Radical scavenging and antibacterial properties of the extracts from different Thymus pulegioides L. chemotypes

Radical scavenging and antibacterial properties of large thyme extracts isolated from five chemotypes of Thymus pulegioides L. growing wild in Lithuania were studied. The chemotypes were defined according to the main essential oil components: linalool (L), geranial/geraniol/neral (G/G/N), thymol (T), carvacrol/γ-terpinene/p-cymene (C/γT/pC) and thymol/carvacrol/γ-terpinene/p-cymene (T/C/γT/pC). The contents of phenolic compounds, flavonoids and flavonols were determined. It was found that the extracts of phenolic chemotypes containing remarkable concentrations of thymol and/or carvacrol were stronger DPPH and ABTS free radical scavengers in the model systems. The antibacterial activity of the extracts depended on the plant chemotype, extract preparation, solvent used and finally the sensitivity of bacteria. Bacillus cereus, Micrococcus luteus, Staphylococcus epidermidis and Staphylococcus aureus were the most sensitive to the all extracts applied, whereas Escherichia coli, Salmonella typhimurium and Enterobacter aerogenes remained resistant.     

Chemical composition and biological activities of Algerian Thymus oils

The compositions of essential oils isolated from nine samples of three Thymus species (Thymus algeriensis, Thymus pallescens and Thymus dréatensis) were analysed by GC and GC–MS, and a total of 114 components were identified. T. pallescens collected from various regions showed a great similarity in their compositions and were characterised by carvacrol (44.4–57.7%), p-cymene (10.3–17.3%) and γ-terpinene (10.8–14.2%) as the major components for four samples; only one sample was thymol-rich (49.3%) with a small amount of carvacrol (9.0%). On the other hand, T. algeriensis showed a chemical polymorphism, even for samples from the same location, and two new chemotypes for this species were proposed. Oxygen-containing monoterpenes were the predominant class (76.3%) in T. dreatensis oil, with linalool (30.4%), thymol (20.2%) and geraniol (19.6%) as the principal constituents. The oils were screened for their possible antioxidant activities by four complementary assays, namely DPPH free radical scavenging, hydroxyl radical scavenging, inhibition of lipid peroxidation and reducing power. The two new chemotypes of T. algeriensis exhibited strong hydroxyl radical scavenging (IC₅₀ = 2.2–3.3 μg/ml), but were not or only slightly active against the other radicals and exhibited a weak reducing power. Despite their chemical similarity, T. pallescens oils sometimes produced significant differences in their antioxidant activities. The essential oils were also screened for their antimicrobial activity against five bacteria (three Gram-positive and two Gram-negative) and one yeast (Candida albicans). The tested essential oils showed antimicrobial activity against the microorganisms used, in particular against two important pathogens, C. albicans and Helicobacter pylori.     

Phytochemical composition of the essential oils from three Apiaceae species and their antibacterial effects on food-borne pathogens

Essential oils (EOs) of three Apiaceae species, including Bunium persicum, Cuminum cyminum and Carum copticum, extracted by hydrodistillation, were analyzed by gas chromatography (GC) and GC/mass spectrometry. The main components of EOs of B. persicum were γ-terpinene (44.2%), cuminaldehyde (16.9%), γ-terpinen-7-al (10.5%), and ρ-cymene (8%). The major constituents of Cu. cyminum were cuminaldehyde (30.2%), ρ-cymene (14.1%), γ-terpinene (12.8%), and safranal (9.4%), while those of Ca. copticum were thymol (48.4%), ρ-cymene (21.8%) and γ-terpinene (21.3%). The antibacterial effects of the EOs were assessed on several food-borne pathogens, namely Staphylococcus aureus, Bacillus cereus, Escherichia coli O157:H7, Salmonella enteritidis, and Listeria monocytogenes. The ranges of minimum inhibitory concentration (MIC) of the oils were 0.03–0.5, 0.18–3.0, and 0.37–3.0 mg/ml, respectively, for Ca. copticum, B. persicum and Cu. cyminum. Moreover, the combination of B. persicum and Cu. cyminum EOs confirmed synergistic and additive activities against the pathogens.     

