TY - JOUR
T1 - Development of a high-performance liquid chromatography-based assay for carotenoids in human red blood cells
T2 - Application to clinical studies
AU - Nakagawa, Kiyotaka
AU - Kiko, Takehiro
AU - Hatade, Keijiro
AU - Asai, Akira
AU - Kimura, Fumiko
AU - Sookwong, Phumon
AU - Tsuduki, Tsuyoshi
AU - Arai, Hiroyuki
AU - Miyazawa, Teruo
N1 - Funding Information:
Financial support was provided by Project M Company (Sendai, Japan). This work was also supported by KAKENHI (20228002) of the Japan Society for the Promotion of Science (JSPS). We thank A. Nagao (National Food Research Institute, Tsukuba, Japan) for the gift of echinenone, and we thank K. Osada (Faculty of Agriculture, Meiji University, Kawasaki, Japan) for helpful discussions and technical advice regarding RBC carotenoid identification by APCI/MS.
PY - 2008/10/1
Y1 - 2008/10/1
N2 - 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.
AB - 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.
KW - Carotenoid
KW - Dementia
KW - HPLC-DAD-APCI/MS
KW - Lipid peroxidation
KW - Red blood cells
KW - Xanthophyll
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U2 - 10.1016/j.ab.2008.06.038
DO - 10.1016/j.ab.2008.06.038
M3 - Article
C2 - 18638443
AN - SCOPUS:49049092325
SN - 0003-2697
VL - 381
SP - 129
EP - 134
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -
