Thermo finnigan surveyor hplc system

The optical rotations were measured using a Bellingham+Stanley (Berlin, Germany), model ADP410 Polarimeter with a 5 dm cell. UV/VIS spectra were recorded using an Ultraspect 3100 Pro Amersham Bioscienses spectrophotometer (Champaign, IL, USA) with a path length of 1 cm, and IR spectra were recorded on a Perkin Elmer Spectrum Two FT-IR Spectrometer (Santa Clara, CA, USA). ¹H and 2D NMR spectral data were recorded at 400 or 600 MHz in CDCl₃ containing Me₄Si as internal standard on Bruker Advance and Bruker BioSpin spectrometers respectively (Ettlingen, Germany). ¹³C NMR spectra were acquired at 100 MHz on a Bruker Advance spectrometer. High resolution ESI-TOF mass spectra were obtained through acquired services using an Agilent 6230 Accurate-Mass TOFMS spectrometer (Santa Clara, CA, USA) by the mass spectrometry facility at the Department of Chemistry and Biochemistry at the University of California, San Diego, La Jolla, CA. Low-resolution LC/MS data were measured through acquired services at MARINOVA, CIIMAR, Portugal, using a Thermo Finnigan Surveyor HPLC System (Thermo Fisher Scientific, Needham, MA, USA), coupled with Mass Spectrometry LCQ Fleet™ Ion Trap Mass Spectrometer (Thermo Fisher Scientific, Needham, MA, USA), with reversed-phase C₁₈ column (Phenomenex Luna, 100 mm × 1.0 mm, 5 µm), ACN:H₂O 10–100% gradient, with 0.1% formic acid, at a flow rate of 0.7 mL/min.

The anthocyanins identification from untreated skin extracts were determined using a procedure performed by Turturica et al. [14 (link)] using a Thermo Finnigan Surveyor HPLC system (Thermo Scientific, Waltham, MA, USA).

For extraction of sugars and organic acids, 0.05 g of powder was weighed and poured over with 2 ml of bidistilled water. Only edible parts (pericarp and placenta) were used for total sugars and organic acids analysis. Samples were then shaken on an orbital shaker for 30 min. After shaking, they were placed in a cooled centrifuge in which the samples were rotated at 10,000 rpm for 8 min. Samples were filtered through 25 µl cellulose filters (Chromafil A-25/25; Macherey–Nagel, Düren, Germany) and saved at – 20 °C until analysis on the Thermo Finnigan Surveyor HPLC system (Thermo Scientific, San Jose, USA). Two columns were used for the analysis: for sugars (Rezex RCM-monosaccharide (30 × 0.78 cm; Thermo Scientific, San Jose, USA)) and for organic acids (Rezex ROA organic acid column (30 × 0.78 cm; Phenomenex, Torrance, USA). Extraction method and HPLC settings were based on Zamljen et al.^{12 (link)} The results are reported in g/kg dry weight (DW) for sugars, citric, malic and quinic acid and in mg/100 g DW for succinic, fumaric and oxalic acid. All sugars and organic acids (including ascorbic acid) were determined only in edible parts (pericarp and placenta).