Mycobacterium tuberculosis is the causative agent of tuberculosis, an infectious bacterial disease, which most commonly affects the lungs. In the search for novel active compounds or medicines against tuberculosis, an ethnopharmacological survey combined with a host-pathogen assay has recently highlighted the potency of an aqueous extract of Combretum aculeatum. C. aculeatum is used in traditional medicine and has demonstrated a significant in vitro antimycobacterial activity. Punicalagin, an ellagitannin, was isolated and found to be related to the biological activity of the extract. An analytical method for the evaluation of punicalagin in C. aculeatum was developed by capillary electrophoresis. After method optimization, the quantification of punicalagin was achieved for the evaluation of various plant extracts to determine the content of punicalagin related to the extraction modes and conditions, origin of the plant material, and harvesting period. The developed method demonstrated that the leaves presented the highest punicalagin content compared to the seeds and stems. A decoction of 30 min in boiling water was found to be the best extraction mode of C. aculeatum.