Alain Foucault

Intensified extraction of ionized natural products by ion pair centrifugal partition extraction

The potential of centrifugal partition extraction (CPE) combined with the ion-pair (IP) extraction mode to simultaneously extract and purify natural ionized saponins from licorice is presented in this work. The design of the instrument, a new laboratory-scale Fast Centrifugal Partition Extractor (FCPE300(®)), has evolved from centrifugal partition chromatography (CPC) columns, but with less cells of larger volume. Some hydrodynamic characteristics of the FCPE300(®) were highlighted by investigating the retention of the stationary phase under different flow rate conditions and for different biphasic solvent systems. A method based on the ion-pair extraction mode was developed to extract glycyrrhizin (GL), a biologically active ionic saponin naturally present in licorice (Glycyrrhiza glabra L., Fabaceae) roots. The extraction of GL was performed at a flow rate of 20 mL/min in the descending mode by using the biphasic solvent system ethyl acetate/n-butanol/water in the proportions 3/2/5 (v/v/v). Trioctylmethylammonium with chloride as a counter-ion (Al336(®)) was used as the anion extractant in the organic stationary phase and iodide, with potassium as counter-ion, was used as the displacer in the aqueous mobile phase. From 20 g of a crude extract of licorice roots, 2.2g of GL were recovered after 70 min, for a total process duration of 90 min. The combination of the centrifugal partition extractor with the ion-pair extraction mode (IP-CPE) offers promising perspectives for industrial applications in the field of natural product isolation or for the fractionation of natural complex mixtures.

Centrifugal partition extraction, a new method for direct metabolites recovery from culture broth: case study of torularhodin recovery from Rhodotorula rubra.

Centrifugal partition extraction (CPE), close to centrifugal partition chromatography, put in contact in a continuous way two immiscible liquid phases. This work presents early experiments on CPE use for solid-liquid-liquid extraction. It was applied to the direct treatment of culture broth for metabolites recovery. Torularhodin is one of the carotenoid pigments produced by the yeast Rhodotorula sp., with a terminal carboxylic group considered nowadays as a powerful antioxidant to be included in food and drugs formulations. Torularhodin was extracted from Rhodotorula rubra ICCF 209 cells by CPE. The recovery of torularhodin reaches 74 μg/g of biomass i.e. 294 μg/L of culture medium. The efficiency of the extraction step increased with the operating flow rate. The extraction yield could reach 91% with a contact time lower than 2 min. A 300 mL apparatus allowed a feed at 90 mL/min. The technique is proposed for extraction or sample preparation before analysis.

Purification of a modified cyclosporine A by co-current centrifugal partition chromatography: process development and intensification.

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.

Chapter 5 - Centrifugal partition chromatography: an engineering approach

A centrifugal partition chromatography (CPC) is a one-axis centrifugal system operating with two rotary seals. A CPC column is a series of cells linked in a cascade by ducts and regularly disposed around a rotor as cartridges or disks, and submitted to a constant centrifugal acceleration generated by a rotation. This chapter presents the first assumptions on flow patterns. While the rotor of the centrifuge is in motion, the mobile liquid phase is pumped into the cells and “bubbles down” if heavier, or “bubbles up” if lighter, through the stationary phase by the action of the centrifugal force. The “true” flow rate of the bubbles of the mobile phase through the stationary phase is determined by the strength of the applied centrifugal force. As long as the pumping rate (the apparent flow rate) does not exceed the “true” flow rate, it does not affect the resolution of the peaks. This means that the separation can be completed in a short time provided a sufficient centrifugal force is applied.