Abstract: The present study checks the variation in tannin`s contents of Acacia nilotica in the Sudan according to subspecies, tissues, pod`s developmental stage and site of growth. Samples were collected from six different sites and Hyde powder method was used for the determination of the percentages of total tannin`s contents, while thin layer chromatography test (a polyamide paper and methanol and water 40:60 solvent) was carried for qualitative comparison. The study revealed that tannin`s contents varied significantly among the three subspecies in the Sudan, especially at Elfasher, Elgazaire and Elfaw sites, where subspecies nilotica being best followed by tomentosa and then adansonii. Pods were generally significantly better than bark in their tannin`s contents and the premature pods were always better than the mature ones. The highest amount (54%) was recorded form the premature pods of subspecies nilotica at Elfaw site. Qualitatively, there were variations in the fractionation ratio (Rf) values of the tannin compounds among the three subspecies according to tissue and pod`s developmental stage. The values were increasing from initial to the premature stage of pods, while the sites were generally in the order: Shambat; Elgazaire, Elfaw and Nayala and then Elfasher, for the compounds of mature pods.