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The HPLC gradient grade acetonitrile (ACN), used for chromatographic analysis, hydrochloric acid utilized for the standard solution preparation, sodium hydrogen phosphate heptahydrate (Na 2 HPO 4 7H 2 O), sodium dihydrogen phosphate dihydrate (NaH 2 PO 4 2H 2 O) and sodium hydroxide were purchased from J.T