Optimization of carvacrol,rosmarinic, oleanolic and ursolic acid extraction from oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.)

The aim of our study was to increase the extraction efficiency of carvacrol, rosmarinic, oleanolic and ursolic acid from the different species of oregano herbs (Origanum onites L., Origanum vulgare spp. hirtum and Origanum vulgare L.). Various extraction methods (ultrasound-assisted, heat-reflux, continuous stirring, maceration, percolation) and extraction conditions (different solvent, material:solvent ratio, extraction temperature, extraction time) were used, and the active substances were determined by HPLC. The lowest content of carvacrol, rosmarinic, oleanolic and ursolic acid was obtained by percolation. During heat-reflux extraction, the content of active substances depended on the solvent used: ethanol/non-aqueous solvent (glycerol or propylene glycol) mixture was more effective compared with ethanol alone. The results showed that for each species of oregano the most optimal extraction method should be selected to maximize the content of biologically active substances in the extracts.     

Antioxidant capacity of Ocimum basilicum L. and Origanum vulgare L. extracts

The antioxidant properties of five different extracts (Et?O, CHCl?, EtOAc, n-BuOH, and H?O) of Ocimum basilicum L. and Origanum vulgare L. were studied. Antioxidant activity was assessed in six different model systems. Free radical scavenging capacity (RSC) was evaluated by measuring the scavenging capacity of extracts on DPPH, NO, O??? and OH radical, as well as on hydrogen peroxide (H?O?). In addition, the protective effects on lipid peroxidation in liposomes (LPx) were evaluated by TBA-assay using the Fe2?/ascorbate induction system. The amount of total phenolic compounds and content of total flavonoids was also determined. EtOAc, n-BuOH and H?O extracts of O. basilicum and O. vulgare expressed very strong scavenger activity. Furthermore, the mentioned extracts showed notable inhibition of LPx. On the other hand, Et?O and CHCl? extracts showed much weaker effect in the neutralization of DPPH, NO and O??? radicals and the neutralization of H?O?. When examining the production of OH radicals and inhibition of LPx, the Et?O and CHCl? extracts showed weak prooxidative properties. The observed differences in antioxidant activity could be partially explained by the levels of phenolics and flavonoids in the investigated O. basilicum and O. vulgare extracts.     

Chemical markers in Origanum vulgare L. from Kumaon Himalayas: a chemosystematic study

The essential oils of four wild growing Origanum vulgare L. (family Lamiaceae) collected from different locations in Kumaon region (Uttarakhand, India) were analysed by capillary GC and GC/MS. The comparative results of O. vulgare L. collected from four different regions showed differences in the chemical constituents of the essential oils. The oil of O. vulgare L. collected from Dhoulchina and Champawat (chemotype I) shows p-cymene (6.7-9.8%), γ-terpinene (12.4-14.0%), thymol (29.7-35.1%) and carvacrol (12.4-20.9%) as major constituents while the oil from Kilbury and Rushi village (chemotype II) shows linalool (6.7-9.7%), bornyl acetate (12.6-16.8%), β-caryophyllene (10.5-13.8%) and germacrene D (6.3-11.3%) as the major constituents. These features highlight the chemosystematics of this genus.     

Variation in the volatile constituents of Artemisia annua var. CIM-Arogya during plant ontogeny

The essential oils yield and composition of the aerial parts of A. annua var. CIM-Arogya grown in Uttarakhand, India were analyzed and compared by capillary GC and GC-MS at different stages of development. The analysis led to the identification of 81 constituents forming 91.0%-97.1% of the essential oils compositions. The essential oil content of the aerial parts was found to vary from 0.3% to 0.7% at different stages of growth. A. annua crop harvested at full flowering and seed setting stage gave higher yield of essential oil (0.6%, 0.7%) than that harvested at pre flowering (0.5%), late vegetative (0.4%, 0.5%), mid vegetative (0.4%, 0.4%) and early vegetative stages (0.3%, 0.3%). The essential oils at different stages of growth showed monoterpenoids (38.5%-72.0%) and sesquiterpenoids (22.2%-48.2%) as major grouped constituents. The major constituents identified were camphor (22.8%-42.6%), 1,8-cineole (3.7%-8.4%), linalool (<0.1%-11.9%), beta-caryophyllene (2.0%-9.2%), (E)-beta-farnesene (1.3%-8.5%), germacrene D (0.5%-7.3%) and 1-epi-cubenol (0.7%-5.2%) in essential oil samples collected at different crop stages.     

