Xbridge prep beh c18

Semiprep-RP-HPLC-MS was used to purify the peptides, formed by a binary gradient Waters 2545, a Waters Alliance 2767 sample manager module and an automatic fraction collector coupled to a dual UV-vis λ Absorbance Detector 2487 and a mass spectrometer Micromass ZQ. An XBridge^® Prep BEH C18 (19 × 100 mm, 5 μm) column (Waters, Cerdanyola del Vallès, Spain) was used. The eluents were A: ACN/HCOOH (99.93:0.07, v/v) and B: H₂O/HCOOH (99.9:0.10, v/v). The column was preconditioned by washing with buffer A for 10 min. Gradient conditions are described below.

Crude peptides were purified by semipreparative RP-HPLC-UV-MS using a system composed by a binary gradient Waters 2545, a Waters Alliance 2767 sample manager module and an automatic fraction collector coupled to Waters 2487 dual UV-vis absorbance detector and an electrospray ion source (ESI-MS) Micromass ZQ mass spectrometer detector. The chromatographic separation of the peptides was achieved using a semipreparative column XBridge^® Prep BEH C₁₈ (19 × 100 mm, 5 µm) (Waters, Cerdanyola del Vallès, Spain) with dual solvent system formed by A: 0.1% TFA in H₂O and B: 0.1% TFA in CH₃CN at a flow rate of 25 mL/min. λ = 220 nm. Elution gradient was specific for each peptide.