GC separation of amino acid enantiomers via derivatization with heptafluorobutyl chloroformate and Chirasil-L-Val column.
Heptafluorobutyl chloroformate (HFBCF), a recently introduced derivatization reagent, was examined in enantioseparation of amino acids (AAs) by GC. Twenty proteinogenic AAs, plus ornithine, cystine and 4-fluorophenylalanine (internal standard) were treated with the reagent and separation properties of the derivatives were assessed on a Chirasil-Val capillary column. Nineteen AA enantiomers were efficiently separated in 43 min except proline, arginine and cystine. The HFBCF derivatives of the studied DL-AAs show improved separation over other chloroformate-based derivatives hitherto reported. A combination of the improved and faster separation with a simple derivatization protocol, involving an immediate one-step reaction-extraction in two-phase aqueous-organic medium, and low elution temperatures extend application of HFBCF to chiral AA analysis.
Department of Analytical Biochemistry, Biology Centre, Academy of Sciences of the Czech Republic, Ceské Budejovice, Czech Republic. helenaz@bclab.eu
Helena
H
Zahradnícková
Petr
P
Husek
Petr
P
Simek
D002849:Chromatography, Gas / Q000379:methods*; D005466:Fluorocarbons / Q000737:chemistry*; D005561:Formates / Q000737:chemistry*; D057230:Limit of Detection; D015203:Reproducibility of Results
D005466:Fluorocarbons; D005561:Formates; C033094:perfluorobutyric acid; C083354:chlorocarbonic acid