1. Crude extract Purified compound Starting materials Final products F ast C entrifugal P artition C hromatograph (FCPC ® ) KROMATON, France

2. Pr Jean Hugues Renault Methodology, process Worldwide specialist in CPC and mass transfer (ion exchange) Luc Marchal Process optimization: cell design, sample analyses Industrialization / Scale-up Worldwide specialist in centrifuges and liquid / liquid extractors for the chemical and pharmaceutical industries 55 employees 7 employees in engineering and design Design, fabrication, commercial, after-sales

3. R EVIEW OF C ENTRIFUGAL P ARTITIONING C HROMATOGRAPHY  Liquid / liquid partitioning chromatography: o Two immiscible liquid phases o Achieved by mixing at least two solvents or solutions  Stationary phase maintained by centrifugal force (rotation)  Mobile phase is pumped through the stationary phase  Separation of the solutes is achieved as a function of the partition coefficient K D between the mobile and stationary phases.

4. Seals between the discs Inlet Outlet Ascending ModeDescending Mode CPC M ECHANISM … Movie FcFc FcFc FcFc Light phase Heavy phase

5. FCPC R ANGE All stainless steel, corrosion resistant Quiet operation Easy access to rotors Interchangeable rotors with different capacities 50 mL200 mL 1000 mL

6. FCPC R ANGE Unique 25 ml capacity rotor Reduced weight and footprint P max = 80 bars (1176 psig) Rotation 3000 rpm Milligram scale samples in minutes

7. Separation of 2 proteins: myoglobin and lysozyme ProteinKDKD V r (ml)t r (min) Myoglobin1.233.88.4 Lyzozyme5.875.018.7 Column volume32 ml Solvent system ATPS PEG 1000/K 2 HPO 4 Descending mode Injected volume 1 ml d’une solution Myoglobine 4,3 g/L Lyzozyme 8,3 g/L Flow rate4 ml/min Rotation2000 rpm Pressure27 bars (397 psig) Stationary phase retention 28 %

8. polar AQUEOUS SYSTEM BuOH AcOH/Water TFA/Water Water BuOH MtBE EtOAc Water PEG Buffer + + MtBE EtOAc Water ARIZONA RANGE System A ACN MeOH Hept MtBE EtOAc Water ACN MeOH Hept System Z TERNARY SYSTEMS Simple Lipids Phospholipids Glycolipids SterolsProteins Peptides Polyphenols Anthocyanins Polysaccharides Various organic acids Inorganic elements FlavonidsAlkaloids  -amino acids non- polar A PPLICATION E XAMPLES

9. Crude extract Purified Compound Starting materials Final products Plus d’informations sur WWW.KROMATON.COM KROMATON ROUSSELET 45, Avenue Rhin et Danube – 07100 Annonay - FRANCE Tel. : +33 (0)475 692 211 Fax : +33(0)4 75 67 69 80 j.meucci@kromaton.com

10. Thank you for your attention…