Antibacterial activity of some Lamiaceae essential oils using resazurin as an indicator of cell growth

The aim of this work was to investigate the antibacterial activity of six Lamiaceae essential oils, against pathogenic and food spoilage bacteria. The chemical profiles of essential oils were evaluated by the means of GC and GC-MS. The major constituents of the oils were 1,8-cineol (29.2%), camphor (17.2%), α-pinene (11.5%) in Rosmarinus officinalis, citronellal (20.5%), β- geraniol (17.0%), β-citronellol (11.5%) in Melissa officinalis, 1,8-cineol (27.4%), α-thujone (16.3%), β-thujone (11.2%), borneol (10.4%), camphor (7.98%) in Salvia officinalis, linalool (25.1%), linalyl acetate (22.5%) in Lavandula angustifolia, thymol (52.4%), p-cymene (17.9%) in Thymus vulgaris and Patcholene alcohol (22.7%), α-bulnesene (17.1%), α-guaine (13.8%) in Pogostemon cablin. On quantitative basis, the amounts of 1,8-cineol, citronellal, 1,8-cineol, linalool, thymol and patchouli alcohol, calculated using calibrated curve with pure standard compounds, in the respective essential oils were found to be 28.4, 19.0, 26.7, 23.3, 51.1 and 21.1 g/100 g of oil, respectively. The modified resazurin microtitre-plate assay was used to evaluate the antibacterial activity of the essential oils and their principal components. All the essential oils analyzed presented inhibitory effects on most of the strains tested. Thymus vulgaris essential oil showed the highest inhibition. It was concluded that modified resazurin assay could be effectively used for reliable assessment of antibacterial activity of the tested essential oils against several Gram positive and negative bacterial taxa. The present results also demonstrated that Lamiaceae essential oils exhibiting higher antibacterial activity were generally rich in oxygenated monoterpens.     

Chemical composition and antioxidant and antimicrobial activity of two Satureja essential oils

Hydro-distilled volatile oils from the aerial parts of Satureja montana L., and Satureja subspicata Bartl. ex Vis., growing wild in Bosnia and Herzegovina, were analyzed by GC/MS. More than one hundred compounds were identified in both plant oils, representing 92.4–98.1% of the total oil. The major constituents of essential oils obtained from the plant material of S. montana, collected from two different localities, were thymol (31.7%), and geraniol (22.3%), respectively. The most abundant compounds in essential oils of S. subspicata, collected at two different stages of development, were thymol (28.6%), and spathulenol (37.6%), respectively. The screening of antimicrobial activity of essential oil samples was individually evaluated against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Bacillus subtilis using a paper disc diffusion method. All tested microorganisms were inhibited by essential oil samples. Antioxidant activity was tested using the DPPH radical-scavenging method. All samples showed activity comparable to thymol, which was used as a positive probe.     

Phytochemical composition and antimicrobial activities of the essential oils from Satureja subspicata Vis. growing in Croatia

The present study describes the phytochemical profile and antimicrobial activity of Satureja subspicata Vis. essential oils, collected in Dalmatia (Croatia). Three samples of essential oils were obtained from the aerial parts of the plant by hydrodistillation and analyzed by GC–MS. From the 24 compounds representing 97.47% of the oils, carvacrol (16.76%), α-pinene (13.58), p-cymene (10.76%), γ-terpinene (9.54%) and thymol methyl ether (8.83%) appear as the main components. The oils also contained smaller percentages of myrcene, linalool, β-caryophyllene, limonene, geranyl acetate, 1-Octen-3-ol, nerol, thymol and borneol. Furthermore, antimicrobial activity of the oil was evaluated using agar diffusion and broth microdilution methods. The antimicrobial test results showed that the oils had a great potential antimicrobial activity against all 13 bacteria and 9 fungal strains. Gram-positive bacteria are more sensitive to the investigated oil, with a range of 0.09 to 6.25 μl/ml than Gram-negative bacteria in the range which is significantly higher from 1.56 to 25.00 μl/ml. Results presented here may suggest that the essential oil of S. subspicata possesses antimicrobial properties, and is therefore a potential source of antimicrobial ingredients for the food and pharmaceutical industry.     

