Comparison of the Microbiological Activities of Different Varieties of Hop (Humulus lupulus) Extracts by Thin-Layer Chromatography—Direct Bioautography

JPC – Journal of Planar Chromatography – Modern TLC - Commercially available hop pellets of different origins were extracted by use of ethanol and water, chromatographed on silica...     

Investigation of Different Extraction Methods on the Content and Biological Activity of the Main Components in Coffea arabica L. Extracts

JPC – Journal of Planar Chromatography – Modern TLC - Coffee, due to its common consumption, is one of the main sources of polyphenols in human diet. Coffee species and coffee-related...     

Screening of Antibacterial Compounds in Thymus vulgaris L. Tincture Using Thin-Layer Chromatography—Direct Bioautography and Liquid Chromatography—Tandem Mass Spectrometry Techniques

JPC – Journal of Planar Chromatography – Modern TLC - In this study, thin-layer chromatography—direct bioautography (TLC—DB) was used for guiding the isolation and...     

TLC–direct bioautography as a method for evaluation of antibacterial properties of Thymus vulgaris L. and Salvia officinalis L. essential oils of different origin

ABSTRACT

The dot-blot bioautography was used to evaluate the antibacterial properties of Thymus vulgaris L. and Salvia officinalis L. essential oils (EOs) produced by three different manufacturers. The whole samples were applied at three concentrations on thin-layer chromatography (TLC) plates which were then subjected to bioautography against Bacillus subtilis. The samples of the highest activity were found. Then, they were separated using TLC and once again subjected to bioautography against B. subtilis. As was proved, only the essential oils of T. vulgaris L. possessed strong antibacterial properties for which mostly thymol and carvacrol were responsible. Their contents were calculated using TLC–UV densitometry. The highest contents were found in the essential oils of the highest antibacterial activity revealed in the dot-blot test. It means that a dot-blot test can be used for simple and fast evaluation of antibacterial properties of essential oils.     

Development of a novel direct bioautography-thin-layer chromatography test: optimization of growth conditions for gram-negative bacteria, Escherichia coli

With the aim of developing a TLC-direct bioautography assay using Escherichia coli as test bacteria, various parameters influencing the viability of microorganisms on TLC plates were examined and checked for flumequine standards. The optimal times for preincubation and incubation of bacterial broth were 20 h at 37 degrees C and 2 h at 37 degrees C, respectively. The optimal viscosity of the broth was obtained for 0.05% agarose solution in Mueller-Hinton broth. Various incubation times of the seeded TLC plates were also tested (5 h proved to be optimal). After incubation, the plates were sprayed with 0.2% aqueous [3-(4,5-dimethyldiazol-2-yl)-2,5 diphenyltetrazolium bromide] (MTT) solution and incubated for 0.5 h at 37 degrees C. The precision of the method was evaluated by the repeatability (intraday assay) and intermediate precision (interday assay). The regression coefficients were 0.9977 and 0.9968, respectively, for intraday and interday curves. The calibration curves show good linearity in the range of 0.005-0.50 microg (0.5-50.0 microg/mL). The established LOD of flumequine equaled 0.5 microg/mL, i.e., 5 ng flumequine in the spot. The developed direct bioautography test significantly enhances the sensitivity of the TLC method.