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 the essential oil and various extracts of Nepeta flavida Hub.-Mor. from Turkey

This study was designed to examine the chemical composition and in vitro antioxidant activity of the essential oil and various extracts (hexane, dichloromethane and methanol sub-fractions) of Nepeta flavida. GC and GC–MS analyses of the essential oil resulted in the identification of 68 compounds, representing 96.4% of the oil; 1,8-cineole (38.9%) and linalool (25.1%) were the main components, comprising 64.0% of the total oil. The samples were subjected to a screening for their possible antioxidant activities by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene-linoleic acid assays. In the first case, the IC₅₀ value of the N. flavida essential oil was determined to be 42.8 ± 2.19 μg/ml. Among the extracts, the strongest activity was exhibited by the polar sub-fraction of the methanol extract with an IC₅₀ value of 63.2 ± 1.75 μg/ml. In the β-carotene-linoleic acid system, N. flavida essential oil exhibited 86.3% ± 1.69 inhibition against linoleic acid oxidation. Among the extracts prepared with various solvents, a correlation was observed between the polarity and antioxidant activity. The extracts exhibited the same activity pattern in this system the most active one is the polar sub-fraction, 79.7% ± 0.89. On the other hand, 1,8-cineole, a major compound of the essential oil, exhibited marked antioxidant activity in both systems, whereas the other compound, linalool, did not show any activity. The amount of total phenolics was highest in the polar and non-polar sub-fractions. Particularly, a positive correlation was observed between the total phenolic content and the antioxidant activity of the extracts. As estimated from the results, amounts of phenolic compounds were less in hexane and dichloromethane extracts than in the others. In conclusion, antioxidant potentials of polar and non-polar methanol sub-fractions could be attributed to their high phenolic contents. In both systems, antioxidant capacities of BHT, ascorbic acid, curcumin and α-tocopherol were also determined in parallel experiments.     

Determination of antioxidant activities of various extracts and essential oil compositions of Thymus praecox subsp. skorpilii var. skorpilii

The aim of this work is to examine the chemical composition and antioxidant activity of separated essential oils and different solvent extracts of Thymus praecox subsp. skorpilii var. skorpilii (TPS). The ethanol, acetone, methanol, hexane, aqueous extracts and separated essential oils of TPS were assessed for their antioxidant activities. Antioxidant activities were evaluated by reduction of Mo(VI) to Mo(V), reducing power, superoxide scavenging activity, free radical-scavenging activity, metal chelating activity, linoleic acid peroxidation, hydrogen peroxide scavenging activity and peroxide scavenging activity. Essential oils were characterized in total to be 41 components, whereas 9 components were isolated by column chromatography for antioxidant activity. TPS essential oil was found to contain thymol (40.31%) and o-cymene (13.66%) as the major components. The ethanol, methanol and water extracts exerted significant free radical-scavenging activity. The methanol and water extracts displayed highest superoxide scavenging activity. The water extract has the highest total phenolics (6.211 mg gallic acid (GAE)/g DW) and flavonoids (0.809 mg quercetin/g DW).     

Essential oil composition and antibacterial activity of Thymus caramanicus at different phenological stages

Thymus species are well known as medicinal plants because of their biological and pharmacological properties. Thymus caramanicus is an endemic species grown in Iran. Variation in the quantity and quality of the essential oil of wild population of T. caramanicus at different phenological stages including vegetative, floral budding, flowering and seed set are reported. The oils of air-dried samples were obtained by hydrodistillation. The yields of oils (w/w%) at different stages were in the order of: flowering (2.5%), floral budding (2.1%), seed set (2.0%) and vegetative (1.9%). The oils were analyzed by GC and GC–MS. In total 37, 37, 29 and 35 components were identified and quantified in vegetative, floral budding, full flowering and seed set, representing 99.3, 98.6, 99.2 and 97.8% of the oil, respectively. Carvacrol was the major compound in all samples. The ranges of major constituents were as follow: carvacrol (58.9–68.9%), p-cymene (3.0–8.9%), γ-terpinene (4.3–8.0%), thymol (2.4–6.0%) and borneol (2.3–4.0%). Antibacterial activity of the oils and their main compounds were tested against seven Gram-positive and Gram-negative bacteria by disc diffusion method and determining their minimum inhibitory concentration (MIC) values. The inhibition zones (IZ) and MIC values for bacterial strains, which were sensitive to the essential oil of T. caramanicus, were in the range of 15–36 mm and 0.5–15.0 mg/ml, respectively. The oils of various phenological stages showed high activity against all tested bacteria, of which Bacillus subtilis and Pseudomonas aeruginosa were the most sensitive and resistant strains, respectively. Thus, they represent an inexpensive source of natural antibacterial substances that exhibited potential for use in pathogenic systems.     

