Acetone

The graphite (Grafine 996100) was acquired from the company Nacional de Grafite (Salto da Divisa, Brazil). H₂SO₄, H₃PO₄, HCl, and acetone were purchased from Labsynth (Diadema, Brazil), while KMnO₄ and H₂O₂ used in the synthesis of GO were acquired from Sigma-Aldrich (São Paulo, Brazil). The GO-based nanocomposites were prepared using a standard rigid acrylic photosensitive resin obtained from Shenzhen Creality 3d Technology Co. (Shenzhen, China). This acrylic resin is transparent, with a dynamic viscosity of 150–200 mPa.s, density of 1.15 g.cm^–3, and an absorption band between 355 and 410 nm [37 (link)]. All reagents were used as purchased.

HPLC-grade methanol, isopropanol, and methyl tert-butyl ether (MTBE) were purchased from J. T. Baker (Phillipsburg, NJ, USA). Acetone was acquired from Labsynth (Diadema,SP, Brazil), and all ultrapurified water was generated on-site using a Milli-Q 3 Ultrapure Water System (Merck-Millipore, Darmstadt, Germany). The steroid standards were acquired from Sigma-Aldrich (St. Louis, MO, USA). The internal standards cortisol-d₄, testosterone-¹³C₃, and estradiol-d₂ were purchased from Sigma Aldrich (St. Louis, MO, USA) and progesterone-d₉ was purchased from Steraloids Inc (Newport, RI, USA).
Standards’ stock solutions for all of the steroids and their respective internal standards were gravimetrically prepared for calibration. A mass of 1.0 to 3.0 mg of each hormone was accurately weighed and dissolved in 15.0 mL of methanol to prepare solutions ranging from 24.63 to 70.42 μg g^-1. The working and calibration solutions were prepared accordingly by dilutions with methanol. Six different concentration levels were used for the calibration curves, and three recovery levels were prepared daily and analyzed during three sequential days to evaluate the method recovery and performance.

The following reagents were employed: acetone was acquired from Labsynth (Diadema, SP, Brazil), hexane, ethanol, methanol and methyl tert-butyl ether (MTBE) were acquired from Merck (Darmstadt, Germany), vanillin and sulfuric acid (H₂SO₄) were obtained from Dinâmica Química Contemporânea (Diadema, SP, Brazil), and the Folin–Ciocalteu reagent, sodium carbonate Na₂CO₃, dithiothreitol (DTT), metaphosphoric acid, potassium chloride (KCl), Tris (hydroxymethyl) aminomethane, potassium peroxydisulfate, 2,20-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), monobasic and dibasic potassium phosphate, fluorescein sodium salt, 2,20-azobis(2-methylpropionamidine) dihydrochloride (AAPH), sodium hypochlorite solution (NaOCl) and rhodamine 123 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Gallic acid, ascorbic acid, lutein, zeaxanthin, β-cryptoxanthin, α-carotene β-carotene, catechin, (±)-6-hydroxy-2,5,7,8-tetramethylchromane-2-car- boxylic acid (Trolox) were used as standards and purchased from Sigma-Aldrich (St. Louis, MO, USA). Ultrapure water was obtained from a Millipore Milli-Q System (Merck Millipore, Steinfurt, Germany).

All aqueous solutions were prepared using ultrapure deionized water (18.2 M.Ω.cm, Gehaka, Master WFI, São Paulo, SP, Brazil). Dichloromethane and acetone were purchased from LabSynth, Diadema, SP, Brazil. Gelatin, Tween 80^®, PCL, sorbitan stearate (Span 60), and caprylic/capric acid triglyceride (TAAC) were purchased form Sigma-Aldrich, Barueri, Brazil. Transglutaminase was purchased from GastronomyLab, Brasília, Brazil.

All aqueous solutions were prepared using ultrapure deionized water (18.2 M.Ω.cm, Master WFI, Gehaka, São Paulo, Brazil). Acetone, diiodomethane, formamide and ethylene glycol were purchased from LabSynth, Diadema—São Paulo, Brazil. Polyvinyl alcohol (PVA), poly (D, L-lactic-co-glycolic acid) [(50:50); Mw = 30,000–60,000 g·mol⁻¹] and Kollisolv^® PEG400 (polyethylene glycol 400; Mw = 380–420 g·mol⁻¹) were purchased from Sigma-Aldrich (Burlington, MA, USA). Aminolevulinic acid hydrochloride [95% and 99% (5-ALA)] were purchased from PDTPharma (Cravinhos, São Paulo, Brazil).