Optimized Enantioselective Derivatization of Amino Acids with Marfey’s Reagent for Reversed-Phase UHPLC–HRMS Determination

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Abstract
The determination of amino acid enantiomers plays an important role in addressing a wide range of analytical challenges. Chiral chromatographic columns are commonly used for this purpose; however, they are expensive and require highly qualified personnel. This study optimizes the derivatization conditions using the chiral derivatizing reagent Marfey’s reagent, which enables the determination of amino acid enantiomer derivatives by reversed-phase UHPLC–HRMS with a C18 column. The D- and L-forms of the analytes exhibited identical ionization efficiencies, an important prerequisite for accurate quantitative chemical analysis. The results also demonstrate that gabapentin can serve as an internal standard instead of isotope-labeled analytes. Under the optimized conditions, the method achieved detection limits of 20 ng/mL for the compounds investigated.

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