Artifacts associated with the fractionation of colloids in a freshwater sample were investigated for conventional membrane filtration (0.45 micron cutoff), and two tangential flow ultrafiltration cartridges (0.1 micron cutoff and 3000 MW cutoff). Membrane clogging during conventional filtration removed some colloids smaller than 0.1 micron in diameter, much smaller than the nominal size of the filter. For certain constituents (e.g., Fe), filter clogging had a significant effect on filtrate concentrations, while artifacts associated with tangential flow ultrafiltration using 0.1 micron cutoff were minimal. Artifacts occurred during tangential flow ultrafiltration with a 3000 MW cutoff, but did not deviate from predicted changes in concentration based on a standard permeation model. Comparison of filtrate concentrations for membrane filtration (at 1.0 and 0.45 micron) and tangential flow ultrafiltration (at 0.1 micron) for a large number of samples from Connecticut rivers shows that significant and consistent differences exist between their separation characteristics. Results for organic carbon, Fe, Mn, Al, Cu, and Pb demonstrate the magnitude of the effects of the fractionation technique on filtrate element concentration, show variability by element and flow condition, and highlight the importance of larger colloids to freshwater metal speciation. One implication of the research is that tangential flow ultrafiltration with large size cutoff membranes (e.g., 0.1 micron) may be superior to conventional filtration with filters in the same size discrimination range, and potentially more appropriate for the fractionation of natural water samples.