Selection for tolerance to volatile monoterpenoids in Sitophilus oryzae (L.), Rhyzopertha dominica (F.) and Cryptolestes pusillus (Schönherr)

Selection for tolerance to 8 monoterpenoids (linalool, camphor, γ-terpinene, S-carvone, geraniol, estragole, E-anethole, and fenchone) in three pests of stored rice, Sitophilus oryzae, Rhyzopertha dominica and Cryptolestes pusillus, as well as the metabolic mechanisms implicated, was studied. Strains were selected for tolerance towards each monoterpenoid from susceptible insect populations for each of seven generations.After selection, tolerance was observed towards linalool, S-carvone and estragole in S. oryzae and towards camphor in C. pusillus. Populations of R. dominica were not observed to develop tolerance to monoterpenoids.The activity of three enzymatic systems detoxifying these monoterpenoids, microsomal P-450 monooxygenase, esterase, and glutathione S-transferase (GST), were analyzed to study the metabolic mechanisms involved. Esterases could be involved in S. oryzae tolerance whereas P-450 monooxygenases could be involved for most monoterpenoids in selected populations of R. dominica and C. pusillus.     

Composition and antimicrobial activity of the essential oils of three Satureja species growing in Tanzania

Hydro-distilled volatile oils from the aerial parts of three Satureja species were investigated, mainly by a combination of GC and GC/MS. One hundred and thirteen compounds were identified, representing 82.9–92.0% of the total oil. Among the identified components, spathulenol, cis-piperitone oxide, α-bisabolol oxide-B, terpinen-4-ol, linalool, bornyl acetate, β-bourbonene, isomenthone, thymol, neoisomenthol and menthone were found as the main components. Furthermore, the essential oils were investigated for their antimicrobial activity, by the agar dilution technique. The antimicrobial test results showed that the oils had a high antimicrobial activity against two Gram-positive and four Gram-negative bacteria, two oral pathogens and three pathogenic fungi. Gram-positive bacteria were more sensitive to the investigated oils than were Gram-negative bacteria. These results could support the suggestion of Satureja species as a source of antimicrobial ingredients for the food industry.     

Monoterpene alcohols release and bioconversion by Saccharomyces species and hybrids

Terpene profile of Muscat wines fermented by Saccharomyces species and hybrid yeasts was investigated. The amount of geraniol decreased in most wines with respect to the initial must except for Saccharomyces bayanus wines. On the other hand, alpha-terpineol amount was higher in wines fermented by Saccharomyces cerevisiae and hybrid yeasts. The amount of linalool was similar in all wines and comparable to the amount in the initial must. Lower levels of beta-d-glucosidase activity were found in the hybrid yeasts with respect to S. cerevisiae. Moreover, no relationship between beta-d-glucosidase activity and terpenes profile in Muscat wines fermented with Saccharomyces species and hybrids was found. Growth of yeasts on minimum medium supplemented with geraniol showed bioconversion of geraniol into linalool and alpha-terpineol. Percentages of geraniol uptake and bioconversion were different between Saccharomyces species and hybrids. Strains within S. bayanus, Saccharomyces kudriavzevii and hybrids showed higher geraniol uptake than S. cerevisiae, whereas the percentage of produced linalool and alpha-terpineol was higher in S. cerevisiae and hybrid yeasts than in S. bayanus and S. kudriavzevii. The relationship between geraniol uptake and adaptation of Saccharomyces species to grow at low temperature is discussed.     

