Temporal changes in mating behavior and daily sperm movement were examined in adult males of the butterfly Polygonia c‐aureum L. (Lepidoptera: Nymphalidae), reared under L15:D9 photoperiod at 21 °C. In this butterfly, when extra sperm is present in the duplex (storage organ) at mating, sperm reflux occurs: extra sperm are moved from the duplex to the lower and middle portions of the vasa deferentia by peristaltic movement of the lower portion of the vasa deferentia. It is known in several moth species that daily sperm movement from the testis to the upper portion of the vasa deferentia and from the upper vasa deferentia to the duplex occurs in a circadian manner. If sperm reflux and daily sperm movement occur at the same time, it could create confusion, because these sperm movements are directed oppositely. In this study, we examined the temporal changes of sperm movement and mating behavior in P. c‐aureum in order to clarify whether the two sperm movements are separated in time. Our results showed that most matings occurred during the second half of the photophase, whereas daily eupyrene (nucleate) sperm movement from the upper to the middle portion of the vasa deferentia occurred immediately after light on (early photophase), indicating the presence of a time lag between mating and daily sperm movement. As sperm reflux occurs during mating, these two oppositely directed sperm movements could be performed without conflict.