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Reversed-phase (RP) HPLC typically uses octadecyl silica (C18) columns in combination with acidic mobile phases, composed of acetonitrile/water [32, 41,42,43], with additives like formic acid (FA) [41, 43, 45, 46], ammonium formate [42, 45, 47, 48], acetic acid (AcA) [49], ammonium acetate [32, 50], trichloroacetic acid (TCA) [38, 51], trifluoroacetic acid (TFA) [32, 50, 52], or ammonium phosphate [53]

Ammonium persulfate (100 L, 20%, Sigma) was added while vortexing and continued for 1 min