Highly sensitive quantification of key regulatory oxysterols in biological samples by LC-ESI-MS/MSs⃞

Akira Honda; Kouwa Yamashita; Takashi Hara; Tadashi Ikegami; Teruo Miyazaki; Mutsumi Shirai; Guorong Xu; Mitsuteru Numazawa; Yasushi Matsuzaki

Journal of Lipid Research (Feb 2009)

Highly sensitive quantification of key regulatory oxysterols in biological samples by LC-ESI-MS/MSs⃞

Akira Honda,
Kouwa Yamashita,
Takashi Hara,
Tadashi Ikegami,
Teruo Miyazaki,
Mutsumi Shirai,
Guorong Xu,
Mitsuteru Numazawa,
Yasushi Matsuzaki

Affiliations

Akira Honda: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Kouwa Yamashita: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Takashi Hara: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Tadashi Ikegami: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Teruo Miyazaki: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Mutsumi Shirai: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Guorong Xu: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Mitsuteru Numazawa: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103
Yasushi Matsuzaki: Center for Collaborative Research, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Faculty of Pharmaceutical Science, Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, Japan; Ibaraki Prefectural Institute of Public Health, Mito, Ibaraki 310-0852, Japan; Department of Gastroenterology, Tokyo Medical University, Kasumigaura Hospital, Ami, Ibaraki 300-0395, Japan; Department of Medicine, University of Medicine and Dentistry of New Jersey-New Jersey Medical School, Newark, NJ 07103

Journal volume & issue: Vol. 50, no. 2
pp. 350 – 357

Abstract

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We describe a highly sensitive and specific method for the quantification of key regulatory oxysterols in biological samples. This method is based upon a stable isotope dilution technique by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After alkaline hydrolysis of human serum (5 μl) or rat liver microsomes (1 mg protein), oxysterols were extracted, derivatized into picolinyl esters, and analyzed by LC-MS/MS using the electrospray ionization mode. The detection limits of the picolinyl esters of 4β-hydroxycholesterol, 7α-hydroxycholesterol, 22R-hydroxycholesterol, 24S-hydroxycholesterol, 25-hydroxycholesterol, 27-hydroxycholesterol, and 24S,25-epoxycholesterol were 2–10 fg (5–25 amol) on-column (signal-to-noise ratio = 3). Reproducibilities and recoveries of these oxysterols were validated according to one-way layout and polynomial equation, respectively. The variances between sample preparations and between measurements by this method were calculated to be 1.8% to 12.7% and 2.9% to 11.9%, respectively. The recovery experiments were performed using rat liver microsomes spiked with 0.05 ng to 12 ng of oxysterols, and recoveries of the oxysterols ranged from 86.7% to 107.3%, with a mean recovery of 100.6%. This method provides reproducible and reliable results for the quantification of oxysterols in small amounts of biological samples.

Published in Journal of Lipid Research

ISSN: 0022-2275 (Print); 1539-7262 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Chemistry: Organic chemistry: Biochemistry
Website: https://www.journals.elsevier.com/journal-of-lipid-research

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