Semi-continuous monitoring of HMWPAH in microalgae cultures by PT-SPE/HPLC/FD-UV: Estimation of the degradation constant.

A degradation study has been performed with Selenastrum capricornutum incubated with benzo[a]anthracene and benzo[a]pyrene at 50, 100 and 266 ng mL in liquid cultures. After incubation, these high molecular weight polycyclic aromatic hydrocarbons (HMW PAH) were extracted from both, the medium and biomass in a single step, and then quantified by a sensitive and validated analytical methodology based on pipette-tip SPE and HPLC with fluorescence and UV detection (PT-SPE/HPLC/FD-UV). The methodology presented good linearity r > 0.99, LOD of 0.9 and 0.7 ng mL for BaA and BaP, respectively. A fast and semi-continuous appreciation of the degradation behavior was achieved. The pollutants were monitored at different times (0.5-18 h) in the same culture flask, with sampling volume of 1 mL. Biodegradation percentages close to-90% were observed at 18 h. The degradation curves were fitted to the first order reaction (r > 0.95) and the degradation rate constants were similar in all bioassays (0.1 h) and independent of concentration and compound. The degradation pathways of HMW PAH by microalgae and their enzyme are poorly known but the hypothesis of the degrading enzyme proportionally activated according to the PAH concentration is supported by this result. The early emergence dihydrodiol-type metabolites were detected.