Whole grain consumption has been associated with reduced risk of chronic diseases, such as cardiovascular diseases and cancer. These beneficial effects have been attributed to the unique phytochemicals of grains that complement those found in fruits and vegetables. Wheat is one of the major grains in the human diet; however, little is known about the inherent varietal differences in phytochemical profiles, total phenolic and carotenoid contents, or total antioxidant activities of different wheat varieties, which ultimately influence the associated nutritional and health benefits of wheat and wheat products. The objectives of this study were to determine the phytochemical profiles and total antioxidant activity for 11 diverse wheat varieties and experimental lines. The profiles included free, soluble-conjugated, and insoluble-bound forms of total phenolics, flavonoids, and ferulic acids and carotenoid content including lutein, zeaxanthin, and beta-cryptoxanthin. The results showed that total phenolic content (709.8-860.0 micromol of gallic acid equiv/100 g of wheat), total antioxidant activity (37.6-46.4 micromol of vitamin C/g), and total flavonoid content (105.8-141.8 micromol of catechin equiv/100 g of wheat) did not vary greatly among the 11 wheat lines. However, significant differences in total ferulic acid content (p < 0.05) and carotenoid content (p < 0.05) among the varieties were observed, with carotenoid content exhibiting the greatest range of values. Carotenoid content among the 11 wheat varieties exhibited 5-fold, 3-fold, and 12-fold differences in lutein, zeaxanthin, and beta-cryptoxanthin, respectively. A synthetic wheat experimental line, W7985, gave the lowest carotenoid concentrations of any of the genotypes in this study. Such large genotypic differences in carotenoid content may open up new opportunities for breeding wheat varieties with higher nutritional value.