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This paper focuses on the development of an enhanced LC/ESI-MS method for the quantification of ketoprofen(KET)enantiomers.KET was first derivatized with(S)-1-methyl-4-(5-(3-aminopyrrolidin-1-yl)-2,4-dinitro-phenyl)piperazine(APy-PPZ),forming(KET-Apy-PPZ)derivative,and then was further converted to(KET-Apy-MPPZ)iodide or(d3-KET-Apy-MPPZ)iodide by reacting with CH3I or CD3I in one-pot.