An approach based on the coupling HPLC-UV-CV/HG-mAFS-AFS has been developed for the simultaneous determination of mercury and arsenic species. A home modified AFS detector has been used for the analysis of mercury coupled in series with another similar detector for arsenic determination. The determined species were Hg2+, methylmercury (MeHg+), As(III), As(V) and monomethylarsinate (MMA). A critical aspect is the chromatographic separation, which was carefully optimized for the separation of all the species. The detection limits for these species are 11, 8, 17, 17 and 3 ng ml(-1), respectively. Linear curves for MMA were obtained between 10 and 200 ng ml(-1). The linear dynamic range for all the other species was comprised between the detection limit and 500 ng ml(-1). The influence of cations and anions at the concentration usually present in natural freshwater was studied. The procedure was validated by application to spiked natural freshwater samples from the south-west Spain, and it can be considered for routine analysis of polluted sites.