Multidisciplinary Studies of Folk Medicine “Five Thieves’ Oil” (Olejek Pięciu Złodziei) Components

To meet the growing interest in natural antibacterial agents, we evaluated the physicochemical and biological properties of the folk medicine known as “five thieves’ oil” (Polish name: olejek pięciu złodziei). Five thieves’ oil consists of a mixture of five oils: rosemary, lemon, clove, eucalyptus, and cinnamon. In this study, we performed gas chromatography, FTIR, and UV–vis spectroscopic analysis, as well as L-a-b color tests, contact angle determination, and surface tension determination. To verify its antibacterial activity, the metabolic activity and changes in cell membrane permeability of bacteria of the genus Pseudomonas were studied. As a result, it was found that among the constituent oils, the oils of clove and cinnamon were the least volatile and, at the same time, had the strongest antibacterial activity. However, a mix of all the oils also showed comparable activity, which was even more pronounced for the oils after 4 weeks of aging. This effect can be linked to the high content of terpene derivatives such as eugenol and cinnamaldehyde, which can cause changes in bacterial membrane permeability, affecting cell activity and survival. This study is the first to characterize the constituents of the popular folk medicine five thieves’ oil, confirming and explaining its strong antibacterial activity, thus constituting a significant contribution to contemporary health education.     

Chemical Composition and Antibacterial Activity of Liquid and Volatile Phase of Essential Oils against Planktonic and Biofilm-Forming Cells of Pseudomonas aeruginosa

Pseudomonas aeruginosa is an opportunistic pathogen causing life-threatening, hard-to-heal infections associated with the presence of a biofilm. Essential oils (EOs) are promising agents to combat pseudomonal infections because of the alleged antimicrobial activity of their volatile fractions and liquid forms. Therefore, the purpose of this paper was to evaluate the antibacterial efficacy of both volatile and liquid phases of seven EOs (thyme, tea tree, basil, rosemary, eucalyptus, menthol mint, lavender) against P. aeruginosa biofilm and planktonic cells with the use of a broad spectrum of analytical in vitro methods. According to the study results, the antibacterial activity of EOs in their liquid forms varied from that of the volatile fractions. Overall, liquid and volatile forms of rosemary EO and tea tree EO displayed significant antibiofilm effectiveness. The outcomes indicate that these particular EOs possess the potential to be used in the therapy of P. aeruginosa infections.     

Correlation between chemical composition and antibacterial activity of essential oils from fifteen Eucalyptus species growing in the Korbous and Jbel Abderrahman arboreta (North East Tunisia)

The essential oils of fifteen Eucalyptus species harvested from the Jbel Abderrahman and Korbous arboreta (North East Tunisia) were screened for their antibacterial activities by the agar disc diffusion method. Eighteen major components as identified by GC/FID and GC/MS were selected for a study of the chemical and biological activity variability. The main one was 1,8-cineole, followed by spathulenol, trans-pinocarveol, α-pinene, p-cymene, globulol, cryptone, β-phellandrene, viridiflorol, borneol, limonene and isospathulenol. The chemical principal component analysis identified five species groups and subgroups, where each group constituted a chemotype, however that of the values of zone diameter of the inhibition (zdi) identified six groups of Eucalyptus oils, characterized by their antibacterial inhibition ability. The strongest activity was shown by E. platypus oil against Enterococcus faecalis and by E. lamannii oil against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli. A correlation between the levels of some major components and the antibacterial activities was observed.     

