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51 References


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Figure 5. Chromatographic traces of the separation in HPSFC/UHPSFC of the enantiomers of trans-stilbene oxide on Regis-Whelk-O1 150x4.6 mm 5 µm (Black lines), Regis-Whelk-O1 150x4.6 mm 3.5 µm (Red lines) and UHPC-Whelk-O1 1.8 µm (Green lines) at their optimal flow-rates (1.5, 2.5 and 3.7 mL/min respectively). CO2/ MeOH 80:20. ABPR: 1800 psi. T: 35°C. (For interpretation of the colours in this legend, please refer to the web version of this article)


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Figure 6. Summarising results of large library screening of 129 racemates in UHPSFC on 50x4.6 mm L.xI.D. Whelk-O1 1.7 µm. Gradient table and analytical conditions are reported in figure.


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gradient with 0.1% TFA or 0.1% NH3 added for acidic and basic compounds, respectively. A flow rate of 3.5 mL/min allowed very fast and highly efficient separations, most of samples were eluted and successfully separated in less than 4 min. As expected, the best results were obtained concerning acid and neutral samples, but this CSP showed great results also on basic analytes: 47% of basic racemates were separated with a resolution value higher than 0.3, demonstrating the possibility to use this selector for acid, neutral and also basic chiral molecules.


CO2 Conclusions


A complete characterisation from the kinetic point of view of the UHPC-Whelk-O1 1.8 µm was performed and compared to those of the equivalent 3.5 and 5 µm commercial Whelk-O1 columns, with great results: efficiencies higher than 260,000 N/m on trans- stilbene oxide at a high optimal flow-rate of 1.8 mL/min were reached. The resulting separation was completed in less than 2


Acknowledgment


The authors would like to thank Professor Francesco Gasparrini for his support and for his invaluable advice, Waters s.p.a. (Milano, Italy) for providing UPC2 instrumentation and Dr E. Bianchini, Department of Chemistry and Pharmaceutical Sciences, University of Ferrara, for elemental analysis measurements.


minutes with a reduction of the analysis time of more than four times in comparison with the 5 µm column. The UHPC-Whelk-O1 was also used in UHPSFC maintaining the same trend of UHPLC with very high efficiencies and very high optimal linear velocities. The separation of the two enantiomers of trans- stilbene oxide was completed in just 1 minute.


This CSP was packed into a 5cm long column and was used to screen 129 chiral compounds in sub-critical fluid chromatography with a 9-min screening method. 63% of this library was resolved in just ‘one working day’.


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