Composition and antioxidant and antimicrobial activity of the essential oil and extracts of Stachys inflata Benth from Iran

Composition and antioxidant and antimicrobial activities of essential oil and methanol extract polar and nonpolar subfractions of Stachys inflata were determined. GC and GC/MS analyse of the essential oil showed 45 constituents representing 95.46% of the oil, the major components linalool (28.55%), α-terpineol (9.45%), spathulenol (8.37%) and (2E)-hexenal (4.62%) constituted 50.99% of it. Essential oil and extracts were also tested for their antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and β-carotene/linoleic acid assays. In the DPPH test, IC₅₀ value for the polar subfraction was 89.50 μg/ml, indicating an antioxidant potency of about 22% of that of butylated hydroxytoluene (IC₅₀ = 19.72 μg/ml) for this extract. In β-carotene/linoleic acid assay, the best inhibition belonged to the nonpolar subfraction (77.08%). Total phenolic content of the polar and nonpolar extract subfractions was 5.4 and 2.8% (w/w), respectively. The plant also showed a week antimicrobial activity against three strains of tested microorganisms. Linalool and α-terpineol were also tested as major components of the oil and showed no antioxidant but considerable antimicrobial activities.     

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.     

Chemical composition,cytotoxic and antioxidant activity of the leaf essential oil of Photinia serrulata

The leaf essential oil of Photinia serrulata was obtained by hydrodistillation and analyzed by GC and GC-MS. Seventy-one components were identified in the essential oil and the main components of the oil were 10-epi-γ-eudesmol (12.72%), pinene (6.85%), sabinene (5.93%), α-humulene (5.87%) and α-thujene (5.47%). The in vitro cytotoxicity of the oil on human cancer cell lines Hela, A-549 and Bel-7402 was examined. The oil was found to be very active against all the three human tumor cell lines tested with low IC₅₀ of 0.0427 μl/ml (Hela), 0.0219 μl/ml (A-549) and 0.0301 μl/ml (Bel-7402). The oil was also found to possess antioxidant activity as demonstrated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method.     

Salinity impact on fruit yield,essential oil composition and antioxidant activities of Coriandrum sativum fruit extracts

This study is designed to examine the fruit essential oil composition, the total phenolic amounts and the antioxidant activities in methanolic extracts of Coriandrum sativum under saline conditions. Increasing NaCl levels to 75 mM reduced significantly the fruit yield by 36%. The essential oil yield was 0.30%, based on the dry weight; it increased by 77% and 84% at 50 and 75 mM NaCl, respectively, in comparison to the control. The major constituents were linalool and camphor, whose amounts increased with increasing NaCl concentrations. Antioxidant activities of the methanol extracts were determined by three different test systems, namely DPPH, β-carotene/linoleic acid and reducing power assays. In these three test systems, the highest activity was exhibited in control plants and was reduced significantly with increasing NaCl levels. In control plants, the total phenolic amount was 1.04 mg GAE/g DW which decreased by 43% and 66% at 50 and 75 mM NaCl, respectively.     

Antioxidant and antimicrobial activity evaluation and essential oil analysis of Semenovia tragioides Boiss. from Iran

This study reports the essential oil composition, antioxidant and antimicrobial activity of the essential oil and methanol extract of aerial parts of Semenovia tragioides. GC and GC/MS analysis identified 17 compounds representing 99.4% of the oil. The main components comprising 61.9% of the oil were lavandulyl acetate (25.5%), geranyl acetate (12.5%), trans-β-ocimene (8.8%), p-cymene (7.7%) and γ-terpinene (7.4%). The samples were screened for their antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and β-carotene/linoleic acid assay methods. None of the plant samples showed appreciable antioxidant activity in DPPH test. However, methanol extract exhibited considerable linoleic acid oxidation inhibition (77.4%) in the β-carotene/linoleic acid test, a value near to that of butylated hydroxytoluene (BHT, 95.6%). Total phenolic content of the plant extract as gallic acid equivalents was 7.5 μg/mg. The essential oil exhibited strong antimicrobial activity against all but one of the tested microorganisms while the plant extract only inhibited two of them weakly.     

