Over the past quarter century, petroleum biomarkers have persisted in sequestered Exxon Valdez oil in Prince William Sound and the Gulf of Alaska (USA), and hence the oil has remained identifiable. These biomarkers are molecular fossils derived from biochemicals in previously living organisms. Novel pattern matching indicated the presence of Alaska North Slope crude oil (ANSCO) over the entire observation period at most sites (7 of 9) and distinguished this source from several other potential sources. The presence of ANSCO was confirmed with Nordtest forensics, demonstrating the veracity of the new method. The principal advantage of the new method is that it provides sample-specific identification, whereas the Nordtest approach is based on multisample statistics. Biomarkers were conserved relative to other constituents, and thus concentrations (per g oil) in initial beach samples were greater than those in fresh oil because they were lost more slowly than more labile oil constituents such as straight-chain alkanes and aromatic hydrocarbons. However, biomarker concentrations consistently declined thereafter (1989-2014), although loss varied substantially among and within sites. Isoprenoid loss was substantially greater than tricyclic triterpane, hopane, and sterane loss. Environ Toxicol Chem 2016;35:2683-2690. © 2016 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC. This article is a US government work and as such, is in the public domain in the United States of America.