Luna omega ps

Polyphenols profile detection and quantification were performed according to the protocol described in detail by El-Nakhel et al. [32 (link)]. Briefly, 5 μL of the extracted samples according to the procedure described by Vallverdú-Queralt et al. [33 (link)], were analyzed using a Dionex Ultimate 3000 ultra-high-pressure liquid chromatography (UHPLC) system (Thermo Fisher Scientific^TM, Waltham, MA, USA) coupled to an Orbitrap high resolution mass spectrometry (HRMS) (Thermo Fisher Scientific^TM, Waltham, MA, USA). The chromatographic separation of polyphenols was carried out with a Luna Omega PS (1.6 μm, 50 × 2.1 mm, Phenomenex, Torrance, CA, USA) thermostated column (T = 25 °C). The mobile phase consisted of a two-phase solution: water (phase A) and acetonitrile (phase B). Both mobile phases contained 0.1% formic acid (v/v). An ESI source (Thermo Fisher Scientific^TM, Waltham, MA, USA) was used in negative ion mode (ESI–), setting two scan events (Full ion MS and All ion fragmentation, AIF) for all compounds of interest. Data processing was performed with Quan/Qual Browser Xcalibur software, v. 3.1.66.10 (Thermo Fisher Scientific^TM, Waltham, MA, USA). Polyphenols were expressed as μg 100 g⁻¹ fw.

Quantification and separation of phenolic compounds were performed by UltraHigh-Pressure Liquid Chromatography (Dionex UltiMate 3,000 UHPLC, Thermo Fisher Scientific, Waltham, MA, United States) coupled to the Q Exactive Orbitrap LC–MS/MS Mass Spectrometer (Thermo Fisher Scientific, Waltham, MA, United States) as described by El-Nakhel et al. (2021) (link). The polyphenols were separated by using a Luna Omega PS (1.6 m, 50 × 2.1 mm, Phenomenex, Torrance, CA, United States) at 25 ° C. The mobile phase was a two-phase solution containing water (phase A) and acetonitrile (phase B). Both mobile phases contained 0.1% formic acid (v/v). Polyphenolic compounds were eluted using the following gradient schedule: 0–1.3 min 5% B, 1.3–9.3 min 5–100% B, 9.3–11.3 min 100% B, 11.3–13.3 min 100–5% B, 13.3–20 min 5% B. The flow rate was 0.2 ml min⁻¹. For all compounds of interest, an ESI source (Thermo Fisher Scientific, Waltham, MA, United States) was used in negative ion mode, with full ion (MS) and all ion fragmentation (AIF) scanning events. Data acquisition and processing were performed with Quan/Qual Browser Xcalibur software, v. 3.1.66.10 Xcalibur, Thermo Fisher Scientific, (Thermo Fisher Scientific, Waltham, MA, United States). Polyphenols were expressed as μg g⁻¹ dw.