title: Purification of a modified cyclosporine A by co-current centrifugal partition chromatography: process development and intensification.
abstract: Synthetic hydrophobic non-ionizable peptides are not soluble in most common solvents and are thus difficult to purify by preparative reversed-phase HPLC, normally used for industrial production. The challenge exists to develop alternative purification chromatographic processes using suitable solvents and providing good yields, high purity and sufficient productivity. A 11mer hydrophobic synthetic modified cyclosporine, showing an anti-HIV activity, was successfully purified by centrifugal partition chromatography using the biphasic solvent system heptane/ethyl acetate/acetone/methanol/water (1:2:2:1:2, v/v). A 5% co-current elution - made possible by the liquid nature of the two phases - has been used in order to avoid hydrodynamic instabilities mainly due to the physico-chemical properties of the target peptide. This original solution was developed after the study of the effect of the peptide on the hydrodynamic behavior of the two phases during the separation, and the visualization of the flow patterns using the Visual-CPC device. Critical impurities were efficiently eliminated and the peptide was recovered in high yield and high productivity achieving the specifications requirements.
authors.affiliation: ICMR UMR CNRS 7312, SFR CAP'SANTE, Université de Reims Champagne-Ardenne, Bât. 18, Moulin de la Housse, B.P. 1039, 51687 Reims cedex 2, France.
authors.forename: Nassima
authors.initials: N
authors.lastname: Amarouche
authors.forename: Leslie
authors.initials: L
authors.lastname: Boudesocque
authors.forename: Charlotte
authors.initials: C
authors.lastname: Sayagh
authors.forename: Matthieu
authors.initials: M
authors.lastname: Giraud
authors.forename: John
authors.initials: J
authors.lastname: McGarrity
authors.forename: Alessandro
authors.initials: A
authors.lastname: Butte
authors.forename: Luc
authors.initials: L
authors.lastname: Marchal
authors.forename: Alain
authors.initials: A
authors.lastname: Foucault
authors.forename: Jean-Hugues
authors.initials: JH
authors.lastname: Renault
mesh_terms: D000085:Acetates / Q000737:chemistry; D019380:Anti-HIV Agents / Q000737:chemistry / Q000302:isolation & purification*; D002498:Centrifugation / Q000295:instrumentation*; D002853:Chromatography, Liquid / Q000295:instrumentation*; D016572:Cyclosporine / Q000737:chemistry / Q000302:isolation & purification*; D004867:Equipment Design; D000432:Methanol / Q000737:chemistry; D012997:Solvents / Q000737:chemistry
chemical_list: D000085:Acetates; D019380:Anti-HIV Agents; D012997:Solvents; C007650:ethyl acetate; D016572:Cyclosporine; D000432:Methanol
keywords: Biphasic solvent systems; Centrifugal partition chromatography; Co-current chromatography; Intensification; Peptides; Preparative purification