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Isotopic ratios of strontium and sulfur in six layers across a horizontal section of a hydrothermal barite–silica chimney from Franklin Seamount of western Woodlark Basin have been investigated. Sr-isotopic ratios in barite samples (87Sr/86Sr = 0.70478–0.70493) are less radiogenic than seawater (87Sr/86Sr = 0.70917) indicating that substantial leaching of sub-seafloor magma was involved in the genesis of hydrothermal fluid. The SO2 of magma likely contributed a considerable amount of lighter S-isotope in fluid and responsible for the formation of barite, which is isotopically lighter (δ34S = 19.4–20.5 ‰) than modern seawater (δ34S ~ 21 ‰). The systematic changes in isotopic compositions across the chimney wall suggest temporal changes in the mode of mineral formation during the growth of the chimney. Enrichment of heavy S- and Sr-isotopes (δ34S = 20.58 ‰; 87Sr/86Sr = 0.70493) in the outermost periphery of the chimney indicates that, at the initial stage of chimney development, there was a significant contribution of seawater sulfate during barite mineralization. Thereafter, thickening of chimney wall occurred due to precipitation of fluid carrying more magmatic components relative to seawater. This led to a gradual enrichment of lighter isotopes (δ34S = 20.42–19.48 ‰; 87Sr/86Sr = 0.70491–0.704787) toward the inner portion of the chimney wall. In contrast, the innermost layer surrounding the fluid conduit is characterized by heavier and more radiogenic isotopes (δ34S = 20.3 ‰; 87Sr/86Sr = 0.7049). This suggests there was increasing influence of percolating seawater on the mineral paragenesis at the waning phase of the chimney development. © 2016, Springer-Verlag Berlin Heidelberg.