DETERMINATION OF UBIQUINONE IN SERUM AND LIVER BY HIGH-SPEED LIQUID CHROMATOGRAPHY

• Published in: Journal of Nutritional Science and Vitaminology 1978, Vol.24(6), pp.555-567
• Main Authors: ABE, Kouichi, ISHIBASHI, Kyoko, OHMAE, Masahiko, KAWABE, Kiyoshi, KATSUI, Goichiro
• Format: Article
• Language: English
• Subjects: Animals; Chromatography, High Pressure Liquid - methods; Fluorometry; Humans; Liver - analysis; Mice; Ubiquinone - analysis; Ubiquinone - blood; Ultraviolet Rays
• ISSN: 0301-4800; 1881-7742
• DOI: 10.3177/jnsv.24.555

Two kinds of high-speed liquid chromatographic methods using either ultraviolet or a fluorometric detector (HSLC-UV and HSLC-fluorometric methods) are described for the determination of ubiquinones (UQ). HSLC-UV method: Serum or liver was saponified with methanolic alkaline solution containing pyrogallol as an antioxidant and 2, 3, 6trimethyl-5-nonaprenyl-l, 4-benzoquinone as an internal standard and then extracted with n-hexane. The extract was evaporated and then dissolved in dioxane. The resulting solution was applied to a HSLC system using a Permaphase ODS column, ethanol-water (80:20) as a mobile phase and a high sensitivity UV detector (275 nm). HSLC-fluorometric method: Serum was deproteinized with ethanol and then extracted with n-hexane. The resulting extract was reacted with alkaline ethylcyanoacetate (ECA) reagent to give fluorescence. The reacted solution was applied to a HSLC system using a Permaphase ODS column, ethanol-water (75:25) as a mobile phase and a spectrofluorometer (Ex. 430 nm, Em. 530 nm). Neither of the proposed HSLC methods were affected by other fat-soluble substances and seemed to be more simple, sensitive and specific than other conventional determination methods.