G1315d dad

The high-performance liquid chromatography (HPLC) system consisted of an Agilent infinity series 1260 liquid chromatography system with a G1311B quaternary pump, G1329B autosampler, G1316A column oven and G1315D DAD detector, connected to Agilent ChemStation software (Agilent, Waldbronn, Germany). The separation was conducted with a YMC-PACK ODS column (4.6 mm × 250 mm, 5 μm). The gradient profile was as follows: 0–5 min, initial mobile phase of 0.1% formic acid prepared in acetonitrile and 0.1% formic acid prepared in water (10:90); 5–45 min, linear gradient of 70:30; and 45–50 min, isocratic 70:30. The flow rate was 1.0 ml/min, and detection was at 254 nm. The sample concentration was 10 mg/min in MeOH, and the injection volume was 10 μl. The HPLC chromatogram of SCE is shown in Fig. 1B.

The monosaccharide compositions of GLP-1 and GLP-2 were analyzed by high-performance liquid chromatography (HPLC), as described in a previous study [23 (link)], but with some modifications. A polysaccharide sample (2 mg) was hydrolyzed with 2 M trifluoroacetic acid (1 mL) at 110 °C for 6 h, followed by derivatization with 0.5 M PMP. The PMP derivatives were analyzed on an Agilent 1200 Series HPLC system (G1322A Degasser, G1311A Quat Pump, G1329A ALS, G1315D DAD, Agilent Technologies, Inc., Santa Clara, CA, USA) equipped with an Eclipse XDB-C18 column (250 mm × 4.6 mm × 5 µm, Agilent, Santa Clara, CA, USA) at 30 °C. The detection wavelength was set at 250 nm, and the flow rate was 0.8 mL/min. The mobile phase was a mixture of phosphate-buffered saline (0.1 M, pH 6.5) and acetonitrile (84:16, v/v). Rhamnose, ribose, fucose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid, and galacturonic acid were used as standards.