Development of the separation-oriented derivatization method in HPLC analysis using pentafluorophenyl moiety

Abstract

We have developed a new separation-oriented derivatization method based on the affinity of the interaction between the pentafluorophenyl (PFP) group for the HPLC determination of native fluorescent non-steroidal anti-inflammatory drugs (NSAIDs). This method involves the derivatization of NSAIDs with pentafluorobenzyl bromide and the separation of derivatives using a PFP-modified stationary phase column. In this method, the PFP-derivatized NSAIDs were selectively retained on the PFP column, followed by fluorescence detection, whereas the derivatives were not retained on the ODS-, phenyl-, and perfluoroalkyl-modified HPLC columns. The detection limits (signal-to-noise ratio of 3) of the NSAID standards were found to be 44 - 84 fmol on column for a 20 μL injection, and the repeatability of this method was within 7.1% as relative standard deviation. We show that the proposed method can be used to analyze NSAIDs in human plasma samples without the interference of endogenous matrices. © 2013 The Japan Society for Analytical Chemistry.