Methodology for Quantitative Determination of Polycyclic Aromatic Hydrocarbons in Protozoa Cultures

Two simple and sensitive analytical methods based on ma- trix solid phase dispersion (MSPD) and solid phase extraction (SPE), both followed by high performance liquid chromatography (HPLC) with fluorescence detection (FL) were developed for determining the high molecular weight polycyclic aromatic hydrocarbons (PAH) benzo(a)anthracene, benzo(b)fluoranthene and benzo(a)pyrene, in liquid cultures of the ciliate protozoan Colpoda cucullus. The first method employed MSPD technique for extraction of the analytes from a very small amount of microorganisms and insoluble organic matter. The second method used SPE to extract analytes from the liquid me- dium. Validation parameters indicated good linearity (r 2 > 0.99) and precision (inter-day RSDs 90% for MSPD and 70% for SPE with limits of detection between 0.02 and 0.03 µg/g for MSPD-HPLC and between 0.03 and 0.04 µg/L for SPE-HPLC. The developed methodology was success- fully used for the quantitative determination of PAH in microorganism cultures for the first time.