Xbridge beh amide hplc column

Glucose, fructose, and galactose in enteric tissue samples were detected using UPLC-Q-TOF/MS (Waters). Briefly, jejunum, ileum, or colon tissue (30 mg) was transferred to homogenizer (pre-cooled at −20 °C) and 100 µL of H₂O (pre-cooled at 4 °C) was added for homogenization for 1 min. Next, 800 µL methanol solution containing 4-chlorophenylalanine (1 μg/mL) were added for protein precipitation and vortexed for 10 min. The sample lysates were centrifuged at 30,000×g for 10 min at 4 °C and dried under vacuum. It was reconstituted with 100 μL of methanol: water solution (50:50, v/v) before analysis. An XBridge BEH Amide HPLC column (100 mm × 4.6 mm, 3.5 μm; Waters) was used. The column was maintained at 40 °C. The injection volume was 10 μL and the flow rate was 0.6 mL/min. The mobile phase A consisted of 95% of 5 mM ammonium acetate buffer, and 5% acetonitrile (pH = 9), and B was acetonitrile. The gradient was as 0–3 min, 85% B; 3–6 min, 85~30% B; 6–11 min, 30~2% B; 11–13 min, 2% B; 13–14 min, 2~85% B; and 14–21 min, 85% B. The glucose, fructose, and galactose in samples were detected using QTOF-MS (Xevo G2-XS, Waters) operating in negative ion mode. Raw data for glucose, fructose, and galactose quantification were obtained with MassLynx v4.1 and analyzed by Quanlynx v4.1(Waters, Milford, MA).