Vydac 218tp1022 c18 column

All peptides and derivatives were synthesized by the solid phase methodology on Rink amide MBHA resin using a Liberty microwave-assisted automated peptide synthesizer (CEM, Saclay, France) and the standard manufacturer’s procedures at 0.1 mmol scale as previously described [54 (link)]. All Fmoc-amino acids and building blocks (0.5 mmol, 5 equiv) were coupled by in situ activation with HBTU (0.5 mmol, 5 equiv) and DIEA (1 mmol, 10 equiv). Peptides and derivatives were deprotected and cleaved from the resin by adding a TFA/TIS/H₂O (99.5/0.25/0.25) mixture for 120 min at room temperature. After filtration, crude peptides were precipitated by addition of TBME, centrifuged and recovered by elimination of the supernatant (3 folds). Peptides and derivatives were purified by reversed-phase HPLC (RP-HPLC) on a 2.2 × 25 cm Vydac 218TP1022 C₁₈ column (Grace, Epernon, France) using a linear gradient (10–40%, 10–50%, 10–60%, 20–40%, 20–50% or 20–60% over 45 min) of acetonitrile/TFA (99.9/0.1) at a flow rate of 10 mL/min. The purified peptides were then characterized by MALDI-TOF mass spectrometry on a UltrafleXtreme (Bruker, Strasbourg, France) using CHCA as a matrix. Analytical RP-HPLC, performed on a 0.46 × 25 cm Vydac 218TP54 C₁₈ column (Grace), showed that the purity of all compounds was >97.3%.