Environmental factors affecting nutrient availability during development can have long-term effects including predisposition to diseases later in life. In order to identify chemicals in the environment capable of altering nutrient mobilization, we analyzed yolk malabsorption in the zebrafish embryo, which relies on maternally-derived yolk for nutrition during the first week of life. Embryos of the transgenic zebrafish line HGn50D, which fluoresce in the yolk syncytial layer, the heart, and the eyes of the zebrafish, were exposed from two to five days post fertilization to nine chemicals, including pesticides (prochloraz, imazalil, and butralin), pharmaceuticals (clofibrate and gemfibrozil), flame retardants (tetrabromobisphenol A, and tetrachlorobisphenol A), a surfactant (perfluorooctanoic acid) and a biocide (tributyltin). We developed a software package named ZebRA to automatically and accurately segment and quantify the area of the fluorescing yolk in images captured at the end of the treatment period. Based on this quantification, we found that prochloraz decreased yolk absorption, while butralin, tetrabromobisphenol A, tetrachlorobisphenol A and tributyltin, at the higher concentrations tested, increased yolk absorption. Given the number and variety of industrial chemicals in commerce today, development of automated image processing allowing for high-speed analysis and accurate quantification of biological effects is an important step for enabling high throughput screening to identify chemicals adversely impacting nutrient absorption.