Rapid Determination of Anthracene and Benzo(a)pyrene by High-Performance Liquid Chromatography with Fluorescence Detection

Abstract

ABSTRACTA rapid and simple method was optimized and validated for the separation and quantification of anthracene and benzo(a)pyrene, two of the most toxic polycyclic aromatic hydrocarbons, at ultratrace levels in aqueous samples by direct injection. The determination of anthracene and benzo(a)pyrene was performed by reverse-phase high-performance liquid chromatography (HPLC) with fluorescence detection. A fractional factorial matrix and a Box–Behnken design were chosen for screening and optimization purposes, respectively. The optimized parameters that significantly influenced the system were the flow rate (1 mL min−1), mobile-phase strength (90% acetonitrile:10% deionized water), column temperature (35°C), and excitation wavelength (254 and 267 nm for anthracene and benzo(a)pyrene, respectively). The injection volume and emission wavelength were fixed at 100 µL and 416 nm, respectively. The quantification limits were 75 ng L−1 for anthracene and 30 ng L−1 for benzo(a)pyrene. The relative standard deviations for the recovery and intra and interday precision values were lower than 20%. The method allows the analysis of aqueous samples with a good resolution for anthracene and benzo(a)pyrene below values permitted by the recently developed European Directive 2013/39/EU for inland surface waters. In addition, this work provides guidelines for the simultaneous optimization of several parameters by experimental design.