An electrochemical sensor based on molecularly imprinted poly(o-phenylenediamine) for the detection of thymol.

A molecularly imprinted electrochemical sensor was facilely fabricated for the detection of thymol (THY). o-Phenylenediamine (oPD) was used as the functional monomer and electropolymerized on the surface of the glassy carbon electrode (GCE) by using THY as the templates. After the THY templates were removed with 50 % (v/v) ethanol, imprinted cavities complementary to the templates were formed within the poly(o-phenylenediamine) (PoPD) films. The resultant molecularly imprinted PoPD/GCE (MI-PoPD/GCE) was used for the detection of THY, and a wide linear range from 0.5 to 100 µM with a low limit of detection (LOD) of 0.084 µM were obtained under the optimal conditions. The developed MI-PoPD/GCE also displays high selectivity, reproducibility and stability for THY detection. Finally, the content of THY in the real samples was accurately determined by the as-fabricated MI-PoPD/GCE, demonstrating its high practicability and reliability.

Evaluation of Different Drying Treatments with Respect to Essential Oil Components, Phenolic and Flavonoid Compounds, and Antioxidant Capacity of Ajowan (Trachyspermum ammi L.).

Ajowan (Trachyspermum ammi L.) is considered a valuable spice and medicinal herb. In this study, the essential oil content and composition of the aerial parts of ajowan were investigated under different drying treatments (sun, shade, oven at 45 °C, oven at 65 °C, microwave, and freeze drying). Moreover, the phenolic content, flavonoid content, and antioxidant capacity of samples were also assessed. Fresh samples produced the highest essential oil content (1.05%), followed by those treated under sun (0.7%) and shade drying (0.95%). Based on gas chromatography-mass spectrometry (GC-MS), thirty compounds were determined in which thymol (34.84-83.1%), carvacrol (0.15-32.36%), p-cymene (0.09-13.66%), and γ-terpinene (3.12-22.58%) were the most abundant. Among the drying methods, freeze drying revealed the highest thymol content, followed by drying in a 45 °C oven. The highest TPC (total phenolic content) and TFC (total flavonoid content) were obtained in the fresh sample (38.23 mg TAE g-1 dry weight (DW)) and in the sample oven-dried at 45 °C (7.3 mg QE g-1 DW), respectively. Based on the HPLC results, caffeic acid (18.04-21.32 mg/100 gDW) and ferulic acid (13.102-19.436 mg/100 g DW) were the most abundant phenolic acids, while among flavonoids, rutin constituted the highest amount (10.26-19.88 mg/100 gDW). Overall, freeze drying was the most promising method of drying for preserving the phenolic (TPC) and flavonoid (TFC) compounds and oil components.

Chemical and Olfactory Analysis of the Volatile Fraction of Ocimum gratissimum Concrete from Madagascar.

The chemical composition of the volatile fraction of Ocimum gratissimum concrete (romba) from Madagascar has been determined for the first time by GC/MS and GC-FID. A methyl cinnamate chemotype has been determined for this material, along with a set of compounds typical in essential oils and extracts from plants of the Ocimum genus. Variability was mostly observed on terpenes and terpenoids components. GC-O-MS was also used for a sensory evaluation of this material performed by a master perfumer. The chemical composition of this O. gratissimum extract was then compared with literature data to assess subtle differences between chemotypes of the same species and other species of the same genus within natural variability. A mapping illustrates the occurrence of the cinnamate chemotype in Eastern Africa, India and now Madagascar, while other origins generally present eugenol, thymol, camphor, or linalool chemotypes.

Volatile Oil Yield and Composition, Total Phenolic Content, Antioxidant Activity and Secondary Metabolite Content of Collected Thymus praecox Species in Rize.

