Sorghum is the fifth most important cereal in the world in terms of the cropped area. It is mainly grown for feeding animals and it is also used in the food industry. Sorghum grain is generally a rich source of antioxidants such as polyphenols and carotenoids. For this reason, it is considered as a good source of bioactive food components and it has health-promoting properties. Sorghum is a gluten-free cereal grown in many regions worldwide, primarily in the tropical and subtropical regions. However, new hybrids and forms of sorghum are capable to produce seeds also in temperate climate. The aim of this study was to conduct the influence of weather conditions on bioactive compound content in sorghum grain. The quantitative analysis of selected bioactive compounds, such as phenolic acids, flavonoids, carotenoids, and phytosterols, was carried out. The tested material comprised grain of two varieties: ‘Sweet Susana’ and ‘Sweet Caroline’, which have different color of grain: red and white. The research material was obtained from growing seasons 2016–2018. Quantitative analysis of free phenolic acids, total carotenoids, and total phytosterols was performed by ultra-performance liquid chromatography (UPLC) after prior basic hydrolysis followed by acid. An ultra-efficient liquid chromatograph coupled with an absorption-based detector (UPLC-PDA) was used for these analyses. The results showed the variability of the content of bioactive compounds depending on weather conditions.