1f fructofuranosylnystose gf4

FOS standards 1-kestose (GF2), 1-nystose (GF3), and 1F-fructofuranosylnystose (GF4) were purchased from Wako Pure Chemical Industries, Ltd. (Japan Company). Fructose (F), glucose (G), and sucrose (S) standards were obtained from Sigma Aldrich (St. Louis, MO, USA). A Trolox standard was obtained from Sigma Aldrich (St. Louis, MO, USA). Aguamiel was obtained from “Ejido Las Mangas” (long. −101.102500, Lat. 24.906111), Saltillo, Coahuila, Mexico from Agave salmiana in December 2017. It was sterilized by membrane filtration (0.45 μm) before its use. Orafti HSI® was obtained from Beneo Company (Belgium). Agavins were obtained from Megafarma® (Durango, Mexico). Protein marker Prestained SDS-PAGE standard #1610318 was obtained from Bio-Rad (Hercules, CA, USA). Silver BULLit™ silver stain kit was obtained from AMRESCO (Radnor, PA, USA).

Samples were filtered through 0.22 μm nylon filters (Millipore, St. Louis, MO, USA) and placed in special vials for HPLC. The analysis was performed on a Varian Pro-Star330 HPLC instrument under the following conditions: Carbohydrate Prevail ES column (5 μm, 250 mm × 4.6 mm) at 30 °C using a refractive index (RI) detector at the same temperature. As a mobile phase, an acetonitrile/water solution (70/30 v/v) with 0.04% NH₄OH in water was used at a 1.0 mL/min flow. Standard curves were made with known concentrations of fructose (F), glucose (G), and sucrose (GF) (Sigma Aldrich, St. Louis, MO, USA) and FOS 1-kestose (GF₂), 1-nystose (GF₃), and 1F-fructofuranosylnystose (GF₄) (Wako Pure Chemical Industries, Ltd., Osaka, Japan) [48 (link)].