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The relationship between groundwater and surface water in the Lake Bosumtwi impact crater has been assessed using hydrochemical data and stable water isotopes of δ18O and δD. This study aimed to define likely groundwater flow and recharge zones, estimate the rate of evaporation, and examine the relationship between the lake and groundwater in the study area. The results of Q-Mode hierarchical cluster analysis (HCA) clearly differentiate the lake water from the groundwater based on their spatial relationship. These preliminary results indicated that groundwater recharge occurs on the hilltops of the crater, where it is slightly acidic with low levels of dissolved minerals, characterized by short residence time and rapid unrestricted vertical infiltration and recharge. The groundwater becomes more mineralized with longer contact times and deeper circulation with the host rock, while it flows from the recharge areas towards the lake at lower elevations. Analyses of stable water isotopes of δ18O and δD showed a high evaporation rate on the lake surface, of ~90% with a relatively significant evaporative enrichment, whereas groundwater showed a relatively lower evaporation rate ranging between 54-60%. Both reservoirs do not appear to be hydraulically connected, and where such a connection exists, it is expected to favour the lake.