Bioassay-Guided Isolation and Identification of Antimicrobial Compounds from Thyme Essential Oil by Means of Overpressured Layer Chromatography,Bioautography and GC–MS

A simple method is described for efficient isolation of compounds having an antibacterial effect. Two thyme (Thymus vulgaris) essential oils, obtained from the market, were chosen as prospective materials likely to feature several bioactive components when examined by thin layer chromatography coupled with direct bioautography as a screening method. The newly developed infusion overpressured layer chromatographic separation method coupled with direct bioautography assured that only the active components were isolated by means of overrun overpressured layer chromatography (OPLC) with on-line detection and fractionation. Each of the 5 collected fractions represented one of the five antimicrobial essential oil components designated at the screening. The purity and the activity of the fractions were confirmed with chromatography coupled with various detection methods (UV, vanillin–sulphuric acid reagent, direct bioautography). The antibacterial components were identified with GC–MS as thymol, carvacrol, (−)-linalool, diethyl-phthalate, and α-terpineol. The oil component diethyl-phthalate is an artificial compound, used as a plasticizer or detergent base in the industry. Our results support that exploiting its flexibility and the possible hyphenations, OPLC is especially attractive for isolation of antimicrobial components from various matrixes.

     

Essential Oils as In Vitro Ruminal Fermentation Manipulators to Mitigate Methane Emission by Beef Cattle Grazing Tropical Grasses

There is increasing pressure to identify natural feed additives to mitigate methane emissions from livestock systems. Our objective was to investigate the effects of essential oils (EO) extracts star anise (Illicium verum), citronella (Cymbopogon winterianus), clove bud (Eugenia caryophyllus), staigeriana eucalyptus (Eucalyptus staigeriana), globulus eucalyptus (Eucalyptus globulus), ginger (Zingiber officinale), ho wood (Cinnamomum camphora), melaleuca (Melaleuca alternifolia), oregano (Origanum vulgare) and white thyme (Thymus vulgaris) on in vitro methane emissions from four rumen-cannulated Nellore cattle grazing a tropical grass pasture as inoculum donors. The semi-automated gas production technique was used to assess total gas production, dry matter degradability, partitioning factor, ammoniacal nitrogen, short-chain fatty acids and methane production. All essential oils were tested in four doses (0, 50, 250 and 500 mg/L) in a randomized block design, arranged with four blocks, 10 treatments, four doses and two replicates. Within our study, oregano and white Thyme EO reduced net methane production at 250 mg/L, without affecting substrate degradation. Essential oils from oregano and white thyme have the potential to modify ruminal fermentation and suppress rumen methanogenesis without negative effects on feed digestibility, indicating promise as alternatives to ionophores for methane reduction in beef cattle.     

ATR-FTIR-MIR Spectrometry and Pattern Recognition of Bioactive Volatiles in Oily versus Microencapsulated Food Supplements: Authenticity,Quality, and Stability

Fourier transform infrared spectroscopy on the middle infrared region (ATR-FTIR-MIR) proved to be a convenient and reliable technique to evaluate foods’ quality and authenticity. Plants’ essential oils are bioactive mixtures used as such or in different oily or microencapsulated formulations, beneficial to human health. Six essential oils (thyme, oregano, juniperus, tea tree, clove, and cinnamon) were introduced in three oily formulations (Biomicin, Biomicin Forte, and Biomicin urinary) and these formulations were microencapsulated on fructose and maltodextrin matrices. To study their stability, the microencapsulated powders were kept under light irradiation for 14 days at 25 °C or introduced in biopolymer capsules. All variants were analysed by ATR-FTIR-MIR, recording wavenumbers and peak intensities (3600–650 cm⁻¹). The data were processed by Unscrambler and Metaboanalyst software, with specific algorithms (PCA, PLSDA, heatmaps, and random forest analysis). The results demonstrated that ATR-FTIR-MIR can be successfully applied for fingerprinting and finding essential oil biomarkers as well as to recognize this pattern in final microencapsulated food supplements. This study offers an improved ATR-FTIR-MIR procedure coupled with an adequate chemometric analysis and accurate data interpretation, to be applied for the evaluation of authenticity, quality, traceability, and stability during storage of essential oils incorporated in different matrices.     

