The neutralized extracts were applied to an anion-exchange Partisil-10 SAX column and eluted at a flow rate of 1.5 ml/min with a 50-min concave gradient (curve 7; Waters 600E System Controller; Waters Corp).
The HPLC system was equipped with a Waters 600 Controller, Waters Delta 600 pump, and a Shimadzu SPD-M10A VP Diode Array Detector and connected to a Waters Fraction Collector (Waters Co., Milford, MA, USA).
Sugars were quantified by high-pressure liquid chromatography (HPLC) using a Waters 600E system controller (Waters Corp. Milford, MA, USA) equipped with a refraction index detector (Waters 410) using a Prevail Carbohydrate ES column (250 × 4.6 mm) at 30°C.
Peptide fragments were subjected to liquid chromatography/MASS analysis (Q-tof Ultra global), which was equipped with a three-pumping Waters nano-LC system, a stream selection module and MassLynx 4.0 controller (Waters, UK).
Xylose, fructose, glucose, sucrose and fructosyl-xyloside were quantified by HPLC in a Waters 600E system controller (Waters Corp. Milford, MA) equipped with a refractive index detector (Waters 410), and a Carbohydrate (4.6 × 250 mm) column kept at 35°C, using acetonitrile/water (75:25) as the mobile phase at 1.2 mL/min.
It includes a Pyrex glass reactor (Savat di Rasetti Giuseppe & C. S.a.s, Torino, Italy), connectors, mass flow controllers, water bubbler, and a UV lamp (300 W, Osram Ultra-Vitalux, Munich, Germany).
HPLC separation was performed on an instrument with a Waters 600 controller, a Waters 600 pump, and a Waters 2487 dual λ absorbance detector (detecting wavelength: 230 nm) on a Grace (250 mm×10 mm, i.d).
HPLC separation was performed on an instrument consisting of a Waters 600 controller, a Waters 600 pump, and a Waters 2487 dual λ absorbance detector (Waters Corporation, Milford, USA), with an Prevail (250 mm×10 mm i.d).
The peptide was purified by semipreparative RP-HPLC [Waters 2487 Dual Absorbance Detector equipped with a Waters 2700 Sample Manager, a Waters 600 Controller, a Waters Fraction Collector, a Symmetry column (C18, 5 μm, 30 × 100 mm2), and Millennium software].
KSL-W content of microspheres was determined by reverse-phase high-performance liquid chromatography (HPLC) using a Waters 600 E multisolvent delivery system consisting of a Waters 600 pump and controller, a Waters 717 plus autosampler, and a Waters 996 photodiode array detector.
