Mitochondrial deficiency is the cause of many diseases and the determination of changes in metabolic rates usually requires lysing of the mitochondria and isolating individual mitochondrial proteins. Alternatively, mitochondria can be immobilized on electrode surfaces to utilize electroanalytical evaluation of metabolic rates of intact mitochondria. However, the redox mechanisms are still poorly understood. In this paper, the riboflavin cycle of mitochondria is studied electrochemically and its impact on mitochondrial voltammetry is discussed. The inhibition mechanism of mitochondria by three different inhibitors (rotenone, carboxin, and permethrin) is discussed and it is found that the inhibition behavior observed electrochemically is due to not only ubiquinone, which is the electrochemical communicating species of mitochondrial electrochemistry. It is also shown that riboflavin derivatives interact with ubiquinone leading to a change in the intensity of ubiquinone voltammetry peaks. This interaction is affected by altering the choice of solvent used during the electrode preparation process. Finally, it is concluded that the observed voltammetry of mitochondrial inhibition is due to a change in riboflavin metabolism within the intact mitochondria immobilized on carbon electrodes. (C) The Author(s) 2016. Published by ECS. All rights reserved.