Forty-five cultivars of lettuce (Lactuca sativa) were field-grown using best management practices at the Louisiana State University Agricultural Center (LSU AgCenter) Botanic Gardens in Baton Rouge during the Fall 2011 and Fall 2012 seasons. Recommended cultivars were selected for commercial production in Louisiana based on fresh weight and lettuce size (width and height). Nitrate (NO3–) concentration was analyzed for each cultivar, as lettuces are known to accumulate and concentrate NO3–, and were then compared with the U.S. Environmental Protection Agency’s (EPA) oral reference dose (RfD—the EPA’s maximum acceptable oral dose of a toxic substance) of 1.6 mg NO3-nitrogen (N) per kilogram body weight per day. Recommended butterhead cultivars were Caliente and Harmony (21.6 and 13.9 ppm NO3–, respectively); recommended green-leaf cultivars were Salad Bowl and Tango (10.6 and 4.6 ppm NO3–, respectively); recommended red-leaf cultivars were Red Salad Bowl, Red Sails, and New Red Fire (15.2, 15.4, and 24.0 ppm NO3–, respectively). The only recommended romaine cultivar was Green Towers (11.2 ppm NO3–), and recommended crisphead cultivars included Raider and Ithaca (17.6 and 14.9 ppm NO3–, respectively). Of the highest yielding cultivars, New Red Fire accumulated the greatest NO3– concentration: 24.0 ppm in both years 1 and 2. The NO3– concentration is less than the levels of concern for both men and women 20 to 74 years old, 3.9% of the RfD for men and 4.59% of the RfD for women.