Omega-3 fatty acids (O3FA) are essential for normal human growth, development, and disease prevention. Purslane (Portulaca oleraceaeL.) is an excellent source of the O3FA α-linolenic acid (LNA)—with higher concentrations than any green leafy-vegetable examined to date—and is being considered for cultivation (by USDA-ARS) in an effort to improve the balance of essential fatty acids in the western diet. Twenty-fi ve-day-old seedlings of both a green-leafed and a golden-leafed type of purslane were transplanted into a closed hydroponic system. Nitrogen, at 200 ppm, was provided as NO3 and NH4 forms to yield NO3: NH4 ratios of 1:0, 0.25:0.75, 0.5:0.5, and 0.75:0.25. Treatments were arranged in a randomized complete-block design with five replications. The experiment was repeated. Young, fully expanded leaves were harvested 18 days after treatment initiation, frozen (–60°C), and then analyzed for fatty acids using gas chromatography. Although the two types of purslane did not differ in LNA concentration, the green-leafed purslane produced greater total dry mass than the golden-type. On a leaf dry mass basis, plants grown with a NO3:NH4 ratio of 0.5:0.5 produced 241% and 53% greater LNA than plants grown with NO3:NH4 ratios of 1:0 and 0.75:0.25, respectively. Plants grown with NO3:NH4 ratios of 1:0 and 0.25:0.75 produced similar leaf LNA concentrations. Total dry mass was not affected by the nitrogen treatments.