In this study, the volatile oil yield (Clevenger), volatile oil (VO) composition (Gas Chromatography), phenolic contents (UV-VIS Spectrophotometer), antioxidant activities (UV-VIS Spectrophotometer) and secondary metabolite content (High Pressure Liquid Chromatography) of 11 Thymus praecox subspecies were evaluated. The most detected chemical class were oxygenated monoterpenes (55.18-86.1 %) in investigated samples. In the present study rosmarinic acid, isoquercitrin, gallocatechin and thymol could be detected in high amounts. The min. and max. content values of Flora/Field Samples were 1543.241 and 890.3-1425.3 for rosmarinic acid, 139.44-287.894 and 129.9-312.2 for thymol, 38.619-121.424 and 26.3-112.9 for gallocatechin as mg/g DW. Principal Component Analysis was used to differentiate Thymus praecox species regarding volatile oil composition and secondary metabolite content. The results demonstrated that T. praecox collected from the Rize flora and cultivated afterwards showed variability based on investigated characteristics. Finally, the Thymus praecox samples displaying high bioactive compounds present useful information for further investigations and applications.

Anti-Trypanosomatidae Activity of Essential Oils and Their Main Components from Selected Medicinal Plants.

Kinetoplastida is a group of flagellated protozoa characterized by the presence of a kinetoplast, a structure which is part of a large mitochondria and contains DNA. Parasites of this group include genera such as Leishmania, that cause disease in humans and animals, and Phytomonas, that are capable of infecting plants. Due to the lack of treatments, the low efficacy, or the high toxicity of the employed therapeutic agents there is a need to seek potential alternative treatments. In the present work, the antiparasitic activity on Leishmania infantum and Phytomonas davidi of 23 essential oils (EOs) from plants of the Lamiaceae and Asteraceae families, extracted by hydrodistillation (HD) at laboratory scale and steam distillation (SD) in a pilot plant, were evaluated. The chemical compositions of the EOs were determined by gas chromatography-mass spectrometry. Additionally, the cytotoxic activity on mammalian cells of the major components from the most active EOs was evaluated, and their anti-Phytomonas and anti-Leishmania effects analyzed. L. infantum was more sensitive to the EOs than P. davidi. The EOs with the best anti-kinetoplastid activity were S. montana, T. vulgaris, M. suaveolens, and L. luisieri. Steam distillation increased the linalyl acetate, ß-caryophyllene, and trans-α-necrodyl acetate contents of the EOs, and decreased the amount of borneol and 1,8 cineol. The major active components of the EOs were tested, with thymol being the strongest anti-Phytomonas compound followed by carvacrol. Our study identified potential treatments against kinetoplastids.

Thymus richardii subsp. nitidus (Guss.) Jalas Essential Oil: An Ally against Oral Pathogens and Mouth Health.

The genus Thymus L., belonging to the Lamiaceae family, contains about 220 species with a distribution that mainly extends in Europe, northwest Africa, Ethiopia, Asia, and southern Greenland. Due to their excellent biological properties, fresh and/or dried leaves and aerial parts of several Thymus ssp. have been utilized in the traditional medicine of many countries. To evaluate not only the chemical aspects but also the biological properties, the essential oils (EOs), obtained from the pre-flowering and flowering aerial parts of Thymus richardii subsp. nitidus (Guss.) Jalas, endemic to Marettimo Island (Sicily, Italy), were investigated. The chemical composition of the EOs, obtained by classical hydrodistillation and GC-MS and GC-FID analyses, showed the occurrence of similar amounts of monoterpene hydrocarbons, oxygenated monoterpenes, and sesquiterpene hydrocarbons. The main constituents of the pre-flowering oil were ß-bisabolene (28.54%), p-cymene (24.45%), and thymol methyl ether (15.90%). The EO obtained from the flowering aerial parts showed as principal metabolites ß-bisabolene (17.91%), thymol (16.26%), and limonene (15.59%). The EO of the flowering aerial parts, and its main pure constituents, ß-bisabolene, thymol, limonene, p-cymene, and thymol methyl ether were investigated for their antimicrobial activity against oral pathogens and for their antibiofilm and antioxidant properties.

Optimizing the Distillation of Greek Oregano-Do Process Parameters Affect Bioactive Aroma Constituents and In Vitro Antioxidant Activity?

