: An alternative plasticizer, tris (2-ethylhexyl) trimellitate (TOTM), was developed from di-(2-ethylhexyl) phthalate (DEHP) for use in medical tubing. However, little is known about the biological effects of TOTM and thus its safety is not yet well-established. We investigated the leachability of TOTM from TOTM-plasticized Polyvinylchloride (PVC) feeding tubes. Furthermore, we studied whether TOTM influences the cell toxicity of human leukemia (HL)-60 cells, the cell proliferation of human breast cancer MCF-7 cells, and the binding affinity for human estrogen receptor α.