Spinach (Spinacia oleracea) cv Whitney was tested for iron bioavailabilty using an in vitro human intestinal cell culture ferritin bioassay technique previously developed. Spinach was cultured in a growth chamber for 33 days, harvested, and freeze-dried. Total iron in the samples was an average of 71 µg/g dry weight. Spinach was digested in vitro (pepsin and 0.1 M HCl followed by pancreatin and 0.1 M NaHCO 3 ) with and without the addition of supplemental ascorbic acid. Caco-2 cell cultures were used to determine iron bioavailability from the spinach mixtures. Production of the iron-binding protein ferritin in the Caco-2 cells showed the supplemental ascorbic acid doubled bioavailability of iron from spinach. The data show fresh spinach is a poor source of iron, and emphasize the importance of evaluation of whole meals rather than single food items. The data support the usefulness of the in vitro/Caco-2 cell ferritin bioassay model for prescreening of space flight diets for bioavailable iron. tent of the food, the health and iron status of the individual, the chemical form of the iron, the presence of iron absorption enhancers or inhibitors in the meal, and the individual's genetic predisposition (11). Thus, total iron content measurements of a food do not indicate actual iron bioavailability or iron absorption. Heme-bound iron, as found in meats and fish, is more readily absorbed (more bioavailable) than nonheme iron. The predominant form of iron in vegetables is nonheme iron. Absorption of nonheme iron is enhanced by the presence of ascorbic acid (vitamin C)