Characterization and Comparison of Tea Tree and Lavender Oils by Using Comprehensive Gas Chromatography

The essential oils from French lavender (Lavandula angustifolia) and tea tree (Melaleuca alternifolia) were separated by the two‐dimensional GC technique of comprehensive gas chromatography. A coupled column combination of non‐polar (5% phenyl equivalent) and polyethylene glycol phase columns was used to provide the desired resolution performance. By using a range of known standards, some of the peaks in lavender oil can be assigned. Some of these also occur in tea tree oil; however, from our knowledge of the major constituents in this oil and their relative retention behaviour, most of the major peaks may be tentatively assigned within the 2‐dimensional separation space. There appear to be elution patterns within the 2‐D space which should be useful in correlating retention with chemical and structural properties of the components, although this will require further evaluation. A range of coeluting peaks, which may not be so readily separated by using a single column capillary GC analysis, are resolved in the experiment described.     

Chemical composition, and antioxidant and antimicrobial activities of essential Oil of Phyllostachys heterocycla cv. Pubescens varieties from China

The essential oils of Phyllostachys heterocycla cv. Pubescens, Phyllostachys heterocycla cv. Gracilis, Phyllostachys heterocycla cv. Heterocycla and Phyllostachys kwangsiensis leaves were obtained by steam distillation. Their chemical components were separated and identified by gas chromatography/mass spectrometry (GC/MS). Meanwhile, the effect of scavenging free radicals of essential oil was assayed by using the DPPH·method with Trolox? as control to evaluate their antioxidant capacities. Gram-positive (Staphyloccocus aureus) and Gram-negative (Escherichia coli) were selected as the indicator microorganisms to evaluate the antimicrobial activity. Antimicrobial properties were estimated by the agar diffusion method. The results show that 63 components were separated and identified by GC/MS from these varieties of bamboo leaves. cis-3-Hexenol, whose content in cv. Pubescens, Gracilis, Heterocycla and Ph. kwangsiensis was 27.11%, 24.62%, 30.51% and 34.65%, respectively, was the main constituent. The relative content of alcohol compounds in these varieties of essential oils ranged from 39.8% to 46.64%. All of the bamboo leaf essential oils possessed certain antioxidant capacity; the corresponding IC?? values were 3.1622, 4.9353, 4.2473, and 5.4746 μL/mL, respectively. Essential oils of all tested bamboo spp. were active against Staphylococcus epidermidis and E. coli, showing a positive correlation with the essential oil concentration of 50.42-300 μL/mL. The results indicated there were no significant differences among three varieties and the related species with respect to their antioxidant and antimicrobial activities. This paper provides evidence for studying the essential composition from different varieties of bamboo leaves.     

Investigation of the chemical composition–antibacterial activity relationship of essential oils by chemometric methods

The antibacterial effects of Thymus vulgaris (Lamiaceae), Lavandula angustifolia (Lamiaceae), and Calamintha nepeta (Lamiaceae) Savi subsp. nepeta var. subisodonda (Borb.) Hayek essential oils on five different bacteria were estimated. Laboratory control strain and clinical isolates from different pathogenic media were researched by broth microdilution method, with an emphasis on a chemical composition–antibacterial activity relationship. The main constituents of thyme oil were thymol (59.95%) and p-cymene (18.34%). Linalool acetate (38.23%) and β-linalool (35.01%) were main compounds in lavender oil. C. nepeta essential oil was characterized by a high percentage of piperitone oxide (59.07%) and limonene (9.05%). Essential oils have been found to have antimicrobial activity against all tested microorganisms. Classification and comparison of essential oils on the basis of their chemical composition and antibacterial activity were made by utilization of appropriate chemometric methods. The chemical principal component analysis (PCA) and hierachical cluster analysis (HCA) separated essential oils into two groups and two sub-groups. Thyme essential oil forms separate chemical HCA group and exhibits highest antibacterial activity, similar to tetracycline. Essential oils of lavender and C. nepeta in the same chemical HCA group were classified in different groups, within antibacterial PCA and HCA analyses. Lavender oil exhibits higher antibacterial ability in comparison with C. nepeta essential oil, probably based on the concept of synergistic activity of essential oil components.     

