Preventive approaches against cancer have not been fully developed and applied.For example,the incidence of some types of cancer,including colon cancer,is highly dependent upon lifestyle,and therefore,amenable to prevention.Among the lifestyle factors,diet strongly affects the incidence of colon cancer; however,there are no definitive dietary recommendations that protect against this malignancy.The association between diet-derived bioactives and development of colonic neoplasms will remain ill defined if we do not take into account:(1) the identity of the metabolites present in the colonic lumen;(2) their concentrations in the colon; and(3) the effect of the colonic contents on the function of individual bioactives.We review two approaches that address these questions:the use of fecal water and in vitro models of the human colon.Compared to treatment with individual diet-derived compounds,the exposure of colon cancer cells to samples from fecal water or human colon simulators mimics closer the in vitro conditions and allows for more reliable studies on the effects of diet on colon cancer development.The rationale and the advantages of these strategies are discussed from the perspective of a specific question on how to analyze the combined effect of two types of bioactives,butyrate and polyphenol metabolites,on colon cancer cells. Preventive approaches against cancer have not been fully developed and applied. For example, the incidence of some types of cancer, including colon cancer, is highly dependent upon lifestyle, and therefore, amenable to prevention. Among the lifestyle factors, diet virtually affects the incidence. of colon cancer; however, there are no definitive dietary recommendations that protect against this malignancy. The association between diet-derived bioactives and development of colonic neoplasms will remain ill defined if we do not take into account: (1) the identity of the metabolites present in the colonic lumen; (2) their concentrations in the colon; and (3) the effect of the colonic contents on the function of individual bioactives.We review two approaches that address these questions: the use of fecal water and in vitro models of the human colon. Compared to treatment with individual diet-derived compounds, the exposure of colon cancer cells to samples from fecal water or human colon simulators mimics cl oser the in vitro conditions and allows for more reliable studies on the effects of diet on colon cancer development. rationale and the advantages of these strategies are discussed from the perspective of a specific question on how to analyze the combined effect of two types of bioactives , butyrate and polyphenol metabolites, on colon cancer cells.