Comparison of polarimetry and crown ether-based HPLC chiral stationary phase method to determine (l)-amino acid optical purity
2012
Abstract Although various pharmacopoeias provide titration methods to assay ( l )-amino acid content, none of these methods distinguish between ( l )- and ( d )-amino acids and do not consider the presence of enantiomeric impurities. Consequently, these methods are limited in scope to describe the relationship between content and specific rotation, [α]. In this study, the US Pharmacopoeia method was compared with the crown ether-based high performance liquid chromatographic (HPLC) chiral stationary phase (CSP) method to determine ( l )-amino acid content and specific rotation. The ( l )-amino acid content specified by the US Pharmacopoeia method was not consistent with the specific rotation in the presence of enantiomeric impurities, whereas the HPLC-CSP method was very effective for determining the ( l )-amino acid content and the optical purity. The other advantage is that the HPLC-CSP method requires amino acid samples of quite low concentration (as low as 1 μg/mL), whereas the pharmacopoeia method requires higher concentrations (20–110 mg/mL).
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