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Mitochondrial Disorders

Volume 837 of the series Methods in Molecular Biology pp 149-168

Date:

Measurement of Oxidized and Reduced Coenzyme Q in Biological Fluids, Cells, and Tissues: An HPLC-EC Method

  • Peter H. TangAffiliated withDivision of Pathology and Laboratory Medicine, Cincinnati Children’s Hospital Medical Center
  • , Michael V. MilesAffiliated withDivision of Pathology and Laboratory Medicine, Departments of Pediatrics and Pathology & Laboratory Medicine, Cincinnati Children’s Hospital Medical Center and University of Cincinnati College of Medicine Email author 

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Abstract

Direct measure of coenzyme Q (CoQ) in biological specimens may provide important advantages. Precise and selective high-performance liquid chromatography (HPLC) methods with electrochemical (EC) detection have been developed for the measurement of reduced (ubiquinol) and oxidized (ubiquinone) CoQ in biological fluids, cells, and tissues. EC detection is preferred for measurement of CoQ because of its high sensitivity. Reduced and oxidized CoQ are first extracted from biological specimens using 1-propanol. After centrifugation, the 1-propanol supernatant is directly injected into HPLC and monitored at a dual-electrode. The EC reactions occur at the electrode surface. The first electrode transforms ubiquinone into ubiquinol, and the second electrode measures the current produced by the oxidation of the hydroquinone group of ubiquinol. The methods described provide rapid, precise, and simple procedures for determination of reduced and oxidized CoQ in biological fluids, cells, and tissues. The methods have been successfully adapted to meet regulatory requirements for clinical laboratories, and have been proven reliable for analysis of clinical and research samples for clinical trials and animal studies involving large numbers of specimens.

Key words

Coenzyme Q Ubiquinol Ubiquinone Redox HPLC Coulometric Electrochemical Analysis Plasma Milk Platelet Muscle Liver Brain