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The use of nanoparticle tracking analysis (NTA) for size determination of nanocatalysts dispersed in bitumen or heavy oil fractions was investigated. A method for sample preparation is proposed, and comments on adaptation of the technique and troubleshooting are addressed and discussed. The NTA was able to measure the particle size of a trimetallic catalyst dispersed in bitumen obtaining a mode size of 111 nm, with particles ranging from 40 to 1000 nm and 80% of them between 57 and 176 nm. NTA data was compared to the particle size obtained by depositing the same catalyst on sand and analyzing it through SEM-EDX, obtaining the same particle size range. Refinement of the sample preparation method and measuring parameters are suggested.