AbstractThe sp² carbon nanomaterials have fantastic properties and hold great potential in diverse areas, including electronics, energy, environment, biomedicine, and so on. The wide applications of sp² carbon nanomaterials require the thorough investigations on their biosafety. The quantification of sp² carbon nanomaterials is the first and crucial step in the biosafety evaluations. In this review, we summarized the quantification technologies for sp² carbon nanomaterials and compared the advantages/disadvantages of these technologies. The pharmacokinetics, the biodistribution, and the ecological uptake of sp² carbon nanomaterials were achieved by using the quantification technologies. Furthermore, the influence factors such as surface modification, size, shape, and exposure pathway were concerned, and the general rules in the biological behaviors of sp² carbon nanomaterials were proposed. The implications to the biomedical applications and biosafety evaluations of sp² carbon nanomaterials are discussed.