Polycyclic aromatic hydrocarbons (PAHs) substituted with a ketone or quinone functionality (OPAHs) may be important environmental contaminants. The OPAHs from environmental samples have demonstrated toxicity and may be more harmful than PAHs. Knowledge gaps concerning the occurrence of OPAHs in the total environment arise from analytical difficulties, as well as limited standards and methodologies. An optimized method was developed to quantify five ketone and four quinone OPAHs from matrices ranging from biological tissue to diesel particulates. Five National Institute of Standards and Technology Standard Reference Materials (SRMs) were analyzed. This is the first report of OPAH quantitation in SRM 2977 (mussel tissue), SRM 1944 (New York/New Jersey, USA waterway sediment), SRM 1975 (diesel extract), and SRM 1650b (diesel particulate matter) and among the few to report concentrations from SRM 1649 (urban dust). Furthermore, this is one of the first reports of OPAHs in biological tissue. Σ₉OPAHs were 374 ± 59 mg/kg (mussel tissue), 5.4 ± 0.5 mg/kg (sediment), 16.9 ± 1.6 mg/kg (urban dust), 33.4 ± 0.4 mg/kg (diesel extract), and 150 ± 43 mg/kg (diesel particulate matter). In all SRMs, the levels of OPAHs were similar to or exceeded levels of PAHs. Of the OPAHs tested, the most frequently occurring in the environmental matrices were 9-fluorenone, 9,10-anthraquinone, benzofluorenone, and 7,12-benz[a]anthracenequinone.