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The derivatized free amino acids were applied to a 30-cm Pico-Tag free-amino-acid-analysis column (3.9300 mm) equilibrated with buffer A equipped with a Waters HPLC system (510 HPLC pump, 717 automatic sampler, 996 photodiode array detector, and Millennium 32 chromatography manager) and eluted with a linear gradient composed of buffer B (0%, 14%, 20%, 46%, and 100%) at a flow rate of 1 mL/min at 46