Abietane diterpenoids from Isodon lophanthoides var. graciliflorus and their cytotoxicity

Seven new (1–7) and three known (8–10) abietane diterpenoids were isolated from the methanolic extract of the aerial parts of Isodon lophanthoides var. graciliflorus (Lamiaceae), a folk Chinese medicine and an herb for functional beverages. They were identified as 16-acetoxylsugiol (1), graciliflorin E (2), graciliflorin F (3), 15-O-methylgraciliflorin F (4), 15-hydroxy-20-deoxocarnosol (5), 3β-hydroxysempervirol (6), 15-hydroxy-1-oxosalvibretol (7), abieta-8,11,13-triene-14,19-diol (8), 6,12,15-trihydroxy-5,8,11,13-abietatetraen-7-one (9), and 3α-hinokiol (10) based on the spectroscopic data including COSY (correlated spectroscopy), HMBC (heteronuclear multiple bond correlation), and HR-ESI-MS (high-resolution electrospray ionization mass spectrometry). All the compounds except 10 were obtained from I. lophanthoides for the first time. Compounds 1, 2, 5, 6, 8, and 9 exhibited in vitro cytotoxicity against A549 (human lung adenocarcinoma), MCF-7 (human breast adenocarcinoma), and HeLa (human cervical carcinoma) cell lines with the IC₅₀ values of 1.79–52.67 μM.     

Chemical and biological characteristics of Cuminum cyminum and Rosmarinus officinalis essential oils

Essential oils extracted by hydrodistillation from Cuminum cyminum and Rosmarinus officinalis were characterized by means of GC and GC–MS. C. cyminum and R. officinalis contained α-pinene (29.1%, 14.9%), 1,8-cineole (17.9%, 7.43%) and linalool (10.4%, 14.9%), respectively, as the major compounds. C. cyminum oil exhibited stronger antimicrobial activity than did R. officinalis oil against E. coli, S. aureus and L. monocytogenes. Complete death time on exposure to Cuminum cyminum L. and Rosmarinus officinalis L. oils were 20 and 25 min 180 and 240 min and 90 and 120 min for E. coli, S. aureus and L. monocytogenes, respectively. Radical-scavenging and antioxidant properties were tested by means of 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and the β-carotene bleaching test. These properties were compared to those of Thymus x-porlock essential oil, used as a reference ingredient. The radical scavenging performance of the rosemary oil was better than that of C. cyminum. Results from the antioxidant test were better than those provided by the radical-scavenging activity. C. cyminum and R. officinalis essential oils may be considered as potent agents in food preservation.     

Phytochemical profile of Rosmarinus officinalis and Salvia officinalis extracts and correlation to their antioxidant and anti-proliferative activity

The goal of this study was to monitor the anti-proliferative activity of Rosmarinus officinalis and Salvia officinalis extracts against cancer cells and to correlate this activity with their phytochemical profiles using liquid chromatography/diode array detection/electrospray ion trap tandem mass spectrometry (LC/DAD/ESI-MSⁿ). For the quantitative estimation of triterpenic acids in the crude extracts an NMR based methodology was used and compared with the HPLC measurements, both applied for the first time, for the case of betulinic acid. Both extracts exerted cytotoxic activity through dose-dependent impairment of viability and mitochondrial activity of rat insulinoma m5F (RINm5F) cells. Decrease of RINm5F viability was mediated by nitric oxide (NO)-induced apoptosis. Importantly, these extracts potentiated NO and TNF-α release from macrophages therefore enhancing their cytocidal action. The rosemary extract developed more pronounced antioxidant, cytotoxic and immunomodifying activities, probably due to the presence of betulinic acid and a higher concentration of carnosic acid in its phytochemical profile.     

Antimicrobial activities of methanol extracts and essential oils of Rosmarinus officinalis, depending on location and seasonal variations

Rosmarinus officinalis is widely found in the lands of Aegean and Mediterranean regions of Turkey. The goal of this work was to test the antimicrobial activity of the essential oils and methanolic extracts of R. officinalis collected from three different regions at four different time intervals of the year against Staphylococcus aureus, Proteus vulgaris, Pseudomonas aeruginosa, Klebsiella pneumonia, Enterococcus feacalis, Escherichia coli, Staphylococcus epidermidis, Bacillus subtilis and Candida albicans. Essential oils were obtained from the aerial parts of the plant by using a Clevenger apparatus, for 4 h. After distillation, the distillates were filtered, air-dried and then extracted by using a Soxhlet apparatus for 9 h to obtain the methanolic extracts. The antimicrobial activities of the methanolic extracts were tested by the disc diffusion technique. The antimicrobial activities of the essential oils obtained from R. officinalis were determined by minimum inhibitory concentration (MIC).The results indicated that the tested bacteria were sensitive to the essential oils and partially to the methanolic extracts. The antimicrobial activities of the essential oils against the tested bacteria differed, depending on location and seasonal variations.     

