Hydroxylated polychlorinated biphenyls (OH-PCBs) were measured in surficial sediment from Indiana Harbor and Ship Canal (IHSC), East Chicago, IN and five original Monsanto Aroclors. These compounds were measured using gas chromatography with tandem mass spectrometry (GC-MS/MS) and certified standards that allowed us to identify 65 individual or coeluting congeners. Concentrations in the sediment ranged from 0.20 to 26 ng/g dry weight. Profiles of most samples were similar and were dominated by mono- to penta-chlorinated OH-PCBs. Interestingly, most of the samples strongly resembled the OH-PCB profiles of Aroclors 1221, 1242, 1248, and 1254, yet 25% of OH-PCBs measured in the sediment were not detected in Aroclors. A strong positive correlation was found between ΣOH-PCB and ΣPCB (p < 0.0001) and also between many individual OH-PCB:PCB pairs (p < 0.05). Analysis of OH-PCB:PCB pairs suggest PCB degradation is unlikely as a source of OH-PCBs in IHSC sediment. We are the first to report levels of OH-PCBs in sediment and Aroclors, and our discovery is significant because it is likely that OH-PCB contamination exists in sediment anywhere that PCB contamination from Aroclors is present.