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The aquatic humic substances (AHS) investigated in this study were conventionally isolated from Rio Negro waters - Amazonas State/Brazil by means of the collector XAD 8. A special five-stage tangential-flow ultrafiltration device was used for analytical fractionation of AHS. The fractionation patterns (6 fractions each) showed that metal traces remaining in AHS after their XAD 8 isolation have different size distributions. For instance, the major percentage of traces of Ni, Cu, Zn, Cd and Pb (determined using ICP-AES) was preferably complexed by molecules with relatively high molecular size (30-100 kDa) and the following complexation order was characterized: F2 >> F1 = F4 = F5 > F3 > F6. Moreover, the species formed between AHS and metals prepared by spiking, showed distribution patterns changing as a function of the complexation time (ageing process), indicating a slow transformation process and an inner rearrangements in the binding sites within the AHS molecules.

A sequential system for fractionation by ultrafiltration (SSFU) equipped with advanced membranes filters (molecular size cut-off: 5, 10, 30, 50 and 100 kDalton) of the polyethersulfone type was developed for analytical fractionation of humic substances (HS) extracted from aquatic systems or soils. The device consists of five membrane filters (Sartocon® Micro) operated by a multi-channel peristaltic pump, enabling an easy handling, working in a closed system and with simple collection of the six obtained fractions (F1>100; F2: 100-50; F3: 50-30; F4:30-10; F5: 10-5 and F6