Among natural products, essential oils from aromatic plants have been reported to possess potent anticancer properties. In this work, we aimed to perform the cytotoxic concentration range screening and antiproliferative activity screening of chemically characterized Thymus vulgaris L. essential oil. In vivo bioassay was conducted using the brine shrimp lethality test (BSLT). In vitro evaluation of antiproliferative activity was carried out on three human tumor cell lines: breast adenocarcinoma MCF-7, lung carcinoma H460 and acute lymphoblastic leukemia MOLT-4 using MTT assay. Essential oil components thymol (36.7%), p-cymene (30.0%), γ-terpinene (9.0%) and carvacrol (3.6%) were identified by gas chromatography/mass spectrometry. Analyzed essential oil should be considered as toxic/highly toxic with LC50 60.38 µg/mL in BSLT and moderate/weakly cytotoxic with IC50 range 52.65–228.78 µg/mL in vitro, according to evaluated cytotoxic criteria. Essential oil induced a dose-dependent inhibition of cell proliferation in all tested tumor cell lines and showed different sensitivity. Dose dependent toxicity observed in bioassay as well as the in vitro assay confirmed that brine shrimp lethality test is an adequate method for preliminary toxicity testing of Thymus vulgaris L. essential oil in tumor cell lines.
Five methanolic extracts obtained from different parts of birch, Betula pendula, Roth. (external and internal bark, flowers, leaves and buds), were evaluated for their antibacterial activity in this study. Triterpene compounds, betulin, betulinic acid, oleanolic acid and lupeol, were isolated from the external parts of birch bark using the method of dry column chromatography (DCC) as well as preparative thin layer chromatography (TLC). These compounds were also investigated for their antibacterial activity. Taking into account that decoction is the most commonly used pharmaceutical form of herbal drug, decoctions made from external bark, leaf, flower and bud were investigated for their antibacterial activity. Antibacterial screening, against selected Gram-positive bacteria, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 6538P and Gram-negative bacteria, Escherichia coli ATCC 8739 and Pseudomonas aeruginosa ATCC 9027 was carried out. The methods of diffusion and dilution were used for this investigation according to European Pharmacopoea, 1996. The most prominent antibacterial activity showed oleanolic acid against bacterial species Staphylococcus aureus, expressed as minimal inhibitory concentration (MIC): 1.25% and Bacillus subtilis MIC: 0.625%. Escherichia coli showed resistance on all investigated samples.
Betulae cortex, Betula pendula Roth., Betulaceae, comprise triterpene substances which are confirmed to posses very important pharmacological activities such as anti-inflammatory, anticancer and antiviral. In this study, extraction of triterpene substances from both, inner and external birch bark was carried out and after that qualitative analysis on betulin, betulinic acid, oleanolic acid and lupeol was performed by method of thin layer chromatography. By this separation method, applying system for development benzene-ethyl acetate-formic acid (36:12:5), is gained a good separation of examined triterpene substances from methanol extracts of inner and external birch bark as well as used standards. From obtained row triterpene mixtures, certain triterpene substances are isolated using method of dry column chromatography. To those substances infrared (IR) spectra were recorded and compared with IR spectra of adequate standards. The study encloses all obtained IR spectra and interpretations on the basis of which can be concluded that triterpene substances, betulin, betulin acid and lupeol isolated from external birch bark give identical characteristic signals and absorbance as referent standards. Method of dry column chromatography has resulted as simple, efficient, repeatable and economical for laboratory conditions. Beside this, a sufficient quantity of examined triterpene substances is also obtained for continuation of their further analytical analysis.
Introduction: Present study describes the antimicrobial activity and free radical scavenging capacity (RSC) of essential oil from Mentha longifolia (L.) Huds. Aim of this study to investigate the quality, antimicrobial andantioxidant activity of wild species Mentha longifolia essential oil from Bosnia and Herzegovina.Methods: The chemical profi le of essential oil was evaluated by the means of gas chromatography-mass spectrometry (GC-MS) and thin-layer chromatography (TLC). Antimicrobial activity was tested against 6bacterial strains. RSC was assessed by measuring the scavenging activity of essential oils on 2,2- diphenyl-1-picrylhydrazil (DPPH).Results: The main constituents of the essential oil of M. longifoliae folium were oxygenated monoterpenes,piperitone oxide (63.58%) and 1,8-cineole (12.03%). Essential oil exhibited very strong antibacterial activity.The most important antibacterial activity essential oil was expressed on Gram negative strains: Escherichia coli, Pseudomonas aerginosa and Salmonella enterica. subsp.enterica serotype ABONY. Antioxidant activity was evaluated as a RSC. Investigated essential oil was able to reduce DPPH radicals into the neutral DPPHH form (IC50=10.5 μg/ml) and this activity was dose –dependent.Conclusion: The study revealed signifi cant antimicrobial activity of the investigated essential oil. The examined oil exhibited high RSC, which was found to be in correlation to the content of mainly monoterpeneketones and aldehydes. These results indicate that essential oils could serve as safe antioxidant and antiseptic supplements in pharmaceuticals.
TLC-based strategies were proposed in 1979 (Hostettmann et al.) and 2005 (Friesen & Pauli; GUESS method) to minimize the number of partitioning experiments required for countercurrent separation (CCS) solvent system selection. As semi-empirical approaches, both proposed that the K values defining the sweet spot of optimal CCS corresponded to a matching Rf value range from the silica gel TLC plate developed in the organic phase of a biphasic or a corresponding monophasic solvent system. Despite their simplicity, there has been an absence of theoretical support and a deficiency of reported experimental evidence. The present study explores the theory required to develop correlations between Rf and K. All theoretical models surmise that the optimal Rf value range should be centered at 0.5. In order to validate the feasibility of the concept of matching Rf and K values, 43 natural products and six solvent system families were investigated. Out of 62 correlations, 45 resulted in matched Rf and K values. Based on this study, practical guidelines for the TLC-based prediction strategy are provided. These approaches will equip CCS users with an updated understanding of how to apply the TLC-based solvent system selection strategy to accelerate a targeted selection of CCS conditions.
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