Aluminum chloride

For the preparation of solutions, ultrapure water (type I water) was utilized alongside analytical-grade reagents, including sodium hydroxide (Neon, Suzano, SP, Brazil); ammonium hydroxide (Vetec, Duque de Caxias, RJ, Brazil); gelatin powder (Dinâmica, Indaiatuba, SP, Brazil); copper acetate (Êxodo, Sumaré, SP, Brazil); hydrochloric acid (Vetec, Duque de Caxias, RJ, Brazil); and sulfuric acid (Vetec, Duque de Caxias, RJ, Brazil) for the preparation of some solutions. Alcoholic solutions were prepared using ethanol (Dinâmica, Indaiatuba, SP, Brazil) combined with aluminum chloride (Dinâmica, Indaiatuba, SP, Brazil); ferric chloride (Proquímios, Rio de Janeiro, RJ, Brazil); quinine hydrochloride (Dinâmica, Indaiatuba, SP, Brazil); and vanillin (Dinâmica, Indaiatuba, SP, Brazil). Solvents such as hexane (Sigma-Aldrich, Darmstadt, Hesse, Germany); methyl alcohol (Vetec, Duque de Caxias, RJ, Brazil); and acetone (CRQ Química, Diadema, SP, Brazil) were also employed. Phytochemical screening utilized metallic magnesium shavings (Dinâmica, Indaiatuba, SP, Brazil); dried botanical material lavender and wormwood (Produtos Melvina, Goiânia, GO, Brazil); and oregano and sage (Produtos Gizele, Brasília, DF, Brazil), along with commercially procured bar soap.

Dulbecco’s Modified Eagle’s Medium (DMEM) and all cell culture reagents were purchased from Life Technologies (CA, USA). Fetal bovine serum (FBS) and MTT were obtained from Himedia. FITC AnnexinV Apoptosis Detection Kit was bought from Becton Dickinson (NJ, USA). All other chemicals were of analytical grade. A colorimetric assay kit was used to measure caspase-3 activity (R&D Systems, Inc., Minneapolis, MN, USA). Enzyme-linked immunonosorbent assay (ELISA) kits were used to measure Bax and Bcl-2 protein levels (Elabscience, Houston, TX, USA). Ethanol solution (60%), for extraction procedures, was prepared using ethanol PA from Synth (Brazil). Sodium carbonate and aluminum chloride were obtained from Dinâmica (Brazil) and Synth, respectively, while Folin-Ciocalteu was obtained from Merck^®. Gallic acid and quercetin standards were purchased from Sigma (Annapolis, MD, USA). Methanol was used for UPLC/MS procedures, acetonitrile and formic acid were HPLC grade from Merck^®, and sodium formate was prepared from sodium hydroxide (Dinâmica, Brazil) and formic acid (Merck^®).