The aim of the present work was to optimize the conditions of the distillation process at a pilot scale to maximize the yield of specific bioactive compounds of the essential oil of oregano cultivated in Greece, and subsequently to study the in vitro antioxidant activity of these oils. Steam distillation was conducted at an industrial distillery and a Face-Centered Composite (FCC) experimental design was applied by utilizing three distillation factors: time, steam pressure and temperature. Essential oil composition was determined by static headspace gas chromatography-mass spectrometry (HS-GC/MS). To obtain a comprehensive profile of the essential oils, instrumental parameters were optimized, including sample preparation, incubation conditions, sampling process, injection parameters, column thermal gradient and MS conditions. With the applied GC-MS method, more than 20 volatile compounds were identified in the headspace of the oregano essential oils and their relative percentages were recorded. Carvacrol was the most prominent constituent under all distillation conditions applied. Data processing revealed time as the main factor which most affected the yield. The Desired Space (DSc) was determined by conducting a three-dimensional response surface analysis of the independent and dependent variables, choosing yields of thymol and carvacrol as optimization criteria. The in vitro antioxidant activity of the essential oils of all samples was measured in terms of the interaction with the stable free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) after 20 and 60 min. The most prominent essential oils at different distillation conditions were also tested as inhibitors of lipid peroxidation. Higher % values of carvacrol and thymol were correlated to higher antioxidant activity. Evaluating the impact of the distillation conditions on the in vitro results, it seems that lower pressure, less time and higher temperature are crucial for enhanced antioxidant activities.

Chemical characterization, nematicidal and antioxidant activities of Thymus linearis Benth.

Thymus linearis and its essential oil (EO) are used to cure a range of diseases in traditional medicine. GC-MS analysis of Thymus linearis EO revealed the presence of sixty-four components. Thymol (50.62%), carvacrol (13.23 %), carvacrol acetate (7.72%), -bisabolene (5.47%) and o-cymene (5.47%) are the only five basic constituents in the oil which accounts for 82.07% of oil. When compared to other compounds, the EO and its primary components thymol and carvacrol had the largest proportion of mortality in Meloidogyne javanica. Carvacrol has also been proven to be helpful in suppressing the hatching of M. javanica eggs. This is the first time T. linearis EO and its constituents, such as borneol and caryophyllene, have been studied for nematicidal action. The antioxidant activity of EO components and active compounds was assessed using the ABTS radical scavenging method. Thymol and carvacrol were found to exhibit high antioxidant activity. The IC50 of thymol and carvacrol are found to be 38.18 g/ml and 49.65 g/ml, respectively and are comparable to the positive control trolox (47.12 g/ml). Results clearly showed high potency for EO and its constituents, thymol and carvacrol as nematicidal and antioxidant agents.

Effect of Extraction Time on Origanum onites L. Infusions and Essential Oils - Biological Evaluation, Statistical Principal Component and Hierarchial Cluster Analyses.

Origanum onites L. of Lamiaceae, is used as a culinary spice and herbal tea; also for its antioxidant, antimicrobial effects among other activities and effects. This research aims to examine the in vitro antimicrobial and antioxidant activities of the infusions and the essential oils (EOs) obtained using different conditions from the aerial parts of the processed and raw of O. onites herbal material. The EOs from O. onites were distilled at different times and characterized both by GC-FID and GC/MS systems. The resulting main compounds were identified as carvacrol (65.5-91 %), linalool (0.3-17.9 %), γ-terpinene (2.8-4.6 %), and thymol (0.5-1.5 %), respectively. In addition, Origanum infusions were prepared at four different time intervals, where the volatile compounds of the infusions were analysed using the HS-SPME-GC/MS system. Statistical Principal Component (PCA) and Hierarchical Cluster Analyses (HCA) tools were used to demonstrate the composition variations on different times for O. onites infusion and EO samples.

Quantitation of 1,4-Dichlorobenzene and Thymol in Beeswax Using Dynamic Headspace Vacuum Transfer in Trap Extraction Prior to Gas Chromatography-Mass Spectrometry.

