Human biomonitoring studies to determine total titanium (total-Ti) and titanium dioxide (TiO 2 ) particles require very sensitive inductively coupled plasma high resolution mass spectrometry (ICP-HRMS) methods. The result of such studies can only be reliably when used in a risk assessment if the analytical methods are fully validated, as reported here for human liver and spleen. For total-Ti an acidic HNO 3 /HF digestion was applied, followed by ICP-HRMS analysis of the acidic digests. For TiO 2 particles a two-step digestion procedure was developed. In the first step the formaldehyde-fixed tissue was depolymerized and in the second step an enzymatic digestion was performed. Sample digests were analyzed with ICP-HRMS operated in single particle mode to detect TiO 2 particles. Both methods were validated according to NEN 7777 using a scheme of duplicate analysis over an eight day period. Detection limits for total-Ti and TiO 2 particles were 0.01 and 0.005 mg Ti/kg with a particle size quantification limit of 85 nm for TiO 2 particles. Repeatability and reproducibility were 24% and 31% for the total titanium concentration, 22% and 39% for the particle titanium concentration, and 29% and 24% for the titanium dioxide particle size. In total 30 human tissue samples were analyzed with the developed and validated method. The results show a total-Ti content in the range of 0.02-0.09 mg Ti/kg in liver and 0.02-0.4 mg Ti/kg in spleen. The concentrations of particle TiO 2 ranged from 0.01-0.08 mg Ti/kg in liver and from 0.01-0.1 mg Ti/kg in spleen. On the average, particle TiO 2 explained up to 67% of the total-Ti concentrations. The detected TiO 2 particles had a particle size range of 85 to 720 nm.
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