Per- and polyfluoroalkyl substances (PFAS) and co-occurring inorganic and organic contaminants are widely distributed in the environment and understanding their surface water sources is critical for water resource management. In November 2021, 13 sites were sampled along a stream reach in the Neshaminy Creek basin in southeastern Pennsylvania utilizing a time of travel sampling approach. The reach had known potential PFAS source inputs and documented PFAS detections in water samples. This data release describes the sampling approach, sample processing, laboratory analytical methods, and contains the results. Site (Table1_NC_2021_Site_Info) and constituent information (Table2_Constituents) are provided in tab delimited text (txt) files. Results are provided in a machine readable format in Table3_Reporting_Long.txt for the following hydrologic metrics and chemical constituents: (1) field parameters (instantaneous discharge, water and air temperature, barometric pressure, pH, specific conductance, and dissolved oxygen), (2) optical properties (ultraviolet (UV)-visible absorbance, and fluorescence), (3) dissolved organic carbon and total dissolved nitrogen, (4) major anions, (5) major and trace elements, (6) PFAS, (7) pharmaceutical compounds, and (8) volatile organic compounds. Table_4_Wide_Summary.xlxs is a summary file including the reported values, laboratory standard deviations, remark codes, and qualification codes from Table3_Reporting_Long.txt as eight wide format pivot tables.