Previous research has shown that unspiked OECD 309 tests can be used to quantify chemical biodegradation in surface waters, relying on chemical residues already present in the water. Here we test the hypothesis that unspiked OECD 309 tests can quantitatively predict chemical persistence in the environment by comparing chemical half-lives assessed in the laboratory against those measured in the field. The study object was a Swedish lake heavily impacted by treated municipal wastewater. Half-lives in the field were measured by mass balance over 12 weeks. In parallel, half-lives in the lab were determined with an unspiked OECD 309 test run for 60 days. Chemical analysis was conducted using a non-target screening approach. The field study yielded a half-life <100 days for 38 chemicals for which the dominant source was wastewater; 32 of these were also detected in the lab test, whereby 18 had half-lives with a well-constrained uncertainty that did not intersect infinity. For 14 of the 18 chemicals, the field and lab half-lives agreed within a factor 3. In summary, the lab test predicted chemical attenuation in the field well. Limitations of the approach include the need for measurable chemical concentrations in the water body and failure to account for some attenuation mechanisms like phototransformation.