Industrial case study on alkaloids purification by pH-zone refining centrifugal partition chromatography.
The industrial potential of pH-zone refining centrifugal partition chromatography has been evaluated by studying the purification of pharmaceutical ingredients at the pilot scale. For the first time, a scale up methodology based on both column capacity and mass transfer efficiency as invariants was developed. The purification of catharanthine and vindoline from an industrial crude extract of aerial parts of Catharanthus roseus, was used as a case of study. Toluene/CH3CN/water (4/1/5, v/v/v) was selected as biphasic solvent system, triethylamine as retainer in the organic stationary phase and sulphuric acid as displacer in the aqueous mobile phase. The separation intensification was performed on a 36mL CPC column equipped with 832 partition twin-cells. The combined effects of four parameters (displacer and retainer concentrations for intensive parameters, flow rate and rotational speed for extensive parameters) were studied by design of experiment in order to maximize both recoveries and productivities. Then, scale change was done on two larger columns (305mL and 1950mL of capacity) equipped with only 231 and 238 partition cells. For this step, it has been shown that the global mass transfer coefficient k0a (the efficiency of a column design) and the stationary phase retention (the capacity of the column) were relevant and useful scale up invariants. A CPC model based on acid-base equilibriums and interfacial mass transfer in continuously stirred tank reactors in series was used to predict fully separations on larger CPC column at the optimized operating conditions and to guide the CPC user in its scale-up strategy. The experimental validation on pilot CPC column, by injecting up to 150g of Catharanthus roseus crude extract on the 1950mL column highlighted the preservation of the separation quality, the non-linear character of the scale up in centrifugal partition chromatography and that a productivity of about 4kg of processed crude extract per day can be reached by implementing developed methodology.
GEPEA, UMR CNRS 6144, University of Nantes, 37 bd de l'Université 44602 Saint Nazaire Cedex, France; ICMR, UMR CNRS 7312, University of Reims Champagne-Ardenne, France.
Alexis
A
Kotland
ICMR, UMR CNRS 7312, University of Reims Champagne-Ardenne, France.
Sébastien
S
Chollet
Laboratoires Pierre Fabre, Route de Viars, 81600 Gaillac, France.
Catherine
C
Diard
Laboratoires Pierre Fabre, Route de Viars, 81600 Gaillac, France.
Jean-Marie
JM
Autret
Rousselet-Robatel Kromaton, 45 Avenue Rhin et Danube, 07100 Annonay, France.
Jeremy
J
Meucci
ICMR, UMR CNRS 7312, University of Reims Champagne-Ardenne, France. Electronic address: jh.renault@univ-reims.fr.
Jean-Hugues
JH
Renault
GEPEA, UMR CNRS 6144, University of Nantes, 37 bd de l'Université 44602 Saint Nazaire Cedex, France.
Luc
L
Marchal
D000465:Algorithms; D000470:Alkaloids / Q000302:isolation & purification*; D030008:Catharanthus / Q000737:chemistry; D002498:Centrifugation; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution / Q000379:methods; D006863:Hydrogen-Ion Concentration; D018515:Plant Leaves / Q000737:chemistry; D012997:Solvents; D014747:Vinblastine / Q000031:analogs & derivatives / Q000302:isolation & purification; D014748:Vinca Alkaloids / Q000302:isolation & purification
D000470:Alkaloids; D012997:Solvents; D014748:Vinca Alkaloids; C009667:vindoline; D014747:Vinblastine; C017836:catharanthine
Alkaloids; Centrifugal partition chromatography; Hydrodynamics; Modelling; Scale-up; pH-zone refining