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The availability of fresh produce during longer-duration spaceflight missions is being explored as a countermeasure for human biobehavioral health and performance, including how fresh fruits and vegetables can supplement the crew diet with nutrients that are predicted to become deficient throughout the missions. Crop production in the Veggie and Advanced Plant Habitat vegetable production chambers on the International Space Station (ISS) have tested pick-and-eat crops in spaceflight. However, the limited plant cultivation volume of these chambers has restricted the sample sizes and biomass allocated for nutrition assessment. To gain a better understanding of the nutrient composition of pick-and-eat crops, leafy greens and fruiting plants were grown in the EDEN ISS plant cultivation facility near the German Neumayer Station III in Antarctica from March 2021 to January 2022. Target crops for nutrition sampling aligned with cultivars grown in spaceflight, including ‘Outredgeous’ red romaine lettuce, Mizuna mustard, ‘Red Robin’ dwarf cherry tomato, and NuMex ‘Española Improved’ chili pepper. Plants subsamples were taken at harvest; in the event of multiple harvests from the same plants, subsamples were taken at both the first and final harvests. ‘Outredgeous’ and Mizuna were also grown multiple times throughout the year, allowing multiple grow-outs to be analyzed for potential effects across the growing season. Subsamples were first weighed for fresh mass, dried in a dedicated oven at 70°C for 96 h, weighed again, and stored in air-tight containers inside Neumayer Station III. At the beginning of 2022, the samples were shipped to NASA’s Kennedy Space Center, where they are currently being analyzed with ion chromatography for nutrients of interest in the astronaut diet, including calcium, iron, magnesium, and potassium. This study aims to increase our understanding of how the crops that have successfully grown on the ISS can supplement the crew diet, as well as how other factors like plant age and the number of days the facility has been in operation may impact nutrient concentrations. Such findings can improve the crop selection process and how pick-and-eat crops are cultivated during spaceflight missions.