Isaacson, C.W., Usenko, C.Y., Tanguay, R.L., and Field, J.A. (2007) Quantification of Fullerenes by LC/ESI-MS and Its Application to in Vivo Toxicity Assays. Analytical chemistry. 79(23):9091-9097.
With production and use of carbon nanoparticles increasing, it is imperative that the toxicity of these materials be determined; yet such testing requires specific and selective analytical methodologies that do not yet exist. Quantitative liquid-liquid extraction was coupled with liquid chromatography/electrospray ionization mass spectrometry for the quantitative determination of fullerenes from C60 to C98. Isotopically enriched, 13C60, was used as an internal standard. The method was applied to determine the loss of C60 from exposure water solution and uptake of C60 by embryonic zebrafish. The average recovery of C60 from zebrafish embryo extracts and 1% DMSO in aqueous-exposure solutions was 90 and 93%, respectively, and precision, as indicated by the relative standard deviation, was 2 and 7%, respectively. The method quantification limit was 0.40 mug/L and the detection limit was 0.02 mug/L. During the toxicological assay, loss of C60 due to sorption to test vials resulted in the reduction of exposure-solution concentrations over 6 h to less than 50% of the initial concentration. Time-course experiments indicated embryo uptake increased over course of the 12-h exposure. A lethal concentration that caused 50% mortality was determined to be 130 mug/L and was associated with a zebrafish embryo concentration, LD50, of 0.079 mug/g of embryo.