Extraction and Isolation of Compound from Roots of Kniphofia pumila, and Its Antibacterial Activities in Combination with Zinc Chloride

Abstract

Kniphofia pumila has been traditionally used for the treatment of different diseases in Oromiya regional state, western part of Ethiopia. Thus, the main objective of this study was to extract bioactive natural products from the roots of K. pumila and perform in vitro antibacterial activity tests against selected pathogens. Extraction of compound from the roots of K. pumila was carried out by maceration method at room temperature and its antibacterial activity was done using agar disc diffusion method. The crude extracts alone and along with ZnCl₂ were tested against pathogenic bacteria Escherichia coli (ATCC 25722), Klebsiella pneumoniae subsp. pneumoniae (DSM 19613), Staphylococcus aureus (ATCC 25925) and Salmonella Typhimurium (ATCC 13311). Following its promising activity, the acetone crude extract that have shown better antibacterial activity was subjected to column chromatography for isolation of pure compound. Hence, one pure compound (GZ-1) was obtained from acetone crude extract. Meanwhile, the antibacterial activities of the isolated compound alone and in combination with ZnCl₂ were also performed against all aforementioned bacterial strains. Then, isolated compound was characterized by using NMR spectroscopic techniques such as ¹H-NMR, ¹³C-NMR and, 2D NMR, so as to establish its structure as 3’-acetyl-2’,6’-dihydroxy-4-methoxyphenyl-1,8-dihydroxy-3-methylanthraquinone, trivial name knipholone. Finally, GZ-1 was isolated from the roots of K. pumila and it was noted that the antibacterial activities of ZnCl₂ in combination with crude extract as well as with isolated compound against E. coli (ATCC 25722), S. aureus (ATCC 25925) and S. Typhimurium (ATCC 13311) bacterial strains showed remarkable results which were greater than the antibacterial activity of the positive standard drug (gentamycin). To sum up, from the chromatographic isolation of roots acetone extract of K. pumila one pure compound (knipholone) was obtained. Its antibacterial activity was also evaluated in combination with ZnCl₂ salt and alone.