The landmark publication by the National Research Council putting forward a vision of a toxicology for the 21st century in 2007 has created an atmosphere of departure in our field. The alliances formed, symposia and meetings held and the articles following are remarkable, indicating that this is an idea whose time has come. Most of the discussion centers on the technical opportunities to map pathways of toxicity and the financing of the program. Here, the other part of the work ahead shall be discussed, that is, the focus is on regulatory implementation once the technological challenges are managed, but we are well aware that the technical aspects of what the National Academy of Science report suggests still need to be addressed: A series of challenges are put forward which we will face in addition to finding a technical solution (and its funding) to set this vision into practice. This includes the standardization and quality assurance of novel methodologies, their formal validation, their integration into test strategies including threshold setting and finally a global acceptance and implementation. This will require intense conceptual steering to have all pieces of the puzzle come together.