Liposomal incorporation of carvacrol and thymol isolated from the essential oil of Origanum dictamnus L. and in vitro antimicrobial activity

The chemical composition of the essential oils from Origanum dictamnus L. (wild and organic cultivated plant) was analysed by GC–MS. Carvacrol, thymol, p-cymene, and γ-terpinene were identified as major constituents and isolated from both samples. The above components were successfully encapsulated in phosphatidyl choline-based liposomes and the possible improvement of their antioxidant and antimicrobial activities was tested against selected microbia. The antimicrobial properties of the oils were tested by a diffusion technique against four Gram positive and four Gram negative bacteria and three human pathogenic fungi, as well as the food-borne pathogen, Listeria monocytogenes. The percentage of the encapsulated carvacrol, the major component of the oil, was determined by GC–FID. In order to investigate any possible synergistic or antagonistic effect between carvacrol/thymol and carvacrol/γ-terpinene, the antimicrobial activities of the mixtures, were also determined before and after encapsulation in liposomes. All tested compounds presented enhanced antimicrobial activities after the encapsulation. Finally, in all cases, their antioxidant activity using differential scanning calorimetry was studied, in order to gain knowledge about their oxidation stability.     

Antioxidant activity of twenty wild Spanish Thymus mastichina L. populations and its relation with their chemical composition

The antioxidant activity and chemical composition of essential oils and methanolic extracts of twenty Spanish Thymus mastichina L. populations were studied. Both essential oils and methanolic extracts possessed antioxidant properties. However, the total phenol contents of the methanolic extracts varied between 2.90 and 9.15 mg GAE/g_extract and the EC₂₅ values of DPPH free radical scavenging activity between 0.90 and 3.45 mg/mL for the methanolic extracts and 78–241 mg/mL for essential oils, these showing low antioxidant potential. Actually, in essential oils the main compound determined was the 1,8-cineole (56.8–69.6%), whereas thymol, γ-terpinene, terpinolene and geraniol (species with considerable DPPH scavenging activity) were observed in low amounts. Concerning methanolic extracts, rosmarinic acid was the most abundant polyphenol (1.70–9.85 mg/g), followed by methoxysalicylic acid, apigenin, kaempferol and luteolin.     

Antimicrobial activity and chemical composition of Origanum glandulosum Desf. essential oil and extract obtained by microwave extraction: Comparison with hydrodistillation

Origanum glandulosum Desf. essential oils obtained by hydrodistillation (HD), solvent-free microwave extraction (SFME) and the extract obtained by microwave-assisted extraction (MAE) were investigated by capillary gas chromatography and gas chromatography/mass spectrometry. The main components of both oils obtained by HD and SFME were thymol (41.6–81.1%) followed by γ-terpinene (27.0–3.1%), p-cymene (17.1–4.0%) and carvacrol (2.2–4.4%), respectively. In the same way, thymol (65.4%), γ-terpinene (13.1%), p-cymene (7.2%) and carvacrol (3.5%) were the main components of the extract obtained by hexane microwave extraction. The SFME method was most selective for the extraction of thymol. The examination of the antimicrobial activity of both essential oils against 10 bacteria, two yeasts and four moulds revealed that O. glandulosum oil is more antifungal than antibacterial. To our knowledge, the antifungal activity of the O. glandulosum oil obtained by HD and both antimicrobial and antifungal activities of O. glandulosum SFME oil were not yet reported. Our study suggests that O. glandulosum essential oil has the potential to be used as a food preservative and to prevent the growth of nosocomial bacteria.     

Fingerprint of volatile flavour constituents and antioxidant activities of teas from Thailand

The volatile flavour components of different teas growing in Thailand were extracted using the simultaneous distillation and extraction (SDE) technique. These volatiles were investigated by GC–MS. At least 54 components representing 76.51–83.32% of all samples were identified. Hotrienol, geraniol and linalool were found to be the major components in Green Oolong tea. Green Assam tea contained linalool, geraniol and α-terpineol as the key flavour constituents. Chin Shin Oolong tea was dominated by linalool, indole and cis-jasmone whilst the major flavour volatiles of Chin Hsuan Oolong tea were trans-nerolidol, cis-jasmone and geraniol. Indole, geraniol and cis-jasmone were detected as the main constituents in Four Season tea. Change of quality and quantity of volatile flavour components was related to fermentation methods that increased volatiles were illustrated by the semi-fermented tea processing method. Green Assam tea infusion extract was evaluated to have the strongest antioxidant activities with the highest amount of phenol content followed by Four Season tea, Chin Shin Oolong tea, Chin Hsuan Oolong tea and Green Oolong tea, respectively.     