A method based on dynamic headspace vacuum transfer in trap extraction, followed by gas chromatography coupled with a mass spectrometer (DHS-VTT-GC-MS), was validated for the fast quantitation of 1,4-dichlorobenzene (p-dichlorobenzene; PDCB) and thymol residues in beeswax. The quantitation limits (LOQ) were 0.05 mg/kg (PDCB) and 0.25 mg/kg (thymol). Recoveries above 80% were obtained for PDCB concentrations between 0.05 and 10 mg/kg and for thymol concentrations between 0.25 and 200 mg/kg. Analysis of beeswax samples showed a good correlation between the results obtained by DHS-VTT-GC-MS analysis and those of a previous method based on SPE extraction followed by gas chromatography and triple- quadrupole mass spectrometry (GC-MS/MS) (R2 = 0.9770 for PDCB and 0.9666 for thymol). However, the sample preparation and chromatography were much shorter using the DHS-VTT-GC-MS method. Forty comb foundations samples produced in Switzerland in 2019 and 2021 were analysed using DHS-VTT-GC-MS. Fourteen samples contained PDCB above the LOQ of 0.05 mg/kg, ranging up to a maximum of 1.53 mg/kg. The mean concentration of the positive samples was 0.22 mg/kg. All samples contained thymol residues ranging between 3.9 and 84.4 mg/kg with mean and median concentrations of 22.7 mg/kg and 17.4 mg/kg. Residue levels of PCDB and thymol in Swiss beeswax were substantially below those measured 20 (PDCB) and 10 (thymol) years ago, respectively.

Mexican Oregano (Lippia berlandieri Schauer and Poliomintha longiflora Gray) Essential Oils Induce Cell Death by Apoptosis in Leishmania (Leishmania) mexicana Promastigotes.

Leishmaniasis is a neglected vector-borne disease; there are different manifestations of the diseases and species involved, and cutaneous leishmaniasis caused by Leishmania (L.) mexicana is the most prevalent in Mexico. Currently, the drugs available for the treatment of leishmaniasis are toxic, expensive, and often ineffective; therefore, it is imperative to carry out research and development of new therapeutic alternatives, with natural products being an attractive option. In particular, oregano is a plant with worldwide distribution; in Mexico, two species: Lippia berlandieri Schauer and Poliomintha longiflora Gray are endemic. Both essential oils (EO's) have been reported to have antimicrobial activity attributed to their main components, thymol and carvacrol. In this research, the leishmanicidal effect and mechanism of cell death induced by L. berlandieri EO, P. longiflora EO, thymol, and carvacrol in L. mexicana promastigotes were determined in vitro. Additionally, the cytotoxic activity in mammalian cells was evaluated. L. berlandieri EO presented higher leishmanicidal activity (IC50 = 41.78 µg/mL) than P. longiflora EO (IC50 = 77.90 µg/mL). Thymol and carvacrol were the major components of both Mexican oregano EO's. Thymol presented higher leishmanial inhibitory activity (IC50 = 22.39 µg/mL), above that of carvacrol (IC50 = 61.52 µg/mL). All the EO's and compounds evaluated presented lower cytotoxic activity than the reference drug; thymol was the compound with the best selectivity index (SI). In all cases, apoptosis was identified as the main mechanism of death induced in the parasites. The leishmanicidal capacity of the Mexican oregano EO is an accessible and affordable alternative that can be further explored.

Obtaining Aromatic Extracts from Portuguese Thymus mastichina L. by Hydrodistillation and Supercritical Fluid Extraction with CO2 as Potential Flavouring Additives for Food Applications.

Humans often respond to sensory impulses provided by aromas, and current trends have generated interest in natural sources of fragrances rather than the commonly used synthetic additives. For the first time, the resulting aroma of a selected culture of Thymus mastichina L. was studied as a potential food ingredient. In this context, dried (DR) and fresh (FR) samples were submitted to carbon dioxide (CO2) supercritical extraction (SFE) and hydrodistillation (HD) methods. The extracts were characterised according to their volatile composition by GC-MS, cytotoxicity against a non-tumour cell culture, and sensory attributes (odour threshold and olfactive descriptors). The most abundant aromas were quantified, and the analysis performed by GC-MS revealed an abundance of terpenoids such as thymol chemotype, followed by the precursors α-terpinene and p-cymene. DR and FR extracts (EX) obtained from SFE-CO2 show the highest content of thymol, achieving 52.7% and 72.5% of the isolated volatile fraction. The DR essential oil (EO) contained the highest amount of terpenoids, but it was also the most cytotoxic extract. In contrast, SFE-CO2 products showed the lowest cytotoxic potential. Regarding FR-OE, it had the lowest extraction yield and composition in aroma volatiles. Additionally, all samples were described as having green, fresh and floral sensory notes, with no significant statistical differences regarding the odour detection threshold (ODT) values. Finally, FR-EX of T. mastichina obtained by SFE-CO2 presented the most promising results regarding food application.

