Diclofenac sodium salt

Spectra/Por^® 6 regenerated cellulose (RC) dialysis membranes with a molecular weight cut-off (MWCO) of 50 kDa and a flat diameter of 28 mm were purchased from VWR International GmbH (Darmstadt, Germany). A stabilized suspension of reactant-free gold nanoparticles with a diameter of 50 nm (Sigma-Aldrich Chemie GmbH, Munich, Germany) in 0.1 mM phosphate-buffered saline (PBS) was used for the membrane leakage test. Diclofenac diethylamine (DEA) salt (Cayman Chemical, Ann Arbor, MI, USA) was purchased from Biomol GmbH (Hamburg, Germany) and diclofenac sodium salt was obtained from Sigma-Aldrich Chemie GmbH (Munich, Germany). Olfen^® gel (10 mg/g diclofenac sodium, Mepha Pharma AG, Basel, Switzerland), and Voltaren^® Emulgel (11.6 mg/g diclofenac-DEA, GlaxoSmithKline Consumer Healthcare GmbH & Co. KG, Munich, Germany) were obtained from a retail pharmacy. Idebenone was purchased from Rxn Chemicals (Hadapsar, India). All the other chemicals were of analytical grade or equivalent and used as received. An Ultra Clear^® system (Evoqua water technologies, Günzburg, Germany) was used for water purification in all the experiments.

Polypropylene glycol (molecular weight ~2000) and diclofenac sodium salt was purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). All other chemicals used were of analytical grade, obtained from accredited companies and were used as received.

Poly-β-amino-cyclodextrin (PolyCD, Average MW = 25 kDa, CD content 70%) was synthesized at CycloLab (Budapest, Hungary) by cross-linking ad-hoc derivatized β-CD monomer with epichlorohydrin as already reported [45 (link)]. A colorimetric Kaiser test [48 (link),49 (link)] was performed to spectroscopically establish the quantity of amino groups present in PolyCD, which was estimated to be 0.21 mmol/g (see SI (Supplementary Information)). Adamantanyl-Rhodamine conjugate (Ada-Rhod, MW = 735.5 g/mol) was synthesized as previously reported [46 (link)]. Diclofenac sodium salt (DCF, MW = 318.13 g/mol) and all the solvents (analytical grade) were purchased from Sigma-Aldrich (Milano, Italy). All the dispersions used for nanoassemblies preparation and spectroscopic characterizations were prepared in ultrapure water (Fresenius Kabi Italia) or in 10 mM phosphate buffer containing NaCl (137 mM) and KCl (2.7 mM) at pH 7.4 (PBS) at room temperature (r.t. ≅ 25 °C). pH measurements were obtained using an 827 pH Lab pHmeter—Metrohm.

Diclofenac sodium salt (CAS: 15307-79-6) (≥98%) and bisphenol A (CAS 80-05-7) (≥97%) 4,4′-(propane-2,2-diyl) diphenol were purchased from Sigma-Aldrich (St. Louis, MO, USA) and Acros Organics (Geel, Bélgica), respectively. Acetonitrile and water for high-performance liquid chromatography (HPLC) analysis were acquired from Panreac Química SLU ITWReagents^® (Barcelona, Spain) and BDH Prolabo^® (Johannesburg, South Africa), respectively. DFC and BPA stock solutions were prepared at 5 mM and later sterilized by filtration (0.22 µm). The stock solution of DFC and BPA was used at a final concentration of 100 µM in the bioremediation experiments at flask scale.

Potassium peroxymonosulfate (oxone, 2KHSO₅·KHSO₄·K₂SO₄, >98%), iron(iii) chloride hexahydrate (FeCl₃·6H₂O, 98%), cobalt(ii) nitrate hexahydrate (Co(NO₃)₂·6H₂O, 98%), sodium hydroxide (NaOH, >97%), chloroform (CHCl₃, HPLC grade, >99.9%), acetone (CH₃COCH₃, HPLC grade, >99.8%), and N-trimethylsilyl N-methyl trifluoroacetamide (MSTFA, CF₃CON (CH₃)Si(CH₃)₃, >98%) was purchased from Sigma Aldrich. Diclofenac sodium salt (C₁₄H₁₀Cl₂NNaO, 98%) was prepared from Sigma Aldrich as the studied pollutant. Multi-walled carbon nanotubes (MWCNTs, >110 m² g⁻¹, 95%) were purchased from US research nanomaterials (Houston, USA). All solutions were prepared by purified water.

PEA (Levagen^®) was provided by Gencor Pacific Ltd, Hong Kong. Diclofenac sodium salt, MIA, 4',6-diamidino-2-phenylindole (DAPI), and hematoxylin and eosin (H&E) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kits for prostaglandin E2 (PGE₂), leukotriene B4 (LTB₄), tumor necrosis factor-α (TNF)-α, and interleukin (IL)-1β were purchased from R&D Systems (Minneapolis, MN, USA). Fluorochrome-conjugated secondary antibodies (Alexa 488 and 564) were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Safranin O was purchased from ScienCell Research Laboratories (Carlsbad, CA, USA), and anti-aggrecan and anti-collagen type II alpha 1 (COL2A1) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Stock solutions of the target compounds (Atenolol CAS 29122-68-7, Azithromycin CAS 83905-01-5, Caffeine CAS 58-05-2, Erythromycin CAS 114-07-8, Diclofenac sodium salt CAS 15307-79-6, Cyclophosphamide CAS 50-18-0, CiprOfloxacin CAS 85721-33-1, Sulfamethoxazole CAS 723-46-6, Carbamazepine CAS 298-46-4, Ketoprofen CAS 22071-15-4, Trimethoprim CAS 738-70-5, Clarithromycin CAS 81103-11-9, Ofloxacin CAS 82419-36-1, Estrone CAS 53-16-7, 17-beta Estradiol CAS 50-28-2, Ibuprofen CAS 15687-27-1; Sigma-Aldrich, St. Louis, MO, USA) were prepared in UHPLC-grade MeOH, purchased from Sigma-Aldrich, Co. (St. Louis, MO, USA). MilliQ water was obtained from MilliPore (MA, USA), LiChropur Formic Acid 98–100% and LiChropur Ammonia solution 25% for LC-MS were purchased from Merck KGaA (Darmstadt, Germany), Hydrochloric Acid (HCl), and Ethylenediaminetetraacetic acid trisodium salt dihydrate (Na₄EDTA) were obtained from Fluka Analytical (Sigma-Aldrich, MO, USA), while Ammonium acetate for LC-MS was purchased from Fisher Chemical Scientific (Geel, Belgium).