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At present, there is a lack of understanding of the combined ecotoxicity of carbon-based nanomaterials and co-contaminants. In this paper, we report on the toxicity of three carbon nanomaterials (fullerene-soot, multiwall carbon nanotubes, and graphene). Two standardized toxicity bioassays, the immobilization of the invertebrate Daphnia magna and the bioluminescence inhibition of the marine bacteria Vibrio fischeri, have been used. Synergistic and antagonistic effects of binary mixtures composed of fullerene soot and organic co-contaminants as malathion, glyphosate, diuron, triclosan, and nonylphenol were assessed. The isobologram method was used to evaluate the concentrations producing an effect, in comparison to those effects expected by a simple additive approach. In this study, antagonism was the predominant effect. However, synergism was also observed as in the case of D. magna exposed to mixtures of malathion and fullerene soot. D. magna was shown to be the most sensitive assay when carbon nanomaterials were present. Toxicity to D. magna was as follows: fullerene soot > multiwall carbon nanotubes > graphene. These results were proportional to the size of aggregates, smaller aggregates being the most toxic. The vector function of nanomaterials aggregates and the unexpected release inside living organisms was proven for malathion. These results highlight new insights on the risks associated with the release of carbon nanomaterials into the environment.

We propose for the first time methodology for the determination of a number-based concentration of silica (SiO2) nanoparticles (NP) in biological serum using nanoparticle tracking analysis (NTA) as the online detector for asymmetric flow field-flow fractionation (AF4). The degree of selectivity offered by AF4 was found necessary to determine reliably number-based concentration of the measured NP in the complex matrix with a relative measurement error of 5.1% (as relative standard deviation, n = 3) and without chemical sample pretreatment. The simultaneous online coupling to other size and concentration detectors, such as multiangle light scattering (MALS) and ICPMS, for the measurement of the same NP suspension, was used to confirm the particle size determined with NTA and the equivalent particle number determined by AF4/NTA, respectively. The size- and number-based concentration data obtained by independent techniques were in a good agreement. The developed methodology can easily be extended to other types of particles or particle suspensions and other complex matrices provided that the particle size is above the limit of detection for NTA.