Chemical Composition, Antioxidant, and Antimicrobial Activities of Leaves of Ajuga Iva.

The main objective of this research was to study the biological characteristics in terms of antioxidant and antimicrobial activities of Ajuga iva and determine the best analytical and extraction methods applicable to this specie and studied compounds. A short screening of its nutritional value in terms of chemical composition is also included. A. iva leaves were analyzed for crude protein (CP), cell wall [neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin (ADL)], minerals, fatty acids, essential oils, and phenolic compounds. Mature aerial parts of A. iva were randomly collected during the Spring season from Mograne-Zaghouan, Tunisia. Leaves of A. iva contained 13.4 ± 0.4% CP, 26.3 ± 0.35% NDF, 20.2 ± 0.42% ADF, and 5.13 ± 0.21% ADL. Mineral content (13.0 ± 0.45%) was mainly composed of potassium (4.5% g DM) and magnesium (4.25% DM). Leaves of A. iva had linolenic (26.29 ± 0.760%) and linoleic (37.66 ± 2.35%) acids as the main components of the acid profile. Thymol was found to be the most dominant (23.43%) essential oil, followed by 4-vinylguaiacol (14.27%) and linalool (13.66%). HPLC-PDA-ESI-MS/MS analysis pointed out the presence of phytoecdysteroids. Phenolic acids and flavonoids, such as glycosylated derivatives of naringenin, eriodyctiol, and apigenin, were detected in the methanol extract of A. iva leaves. Our results underline the importance of choosing proper extraction methods and solvents to extract and characterize the described compounds profile of A. iva leaves. Results also show A. iva leaves as a potential source of functional ingredients with beneficial health-promoting properties. Overall, leaves of A. iva have low biological activities (antioxidant and antimicrobial activities) with a chemical composition suitable as a feed for ruminants in rangeland pasture. It also has low-grade antibacterial or medicinal characteristics when fed to ruminants.

Antifungal and antibiofilm activities of the essential oil of leaves from Lippia gracilis Schauer against phytopathogenic fungi.

AIMS: This study aimed to evaluate the antifungal and antibiofilm effects of essential oil (EO) from leaves of Lippia gracilis and its major constituents, thymol and carvacrol, against phytopathogenic fungi. METHODS AND RESULTS: The leaves of L. gracilis were hydrodistilled to obtain the EO and the chemical composition was determined by GC/MS analysis. The antifungal activity of EO of L. gracilis was evaluated on the vegetative and mycelial growth of Colletotrichum gloeosporioides, Colletotrichum lindemuthianum, Fusarium oxysporum and Fusarium solani. In addition, the ability of the oil to inhibit fungal biofilm formation was assessed by total biomass quantification using crystal violet staining, analysis of metabolic activity, and scanning electron microscopy (SEM). Moreover the antifungal and antibiofilm activities of the monoterpenes, thymol and carvacrol, present in EO of L. gracilis were evaluated against F. oxysporum. The analysis of the chemical composition of EO extracted from L. gracilis, revealed the presence of monoterpenes (94·13%), which included carvacrol (48·57%) and thymol (7·78%), and 4 sesquiterpenes (3·74%). In general, EO showed significant antifungal activity and inhibited the formation of fungal biofilms. Furthermore, thymol and carvacrol showed significant antifungal and antibiofilm activities against F. oxysporum. SEM images showed structural changes in fungal morphology upon treatment with EO of L. gracilis. CONCLUSION: The results presented in this study showed promising antifungal and antibiofilm effects of EO of L. gracilis and its major components, carvacrol and thymol. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate that the EO extracted from L. gracilis, and the monoterpenes, carvacrol and thymol have a great potential as antifungal and antibiofilm agents. Furthermore, this is the first report of the antibiofilm activity of the EO of L. gracilis and its major components against phytopathogenic fungi.

Voltammetric Determination of Isopropylmethylphenols in Herbal Spices.

