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Each sample (8 μl) was injected onto an Agilent Poroshell 120 EC-C18 (100 × 2.1 mm, 2.7 um) in a Shimadzu Nexera UHPLC system at a flowrate of 0.2 ml/min using a gradient elution created between buffer A (10 mM tributylamine aqueous solution adjusted pH to 4.95 with 15 mM acetic acid) and buffer B of methanol, from 10% to 40% (buffer B) in 30 min at room temperature