Fluorescence quenching of the fluorescence is a useful method to determine copper complexing properties of the dissolved organic matter. This technique provides express results often by choosing one or two excitation-emission fluorescence measurement for a straight range of concentration. That why it is not representative of the whole sample. On another hand, using total luminescence spectra gives lots of information that is difficult to manipulate in term of complexing properties. This work focus on a fluorescence quenching experiment carried out on a filtered black water from Rio Negro from north Brazil (Sao Gabriel da Cachoeira) using excitation emission matrix of fluorescence (EEMF) with a large range of copper concentration. Indeed, metal additions were performed using logarithmic increments of the total copper concentration to cover a wide range of concentration from 1.7 9 10 -9 to 10 -3 mol l -1 . These data were treated by two different ways for comparison: on the one hand, data treatment were computed, as usually, at different fluorescence intensity positions using multi-response wavelength method with two ligand, and, on the another hand, a statistical method, parallel factor analysis (PARAFAC), was applied to extract fluorescent component. Then a fitting was done on these PARAFAC components with one or two complexing sites each one. Results show that PARAFAC enables quantitative evaluation of complexing parameters for copper as good as certain multi-response methods: L 1T 16.2 9 10 -6 mol l -1 (5.0 \ log(K 1 ) \ 5.8) and L 2T 3.0 9 10 -4 mol l -1 (2.8 \ log(K 2 ) \ 3.9). PARAFAC confirms that for this natural sample only two fluorescent ligands are present with two types of site for each component. No residual fluorescence was detected by the statistical treatment. Wall surface interferences were pointed-out as phenomenon to be solve to overcome the limiting efficiency of the total copper concentration range.

In this study, metal contamination experiments were conducted to investigate the effects of copper and cadmium on the growth of the marine toxic dinoflagellate Alexandrium catenella and on the production of dissolved organic matter (Dissolved Organic Carbon: DOC; Fluorescent Dissolved Organic Matter: FDOM). This species was exposed to increasing concentrations of Cu(2+) (9.93 × 10(-10)-1.00 × 10(-7)M) or Cd(2+) (1.30 × 10(-8)-4.38 × 10(-7)M), to simulate polluted environments. The drastic effects were observed at pCu(2+)=7.96 (Cu(2+): 1.08 × 10(-8)M) and pCd(2+)=7.28 (Cd(2+): 5.19 × 10(-8)M), where cyst formation occurred. Lower levels of Cu(2+) (pCu(2+)>9.00) and Cd(2+) (pCd(2+)>7.28) had no effect on growth. However, when levels of Cu(2+) and Cd(2+) were beyond 10(-7)M, the growth was totally inhibited. The DOC released per cell (DOC/Cell) was different depending on the exposure time and the metal contamination, with higher DOC/Cell values in response to Cu(2+) and Cd(2+), comparatively to the control. Samples were also analyzed by 3D-fluorescence spectroscopy, using the Parallel Factor Analysis (PARAFAC) algorithm to characterize the FDOM. The PARAFAC analytical treatment revealed four components (C1, C2, C3 and C4) that could be associated with two contributions: one, related to the biological activity; the other, linked to the decomposition of organic matter. The C1 component combined a tryptophan peak and a characteristic humic substances response, and the C2 component was considered as a tryptophan protein fluorophore. The C3 and C4 components were associated to marine organic matter production.