The current use and distribution of agricultural water resources is highly prone to effects of global climate change due to shifting precipitation patterns. The production of vegetable crops in open field cultivation often requires demanding water applications, being impaired in regions where climate change will increasingly evoke water scarcity. To date, increasingly occurring precipitation-free periods are already leading to moderate water deficits during plant growth, e.g., in southern Europe. Among all vegetable crops, leafy vegetables such as spinach (Spinacia oleracea
L.) are particularly vulnerable to limited water supply, because leaf expansion is highly dependent on water availability. Besides biomass production, water limitation might also affect the valuable nutritional composition of the produce. Therefore, we investigated the impact of moderately reduced water supply on the chemical composition of spinach, cultivated in the open field in three consecutive years. Two different water supply treatments, full and reduced irrigation, were used in a randomized block design consisting of three sets of six plots each. In the reduced water supply treatment, the total amount of supplied water, including both irrigation and natural precipitation, amounted to 90%, 94% and 96% in 2015, 2016 and 2017, respectively, of the full, optimal water supply treatment. Spinach grown under limited water supply showed significantly higher fresh biomass-based contents of polyols (e.g., inositol, glycerol), ascorbic acid, potassium, nitrogen, phosphorous, zinc and manganese, as well as total flavonoids and carotenoids. Increased dry biomass-based levels were found for total inositol, zinc and manganese, as well as decreased levels for malic acid, fumaric acid, phosphate and chloride. Furthermore, we report a high seasonal variation of several minor phytochemicals, such as single flavonoids. Spinacetin derivatives, spinatoside-glucoside as well as a rather unusual hexuronylated methylenedioxy flavonoid showed highest amounts when grown under relatively low irradiation in autumn. Levels of patuletin derivatives tended to increase under high irradiation conditions during spring. In summary, the chemical composition of spinach was shown to be highly sensitive to moderately reduced water supply and seasonal variation, but the overall nutritional quality of fresh marketable spinach was only marginally affected when considering health-related constituents such as minerals, trace elements, flavonoids and carotenoids.