Comparing the selectivity and chiral separation of d- and l- fluorenylmethyloxycarbonyl chloride protected amino acids in analytical high performance liquid chromatography and supercritical fluid chromatography; evaluating throughput, economic and environmental impact.
The chiral separation of d- and l- FMOC amino acids was undertaken using the Lux Cellulose-1 polysaccharide based chiral column in HPLC (normal phase and reverse phase) and SFC conditions. This was done to compare the relative selectivity and separation between the three separation modes and to evaluate the potential benefits of SFC separations with regards to resolution, throughput, economic and environmental impact. It was established that the separation of d- and l- FMOC amino acids in SFC displayed behaviours that were similar to both normal phase and reversed phase, rather than distinctly one or the other. Additionally, although reversed phase conditions yielded significantly higher resolution values between enantiomers across the range of amino acids studied, improvements in selectivity in SFC via the introduction of higher concentrations of formic acid in the mobile phase allowed for better resolution per unit of time. Moreover since the SFC mobile phase is composed mostly of recyclable CO2, there is a reduction in organic solvent consumption, which minimises the economic and environmental costs.
Australian Centre for Research on Separation Science, School of Science and Health, Western Sydney University, Parramatta, NSW 2150, Australia.
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Vera
Phenomenex Australia, Pty Ltd, Lane Cove, NSW 2067, Australia.
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Shock
Australian Centre for Research on Separation Science, School of Science and Health, Western Sydney University, Parramatta, NSW 2150, Australia.
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Dennis
Pfizer Global R&D, La Jolla, CA 92037, United States.
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Farrell
Australian Centre for Research on Separation Science, School of Science and Health, Western Sydney University, Parramatta, NSW 2150, Australia. Electronic address: r.shalliker@uws.edu.au.
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Shalliker
D000596:Amino Acids / Q000737:chemistry* / Q000302:isolation & purification*; D002245:Carbon Dioxide / Q000737:chemistry; D002482:Cellulose / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D025924:Chromatography, Supercritical Fluid / Q000191:economics* / Q000379:methods*; D004777:Environment; D005449:Fluorenes / Q000737:chemistry* / Q000302:isolation & purification; D005561:Formates / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D013237:Stereoisomerism
C496789:9-fluorenylmethoxycarbonyl; D000596:Amino Acids; D002712:Chlorides; D005449:Fluorenes; D005561:Formates; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D012997:Solvents; C030544:formic acid; D002245:Carbon Dioxide; D002482:Cellulose
Amino acids; Chiral chromatography; Normal phase; Reversed phase; Supercritical fluid chromatography