Biosolids from a municipal sewage treatment plant were studied from the standpoint of characterizing the matrix as well as targeting specific substances. Part of this interest stems from the land application of biosolids thatmay contain pharmaceuticals and personal care products (PPCPs). Targeted analytes were subjected to higher specificity than usual, and this was achieved either through multidimensional chromatography or through high resolution mass spectrometry (accurate mass determination) or both. The bacterial endpoints consisting of fecal sterols and fecal steroneswere present as expected and were generally greater than ppm levels meaning that they could pose significant interferences to target analytes at low ppb levels. Thus, one of our goals in characterization was to identify some of these major components as an aid in formulating cleanup strategies for the determination of PPCPs. Target analytes included nonyl phenols, polybrominated diphenyl ethers (PBDEs), and estrogens. These ranged from ppm levels for the first two groups to low or sub ppb levels for the estrogens. The nonyl phenols were further confirmed using high resolution voltage scanning and mass peak profiles at high resolution. The PBDEs were further confirmed using high resolution selected ion recording as were the estrogens.