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3 protocols using hplc solvents

1

Cellular Assay Optimization for Gefitinib Cytotoxicity

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Gefitinib was purchased from Quimigen. Chlorpromazine hydrochloride (CPZ), sodium dodecyl sulphate (SDS), anthracene, human serum albumin (HSA) were purchased from Sigma-Aldrich. For cell culture experiments, HaCaT cells and Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and penicillin–streptomycin (1.0 × 105 U mL−1, 1.0 × 105 μg mL−1) were supplied by invitrogen. Trypsin–EDTA (0.25–0.02%) and glutamine solutions were provided by Cultek. 2,4-Dinitrophenylhydrazine hydrochloride (DNPH) was purchased from Santa Cruz Biotechnology (Dallas, USA). PBS buffer was prepared by dissolving phosphate-buffered saline tablets (Sigma) using ultrapure water from a Millipore (Milli-Q Synthesis) system. Spectrophotometric HPLC solvents were obtained from Scharlab and used without further purification.
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2

Synthesis and Characterization of Putrescine-Ibuprofen Conjugates

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Both 1,4-diaminobutane (putrescine, PTR) and rac-ibuprofen were purchased from TCI Europe (Paris, France). Glutaraldehyde (GA) grade II (25% in H2O) was purchased from Merck Life Science S.L.U. (Madrid, Spain). Bradford reagent from Bio-Rad (Alcobendas, Madrid, Spain). HPLC solvents were purchased from Scharlab S.L. (Colmenar Viejo, Madrid, Spain).
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3

Phenolic Compounds Analysis Protocol

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All the used HPLC solvents were from Scharlab (Barcelona, Spain). All the used phenolic standards were from Sigma-Aldrich (Madrid, Spain), with the exception of malvidin-3-glucoside (enyn-chloride, Extrasynthese, Genay, France). Among the chemicals for spectrophotometric analysis, the Folin–Ciocalteu reagent, Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), gallic acid, and quercetin were supplied by Sigma-Aldrich (Madrid, Spain); glacial acetic acid, anhydrous sodium carbonate, and aluminum chloride 6-hydrate were supplied by PanReac AppliChem (Barcelona, Spain).
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