Boc-protected amino acids, such as Boc-L-valine, are widely used as key starting materials in the synthesis of pharmaceutical drug substances, necessitating efficient and reliable analytical control strategies. In this work, a streamlined reversed-phase chiral HPLC method was developed for the comprehensive characterization of Boc-L-valine without the need for derivatization. The method enables simultaneous determination of chiral and achiral impurities, assay, and compound identification within a single analytical run. This integrated approach simplifies the analytical workflow, increases throughput, and significantly reduces overall method validation efforts. Moreover, the method provides a robust and versatile platform readily applicable to routine quality control and method development for other Boc-protected or Fmoc-protected amino acids.
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