Influence of harvesting time on composition of the essential oil of Thymus capitatus (L.) Hoffmanns. & Link. growing wild in northern Sicily and its activity on microorganisms affecting historical art crafts

One of the main factors affecting historical art crafts material is the biodeterioration performed by bacteria and fungi, in archives, museums or private collections. Several microorganisms cause degradation to the natural organic material such as fibers, woods, and dyes as well as to stone objects. These alterations produce deterioration of physical, chemical, mechanical and esthetic properties. Consequently, in this publication, we report the high antibacterial and antifungal activities of wild thyme essential oil, oil that can be used as an alternative natural tool in the fight against microorganisms affecting historical art crafts.Essential oil of the Thymus capitatus growing wild in northern Sicily has been extracted by hydrodistillation from aerial parts collected at different growth times. The constituents of the essential oil have been characterized by gas chromatography (GC) and GC–mass spectrometry.Fifty-five compounds have been identified. Essential oils were characterized by a high content of carvacrol (81.2–14.2%), γ-terpinene (34.4–2.6%) and p-cymene (22.8–5.0%) of the total oil content. Essential oil yield and composition vary throughout the vegetation time of the plant. The best time to harvest this species of thyme, for phenol content, is during or immediately before the full bloom. The related oils (Tc2 and Tc3) showed a good antibacterial activity against Bacillus subtilis and Staphylococcus epidermidis and excellent antifungal properties against Fusarium oxysporum and Aspergillus niger.     

Application of high-performance liquid chromatography diode array detection and mass spectrometry to the analysis of characteristic compounds in various essential oils

A high-performance liquid chromatography (HPLC) method was established using an analytical reversed-phase column and gradient elution to achieve chromatographic separation of typical compounds in essential oils. For detection, a diode array detector monitoring different wavelengths simultaneously as well as a mass spectrometer (MS) were used. Atmospheric pressure chemical ionization operating in the positive mode turned out to be a suitable tool to detect volatiles of different chemical classes and to identify them in essential oil matrices. Characteristic fingerprints of eucalyptus, lavender, may chang, pine, rosemary, thyme, and turpentine essential oils monitored at a representative wavelength (220 nm) demonstrated the suitability of HPLC in essential oil analysis. Additional monitoring wavelengths (210, 250, and 280 nm) provided useful information about the identity of the specific component and opened the possibility to differentiate presumably coeluting compounds by means of their distinct absorption behavior. Finally, peak assignment in seven essential oils was performed on the basis of characteristic retention times and UV and MS data of a broad set of reference volatiles.     

Identification of potential antioxidant compounds in the essential oil of thyme by gas chromatography with mass spectrometry and multivariate calibration techniques

Thyme species are used in traditional medicine throughout the world and are known for their antiseptic, antispasmodic, and antitussive properties. Also, antioxidant activity is one of the interesting properties of thyme essential oil. In this research, we aim to identify peaks potentially responsible for the antioxidant activity of thyme oil from chromatographic fingerprints. Therefore, the chemical compositions of hydrodistilled essential oil of thyme species from different regions were analyzed by gas chromatography with mass spectrometry and antioxidant activities of essential oils were measured by a 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging test. Several linear multivariate calibration techniques with different preprocessing methods were applied to the chromatograms of thyme essential oils to indicate the peaks responsible for the antioxidant activity. These techniques were applied on data both before and after alignment of chromatograms with correlation optimized warping. In this study, orthogonal projection to latent structures model was found to be a good technique to indicate the potential antioxidant active compounds in the thyme oil due to its simplicity and repeatability.     