Carum copticum Benth. & Hook., essential oil chemotypes

Carum copticum Benth. & Hook is a popular spice and a traditional flavor that is used in Iran. The fruits of C. copticum have several therapeutic effects including carminative, diuretic and anti-vomiting effects. There are some reports on the chemical composition of C. copticum fruits essential oil. In our research the results of GC–MS analyses of the essential oil from C. copticum fruits and differences among various reports were described. Major constitutes of the oil were thymol (54.50%), γ-terpinene (26.10%) and p-cymene (22.10%). Comparison of the result from this study with other reports indicates that C. copticum have thymol and carvacrol chemotypes.     

Chemical composition and antioxidant activity of the essential oil of Clinopodium vulgare L.

This study was designed to examine the chemical composition and in vitro antioxidant activity of the essential oil of Clinopodium vulgare. GC–MS analysis of the oil resulted in the identification of 40 compounds, representing 99.4% of the oil; thymol (38.9%), γ-terpinene (29.6%) and p-cymene (9.1%) were the main components. The samples were subjected to a screening for their possible antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene-linoleic acid assays. In the first case, IC₅₀ value of the C. vulgare essential oil was determined as 63.0 ± 2.71 μg/ml. IC₅₀ value of thymol and γ-terpinene, the major compounds of the oil, was determined as 161 ± 1.3 μg/ml and 122 ± 2.5 μg/ml, respectively, whereas p-cymene did not show antioxidant activity. In β-carotene-linoleic acid system, C. vulgare essential oil exhibited 52.3 ± 1.19% inhibition against linoleic acid oxidation. In both systems, antioxidant capacities of BHT, curcumine and ascorbic acid were also determined in parallel experiments.     

Identification of volatile components in basil (Ocimum basilicum L.) and thyme leaves (Thymus vulgaris L.) and their antioxidant properties

Aroma compounds in the extracts of basil leaves (Ocimum basilicum L.) and thyme leaves (Thymus vulgaris L.) were identified by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). The major aroma constituents of basil were 3,7-dimethyl-1,6-octadien-3-ol (linalool; 3.94 mg/g), 1-methoxy-4-(2-propenyl) benzene (estragole; 2.03 mg/g), methyl cinnamate (1.28 mg/g), 4-allyl-2-methoxyphenol (eugenol; 0.896 mg/g), and 1,8-cineole (0.288 mg/g). The major aroma constituents of thyme were 2-isopropyl-5-methylphenol (thymol; 8.55 mg/g), 4-isopropyl-2-methylphenol (carvacrol; 0.681 mg/g), linalool (0.471 mg/g), α-terpineol (0.291 mg/g), and 1,8-cineole (0.245 mg/g). Twelve aroma constituents of basil and thyme were examined for their antioxidant activities using the aldehyde/carboxylic acid assay. Eugenol, thymol, carvacrol, and 4-allylphenol showed stronger antioxidant activities than did the other components tested in the assay. They all inhibited the oxidation of hexanal by almost 100% for a period of 30 days at a concentration of 5 μg/ml. Their antioxidant activities were comparable to those of the known antioxidants, α-tocopherol and butylated hydroxy toluene (BHT).     

Volatile composition of hybrids Citrus juices by headspace solid-phase micro extraction/gas chromatography/mass spectrometry

The volatile compounds of Citrus juices have been extracted by headspace solid-phase microextraction (HS-SPME) and analysed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). This work deals with the analysis of 65 cross pollinated hybrid fruits and their parents: mandarin (Citrus reticulata Blanco var. Willow Leaf) and clementine (Citrus reticulata × Citrus sinensis var. Commune). Among the 44 components identified which accounted for 90.2 to 99.8% of the volatile fraction, limonene (56.8–93.3%) and γ-terpinene (0.1–36.4%) were the major components in all samples. The clementine juice was characterised by the pre-eminence of limonene (90.0%) and a minor amount of γ-terpinene (1.2%) while the mandarin juice exhibited high amount of limonene (66.3%) and γ-terpinene (21.1%). All hybrid juices showed qualitatively similar composition but differing in the quantitative profile of the couple limonene/γ-terpinene. The principal component analysis (PCA) and the discriminant analysis indicated that hybrids samples were symmetrically distributed around the both parents. If some studies were found in the literature about mandarin juice, to our knowledge, this work is the first study on the volatile compounds of clementine juice and a large amount of hybrids.     

