Rapid Gas Chromatographic Separation of Amino Acid Enantiomers with a Novel Chiral Stationary Phase

Abstract

The use of novel polysiloxanes as stationary phase carrying chiral groups enables the separation of most amino acid enantiomers in a much shorter time than ever reported previously. Phases of this type exhibit very low volatility and high thermal stability and may be used in routine analysis with open tubular columns ant temperatures of at least 175 degrees C. Most protein amino acids are separated in a temperature program between 90 and 175 degrees C, thus obviating the need for multiple injections. Resolution factors are somewhat lower than those of other diamide phases containing the L-valine t-butyl-amide group, but are sufficient for resolution of almost all protein amino acid enantiomers.

Contributions to the theory and practice of the chromatographic separation of enantiomers

“…Subsequently, in a pioneering approach, Frank, Nicholson, and Bayer at Tübingen university chemically linked the diamide selector of Gil-Av and Feibush to a polysiloxane backbone. 3 The resulting chiral polysiloxane containing valine (named Chirasil-Val) combined the enantioselectivity of the amino acid selector with the unique gas-chromatographic properties and thermal stability of silicones. 3 As the scope of the hydrogen-bonding CSPs was more or less limited to amino acids, amines, or hydroxy acids with the requirement of derivatization prior to analysis, the quest for complementary selectorselectand systems, e.g., based on fast and reversible molecular coordination (e.g., for alkenes) or inclusion (e.g., for unfunctionalized alkanes), was warranted.…”

Purification and partial characterization of human lymphoblast interferon.

“…The analyses were done in the laboratory of P. E. Hare, Geophysical Laboratory, Carnegie Institute of Washington, Washington, DC, using high-performance liquid chromatography and fluorescence detection of o-phthalaldehyde amino acid derivatives (16,17). Proline was measured by gas chromatography as described (18), except that trifluoroacetic anhydride was used rather than pentafluoropropionic anhydride. Cysteic acid and methionine sulfone were measured after performic acid oxidation (19,20).…”

Discrimination between fibrin and fibrinogen by a monoclonal antibody against a synthetic peptide.

“…Sequencing of the final hexapeptide confirmed that the desired amino acid sequence was present. The purity of the hexapeptide was evaluated by thin-layer chromatography, and the racemization was tested by gas chromatography (27). The racemization rate of each amino acid was <2%.…”