Polyol-induced partitioning of essential oils in water/acetonitrile solvent mixtures

Polyol-induced extraction (PIE) is applied to the extraction of essential oils, using glycerol as a mass separating agent. In 1:1 acetonitrile (ACN)/water solvent mixtures, two immiscible phases can be generated. PIE as an alternative extraction technique was assessed by the extraction of the main flavor and fragrance compounds that comprise six essential oils. In the extraction of eugenol (4-allyl-2-methoxyphenol) from clove buds, the partition coefficients were determined and the % recovery and thermodynamic data in the temperature range of ?20 to 20°C were calculated. The main components present in each essential oil extract were identified through gas chromatography/mass spectrometry (GC/MS) and the compositional profile was compared to traditional extraction techniques. The optimized extraction conditions (?10°C, 1:1 ACN/water (v/v), 20% glycerol) for eugenol at ?10°C give a partition coefficient (K_PC) of 87 and an extraction efficiency of 97% in the acetonitrile-rich phase. The eugenol migration to the organic phase is a spontaneous process (ΔG°?=??9.3?kJ/mol) and an endothermic process (ΔH°?=?9.2?kJ/mol) with entropy being the driving force behind the reaction (ΔS°?=?70?J/K, TΔS°?=?18.4?kJ). The technique was applied to five other essential oils (cinnamon bark, caraway seed, spearmint leaf, peppermint leaf, and anise seed oils) with similar results.     

Commercial Origanum compactum Benth. and Cinnamomum zeylanicum Blume essential oils against natural mycoflora in Valencia rice

Chemical composition of commercial Origanum compactum and Cinnamomum zeylanicum essential oils and the antifungal activity against pathogenic fungi isolated from Mediterranean rice grains have been investigated. Sixty-one compounds accounting for more than 99.5% of the total essential oil were identified by using gas chromatography (GC) and gas chromatography–mass spectrometry (GC–MS). Carvacrol (43.26%), thymol (21.64%) and their biogenetic precursors p-cymene (13.95%) and γ-terpinene (11.28%) were the main compounds in oregano essential oil, while the phenylpropanoids, eugenol (62.75%), eugenol acetate (16.36%) and (E)-cinnamyl acetate (6.65%) were found in cinnamon essential oil. Both essential oils at 300 μg/mL showed antifungal activity against all tested strains. O. compactum essential oil showed the best antifungal activity towards Fusarium species and Bipolaris oryzae with a total inhibition of the mycelial growth. In inoculated rice grains at lower doses (100 and 200 μg/mL) significantly reduced the fungal infection, so O. compactum essential oil could be used as ecofriendly preservative for field and stored Valencia rice.     

Biological activities,phenolic profiles and essential oil components of Tanacetum cilicicum (BOISS.) GRIERSON

Tanacetum species are consumed as tea in Turkey. We comparatively evaluated the phytochemical potentials and antioxidant activities of the essential oil and methanolic extracts of Tanacetum cilicicum. The chemical constitutes of T. cilicicum essential oil and antioxidant activity of this species was analysed by gas chromatography–mass spectrometry (GC–MS) method. Bicyclo (3,1,1) hept-2-en-4-ol (21.92%), camphor (15.56%) and 1,8-cineole (13.45%) which are oxygenated monoterpenes were found as the major constituents. Phenolic acids and flavonoids were quantificated by HPLC–UV. Catechin was found as the main component. The essential oils and methanolic extracts were evaluated by antioxidant activities. The leaves exhibited significant metal chelation activity with a value of 20.75 ± 4.63 μg/mL.     

Application of Comprehensive Two‐Dimensional Gas Chromatography (GC×GC) to the Qualitative Analysis of Essential Oils

This paper investigates the separation of moderately complex samples by comprehensive two‐dimensional gas chromatography (GC×GC). The analysis of peppermint (Mentha piperita) and spearmint (Mentha spicata) essential oil components, including acetates, alcohols, furans, ketones, sesquiterpenes, and terpenes, was achieved by one‐dimensional gas chromatography with quadrupole mass spectrometry detection (GC/MSD) and GC×GC with flame ionization detection. Peppermint essential oil was found to contain 89 identifiable peaks by GC×GC compared to 30 peaks in the GC/MSD chromatogram. Likewise, 68 peaks were found in the GC×GC chromatogram of spearmint (compared to 28 in GC/MSD). Plots of the first dimension versus second dimension retention times provided a fingerprint of the two essential oils, which revealed 52 similar compounds between the two essential oils as opposed to 18 matches by 1D GC.     