Agronomic,metabolomic and lipidomic characterisation of Sicilian Origanum vulgare (L.) ecotypes

Although Origanum vulgare (L.) has been deeply analysed at phytochemical level, poor knowledge is available regarding non-volatile compounds such as lipids. The aim of this work was to characterise five wild Sicilian Origanum ecotypes from an agronomic, metabolomic and lipidomic perspective. Serradifalco presented higher dry weight and inflorescences/plant than the others while Favara had a significantly higher number of branches per plant and more extensive flowered stratum. Metabolomic analysis, performed with LC-MS-TOF, allowed a preliminary characterisation of the non-volatile metabolome of the five oregano ecotypes Origanum vulgare ssp. hirtum. Twenty-five metabolites were identified belonging to organic acids, amino acids, lysophosphatidylcholines, carnithines, nucleic bases and lysophosphatidylethanolamines. Lipidomic analysis identified 115 polar plant membrane glycerolipid species. Thirteen of them were differentially present in the two chosen ecotypes. The role of these metabolites in plant physiology from a qualitative and pharmacological point of view was discussed.     

Multiple analytical approaches for the organic and inorganic characterization of Origanum vulgare L. samples

Origanum vulgare L. samples, marketed in different geographic locations, were characterized by their organic and inorganic chemical composition. A total of 35 commercial samples were collected from various sites and analyzed to determine the qualitative and quantitative profile of essential oils, phenolic compounds and some inorganic elements. The variation in the content and composition of the essential oil was assessed by GC and GC-MS analyses, the phenolic fraction was investigated by UPLC^®/PDA, and the inorganic elements were determined by ICP-MS. The Principal Component Analysis (PCA) was applied with the aim to sort out the Origanum vulgare L. samples with different composition according to the different belonging origins. The results showed appreciable qualitative and quantitative differences among samples from different geographic origin.     

Spectroscopy,Microscopy and Fluorescence Imaging of Origanum vulgare L. Basis for Nondestructive Quality Assessment

The organs of Origanum vulgare L. plant were examined by optical microscopy, scanning electron microscopy and autofluorescence imaging. The different organs were also studied spectroscopically. Fluorescence emission spectra were recorded for intact inflorescences, leaves and stems. Several fluorescence ratios (Blue/Red, Blue/Far‐red, Green/Red and Green/Far‐red), which varied depending on the considered organ of the plant, were derived. For leaves, a dependence of fluorescence spectra with water content was obtained as well. The intact samples were also analyzed by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy. These spectra were transformed to the Remission function depending on the wavenumber and two absorption bands (811 and 1740 cm^?1), which displayed differences according to the plant organ sampled, were detected. These results were consistent with higher carvacrol content in inflorescences. The spectroscopic results were connected with the microscopic observation and with the presence of relevant nutraceutics contained in the plant. The optical indexes derived in this work may serve as potential indicators to be explored in the development of nondestructive methods for oregano quality assessment.     

The Antimicrobial Activity of Origanum vulgare L. Correlated with the Gastrointestinal Perturbation in Patients with Metabolic Syndrome

(1) The metabolic syndrome (MS) promotes acute and chronic infections, due to the pro-inflammatory condition given by TNFα and IL6 or by affecting the microbiota. MS is also correlated with insulin resistance, causing inflammation and infections throughout the organism. (2) The purpose of this study was to track the effect of using the essential oil of Origanum vulgare L. (EOO) as an antibacterial treatment, compared to allopathic treatment with antibiotics in MS patients. A group of 106 people with MS was divided into four subgroups: L1—staphylococcal infection group, L2—Escherichia coli infection group, L3—streptococcal infection group with EOO treatment, and CG—control group without infection or treatment. (3) EOO is responsible for the antibacterial effect, and reduced minor uncomplicated infections. After a 10-day treatment, intestinal side effects were absent, improving the phase angle. (4) The results suggest that EOO may exhibit an antibacterial effect, similar to the antibiotic treatment, without promoting MS-specific dysbiosis, and it also improves the phase angle in patients, which is used as an index of health and cellular function.     