Solvent-free microwave extraction of essential oil from Dryopteris fragrans and evaluation of antioxidant activity

Solvent-free microwave extraction (SFME) of the essential oil from Dryopteris fragrans and its antioxidant activity were investigated. A central composite design combined with response surface methodology was applied to study the influences of extraction time, irradiation power and humidity (proportion of water pretreatment). A maximal extraction yield of 0.33% was achieved under optimal conditions of extraction time 34 min, irradiation power 520 W and humidity 51%. Sixteen compounds, representing 89.65% of the oil, were identified, of which the major ones, (1R,4S,11R)-4,6,6,11-tetramethyltricyclo[5.4.0.0(4,8)]undecan-1-ol (30.49%), 1R,4S,7S,11R-2,2,4,8-tetramethyltricyclo[5.3.1.0(4,11)]undec-8-ene (22.91%) and, 1,4,4a,5,6,7,8,8a-octahydro-2,5,5,8a-tetramethyl-1-naphthalenemethanol (15.11%), accounted for 68.51% of the oil. The antioxidant activity of the essential oil was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH), β-carotene/linoleic acid, and reducing power assay, the IC₅₀ values were 0.19, 0.09 and 0.18 mg/mL, respectively. All these results suggest that SFME represents an excellent alternative protocol for production of essential oils from plant materials.     

The in vitro and in vivo antioxidant properties of seabuckthorn (Hippophae rhamnoides L.) seed oil

The antioxidant capacity of seabuckthorn (Hippophae rhamnoides L.) seed oil was investigated with a number of established in vitro assays and in an in vivo study of carbon tetrachloride (CCl₄)-induced oxidative stress in mice. The results showed that DPPH radical scavenging activity, ferrous ion chelating activity, reducing power and inhibition of lipid peroxidation activity all increased with increasing concentrations of seabuckthorn seed oil. Moreover, the EC₅₀ values of seabuckthorn seed oil from the hydrogen peroxide, superoxide radical, hydroxyl radical scavenging assays were 2.63, 2.16 and 0.77 mg/ml, respectively. In the in vivo study, seabuckthorn seed oil inhibited the toxicity of CCl₄, as seen from the significantly increased activities of the antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. The GSH content in the liver was also increased, whereas hepatic malondialdehyde was reduced. Taken together, these results clearly indicate that seabuckthorn seed oil has significant potential as a natural antioxidant agent.     

Phytochemical composition and in vitro antimicrobial and antioxidant activities of essential oils of three endemic Soqotraen Boswellia species

The chemical composition of three essential oils obtained from the barks of three endemic Boswellia species namely, Boswellia dioscorides, Boswellia elongata and Boswellia socotrana which were collected from the Soqotra Island (Yemen), was investigated. In parallel to that evaluation of their antimicrobial and antioxidant activities was also carried out. The investigation led to the identification of 72, 70 and 67 constituents for B.dioscorides, B. elongata, and B. socotrana, respectively. The B. dioscorides oil was found to have a high content of monoterpene hydrocarbons (32.8%) with α-thujene (9.3%) and α-pinene (8.3%) as main components. Whereas, the essential oil of B. elongata was characterised by high diterpene content (31.7%), in which incensol (14.8%) was found to be the major constituent. On the other hand, the oil obtained from the B. socotrana was found to be predominated of monoterpene hydrocarbons (26.4%) and oxygenated monoterpenes (30.7%) with p-cymene (13.0%), 2-hydroxy-5-methoxy-acetophenone (16.3%) and camphor (11.6%). All essential oils possessed antimicrobial activity especially against Gram-positive bacteria with MIC-values between 1.8 and 17.2 mg/ml. Furthermore, the DPPH-radical scavenging assay exhibited only weak antioxidant activities (28%) at 1.0 mg/ml.     

Antioxidant activity and rosmarinic acid changes in salicylic acid-treated Thymus membranaceus shoots

In order to characterise the antioxidant activity of Thymus membranaceus Boiss. subsp. membranaceus, an endemic species in Southeast Spain, five different analytical methods were used. Water, methanol and hexane extracts obtained from 60-day-old in vitro-grown shoots were assayed for their antioxidative properties using DPPH (1,1-diphenyl-2-picrylhydrazyl radical), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonate), FRAP (ferric reducing antioxidant power), reducing power and conjugated diene assays. Total soluble phenol content, as well as rosmarinic acid, in the different extracts were also determined. Methanolic extracts exhibited the best antioxidant activity and the highest amounts of total soluble phenolics. A single application of salicylic acid (10 μM) on culture media resulted in an increase in rosmarinic acid and phenolic levels, which in turn improved the extracts’ antioxidant properties. These current findings open new opportunities for obtaining valuable natural antioxidants for commercial exploitation by using tissue culture systems.     