GC/MS analysis and in vitro antioxidant activity of essential oil and methanol extracts of Thymus caramanicus Jalas and its main constituent carvacrol

Chemical composition of the essential oil, antioxidant activity (DPPH and β-carotene/linoleic acid assays), and total phenolic content (Folin–Ciocalteu assay) of aerial parts of Thymus caramanicus were determined. The highest radical-scavenging activity (DPPH test) was shown by the polar subfraction of the methanol extract (IC₅₀ = 43.0 μg/ml) which was also higher than that of butylated hydroxytoluene (BHT, IC₅₀ = 19.7 μg/ml). However, it was the nonpolar subfraction of the methanol extract that showed the highest inhibition (84.4%), as assessed by the β-carotene/linoleic acid assay, which was only slightly lower than that shown by BHT (93.3%). The antioxidant activities of the essential oil main component (carvacrol) were also evaluated for comparison. Total phenolic content of the polar subfraction, as gallic acid equivalents, was 124.3 μg/mg. Essential oil extracted from the aerial parts by hydrodistillation was analysed by GC and GC/MS. Fifteen constituents, representing 99.3% of the oil, were identified, of which the major ones, carvacrol (85.9%), thymol (3.3%), p-cymene (3.2%), γ-terpinene (1.8%) and borneol (1.3%), accounted for 95.6% of the oil.     

Genoprotective Effects of Essential Oil Compounds Against Oxidative and Methylated DNA Damage in Human Colon Cancer Cells

Essential oils (EO) are widely used in foods as flavoring and preservative agents. Many of the biological activities of EO have been attributed to major essential oil compounds (EOC) but their direct interaction with colonic epithelial cells and their genotoxic and genoprotective effects are not well established. In this study, the cytotoxicity and genotoxicity of EOC including nerolidol, thymol, geraniol, methylisoeugenol, eugenol, linalool, and a commercial blend (Agolin) were determined. Furthermore, the genoprotective effects of EOC against oxidative and methylating damage were assessed using the comet assay in HT‐29 colorectal adenocarcinoma cells. The majority of EOC were cytotoxic to HT‐29 cells at or above 250 ppm after 24 hr exposure. At noncytotoxic doses, none of the EOC was genotoxic in the comet assay. Genoprotection against oxidative DNA damage was observed for nerolidol (at 62.5 ppm), thymol (at 12.5 ppm), geraniol, and methylisoeugenol (both at 125 ppm), as well as linalool and Agolin (both at 250 ppm). Thymol was the most protective compound against oxidative DNA damage and geraniol (at 125 ppm) also protected cells against methylating DNA damage. This study highlights the potential of EOC such as thymol to protect the colonic epithelium against oxidative DNA damage and geraniol against methylating DNA damage. Further in vivo studies are needed to confirm these findings for safety and efficacy to exploit their potential pharmaceutical or nutraceutical uses for colonic health.     

Physical and antimicrobial properties of Gelidium corneum/nano-clay composite film containing grapefruit seed extract or thymol

To improve the physical properties of Gelidium corneum (GC) film, Cloisite Na⁺ and 30B were incorporated into the preparation of the film. X-ray diffraction patterns of the films indicated that a degree of exfoliation and intercalation are formed depending on the type of nano-clays and its concentration and the film structure affects the physical properties of the GC/nano-composite films. Tensile strength (TS) of the GC film was increased by the addition of nano-clays. GC film had a TS of 19.59 MPa for the control, while the GC film having 3% Cloisite Na⁺ or 30B had TS of 27.37 and 26.40 MPa, respectively. Elongation at break and water vapor permeability were also improved by the addition of 3% nano-clays. The physical properties of the film were not improved by the addition of GSE or thymol, but the additions did inhibit the growth of Escherichia coli O157:H7 and Listeria monocytogenes. The results suggest that the nano-composite films containing GSE or thymol may extend the shelf life of food.