Dionex ic3000

HMO degradation was investigated using HPAEC-PAD, on Dionex IC3000 equipped with a CarbopacPA20 column (Thermo Fisher Scientific, Reinach, Switzerland) and an electrochemical detector with a gold electrode. Water (A), 200 mM NaOH (B), and 1 M Na-acetate (C) were used as solvents at a flow rate of 0.25 mL min⁻¹. For HMO separation, a gradient of: 0 min 30.4 % B, 1.3 % C, 22 min 30.4 % B, and 11.34 % C followed by washing and regeneration was applied. HMOs and L-fucose were identified using external standards.

HMO profiles before and after milk storage were qualitatively analyzed using high pH anion exchange chromatography with amperometric detection (HPAEC-PAD) on a Dionex IC3000 equipped with a CarbopacPA20 column (Thermo Fisher) and an electrochemical detector with a gold electrode. HMO were analysed directly from milk supernatants without further extraction procedure. Water (A), 200 mM NaOH (B) and 1 M Na-acetate (C) were used as solvents at a flow rate of 0.25 ml/min with the following gradient: 0 min 30.4% B, 1.3% C, 22 min 30.4% B, 11.34% C, followed by washing and regeneration. Main HMO were identified using 2′FL, 3′FL, 3′SL, 6′SL, lacto-N-neotetraose, N-acetylneuraminic acid supplied by Glycom A/S (Denmark) as external standards.