HPLC with diode‐array detection for the simultaneous determination of di(2‐ethylhexyl)phthalate and mono(2‐ethylhexyl)phthalate in seminal plasma

Abstract
A high‐performance liquid chromatographic method for the simultaneous determination of di(2‐ethylhexyl)phthalate (DEHP) and its major metabolite mono(2‐ethylhexyl)phthalate (MEHP) in seminal plasma was developed and validated. The method involves liquid–liquid extraction followed by isocratic reversed‐phase chromatography with diode‐array detection. The recovery, selectivity, linearity, precision and accuracy of the method were evaluated from the analysis of spiked seminal plasma samples. The effect of mobile‐phase composition and pH on the retention of the target analytes was investigated. The limits of detection were 0.010 and 0.015 µg/mL, for DEHP and MEHP, respectively. This method was used to analyze real samples in support of clinical studies on these potential endocrine disruptors. Copyright © 2007 John Wiley & Sons, Ltd.