Direct access to good-quality drinking water in Uzbekistan is becoming a major issue. This challenge is exacerbated by growing demands, driven by population growth, industrial development, and desert land cultivation. In this regard, there is a significant need for a thorough analysis of the existing water resources. This study aims to identify the chemical composition of springs’ water in the Kashkadarya region of Uzbekistan and to assess their suitability as drinking water sources. To analyze the water quality, three springs were selected, which are the most used springs by the population of this region, “Hazrat Bashir”, “Boshmanbulak”, “Chillabulak”, and the artesian waters of the city of Shakhrisabz were also included in the study. The quality of the spring water was evaluated based on the primary water quality indicators, including salt composition, heavy metals, and some essential trace elements such as iron (Fe), chromium (Cr), and selenium (Se) in the selected samples. According to the results, the spring water in Chillabulak was found to be relatively harder than the other samples, with magnesium and calcium concentrations of 28 and 390 mg/l, respectively. However, the study also revealed that the water from the other three examined samples meets sanitary norms and water quality standards, confirming their suitability for human consumption. The results can be used to develop water quality management strategies and environmental protection, as well as to inform the population about the ecological safety of regional aquatic reserves.