385 elsd

Optical rotations were recorded on a Unipol L1000 polarimeter at the sodium D-line (589.3 nm) with a 5 cm cell at 20 °C (Schmidt+Haensch, Berlin, Germany). UV and ECD were recorded on a Chirascan V100 with a 1.0 cm quartz cuvette (Applied Photophysics, Leatherhead, UK). IR data were recorded on a PerkinElmer spectrum 100 FT-IR spectrometer (PerkinElmer, Waltham, MA, USA). NMR experiments were performed on a 500 MHz Varian Inova spectrometer (Agilent, Santa Clara, CA, USA). Chemical shifts (δ in ppm) were referenced to the carbon (δ_C 39.52) and proton (δ_H 2.50) signals of DMSO-d₆. HRESIMS were obtained using an Agilent 6540 Q-Tof mass spectrometer equipped with an Agilent 1290 UPLC and autosampler (Agilent). Preparative and semipreparative HPLC was carried out on a Jasco LC-2000 series equipped with a coupled UV detector. Analytical HPLC was carried out on an Agilent 1260 HPLC system equipped with a DAD detector coupled with an Agilent 385-ELSD. All solvents used for extraction and separation were HPLC grade, and H₂O was Milli-Q (Millipore Ireland B.V., Carrigtwohill, County Cork, Ireland) filtered.

The HPLC equipment was an Agilent system, and included an Agilent 1260 TCC, an Agilent 1260 quat pump, an Agilent 385-ELSD (Evaporative Light Scattering Detector), and an Agilent workstation (Agilent, Palo Alto, CA, USA).
The prebiotics and FOSs purified from the HSCCC separation were analyzed by HPLC-ELSD with a Xamide 100A column (250 × 4.6 mm, id) and a column temperature of 25 °C. The mobile phase consisted of acetonitrile–water (75:25, v/v) at a flow rate of 1.0 mL/min.