Abstract This study describes the determination of Polynuclear aromatic hydrocarbons in water using a new modification for the commercial SPME-HPLC interface which incorporates a dynamic SPME-HPLC extraction and vibration in the desorption chamber during the extraction and desorption steps. Extraction and desorption parameters were investigated using fourteen PAHs of different volatilities (naphthalene, acenaphthalene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo(a)anthracene, chrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene, dibenzo(a,h)anthracene, benzo(g,h,i)perylene) as test compounds and fluorimetric detection. Regression coefficients close to 0.99 with RSD < 8.1% and detection limits in the range 0.004-0.59 µg/L were found. A method was applied to determine the above PAHs in water samples. The results were compared with the 550.1 EPA method at the 0.05 significance level.