Reagents and solvents were used as received, with the exception of dichloromethane, which was distilled after drying with calcium hydride under reflux. Synthesis and characterization of rhodamine B-labeled triglyceride (1) CAO, whose main component is ricinolein (the triglyceride of ricinoleic acid, approximately 90%) [23], was covalently coupled with a fluorescent dye, rhodamine find more B (RhoB). Briefly, rhodamine B (1.91 g) and DMAP (0.49 g) were dissolved

in dry dichloromethane (30 mL) at room temperature under argon. After 40 min of stirring, EDCI.HCl (0.82 g) dissolved in dry dichloromethane (12 mL) was added to the reaction medium cooled in an ice bath. After 40 min under stirring, the CAO (2.08 g) dissolved in dry dichloromethane (4 mL) was then added. The reaction

medium was kept under stirring for 2 days in an argon atmosphere at room temperature. After this period, dichloromethane (30 mL) was added to the organic phase, and the extraction was carried out with aqueous solutions of firstly 1 mol L-1 HCl (3 × 40 mL) and then saturated NaHCO3 (3 × 40 mL). The organic phase was extracted with water (6 × 40 mL), dried under magnesium sulfate anhydrous, filtered, and evaporated under reduced pressure. The fluorescent product ICG-001 was purified by column chromatography using silica gel (60 to 200 mesh) and CHCl3 as eluent. The product 1 was Proteasome inhibitors in cancer therapy obtained as an oil. After purification, the process yielded 1.0 g of product 1. The product 1 was characterized by thin layer chromatography (TLC), Fourier transform infrared spectroscopy (FTIR), proton nuclear magnetic resonance (1H-NMR), size exclusion chromatography (SEC), UV-vis spectroscopy, and spectrofluorimetry. The TLC was performed using dichloromethane/methanol (9:1, v/v) as eluent and an aluminum not sheet (Merck, Whitehouse Station, NJ, USA) covered with silica gel 60 (70 to 230 mesh) as stationary phase. The bands were revealed under UV light at 365 nm (BOIT-LUB01, Boitton, Brazil). FTIR spectra were recorded

on a Varian® 640-IR spectrophotometer (Palo Alto, CA, USA) from 4,000 to 400 cm-1 (100 scans, 2 cm-1 resolution), using sodium chloride crystals. FTIR: 3,390 cm-1 (OH stretching), 2,940 and 2,850 cm-1 (CH2, asymmetric and symmetric stretching), and 1,740 cm-1 [C = O (ester)]. SEC analysis was carried out using a Viscotek® VE 2001 chromatograph with a Viscotek® TDA 302 triple detector and PS/DVB column (Malvern Instruments, Westborough, MA, USA). The purified product 1 and raw castor oil were dissolved in tetrahydrofurane, filtered (0.45 μm), and analyzed using polystyrene as reference. The product 1 was diluted in ACN and the maximum absorption wavelength (λ ab) was evaluated by UV-vis spectroscopy using a spectrophotometer (Shimadzu® UV-1601PC, Nakagyo-ku, Kyoto, Japan). The λ ab value was used to determine the maximum emission wavelength (λ max-em) by fluorimetry with a spectrofluorometer (Cary® 100, Agilent, Santa Clara, CA, USA).