Anticholinesterase Activity of Selected Medicinal Plants from Navarra Region of Spain and a Detailed Phytochemical Investigation of Origanum vulgare L. ssp. vulgare

Alzheimer’s disease is a neurodegenerative disease characterized by progressive memory loss and cognitive impairment due to a severe loss of cholinergic neurons in specific brain areas. It is the most common type of dementia in the aging population. Although many anti-acetylcholinesterase (AChE) drugs are already available on the market, their performance sometimes yields unexpected results. For this reason, research works are ongoing to find potential anti-AChE agents both from natural and synthetic sources. In this study, 90 extracts from 30 native and naturalized medicinal plants are tested by TLC and Ellman’s colorimetric assay at 250, 125 and 62.5 μg/mL in order to determine the inhibitory effect on AChE. In total, 21 out of 90 extracts show high anti-AChE activity (75–100% inhibition) in a dose-dependent manner. Among them, ethanolic extract from aerial parts of O. vulgare ssp. vulgare shows an IC50 value 7.7 times lower than galantamine. This research also establishes the chemical profile of oregano extract by TLC, HPLC-DAD and LC-MS, and twenty-three compounds are identified and quantified. Dihydroxycinnamic acids and flavonoids are the most abundant ones (56.90 and 25.94%, respectively). Finally, total phenolic compounds and antioxidant properties are quantified by colorimetric methods. The total phenolic content is 207.64 ± 0.69 µg/mg of extract. The antioxidant activity is measured against two radicals, DPPH and ABTS. In both assays, the oregano extract shows high activity. The Pearson correlation matrix shows the relationship between syringic acids, a type of dihydroxybenzoic acid, and anti-AChE (r² = −0.9864) and antioxidant activity (r² = 0.9409 and 0.9976). In conclusion, the results of this study demonstrate promising potential new uses of these medicinal herbs for the treatment of Alzheimer’s. Origanum vulgare ssp. vulgare and syringic acids, which have anti-AChE activity and beneficial antioxidant capacity, can be highlighted as potential candidates for the development of drugs for the treatment of Alzheimer’s disease and other diseases characterized by a cholinergic deficit.     

Chemical variability in the essential oil of Cinnamomum tamala L. leaves from India

The chemical composition of the essential oils obtained from the fresh leaves of Cinnamomum tamala Nees et Eberm. was determined by GC and GC-MS. The yield of the oil on a dry weight basis ranged from 1.2% to 3.9% (w/w). Phenyl propanoids constitute the major portion (88.9-95.0%) of the oils. Fifty-four compounds were identified from the oils. Eugenol (91.4-41.8%) was the main compound, followed by eugenyl acetate (0.0-47.1%) and α-phellandrene (0.6-2.5%) in the analysed oils.     

Volatile oils from the plant and hairy root cultures of Ageratum conyzoides L

Two lines of hairy root culture of Ageratum conyzoides L. induced by Agrobacterium rhizogenes ATCC 15834 were established under either complete darkness or 16 h light/8 h dark photoperiod conditions. The volatile oil yields from aerial parts and roots of the parent plant, the hairy root culture photoperiod line and the hairy root culture dark line were 0.2%, 0.08%, 0.03% and 0.02%, (w/w), respectively. The compositions of the volatiles from the hairy roots, plant roots and aerial parts were analysed by GC and GC-MS. The main components of the volatiles from the hairy root cultures were β-farnesene, precocene I and β-caryophyllene, in different amounts, depending on light conditions and also on the age of cultures. Precocene I, β-farnesene, precocene II and β-caryophyllene were the main constituents of the volatile oils from the parent plant roots, whereas precocene I, germacrene D, β-caryophyllene and precocene II were the main constituents of the aerial parts of the parent plant. Growth and time-course studies of volatile constituents of the two hairy root lines were compared. Qualitative and quantitative differences were found between the volatile oils from the roots of the parent plant and those from the hairy roots.     