Thymol and carvacrol-the components of herbal spices-are known for their broad biological activity as antimicrobials and antioxidants. For this reason, it is important to develop new methods for their determination in plant material. A simple, rapid, and sensitive method for determination of total content of these analytes in herbal spices using differential pulse voltammetry (DPV) has been developed. The basis of the research is the oxidation process of isopropylmethylphenols on a platinum microelectrode in glacial acetic acid containing acetonitrile (20%, v/v) and 0.1 mol L-1 sodium perchlorate as the supporting electrolyte. Linear voltammetric responses for thymol and carvacrol were obtained in a wide concentration range from 0.39-1105 and 0.47-640 µg mL-1, with a low detection limit of 0.04 and 0.05 µg mL-1, respectively. The analysis was performed using the multiple standard addition method. The results of the voltammetric determination are in good agreement with the data of the standard chromatographic method. To the best of our knowledge, this is the first presentation of an electrochemical procedure to determine these compounds in these environmental and electrode materials.

Origanum vulgare ssp. hirtum (Lamiaceae) Essential Oil Prevents Behavioral and Oxidative Stress Changes in the Scopolamine Zebrafish Model.

Origanum vulgare ssp. hirtum has been used as medicinal herbs promoting antioxidant, anti-inflammatory, antimicrobial, and neuroprotective activities. We investigated the protective effects and the mechanism of O. vulgare ssp. hirtum essential oil (OEO) on cognitive impairment and brain oxidative stress in a scopolamine (Sco)-induced zebrafish (Danio rerio) model of cognitive impairment. Our results show that exposure to Sco (100 µM) leads to anxiety, spatial memory, and response to novelty dysfunctions, whereas the administration of OEO (25, 150, and 300 µL/L, once daily for 13 days) reduced anxiety-like behavior and improved cognitive ability, which was confirmed by behavioral tests, such as the novel tank-diving test (NTT), Y-maze test, and novel object recognition test (NOR) in zebrafish. Additionally, Sco-induced brain oxidative stress and increasing of acetylcholinesterase (AChE) activity were attenuated by the administration of OEO. The gas chromatography-mass spectrometry (GC-MS) analyses were used to elucidate the OEO composition, comprising thymol (38.82%), p-cymene (20.28%), and γ-terpinene (19.58%) as the main identified components. These findings suggest the ability of OEO to revert the Sco-induced cognitive deficits by restoring the cholinergic system activity and brain antioxidant status. Thus, OEO could be used as perspective sources of bioactive compounds, displaying valuable biological activities, with potential pharmaceutical applications.

A New Method for Determination of Thymol and Carvacrol in Thymi herba by Ultraperformance Convergence Chromatography (UPC2).

Ultraperformance convergence chromatography is an environmentally friendly analytical technique for dramatically reducing the use of organic solvents compared to conventional chromatographic methods. In this study, a rapid and sensitive ultraperformance convergence chromatography method was firstly established for quantification of thymol and carvacrol, two positional isomers of a major bioactive in the volatile oil of Thymi herba, the dried leaves and flowers of Thymus mongolicus or Thymus przewalskii, known in China as "Dijiao." Using a TrefoilTM CEL1 column, thymol and carvacrol were separated in less than 2.5 min and resolution was enhanced. The method was validated with respect to precision, accuracy, and linearity according to the National Medical Products Administration guidelines. The optimized method exhibited good linear correlation (r = 0.9998-0.9999), excellent precision (relative standard deviations (RSDs) < 1.50%), and acceptable recoveries (87.29-102.89%). The limits of detection for thymol and carvacrol were 1.31 and 1.57 ng/L, respectively, while their corresponding limits of quantification were 2.63 and 3.14 ng/L. Finally, the quantities of the two compounds present in 16 T. mongolicus and four T. przewalskii samples were successfully evaluated by employing the developed method. It is hoped that the results of this study will serve as a guideline for the quality control of Thymi herba.

Ultrasound combined with manganese-oxide nanoparticles loaded on activated carbon for extraction and pre-concentration of thymol and carvacrol in methanolic extracts of Thymus daenensis, Salvia officinalis, Stachys pilifera, Satureja khuzistanica, and mentha, and water samples.

