Chromafil xtra 20 13

For the purification of the crude seed extracts, 100 mg Strata-X RP cartridges were used (Phenomenex, Torrance, CA, USA). The seed extracts were initially filtered (0.45 µm pore size syringe filters; Chromafil A-45/25; cellulose acetate, hydrophilic membrane; Macherey-Nagel, Düren, Germany), to remove any solid residue. The cartridges were conditioned with 3.0 mL of methanol followed by 3.0 mL of Milli-Q water. Then, 30 mL of the diluted methanol seed extracts (extract:water, 1:9) or 3.0 mL of the hydrolysed seed extracts were applied to the cartridges, and allowed to pass through them. The phenolic compounds remained bound to the cartridges, and the co-extracted compounds were washed from the columns with 4.0 mL of Milli-Q water. The cartridges were then dried using a vacuum pump. The compounds bound to the cartridges were eluted with 2.0 mL of 70% (v/v) aqueous methanol. The resulting extracts were filtered through 0.20 µm pore size syringe filters (Chromafil Xtra-20/13; cellulose acetate; Macherey-Nagel, Düren, Germany), and then stored at −80 °C until the liquid chromatography–mass spectrometry (LC-MS) analysis.

Crude grain extracts were purified using 100 mg Strata-X RP cartridges (Phenomenex, Torrance, CA, USA) according to a previously described method [7 (link)]. Briefly, 30 mL of the diluted crude methanol extracts (extract:water, 1:9) or 3.0 mL of the hydrolyzed extracts were applied to the SPE cartridges, washed with 4.0 mL of Milli-Q water, and dried with a flow of air. The compounds bound to the cartridges were eluted with 2.0 mL of 70% (v/v) aqueous methanol. The resulting extracts were filtered through syringe filters with a pore size of 0.20 µm (Chromafil Xtra-20/13; cellulose acetate; Macherey-Nagel, Düren, Germany) and then stored at −80 °C until liquid chromatography–mass spectrometry analysis (LC-MS).