Carbon nanomaterial-derived lung burden analysis using UV-Vis spectrophotometry and proteinase K digestion
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RESEARCH
Open Access
Carbon nanomaterial-derived lung burden analysis using UV-Vis spectrophotometry and proteinase K digestion Dong-Keun Lee1, Soyeon Jeon1, Jiyoung Jeong1, Kyung Seuk Song2 and Wan-Seob Cho1*
Abstract Background: The quantification of nanomaterials accumulated in various organs is crucial in studying their toxicity and toxicokinetics. However, some types of nanomaterials, including carbon nanomaterials (CNMs), are difficult to quantify in a biological matrix. Therefore, developing improved methodologies for quantification of CNMs in vital organs is instrumental in their continued modification and application. Results: In this study, carbon black, nanodiamond, multi-walled carbon nanotube, carbon nanofiber, and graphene nanoplatelet were assembled and used as a panel of CNMs. All CNMs showed significant absorbance at 750 nm, while their bio-components showed minimal absorbance at this wavelength. Quantification of CNMs using their absorbance at 750 nm was shown to have more than 94% accuracy in all of the studied materials. Incubating proteinase K (PK) for 2 days with a mixture of lung tissue homogenates and CNMs showed an average recovery rate over 90%. The utility of this method was confirmed in a murine pharyngeal aspiration model using CNMs at 30 μg/ mouse. Conclusions: We developed an improved lung burden assay for CNMs with an accuracy > 94% and a recovery rate > 90% using PK digestion and UV-Vis spectrophotometry. This method can be applied to any nanomaterial with sufficient absorbance in the near-infrared band and can differentiate nanomaterials from elements in the body, as well as the soluble fraction of the nanomaterial. Furthermore, a combination of PK digestion and other instrumental analysis specific to the nanomaterial can be applied to organ burden analysis. Keywords: Carbon black, Nanodiamond, Multi-walled carbon nanotube, Carbon nanofiber, Graphene, Lung burden
Background Inhalation is the most common and hazardous route of exposure to nanomaterials in an occupational setting. Inhalation of nanomaterials produces a higher deposition rate of the micron-sized particles within the alveoli as a result of their size-dependent aerodynamic properties [1–3]. Furthermore, deposited particles exhibit limited clearance rates from the alveoli due to the absence of * Correspondence: [email protected] 1 Lab of Toxicology, Department of Health Sciences, Dong-A University, 37, Nakdong-daero 550 beon-gil, Saha-gu, Busan 49315, Republic of Korea Full list of author information is available at the end of the article
mucociliary clearance. The clearance of these nanomaterials from the alveoli is influenced by the physicochemical properties of the material including size, shape, functionalization, and dissolution [4–6]. Because of the long retention period for nanomaterials in the lungs, the Organization for Economic Cooperation and Development (OECD) testing guidelines call for repeated inhalation studies (i.e., TG 412 and 413) and were revised in 2018 to include lung burden measurements showing l
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