Novel method using hybrid markers: development of an approach for pulmonary measurement of multi-walled carbon nanotubes

Background Multi-walled carbon nanotubes (MWCNT)s are suspected to induce pulmonary and pleural cancers due to their asbestos-like configurations. Therefore, accurate measurement of inhaled nanotubes in target organs is crucial for assessing cancer risk. Conventionally, nanotubes are measured after combustion at high temperature for conversion into CO 2 ; however, the sensitivity is poor and the method lacks versatility. We have therefore developed a novel approach using hybrid markers for nanotube analysis, featuring high sensitivity and the capacity to conduct repeated analyses. The method involves adsorption of markers to nanotubes, followed by their desorption and assessment by means of high performance liquid chromatography (HPLC). Methods Recovery of MWCNT from rat lungs was conducted, and pulmonary MWCNT amounts were determined using rats intratracheally-exposed to MWCNT aerosol at 5 mg/m 3 for 6 hours/day. Results The correlation coefficient for the calibration curve of MWCNT weight and the HPLC area was 0.9991. Consequently, the lower quantitation limit yielded was 0.2 μg. The recovery was 92-98% at approximately 0.4-2.0 μg demonstrating that MWCNTs in the lung could be measured accurately and precisely. Conclusions We have developed a novel method using a hybrid marker approach for nanotube analysis, featuring very high sensitivity and the capacity to conduct repeated analyses. We further confirmed correlations between the amounts of nanotubes and markers and pulmonary nanotube measurement demonstrated that trace amounts could be detected with values closely relating to the administered dose, verifying that the method is sensitive and precise.