Antioxidant activity and chemical constituents of essential oil and extracts of Rhizoma Homalomenae

Antioxidant activity and composition of essential oil and extracts of Rhizoma Homalomenae were determined. The extracts, especially the ethyl acetate extract (QJ4 fraction) of the aqueous residue after oil distillation, had considerable antioxidant potency which was significantly associated with their total phenolic and flavonoid contents, but the essential oil showed only weak or moderate activity. GC–MS analysis of the essential oil (yield: 0.82%, v/w) resulted in the identification of 77 compounds, accounting for 96.5% of the content of the oil. The major components, epi-α-cadinol (14.8%), α-cadinol (14.8%), α-terpineol (13.8%), linalool (11.1%), terpinen-4-ol (4.92%), and δ-cadinene (4.91%) constituted 64.3% of it. LC–MS/MS and HPLC analyses showed seven phenolic compounds (protocatechuic acid, vanillic acid, syringic acid, caffeic acid, p-coumaric acid, ferulic acid and apigenin) with a great amount in the ethyl acetate extract (QJ4 fraction). The strong antioxidant properties of the plant extracts may be attributed to the presence of these phenolics.     

A comparative study of tocopherols composition and antioxidant properties of in vivo and in vitro ectomycorrhizal fungi

In aerobic organisms, the free radicals are constantly being produced during the normal cellular metabolism. The antioxidant properties of many organisms and particularly of wild mushrooms with their content in antioxidant compounds such as tocopherols, can detoxify potentially damaging forms of activated oxygen. Herein, a comparative study of tocopherols composition and antioxidant properties of in vivo (fruiting bodies) and in vitro (mycelia) ectomycorrhizal fungi: Paxillus involutus and Pisolithus arhizus. Tocopherols were determined by high performance liquid chromatography (HPLC) coupled to a fluorescence detector. The antioxidant properties were studied in terms of DPPH radical-scavenging activity, reducing power and inhibition of β-carotene bleaching. Fruiting bodies revealed the highest antioxidant properties, including scavenging effects on free radicals (EC₅₀ = 0.61 and 0.56 mg/ml) and inhibition of lipid peroxidation capacity (EC₅₀ = 0.40 and 0.24 mg/ml for P. involutus and P. arhizus, respectively), than mycelia produced in vitro cultures. Nevertheless, mycelia revealed higher levels of total tocopherols than fruiting bodies, and particularly P. arhizus mycelium proved to be a powerful source of γ-tocopherol (154.39 μg/g dry weight).     

Polyphenolic content and antioxidant activity of Eugenia pollicina leaf extract in vitro and in model emulsion systems

The polyphenolic profile of a leaf extract of the Mauritian endemic plant, Eugenia pollicina, was assessed as a source of natural antioxidants. The amounts of flavan-3-ol derivatives determined by HPLC, were in the order of (−)-epicatechin (EC) > (−)-epigallocatechin gallate (EGCG) > (+)-catechin (C) > (−)-epicatechin gallate (ECG) with the levels of Procyanidin B2 and B1 dimers ranging from 1 to 3 mg g⁻¹ FW. The trolox equivalent antioxidant capacity and ferric reducing antioxidant power values were 796 ??mol g⁻¹ FW and 302 ??mol g⁻¹ FW respectively. E. pollicina extracts also strongly inhibited the FeCl₃ and ascorbate-dependent microsome lipid peroxidation, a function that is linked to their flavonoid contents. The extent of DNA damage induced by the extract under study in the copper-phenanthroline assay was lower than the effect of a reference of 240 ??M ascorbate. E. pollicina extracts also inhibited lipid autoxidation in the 30% (v/v) olive oil and soybean oil oil-in-water (O/W) emulsions and was effective in slowing down the formation of hydroperoxides in the emulsions during 13 days storage at 40 °C as determined by the peroxide, conjugated diene and para-anisidine values. The high levels of total phenolics, flavonoids and procyanidins measured indicate that E. pollicina is a potential source of antioxidants relevant to the maintenance of oxidative stability of the food matrix, cosmetics and/or pharmaceutical preparations.