Amentoflavone and 4′ monomethoxy amentoflavone were previously isolated from Garcinia livingstonei leaves. These compounds had good activities (MIC 6 and 8 μg/ml) against some nosocomial bacteria. In this study, the activity of these purified compounds were tested against fast-growing non-pathogenic Mycobacterium smegmatis. Amentoflavone was the most active compound, with an MIC of 0.60 ± 0.70 mg/ml. The MIC of 4′ monomethoxy amentoflavone and the positive control isoniazid against Mycobacterium smegmatis were similar 1.40 ± 1.56 and 1.30 ± 1.70 mg/ml respectively. Although, Mycobacterium smegmatis is a non-pathogenic fast growing mycobacterium the activities of these compounds may also be useful in combating infections by pathogenic Mycobacterium spp.