Abstract
A highly accurate and sensitive method has been developed for assessing the enantiomeric purity of L-prolinamide hydrochloride using reverse-phase HPLC. The method involves utilizing Marfey’s reagent to derivatize DL-prolinamide, followed by pre-column derivatization into a diastereomer, and subsequent RP-HPLC analysis using Hypersil BDS C18 (4.6 x 250mm, 5.0m). The method allows for excellent separation of the two enantiomers, with a resolution of over 3 and a tailing factor of less than 1.5. Notably, the concentration range for D-prolinamide is 0.15 to 0.75%, with exceptional linearity. Moreover, the limit of detection and limit of quantitation for D-prolinamide are 0.075% and 0.15%, respectively. Overall, this method is highly specific and simple, providing a reliable approach for assessing enantiomeric purity in L-prolinamide hydrochloride.
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