Effects of Fractionation and Combinatorial Evaluation of Tamarindus indica Fractions for Antibacterial Activity

Abstract

Six fractions, named TiA – TiF, were obtained by fractionating the crude ethanol extract of the stem bark of Tamarindus indica using column chromatographic techniques. On TLC, fraction TiB showed five bands, TiC three bands, while TiD and TiE showed two bands each. TiC, TiD and TiE were re-eluted with different solvent systems to yield two fractions each, while TiB yielded four. These subfractions were designated B1-B4; C1-C2; D1-D2 and E1-E2, respectively. Tannins, flavonoids and alkaloids, among other components, were detected, albeit in different proportions with respect to fractions and subfractions and were compartmentalized with respect to the solvent systems used. The in vitro antibacterial activity of fractions and subfractions was tested separately and in combinations using the agar well diffusion technique. The susceptibly of test strains (expressed as %) were: 83.3% (TiA and TiB), 75.0% (crude extract and TiC), 66.7% (TiD), 50.0% (TiE) and 16.7% (TiF) when used singly, whereas in combination, the corresponding susceptibilities were 100% (CE), 83.3% (DE), 66.7% (AB, AF, BC, BD, DE and EF), 50% (AC and CD), 33.3% (BE and BF) and 16.7% (AD) against Gram negative bacteria strains and 100% (EF), 80% (DE), 60% (AB, BC and CE), 40% (AC, BD, BF, CF and DF) and 20% (AE, AF, BE and CD) against Gram positive strains. Percentage susceptibility with combinatorial use of re-fractions ranged from 85.7–57.1% and 60–40% against Gram negative and positive strains (TiB subfractions), respectively, 100–85.7% and 40–0% against Gram negative and positive strains (TiC, TiD and TiE sub-fractions).

Keywords

• combinatorial assay
• thin layer chromatography
• phytochemistry
• subfraction
• antibacterial activity
• solvent system