Determination of 3-nitrobenzanthrone, its metabolites, and 41 polycyclic aromatic compounds (16 PAHs, 19 nitro-PAHs, and 6 oxy-PAHs) in ascidians (Phallusia nigra)

Abstract

This work demonstrates an effective solid–liquid extraction procedure, in combination with a microscale dispersive solid-phase extraction cleanup, for the determination of 44 polycyclic aromatic compounds, including the potent mutagenic compound 3-nitrobenzathrone and its metabolites 3-aminobenzanthrone and 3-acethyl-aminobenzanthrone, in Phallusia nigra. A new customized device was constructed using borosilicate glass to perform the ultrasound-assisted extraction of the compounds. The dispersive solid-phase extraction cleanup was carried out in a microscale device filter and optimized though multivariate statistical tools. The optimum conditions were determined as 93 mg of activated alumina/silica gel (1:1, w/w) and vortex-assisted agitation for 3.50 min. The limit of detection ranged from 5.47 to 9.89 ng g−1, 7.27 to 1,524 ng g−1, and 42.7 to 108 ng g−1 for PAHs, oxy-PAHs, and nitro-PAHs, respectively. The ANOVA test (p < 0.05) indicated no evidence of lack-of-fit for the matrix-matched analytical curves, with R2 > 0.990. The recoveries on the DORM-3 reference standard material ranged from 75.3% to 120%, 77.6% to 117%, and 70% to 124% for PAHs, oxy-PAHs, and nitro-PAHs, respectively. The developed procedure was applied to nine ascidians samples. PAHs, oxy-PAHs, and nitro-PAHs were detected in the samples at concentrations ranging from 5.02 to 489 ng g−1 dw, 28.8 to 6,587 ng g−1, and 32.3 to 1,635 ng g−1, respectively. Thus, the proposed procedure was comprehensive, precise, accurate, and suitable for the determination of these compounds in ascidians samples.