Abstract
Chiral chromatography is a sensitive technique used to separate l- and d-amino acids. Marfey’s reagent (1-fluoro-2,4-dinitrophenyl-5-l-alaninamide, FDAA) has been used as a chiral derivatization reagent to determine amino acid stereochemistry. Replacement of alanylamide in this reagent with leucylamide resulted in the derivative 1-fluoro-2,4-dinitrophenyl-5-l-leucinamide (FDLA), which proved a superior chiral derivatization reagent for the separation of d,l-amino acids due to the improved LC/MS measurement sensitivity. However, FDLA derivatization requires alkaline conditions, achieved with the use of NaHCO3, which necessitates an additional desalting step (e.g., liquid–liquid extraction) before quantitative analysis using LC/MS/MS. To establish a simplified LC/MS/MS measurement method, solid NaHCO3 was replaced with volatile triethylamine (TEA). The 30-min derivatization using FDLA under alkaline conditions with TEA at room temperature allowed for exclusion of the desalting step from the FDLA pre-column derivatization. Furthermore, using TEA, larger peak areas and more effective detection of hydrophilic amino acids could be achieved.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.