Studies relating to the development of a galenical drug with antimicrobial activity from clove buds

The article presents the results of studies on the development of a galenical drug with antimicrobial activity from clove buds. For studies, we used grinded clove buds with particles fraction less than 0.5 mm. As an extractant, we used ethanol with different concentration 26, 43, 56, 72, 82, 97±1% v/v. Methods of extraction: simple maceration and filtration extraction. Methods of analyses: RP HPLC and gravimetry. Standard substances were eugenol and gallic acid. Analytical wave lengths were 370, 282, and 270 nm. For the antibacterial study of extracts, we used the agar well diffusion method. In our research, we used six test-strain microorganisms: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Proteus vulgaris ATCC 4636, Pseudomonas aeruginosa ATCC 27853, Candida albicans ATCC 885/653, and Bacillus subtilis ATCC 6633. Ethanol concentration, which extracts most of the substances was found; minimal eugenol concentration in the extract and tincture for their standardizations was suggested; the level of antimicrobial activity of ethanol-water extracts equivalent to a reference drug was determined. It was found that the major yield among the substances determined is observed with an ethanol concentration of 80±5% v/v. It was found that using filtration extraction method for one hour of the extraction process, we could obtain the liquid extract 1:1 m/v with eugenol concentration of 13.8% m/v and its yield 93%. In two and a half hours of the extraction process, we could obtain the tincture 1:5 m/v with eugenol concentration of 3.0% m/v and its yield 100%. RP HPLC method of analysis determined the presence of the following nine main substances: eugenol, acetyl eugenol, gallic acid, three flavonoids, and three non-identified substances in ethanol-water extracts from clove buds. It was suggested to carry out standardization of the resulting galenical drugs by eugenol. For the extract (1:1 m/v), low limit of eugenol concentration 13.0% m/v was suggested, and for the tincture (1:10 m/v), low limit of eugenol concentration 1.3% m/v was suggested. Antimicrobial activity of ethanol-water solutions of the extract (1:1 m/v) was studied; the lowest eugenol concentration of 0.44% m/v in ethanol-water solutions equivalent in terms of its antimicrobial activity to the reference drug 0.05% chlorhexidine digluconate water solution.

RP HPLC analysis, antimicrobial activity, eugenol, gallic acid, clove buds, tincture, extract, filtration extraction method