Main conclusion Present findings revealed a great variation in protein, oil, starch, tannin and phytic acid-like biochemical attributes of sorghum genotypes. GT bi-plot analysis was used for assessment of biochemical analysis results. Sorghum is quite resistant to negative environmental conditions. Thus, it has become the basic source of nutrient in majority of developing countries. It is also used as a supporting product against several diseases. In this study, biochemical composition of new 154 sorghum lines, selected from Turkish sorghum landraces through morphological, biochemical and molecular studies, was determined. Present findings revealed based on average of two years that crude protein content of 80 lines (> 9.65%), digestible protein content of 94 lines (> 56.25%), crude oil content of 75 lines (> 3.87%), resistant starch ratio of 32 lines (> 3.93%), starch content of two lines (> 77.07%) and amylose content of 10 lines (> %25.26) were greater than standard cultivar with the greatest values and amylopectin contents of 74 lines (< 47.75%) were lower than the standard cultivar with the lowest values. As the average of years, phytic acid contents and condense tannins contents of the lines, respectively, varied between 0.02 (IS 12850/1)-6.49% (PI 177161 03/2) and 0.09 (IS 12819/1)-5.38% (PI 255738 02/3). With this study, several sorghum lines superior than the standard cultivars were identified. Those lines can be used for direct cultivar registration in short run and can be used as parent materials in further breeding studies to be conducted for various purposes in long run.