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4.4. Reversed-Phase High-Performance Liquid Chromatography Analysis
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Crude peptides were purified by semipreparative RP-HPLC using a solvent system of (A) 0.1% TFA and (B) 80% acetonitrile, 0.1% TFA, and a linear gradient from 0 to 30% (B) in 60 min. Purification was performed on a Phenomenex Jupiter Proteo 90 Å column (250 × 10 mm) using a Shimadzu HPLC apparatus (Berlin, Germany). Absorbance was detected at 220 nm. Purity was evaluated by analytical RP-HPLC using a Phenomenex Luna 10 μ C18 100 A column and an Agilent 1100 HPLC instrument (Palo Alto, CA, USA).
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