Nelson 3200 series

Extraction of lycopene was made according to Barba [40 (link)]; 0.3 g of tomato peel and pulp (taken from the pull described previously in Section 2.4) were added to 10 mL of a solvent solution made by hexane/acetone/ethanol (50:25:25 v/v/v) and homogenized with Ultra-Turrax (IKA^®). Subsequently, 1.5 mL of distilled water was added, and the samples were vortexed. The upper layer (1 mL) was dried under vacuum and the dry extract was resuspended in 0.4 mL of tetrahydrofuran (THF)/acetonitrile (ACN)/methanol (15:30:55 v/v/v). The mobile phase for HPLC (Perkin Elmer Nelson 3200 Series) analysis consisted of methanol/ACN (90:10 v/v) and 9 mM triethanolamine (TEA) at a flow rate of 0.9 mL/min, using a RP-C18 column (SUPELCO Kromasil 100A-5u-C18 4.6 mm × 250 mm); the absorbance was set at 475 nm and the run time was 20 min. Quantification was carried out using a standard calibration curve consisting of five points at increasing concentrations (6.25, 12.5, 25, 50, and 100 µg/mL) of lycopene standard (Sigma Chemical, St. Louis, MO, USA). The experiment was conducted in three technical replicates for each sample. Finally, the mean and standard deviation were calculated. To verify the significance of the data obtained, t-tests (* p ≤ 0.05, ** p ≤ 0.01) were carried out.

High-performance liquid chromatography (HPLC—Perkin Elmer Nelson 3200 Series) was used for quantitative and qualitative analysis of vitamin E. One gram of sample (a volume weight of 1 g was taken for the digested liquid) was diluted in 2 mL of pure ethanol, then the sample was homogenized with Turrax (IKA^®-Werke GmbH & Co. KG, Staufen im Breisgau, Germany) for 5 min and finally centrifuged at 4000 rpm (Eppendorf^® 5415D centrifuge, Hamburg, Germany) for 10 min. Subsequently, 200 μL of the supernatant was collected and analyzed by HPLC. The chromatography setup consisted of a Waters 996 photodiode array detector and a 600E pump. A C18 250 × 4.6 mm (5 μm) column (SUPELCO Kromasil 100A-5u-C18 4.6 mm × 250 mm) was used for the analysis of vitamin E at a flow rate of 0.5 mL/min for 40 min. An injection volume of 50 μL was used, and the mobile (isocratic) phase consisted of 90% methanol, 10% acetonitrile, and 9 mM TEA (triethylamine). The concentration of vitamin E was then determined by standard curve calibration. All steps of the experimental procedure, such as plant source selection, pasta preparation and cooking, in vitro digestion, HPLC, and spectrophotometric analysis, are outlined in Figure 2.