The RP chromatographic separation was achieved with a Kinetex™ 1.7 μm C18 100 Å, LC column 100 × 2.1 mm (phenomenex, Torrance, CA, USA). The ESI-MS settings were as follows: capillary voltage

4500 V, nebulizing gas 1.8 bar, and dry gas 9 l/min at 200 °C. The scan range was from mass-to-charge ratio (m/z) 80–1200. The mobile phase was composed of water containing 1% formic acid (A) and acetonitrile containing 5% water and 1% formic acid (B). The flow rate was 0.2 ml/min with a gradient elution of 5–95% B over 35 min, and standing at 95% B for 20 min. The sample injection volume was 2 μl. The column temperature was set at 40 °C. The ESI-MS selleck inhibitor system was calibrated using sodium formate clusters introduced by loop-injection at the beginning of the LC–MS run. The LC–MS data was processed using Data Analysis 4.1 software (Bruker Daltonik, Bremen, Germany). Molecular ions [M+H]+ were extracted from full scan chromatograms and peak areas were integrated. The extraction window of individual ion chromatograms was ±0.05 m/z units. The compounds present in each sample were identified

by comparing their retention times with those of standards, and based on molecular mass and structural information from the MS detector. The protein content was determined by the Kjeldahl method using a conversion factor of 6.25 for cereal foods (AOAC method 920.87, 1995). The analysis of the fatty acid methyl esters of the oils used in the muffin preparation was carried out using a HTS assay Hewlett Packard HP 5890 gas chromatograph equipped with a flame ionisation detector and fitted with a HP-Innowax capillary column (30 m × 0.25 mm i.d. × 0.25 μm df, Hewlett–Packard, Waldbronn, Germany), according to the method described previously (Mildner-Szkudlarz, Zawirska-Wojtasiak, Obuchowski, & Gośliński, 2009). The tocochromanols of oils were analysed by direct injection of the oil samples dissolved in HPLC-grade n-hexane using a Waters

Alliance HPLC System 600 (Milord, MA, USA) with a fluorescence detector (Waters 474), according to the previously published method (Górnaś, Siger, & Seglin, Y-27632 2HCl 2013). The analysis of the glucose content of the white beet sugar and the raw cane sugar used in muffin preparation was performed as in Trinder (1969). The analysis of the elemental content of white (refined) beet sugar and raw (unrefined) cane sugar was carried out using an inductively coupled plasma optical emission spectrometer (ICP-OES) Vista MPX (Australia) after digestion of samples in a microwave oven (CEM MARS 5), according to the method described by Chojnacka, Michalak, Zielińska, Górecka, and Górecki (2010).