HPLC determination of antilipoxygenase activity of a water infusion of Ligustrum vulgare L. leaves and some of its constituents

The aim of the study was a HPLC evaluation of the lipoxygenase activity inhibiting activity of a water infusion of Ligustrum vulgare L. leaves and selected isolates from it. The antiradical activity of the water infusion was determined using DPPH, ABTS and FRAP tests. Oleuropein and echinacoside concentrations in the water infusion were determined by HPLC. Water infusion, echinacoside and oleuropein were used for an antilipoxygenase activity assay using lipoxygenase isolated from rat lung cytosol fraction. Activity of 8-LOX, 12-LOX and 15-LOX were monitored through formation of 8-HETE, 12-HETE and 15-HETE, respectively. The water infusion exhibited the highest activity against all lipoxygenases, followed by oleuropein. Echinacoside was ineffective against LOXs in lower concentrations, while higher concentration showed similar inhibition on 8-LOX and 12-LOX. 15-LOX was affected more and the presence of echinacoside remarkably decreased its activity.     

Antimycobacterial activity of constituents from Foeniculum vulgare var. dulce grown in Mexico

Bioassay guided fractionation of an antimycobacterial extract of Foeniculum vulgare var dulce (Apiaceae) led to the isolation and characterization of 5-hydroxyfurano-coumarin. The chemical structure of this compound was elucidated by 1H and 13C (1D and 2D) Nuclear Magnetic Resonance (NMR) spectroscopy. In addition, the active fractions were analyzed by GC-MS and seventy eight compounds were identified; the major compounds were 1,3-benzenediol, 1-methoxycyclohexene, o-cymene, sorbic acid, 2-hydroxy-3-methyl-2-cyclopenten-1-one, estragole, limonene-10-ol and 3-methyl-2-cyclopenten-1-one. Twenty compounds identified in the active fractions were tested against one sensitive and three MDR strains of Mycobacterium tuberculosis using the Alamar Blue microassay. Compounds that showed some degree of antimycobacterial activity against all strains tested were the following: linoleic acid (MIC 100 μg/mL), oleic acid (MIC 100 μg/mL), 1,3-benzenediol (MIC 100-200 μg/mL), undecanal (MIC 50-200 μg/mL), and 2,4-undecadienal (MIC 25-50 μg/mL), the last being the most active compound. To our knowledge, this is the first report of the presence of 5-hydroxy-furanocoumarin in F. vulgare.     

Crude extract of Origanum vulgare L. induced cell death and suppressed MAPK and PI3/Akt signaling pathways in SW13 and H295R cell lines

Oregano (Origanum vulgare L.) is a common aromatic plant used in Mediterranean and Asian Regions for treating respiratory diseases, painful menstruation, rheumatoid arthritis, etc. Recently its role as an anticancer plant has been suggested, although oregano has been never evaluated into adrenocortical tumour cell models. This study analysed for the first time the anticancer effects of a crude extract of wild mountain oregano (Origanum vulgare L.) in SW13 and H295R cell lines. The crude extract was characterised by GC/MS and the toxic effects of oregano were first analysed by brine shrimp lethality assay. Our findings demonstrated that oregano decreased cell viability, survival, modified cell cycle and induced cell death (through necrotic process) and that the effects can be attributed to a blockade of MAPK and PI3 K/Akt pathways. These results suggest that oregano extract exerts anticancer activities in adrenocortical tumour cell lines, providing evidence for further research in higher models.     

Volatile constituents of essential oil and rose water of damask rose (Rosa damascena Mill.) cultivars from North Indian hills

Rosa damascena Mill. is an important aromatic plant for commercial production of rose oil, water, concrete and absolute. The rose water and rose oil produced under the mountainous conditions of Uttarakhand were investigated for their chemical composition. The major components of rose water volatiles obtained from the bud, half bloom and full bloom stages of cultivar 'Ranisahiba' were phenyl ethyl alcohol (66.2-79.0%), geraniol (3.3-6.6%) and citronellol (1.8-5.5%). The rose water volatiles of cultivar 'Noorjahan' and 'Kannouj' also possessed phenyl ethyl alcohol (80.7% and 76.7%, respectively) as a major component at full bloom stage. The essential oil of cultivar 'Noorjahan' obtained from two different growing sites was also compared. The major components of these oils were citronellol (15.9-35.3%), geraniol (8.3-30.2%), nerol (4.0-9.6%), nonadecane (4.5-16.0%), heneicosane (2.6-7.9%) and linalool (0.7-2.8%). This study clearly showed that the flower ontogeny and growing site affect the composition of rose volatiles. The rose oil produced in this region was comparable with ISO standards. Thus, it was concluded that the climatic conditions of Uttarakhand are suitable for the production of rose oil of international standards.     