Antibacterial effects of the essential oils of commonly consumed medicinal herbs using an in vitro model

The chemical composition and antibacterial activity of essential oils from 10 commonly consumed herbs: Citrus aurantium, C. limon, Lavandula angustifolia, Matricaria chamomilla, Mentha piperita, M. spicata, Ocimum basilicum, Origanum vulgare, Thymus vulgaris and Salvia officinalis have been determined. The antibacterial activity of these oils and their main components; i.e. camphor, carvacrol, 1,8-cineole, linalool, linalyl acetate, limonene, menthol, a-pinene, b-pinene, and thymol were assayed against the human pathogenic bacteria Bacillus subtilis, Enterobacter cloacae, Escherichia coli O157:H7, Micrococcus flavus, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella enteritidis, S. epidermidis, S. typhimurium, and Staphylococcus aureus. The highest and broadest activity was shown by O. vulgare oil. Carvacrol had the highest antibacterial activity among the tested components.     

Potentiality of independent component regression in assessment of the peaks responsible for antimicrobial activity of <Emphasis Type="Italic">Satureja hortensis</Emphasis> L. and <Emphasis Type="Italic">Oliveria decumbens</Emphasis> Vent. using GC–MS

Satureja hortensis L. and Oliveria decumbens Vent. are used in traditional medicine in the world due to its antifungal, antibacterial and antioxidant properties. In this study, the antimicrobial activities in Satureja hortensis L. and Oliveria decumbens Vent. essential oils against three microbial strains were evaluated by two different methods, including: disk diffusion method and micro-broth dilution assay [with minimal inhibitory concentration (MIC) between 0.025 and 0.500 µL/mL, and minimal lethal concentration (MLC) between 0.050 and 1.000 µL/mL] and qualitative analysis was performed by gas chromatography–mass spectrometry technique (GC–MS). The peaks potentially responsible for the antimicrobial activity in essential oils samples were indicated by some linear multivariate calibration techniques with different preprocessing methods. From the studied techniques, independent component regression (ICR) was preferred to exhibit the potential antimicrobial active compounds in Satureja hortensis L. and Oliveria decumbens Vent. essential oils because of its high repeatability, simplicity, and interpretability of the regression coefficients. Independent components (ICs) can give more chemical explanation than principal components (PCs), because independence is a high-order statistic that is a much stronger condition than orthogonality.     

Chemical composition,phytotoxic and antifungal properties of Ruta chalepensis L. essential oils

The chemical composition, and phytotoxic and antifungal activities of the essential oils isolated by using hydrodistillation from the aerial parts of Tunisian rue were evaluated. Significant variations were observed among harvest periods. The analysis of the chemical composition by gas chromatography/mass spectrometry showed that 2-undecanone (33.4–49.8%), 2-heptanol acetate (13.5–15.4%) and α-pinene (9.8–11.9%) were the main components. The antifungal ability of rue essential oils was tested by using disc agar diffusion against ten plant pathogenic fungi. A high antifungal activity was observed for the essential oil isolated at flowering developmental phase. Furthermore, rue essential oils showed high level of herbicidal activity against several weeds.     

Analysis of gin essential oil mixtures by multidimensional and one-dimensional gas chromatography/mass spectrometry with spectral deconvolution

The composition of essential oils and their mixtures used to formulate gin is usually too complex to separate all sample components by standard capillary gas chromatography (GC). In particular, minor constituents that possess important organoleptic properties can be masked by co-elution with major sample components. A solution is provided that combines gas chromatography/mass spectrometry (GC/MS) with "interactive" spectral deconvolution software. Sequential two-dimensional (2D) GC/MS is used to produce a target compound library, with orthogonal GC-GC providing the separation power required to obtain peak retention times and the corresponding mass spectra needed for the deconvolution database. The combination of these two techniques, mass spectral deconvolution and automated sequential 2D-GC/MS, offers a very effective synergy for both identifying key constituents that determine the perception of flavor and aroma and the quality control needed to analyze mixtures of complex essential oils.