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2 protocols using sodium sulphate anhydrous na2so4

1

Synthesis and Characterization of Stimuli-Responsive Polymers

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N-isopropylacrylamide (NIPAM, >99%, Sigma, St. Louis, MO, USA) was recrystallized from n-hexane before use. Furthermore, 2, 2’-Azobisisobutyronitrile (AIBN, 98%) and 4,4’-Azobis(4-cyanovaleric acid) (ACVA, ≥98%) were obtained from Sigma (St. Louis, MO, USA) and recrystallized from methanol prior to use. Stearic acid (>98%) was purchased from TCI America (Tokyo, Japan) and used as received. Additionally, 4-Cyano-4-[(dodecylsulfanylthiocarbonyl)sulfanyl]pentanoic acid (CDP, 97%), dicyclohexylcarbodiimide (DCC, 99%), 4-dimethylaminopyridine (DMAP, 99%), 2-hydroxyethyl methacrylate (HEMA, ≥99%) carbamazepine (CBZ, ≥98%), 1,3,5-trioxane (≥99%), pyrene (98%) and sodium sulphate anhydrous (Na2SO4, 99%) were purchased from Sigma (St. Louis, MO, USA) and used without any further purification. Then 1,4-Dioxane (99%, Fisher Scientific, Waltham, MA, USA) and tetrahydrofuran (THF, ≥99%, Sigma, St. Louis, MO, USA) was purified by being passed through a short alumina column before use. Chloroform-d (99.8 atom %D), dichloromethane (DCM, ≥99.5%) and ethyl acetate (EtOAc, ≥99.5%) were obtained from Sigma (St. Louis, MO, USA), while sodium bicarbonate (NaHCO3, 99.8%) and sodium chloride (NaCl, 99.9%), diethyl ether (≥99%), methanol (99.8%) and hexane (98.5%) were purchased from Fisher Scientific (Waltham, MA, USA).
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2

Determination of Total Hydrocarbons in Sediments

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Total hydrocarbons were extracted from sub-surface sediment following the 3550C EPA (Environmental Protection Agency) procedure. Briefly, a mixture of CH2Cl2:CH3COCH3 (1:1, v/v) was added to sediment samples. The mixture was sonicated for 30 min in ultrasound bath (Branson 1200 Ultrasonic Cleaner, Branson USA). Samples were further shaken at 150 ×g for 30 min, centrifuged for 10 min at 5000 ×g and supernatant was passed through a ceramic column filled with sodium sulphate anhydrous Na2SO4 (Sigma-Aldrich, Milan). Same treatment of pooled and dried sediment was repeated with CH2Cl2 and the obtained extracts were combined and volatilized to dryness. Residues were re-suspended in CH2Cl2 prior to gas chromatography (GC) analysis. All measures were performed using a Master GC DANI Instruments (Development Analytical Instruments), equipped with SSL injector and FID detector. Samples (1 μl) were injected in split less mode at 330 °C. The analytical column was a Restek Rxi-5 Sil MS with Integra-Guard, 30 m × 0.25 mm (ID × 0.25μm film thickness). Helium carrier gas was maintained at a constant flow of 1.5 ml min -1 . Total hydrocarbons were also calculated for each sample (Genovese et al., 2014) . Three subsamples for each station were analyzed.
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