Kinetix xb c18 column

The purified glycosylated reb C sample was prepared in 25% methanol at 1% (w/v). The compound reb C+1G was separated from the glycosylated reb C mixture by an Agilent (Santa Clara, CA, USA) semi-preparative chromatography system equipped with a Phenomenex (Torrance, CA, USA) Kinetix XB-C18 column (150 mm × 21.2 mm, 5 µm), using 0.1% formic acid in water (solvent A) and methanol (solvent B) as mobile phases at a flow rate of 4 mL/min. UV detection at both 210 nm and 254 nm was used. The fraction containing reb C+1G was dried at 70 °C under vacuum, and the end product was a white crystal, which was different than the free-flow white powder form of reb A. Diffusion-ordered spectroscopy (Figure S7) showed highly similar diffusion constants for all signals, indicating that the crystal is likely one molecule, and not a mixture.

High resolution mass spectrometry was performed on either Agilent 6210 TOF LC/MS using ESI-MS, Waters SYNAPT UPLC-ESI-qTOF, or Waters RDa ESI-TOF. High performance liquid chromatography (HPLC) was performed on an Agilent 1200 Series equipped with a variable wavelength UVVis detector (λ = 220 nm) using Phenomenex Kinetix XB-C18 column (4.6 × 250 mm, 5 µm). Mass spectra were processed in Mestrenova (version 14.1).