New Method for Assaying Free and Total Cholesterol in Cultured Cells by High-pressure Liquid Chromatography

Kozo Hayashi; Yoshio Kuga; Yoshifumi Okura; Kouichi Tanaka; Yuji Yasunobu; Katsuhiko Nomura; Yasushi Toyota; Harumi Ohtani; Tetsuji Shingu; Goro Kajiyama

doi:10.5551/jat1994.2.117

Kozo Hayashi, Yoshio Kuga, Yoshifumi Okura, Kouichi Tanaka, Yuji Yasunobu, Katsuhiko Nomura, Yasushi Toyota, Harumi Ohtani, Tetsuji Shingu, Goro Kajiyama

Author information

Kozo Hayashi
First Department of Internal Medicine, Hiroshima University School of Medicine
Yoshio Kuga
First Department of Internal Medicine, Hiroshima University School of Medicine
Yoshifumi Okura
First Department of Internal Medicine, Hiroshima University School of Medicine
Kouichi Tanaka
First Department of Internal Medicine, Hiroshima University School of Medicine
Yuji Yasunobu
First Department of Internal Medicine, Hiroshima University School of Medicine
Katsuhiko Nomura
First Department of Internal Medicine, Hiroshima University School of Medicine
Yasushi Toyota
First Department of Internal Medicine, Hiroshima University School of Medicine
Harumi Ohtani
First Department of Internal Medicine, Hiroshima University School of Medicine
Tetsuji Shingu
First Department of Internal Medicine, Hiroshima University School of Medicine
Goro Kajiyama
First Department of Internal Medicine, Hiroshima University School of Medicine

Keywords: Cholesterol, 4-cholesten-3-one, 7β-hydroxy-4-cholesten-3-one, High-pressure liquid chromatography, Cultured cells

JOURNAL FREE ACCESS

1996 Volume 2 Issue 2 Pages 117-121

DOI https://doi.org/10.5551/jat1994.2.117

Details

Abstract

We developed a simple, sensitive and accurate method for assaying cellular free and total cholesterol by monitoring 4-cholesten-3-one, a conversion product of the cholesterol oxidasecatalyzed oxidation of the free cholesterol that has a strong chromophoric α, β-unsaturated ketone at 20 nm, using a high-pressure liquid chromatographic system. This method measured picomole quantities of free and total cholesterol and precisely determined their concentrations in cells (10⁴ range) in culture using 7β-hydroxycholesterol as an internal standard. J Atheroscler Thromb, 1996 ; 2 : 117-121.

Corresponding author

Register with J-STAGE for free!