Ubiquinone Coenzyme Q10 HPLC Assay

Additional Information

Assay Principle

For the determination of ubiquinone (coenzyme Q10) a precipitation step which removes high molecular substances is performed first. The internal standard reduces the variation in the sample treatment. After centrifugation the supernatant is mixed with an extraction solution and transferred to the organic phase. The organic solvent is evaporated and the sample is re-suspended in ethanol p.A. and injected into the HPLC system.  The isocratic separation via HPLC at 30°C using a “reversed phase“ column needs 15 minutes. The chromatograms are recorded by a UV-detector. The quantification is performed by the delivered calibrator and the concentration is calculated by the “internal standard method” via integration of the peak areas resp. peak heights.

Assay Background

Ubiquinone is a coenzyme of every cell in the body. Ubiquinone is made up by a chinonic ring and an isoprenic side chain. In humans ubiquinone can be synthesized and taken up by nutrition.  Ubiquinone has two different physiological functions.

1. Component of the energy metabolism
   • During the reduction of oxygen in the oxidative phosphorylation, electrons are transferred from NADPH to oxygen via 6 different redox systems. Ubiquinone is the less abundant redox system in the membrane of the mitochondria. Because of the low amount it is the speed controlling redox-system in the energy metabolism. Normally the amount of ubiquinone is sufficient, but with growing age and exposure to sunlight it is reduced to 50 %.
2. Radical scavenger
   • Ubiquinone has a high amount of carbon double bonds and therefore a higher potential of reduction than either vitamin C or vitamin E. Thereby it is the first line of defense against free radicals. Even when other antioxidants are inert, coenzyme Q10 reacts fast and sensitive against free radicals. Therefore ubiquinone is an optimal stabilizer of the ion channels of the membranes.

The Eagle Biosciences Ubiquinone (Coenzyme Q10) HPLC Assay kit makes it possible to determine ubiquinone in an easy, fast and precise method. The Ubiquinone (Coenzyme Q10) HPLC Assay kit includes all reagents ready to use for preparation and separation of the samples with exception of the column (IC1700rp) and the controls (IC1700ko).  Both can be supplied by Eagle Biosciences.  Besides the complete test kit, it is possible to order all components separately. Please request our single component price list.