We have already shown that prolonged supplementation of CoQ10 in humans reduces the wrinkle area rate and wrinkle volume per unit area in the corner of the eye. CoQ10 supplementation is known to increase the CoQ10 level in serum and in many organs; however, the level of CoQ10 in skin has not yet been fully investigated yet. We examined whether CoQ10 intake elevates the CoQ10 and CoQ9 levels in epidermis, dermis, serum and other organs (kidney, heart, brain, muscle and crystalline lens) in 43-week-old hairless male mice. We also established a method using a high performance liquid chromatograph equipped with an electrochemical detector (HPLC-ECD) to simultaneously quantify CCoQ9 and CoQ10 in the tissues. CoQ10 (0, 1, 100 mg/kg p.o.) was administered daily for 2 weeks. CoQ10 supplementation of 100 mg/kg increased the serum and epidermal CoQ10 levels significantly, but did not increase the CoQ10 levels in either dermis or other organs. In conclusion, we showed that CoQ10 intake elevates the epidermal CoQ10 level, which may be a prerequisite to the reduction of wrinkles and other benefits related to the potent antioxidant and energizing effects of CoQ10 in skin.