Lab of Toxicology, Department of Health Sciences, Dong-A University, 37, Nakdong-daero 550 beon-gil, Saha-gu, Busan, 49315, Republic of Korea.
Korea Conformity Laboratories, 8, Gaetbeol-ro 145 beon-gil, Yeonsu-gu, 21999, Incheon, Republic of Korea.
Part Fibre Toxicol. 2020 Sep 11;17(1):43. doi: 10.1186/s12989-020-00377-9.
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.
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.
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.
量化各种器官中积累的纳米材料对于研究其毒性和毒代动力学至关重要。然而，包括碳纳米材料（CNMs）在内的某些类型的纳米材料在生物基质中难以定量。因此，开发用于定量重要器官中 CNMs 的改进方法对于它们的持续修饰和应用非常重要。
在这项研究中，将炭黑、纳米金刚石、多壁碳纳米管、碳纳米纤维和石墨烯纳米片组装在一起作为 CNMs 面板。所有 CNMs 在 750nm 处均显示出显著的吸光度，而其生物成分在该波长处的吸光度最小。研究表明，使用 750nm 处的吸光度定量 CNMs 在所有研究材料中的准确率均超过 94%。用蛋白酶 K（PK）孵育肺组织匀浆和 CNMs 的混合物 2 天，平均回收率超过 90%。使用 30μg/只小鼠的 CNMs 在鼠咽部抽吸模型中证实了该方法的实用性。
我们开发了一种改进的 CNMs 肺负荷测定方法，使用 PK 消化和紫外-可见分光光度法，准确率>94%，回收率>90%。该方法可应用于任何在近红外波段具有足够吸光度的纳米材料，并可将纳米材料与体内元素以及纳米材料的可溶性部分区分开来。此外，PK 消化与针对纳米材料的其他仪器分析的组合可应用于器官负荷分析。
