Water samples for chemical analysis were collected directly from the cylinder outflow at the start of the experiment (day 0) and days 4, 10 (warm treatment only), 18, 28 (warm treatment only), and 56 for each temperature and crude oil concentration. Samples were acidified (15% HCl) and stored in the dark at 4 °C until analysis. The organic phase was extracted from the water by first transferring the entire sample to separatory funnels and rinsing sample bottles with dichloromethane (DCM, 60–90 mL depending on sample size). Surrogate internal standards (25.08 ng naphthalene-d8, 5.00 ng phenanthrene-d10, 4.86 ng chrysene-d12 and 5.08 ng perylene-d12) were added to each sample. The organic and aqueous phase were allowed to separate after shaking, and the organic phase was transferred to a flask with sodium sulfate. This was repeated two more times before the organic phases were combined and concentrated using a TurboVap evaporator. Before analysis, recovery internal standards (fluorene-d10 and acenaphthene-d10) were added to samples. A laboratory blank (MilliQ water) was included with each sample set.

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