Although forward osmosis (FO) membranes have shown great promise for many applications, there are few studies attempting to create a systematization of the testing conditions at a pilot scale for FO membrane modules. To address this issue, hollow fiber forward osmosis (HFFO) membrane modules with different performances (water flux and solute rejection) have been investigated at different operating conditions. Various draw and feed flow rates, draw solute types and concentrations, transmembrane pressures, temperatures, and operation modes have been studied using two model feed solutions—deionized water and artificial seawater. The significance of the operational conditions in the FO process was attributed to a dominant role of concentration polarization (CP) effects, where the selected draw solute and draw concentration had the biggest impact on membrane performance due to internal CP. Additionally, the rejection of the HFFO membranes using three model solutes (caffeine, niacin, and urea) were determined under both FO and reverse osmosis (RO) conditions with the same process recovery. FO rejections had an increase of 2% for caffeine, 19% for niacin, and 740% for urea compared to the RO rejections. Overall, this is the first extensive study of commercially available inside-out HFFO membrane modules.