Broiler nutrition can be affected by many different factors, and diets can be made up of a wide variety of components. Feed enzymes, different Ca sources, and various ingredient particle sizes all contribute to broiler nutritional status and need to be investigated to determine their full effects on the bird. The objective of the first study was to evaluate various Ca sources and microbial phytases using an in vitro assay to simulate broiler digestion. The trial measured total Ca and P, soluble Ca and P, and pH. It involved limestone, dicalcium phosphate, highly soluble Ca (HSC), whey, and a control with no added Ca. Each diet was supplemented with 0 or 500 FTU/kg of phytase 1 or phytase 2. Diets were analyzed for total Ca and P and then were subjected to a gastric phase digestion, including the addition of HCl and pepsin, with incubation times of 5, 10, or 20 min. They were also subjected to a gastric phase and a SI phase digestion, including the addition of HCl, pepsin, NaHCO3, and pancreatin with incubation times of 5, 10, 20, or 60 min. The objective of the second study was to evaluate various particle sizes, Ca sources, and source locations using the same in vitro assay. Large, medium, and fine particle sizes, as well as limestone from 4 different locations and HSC were subjected to the same assays and conditions as in the first trial. These studies suggest that HSC is a highly soluble Ca source, phytase improves P solubility in vitro, fine particle size is more soluble than large particle size in vitro, and location from which the Ca source is obtained can affect Ca and P solubility. The in vitro assay utilized in this study is a fast and cost efficient technique for evaluating and comparing parameters of broiler diets to make recommendations for commercial poultry nutrition.