Determination of ubiquinone in serum and liver by high-speed liquid chromatography.

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Abstract
High-speed liquid chromatographic methods using either UV or a fluorometric detector (HSLC-UV and HSLC-fluorometric methods) are described for the determination of ubiquinones (UQ). In the HSLC-UV method [human] serum or [mouse] liver was saponified with methanolic alkaline solution containing pyrogallol as an antioxidant and 2,3,6-trimethyl-5-nonaprenyl-1,4-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). In the 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 both seemed to be more simple, sensitive and specific than other conventional determination methods.

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