Four late Pleistocene tephra layers—Tane I (Tn1), II (Tn2), III (Tn3), and IV (Tn4) in ascending order—are intercalated between widespread tephras, Kikai-Tozurahara (K-Tz: 95 ka) and Aira-Tn (AT: 30 cal kBP), on Tanegashima Island, in southern Japan. Paleolithic ruins such as the Yokomine C and Tatikiri archaeological sites were excavated from the loam layer between the Tn4 and Tn3 tephras. To refine the chronological framework on the island, we conducted accelerator mass spectrometry (AMS) radiocarbon dating for 2 paleosol and 6 charcoal samples related with the late Pleistocene tephras and the Yokomine C archaeological site. The obtained 14C dates are consistent with the stratigraphy in calendar years, 33 cal kBP for Tn4, 40 cal kBP for Tn3, and >50 cal kBP for Tn2 and Tn1. The charcoal dates from Yokomine C, 32–38 cal kBP, not only constrain the age of Tn4 and Tn3 ashes, but also serve as a possible date for the site. We also measured the major element compositions of volcanic glass shards with EDS-EPMA to characterize these tephras. Although we could not find a possible correlative for Tn3 and Tn4 ashes using major element oxides of the glass shards, i.e. 75–76 wt% in SiO2, the glass chemistry obtained in this study will be valuable in correlating these tephras with their source volcanoes in the near future.