Introduction

Preparative purification in SFC has advantages for high throughput measurements and post-collection fraction treatment. In terms of post-collection fraction treatment, SFC uses CO₂ as mobile phase, so evaporation to dryness can be easily carried out by drying off the co-solvent (typically an alcohol). CO₂ has a low-cost and high purity, which allows measurement with less impurities and reduces solvent costs.

CO₂ has a 500 times adiabatically-expansion when released into atmospheric pressure at exit of pressure adjustment valve, thus it is important to properly control this expansion to maximize sample recovery. Cyclone Separator-87 (CS-87), gas-liquid separator, provides a high recovery rate for analysis at less than 50 mL/min CO₂ flow rates.

In this application, chiral preparative purification of flavanone using the Prep SFC System-M, CS-87, and fraction collector management program for EXTREMA UFC is reported.

Experimental

Experimental Condition
Column: CHIRALPAK IA (20 mmI.D. x 250 mmL, 5 µm)
Eluent: CO₂: 50 mL/min, methanol: 10 mL/min
Column temp.: 40ºC
Wavelength: UV: 210 nm
Back pressure.: 10 MPa
Injection volume: 1000 µL
Standard: 1 mg/mL flavanone in methanol

Schematic diagram

Figure 1 shows the Cyclone Separator-87 (CS-87), gas-liquid separator. CS-87 provides easy installation, repeatedly use and significant increase in fraction recovery.

Fig. 1 Cyclone Separator-87(CS-87)

Keywords

Flavanone, Prep-SFC, UFC, CS-87, ChromNAV Ver.2, Fraction collector management program, Chiral separation

Results

Figure 2 shows the chromatogram of flavanone recovery, and figure 3 shows the re-chromatogram of recovery liquid. As shown in figure 3, contamination is not observed, showing that the CS-87 provides high performance with high recovery.

Fig. 2 Fraction of flavanone

Fig. 3 Re-chromatogram of flavanone recovery liquid

Table 1 shows recovery rate of flavanone. As shown in the table Flavanone-1 and 2 shows high recovery rate.

Table 1  Recovery rate of flavanone