Supercritical Fluid Chromatography SFC Chromatographic Fundamentals Practical Verification of SFC Theoretical Description of SFC / Scale-up SFC on a Preparative Scale: Examples Prostaglandins, Tocopherols DHA / DPA, Phytol On-line Analysis with SFC Continuous Chromatography: SMB Chapter 8 Chromatography with Supercritical Fluids

. Mode of Operation: Elution chromatography

Elution Chromatography: A Chromatogram

Mass transport high Solvent power high Schoenmakers, Uunk 1987 Different Mobile Phases

SFC: Stationary Phases

SFC: Different Gases as Mobile Phase

SFC: Different Modifiers

SFC: Influence of Pressure and Temperature

SFC: Pressure And Density Programming

Overloading by volume Analytical injection Overloading by concentration Concentration Time Chromatograms For Different Amounts of Injection

Adsorption Isotherms And Corresponding Chromatograms

SFC: Flow Scheme of Apparatus

Elution Chromatography: A Chromatogram

Capacity Ratio

Capacity Factors

with n = number of stages for p: Chromatographic Separation

Selectivity Resolution Chromatographic Separation

Van Deemter Chromatographic Separation

SFC Analytical Scale, hp

Influence of temperature Preparative separation Chromatograms of fractions Upnmoor 1992 Separation of Prostaglandins

Separation of Tocopherols

Influence of modifier concentration Separation of Tocopherols

x 4.6 pS 250 x 8.0 pS specific productivity DHA [mg/cm 3 h] Area DHA GC [%] 1mg DHA/(h,cm 3 ) * 500 ml = 0,5 g DHA/h Some kg DHA:Fully automatized plant ! RF=0,842 Productivity: DHA / DPA Separation by SFC

Dynamic axial compressed SFC column; Dimensions: ID = 30 mm, length of packing: 0 to 190 (type I), 0 to 450 mm (type II) P max 400 bar, T max 200 °C. SMB- Plant: Separation Columns

SFC, Preparative Scale

Rotating columnRotating ports Continuous Chromatography

Extract A + D Raffinate B + D Feed A + B + D Desorbent D Zone 1 Purification of Adsorbent Zone 3 Enrichment of B Zone 4 Purification of Desorbent Zone 2 Enrichment of A True Moving Bed (TMB) Process

Principle of Simulated Countercurrent Separation Mazzotti, ETH-Z

Extract A+D Raffinate B+D Feed A+B Desorbens D Concentration A, B Simulated Moving Bed-Process

Gottschall: PREP 95 Performance SMB vs Elution (99.5 % Purity)

Preparative SMB-Plant Depta, 2000

Adsorption isotherms for Phytol cis- and trans- isomer (black lines) and derivatives (red lines). 225 bar, 40 °C, 1.8 mass% isopropanol as modifier. Isotherms exhibit a point of inflection for each isomer. Adsorption Isotherms

Experimental and simulated phytol chromatogramssymbols: experimental data; lines: simulations. Batch-Simulations

Model: equilibrium, axially dispersed plug flow with variable velocity of mobile phase, Pressure drop: Ergun equation, Properties of mobile phase (CO 2 ) calculated with equation of state. SMB process modeled with four key parameters: the net flow ratios m j: Ruthven, Storti. SMB-Simulation

SMB- SFC: Volume-flow is a function of column length. Therefore, net flow ratios are not constant in each zone. New parameter: Representation of SMB-SFC process in a (m 2 * -m 3 * )-plane, solution of mass balance equations with finite difference method [Kniep et al.], adapted to variable velocity of mobile phase. The algorithm is fast enough to calculate the region of complete separation in the (m 2 * -m 3 * )-plane numerically, taking into account: any type of isotherm equation axial dispersion number of used columns change in mobile phase density SMB-Simulation

operating point black triangles:infinite dilution situation and infinite number of theoretical plates same parameter set as operating point in figure 5 Region of complete separation for phytol C feed =5.0 mg/ml 230 bar, no pressure drop, columns: 2/2/2/2; 300 plates per column Columns: 1/1/1/1; 1000 plates per column SMB-Simulation: Phytol Separation

operating point black triangles:infinite dilution situation and infinite number of theoretical plates same parameter set as operating point in figure 5 Region of complete separation for phytol C feed =5.0 mg/ml 230 bar, no pressure drop, columns: 2/2/2/2; 300 plates per column Columns: 1/1/1/1; 1000 plates per column SMB-Simulation: Phytol Separation

Region of complete separation for phytol, infinite dilution, columns: 2/2/2/2; 300 plates per column, 230 bar, no pressure drop Same as in left figure but calculations with pressure drop Pressure drop leads to a shift of the complete separation region to lower values of m 2 * and m 3 * SMB-Simulation: Phytol Separation

low concentration in Feed linear Adsorption isotherm Ideal model Experimental Results of Ibuprofen Separation

140 mg Racemate /min; 2/2/3/1 configuration Separation of Ibuprofen

Verunreinigungen Phytolisomere Conditions of separation: 240 bar, 50°C, column 4 x 250 mm packed with LiChrospher 100 (Silica), flow 2,56 g carbon dioxide / min, modifier 3wt.-%EtOH, productivity 45 mg/(ml, h). 17mg pur 0,85 mg in Hexan OH CH 3 CH 3 CH 3 CH 3 HH CH 3 Phytol Diterpene-alcohol, Intermediate for vitamin E, K1 esterified lipophiliccompound of chlorophyll