeaxanthin, β-cryptoxanthin, α-carotene, β-carotene, and lycopene) were separated, detected by DAD, and concurrently identified based on APCI/MS and UV spectra profiles when an extract from human RBCs was subjected to HPLC-DAD-APCI/MS. The amounts of carotenoids varied markedly (1.3-70.2 nmol/L packed cells), and polar oxygenated carotenoids (xanthophylls) were predominant in RBCs. The HPLC-DAD-APCI/MS method would be a useful tool for clinical studies for evaluating the bioavailability of RBC carotenoids.

Original languageEnglish
Pages (from-to)129-134
Number of pages6
JournalAnalytical Biochemistry
Volume381
Issue number1
DOIs
Publication statusPublished - 2008 Oct 1

Keywords

  • Carotenoid
  • Dementia
  • HPLC-DAD-APCI/MS
  • Lipid peroxidation
  • Red blood cells
  • Xanthophyll

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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