Volatile chemical constituents of Piper aduncum L and Piper gibbilimbum C. DC (Piperaceae) from Papua New Guinea

Exhaustive hydro-distillation of the leaves of Piper aduncum and fruits of Piper gibbilimbum (Piperaceae) afforded colorless and pale orange colored oils in 0.35 and 0.30 % yields, respectively. Detailed chemical analysis by GC/MS indicated the volatile constituents of Piper aduncum to be composed of dill apiole (43.3%), beta-caryophyllene (8.2%), piperitione (6.7%) and alpha-humulene (5.1%), whilst the oil of P. gibbilimbum is dominated by the gibbilimbols A-D (74.2%), with the remaining major constituents being the terpenes camphene (13.6%) and alpha-pinene (6.5%).     

Layered double hydroxide films on nanoporous anodic aluminum oxide/aluminum wire: a new fiber for rapid analysis of Origanum vulgare essential oils

Zn/Al layered double hydroxide (LDH) films were fabricated in situ with anodic aluminium oxide aluminium as both the substrate and the sole aluminium source by means of urea hydrolysis. Headspace solid phase microextraction using LDH fibre in combination with capillary GC–MS was utilised as a monitoring technique for the collection and detection of the volatile compounds of Origanum vulgare. Experimental parameters, including the sample weight, microwave power, extraction time and humidity effect, were examined and optimised.     

Larvicidal activity of plant extracts from Colombian North Coast against Aedes aegypti L. mosquito larvae

The Aedes aegypti L. mosquito is considered the most important vector of arboviruses in the world. Insecticide-resistant phenomenon is a difficult barrier to overcome for government health entities around the planet. This problem forces to increase the concentrations of insecticides in the environment causing environmental pollution and health threat to human beings. Plants have been used to combat pests for centuries and are an eco-friendly source for the search for molecules with larvicidal activity. In this work, 65 ethanol-soluble extracts from 56 plants of the Colombian Caribbean region were evaluated as potential larvicides against Aedes aegypti mosquito, as well as, for their toxic effects on non-target organism. A high larvicidal activity was found for 16 ethanol extracts, however, the most potent activity against the larvae was obtained for the five plant extracts corresponding to the seeds of Annona squamosa L., Annona cherimolia L., Annona muricata L., Tabernaemontana cymosa Jacq. and Mammea americana L., with LC₅₀ value of 58.44, 65.10, 84.92, 25.02 and 38.58 ppm, respectively. T. cymosa seeds extract was selected for bio-guided fractionation due to the high larvicidal activity showed. Through chromatographic techniques and Nuclear Magnetic Resonance (NMR), five indole alkaloids were isolated and characterized from T. cymosa active fraction; among them, voacangine showed activity with a LC₅₀ of 5.13 ppm indicating a high larvicidal potency besides low toxicity against model organism Caenorhabditis elegans. We also report a new indole alkaloid from T. cymosa. Our study demonstrated the potential of the Colombian Caribbean flora as a host of bioactive plants against important vectors such as the A. aegypti mosquito with potential use in controlled environments.     

Changes in Phenols,Polysaccharides and Volatile Profiles of Noni (Morinda citrifolia L.) Juice during Fermentation

The change in phenols, polysaccharides and volatile profiles of noni juice from laboratory- and factory-scale fermentation was analyzed during a 63-day fermentation process. The phenol and polysaccharide contents and aroma characteristics clearly changed according to fermentation scale and time conditions. The flavonoid content in noni juice gradually increased with fermentation. Seventy-three volatile compounds were identified by solid-phase microextraction coupled with gas chromatography–mass spectrometry (SPME-GC-MS). Methyl hexanoate, 3-methyl-3-buten-1-ol, octanoic acid, hexanoic acid and 2-heptanone were found to be the main aroma components of fresh and fermented noni juice. A decrease in octanoic acid and hexanoic acid contents resulted in the less pungent aroma in noni juice from factory-scale fermentation. The results of principal component analysis of the electronic nose suggested that the difference in nitrogen oxide, alkanes, alcohols, and aromatic and sulfur compounds, contributed to the discrimination of noni juice from different fermentation times and scales.