A dispersive micro solid-phase extraction (DMSPE) technique was developed using manganese-oxide nanoparticles loaded on activated carbon (Mn3O4-NPs-AC) as an effective sorbent combined with ultrasound for the extraction and determination of a trace amount of thymol and carvacrol in methanolic extracts of Thymus daenensis, Salvia officinalis, Stachys pilifera, Satureja khuzistanica and mentha, and water samples. Thymol and carvacrol phenolic compounds were extracted from real samples using acetonitrile (ACN) as the desorption solvent. Using central composite design (CCD), the effects of pH, ionic strength (NaCl), nano-sorbent mass, contact time, and desorption volume were investigated. Additionally, based on five-level variables, response surface methodology was used to determine the individual and interactive effects between factors on the process. The optimized extraction conditions included 12 mg of Mn3O4-NPs-AC as the sorbent, 300 µL of ACN as the desorption solvent, pH 3.0, 0.5 w/v% of NaCl, and 4.5 min sonication time. Under the optimized conditions, for all the samples, the limits of detection were 0.054-0.104 ng mL-1 and the limits of quantification were 0.178-0.345 ng mL-1. The correlation coefficients of the calibration curves were >0.985, i.e. in the range of 0.4-6000 ng mL-1. To validate the effects of the matrix, the recovery, reproducibility, repeatability, and overall uncertainty were calculated for the five methanolic extracts, at 50, 100, and 500 ng mL-1. The recovery ranged between 94.5% and 109.0% with a relative standard deviation of <8.0% for the repeatability and reproducibility precision, which strongly supports the favorable repeatability and reproducibility of the method. The presented method also has the excellent sorbent features of NPs for the sorption of the analyte, which is due to the use of ultrasound for dispersion of the material in the sample matrix.

Composition of Essential Oils from Roots and Aerial Parts of Carpesium divaricatum, a Traditional Herbal Medicine and Wild Edible Plant from South-East Asia, Grown in Poland.

Carpesium divaricatum Sieb. and Zucc. has long been used both as traditional medicine and seasonal food. The most extensively studied specialized metabolites synthesized by the plant are sesquiterpene lactones of germacrane-type. Low-molecular and volatile terpenoids produced by C. divaricatum, however, have never been explored. In this work, compositions of essential oils distilled from roots and shoots of C. divaricatum plants, cultivated either in the open field or in the glasshouse have been studied by GC-MS-FID supported by NMR spectroscopy. The analyses led to the identification of 145 compounds in all, 112 of which were localized in aerial parts and 80 in roots of the plants grown in the open field. Moreover, remarkable differences in composition of oils produced by aerial and underground parts of C. divaricatum have been observed. The major volatiles found in the shoots were: α-pinene (40%), nerol (4%) and neryl-isobutyrate (3%), whereas predominant components of the root oil were 10-isobutyryloxy-8,9-epoxythymyl-isobutyrate (29%), thymyl-isobutyrate (6%) and 9-isobutyryloxythymyl-isobutyrate (6%). In the analyzed oils, seventeen thymol derivatives were identified. Among them eight compounds were specific for roots. Roots of the plants cultivated in the glasshouse were, in general, a poor source of essential oil in comparison with those of the plants grown in the open field. Chemophenetic relationships with other taxa of the Inuleae-Inulineae were also briefly discussed.

Effect of thymol on the broiler chicken antioxidative defence system after sustained dietary thyme oil application.

1. The purpose of this study was to examine if the concentration of thymol as the main compound of Thymus vulgaris essential oil (TEO) influenced the antioxidant defence system in broilers. 2. Twenty-four broiler chickens were randomly divided at the day of hatching into three dietary treatment groups (0%, 0.05% and 0.1%, w/w TEO) with eight birds in each and were fed until four weeks of age. 3. Thymol content in plasma, duodenal wall and breast muscle significantly increased when 0.1% of thyme oil was added to the diet (P < 0.05). Thymol concentration in plasma significantly correlated with levels measured in the duodenal wall and feed (rs = 0.7857, P < 0.05; rs = 0.7647, P < 0.05). Superoxide dismutase (SOD) activity increased, and malondialdehyde (MDA) concentration decreased significantly (P < 0.05) in blood from chickens fed 0.1% TEO supplementation. Although the thymol concentration did not significantly decrease MDA amounts in breast muscle, a declining trend was observed. 4. The trial data confirmed the efficient absorption of thymol from the digestive tract into the systemic circulation, but only traces were found in breast muscle. Thymol content was sufficient for expressing its antioxidant properties in blood, but its low content in breast muscle was insufficient to significantly affect lipid oxidation and fatty acid composition.