Analysis of the anti-inflammatory properties of Rosmarinus officinalis L. in mice

The aim of this study was to investigate the anti-inflammatory effects of the crude extract (CE) of Rosmarinus officinalis L. its derived fractions: hexane (HEX), ethyl acetate (AcOEt), and ethanolic (ET), and isolated compounds: carnosol, betulinic acid and ursolic acid, in the mouse pleurisy model induced by carrageenan. Swiss mice were used in the in vivo experiments. The CE and its derived fractions and isolated compounds inhibited leukocytes, exudation, interleukin-1 beta (IL-1β) and tumour necrosis factor-alpha (TNF-α) levels, myeloperoxidase activity (MPO), and nitrite/nitrate production (NO_x) (p < 0.05). R. officinalis L. showed an important anti-inflammatory activity by inhibition not only of leukocytes and exudation, but also of a pro-inflammatory enzyme and mediators (MPO, NO_x, IL-1β, and TNF-α). The present study showed that carnosol, betulinic acid and ursolic acid compounds could be responsible for this anti-inflammatory effect.     

Inhibitory effect of Turkish Rosmarinus officinalis L. on acetylcholinesterase and butyrylcholinesterase enzymes

In the current study, we have tested acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activity of the petroleum ether, ethyl acetate, chloroform, and methanol extracts, rosmarinic acid as well as the essential oil obtained from Rosmarinus officinalis L. growing in Turkey by a spectrophotometric method of Ellman using ELISA microplate-reader at 0.2, 0.5, and 1.0 mg/mL concentrations. In addition, quantification of rosmarinic acid, a common phenolic acid found in rosemary, was carried out by reversed-phase HPLC in the methanolic extract of the plant, which was found to have 12.21 ± 0.95% (122.1 ± 9.5 mg/g extract) of rosmarinic acid. Rosmarinic acid was also tested for its AChE and BChE inhibitory effect and found to cause 85.8% of inhibition against AChE at only 1.0 mg/mL. Besides, the essential oil was analyzed by GC–MS technique, which was shown to be dominated by 1,8-cineol (44.42%) and followed by α-pinene (12.57%).     

In vitro antioxidant activities of Rosmarinus officinalis extracts treated with supercritical carbon dioxide

The leaves of Rosmarinus officinalis (rosemary) were subjected to supercritical CO₂ extraction (SFE). Different sources of variability, including location (Izmir, Canakkale and Mersin) and harvesting time (December, March, June and September), were considered. Among active constituents of rosemary, carnosic acid, carnosol and rosmarinic acid were analyzed by HPLC. Variability of the amounts of active constituents appears to be due to different geographical locations of growth and seasonal variations. The levels of the constituents were higher in the months of December 2003 and September 2004. In addition to this, 12 SFE extracts were screened for their radical-scavenging capacities and antioxidant activities by various in vitro assays, namely total phenol assay, DPPH radical-scavenging activity and trolox equivalent antioxidant capacity (TEAC).     

Antioxidative activity of Rosmarinus officinalis L. essential oil compared to its main components

This study was designed to examine the in vitro antioxidant activities of Rosmarinus officinalis L. essential oil compared to three of its main components (1,8-cineole, α-pinene, β-pinene). GC–MS analysis of the essential oil resulted in the identification of 19 compounds, representing 97.97% of the oil, the major constituents of the oil were described as 1,8-cineole (27.23%), α-pinene (19.43%), camphor (14.26%), camphene (11.52%) and β-pinene (6.71%). The oil and the components were subjected to screening for their possible antioxidant activity by means of 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and β-carotene bleaching test. In the DPPH test system, free radical-scavenging activity of R. officinalis L. essential oil, 1,8-cineole, α-pinene and β-pinene were determined to be 62.45% ± 3.42%, 42.7% ± 2.5%, 45.61% ± 4.23% and 46.21% ± 2.24% (v/v), respectively. In the β-carotene bleaching test system, we tested series concentration of samples to show the antioxidant activities of the oil and its main components, whereas the concentrations providing 50% inhibition (IC₅₀) values of R. officinalis L. essential oil, 1,8-cineole, α-pinene and β-pinene were 2.04% ± 0.42%, 4.05% ± 0.65%, 2.28% ± 0.23% and 2.56% ± 0.16% (v/v), respectively. In general, R. officinalis L. essential oil showed greater activity than its components in both systems, and the antioxidant activities of all the tested samples were mostly related to their concentrations. Antioxidant activities of the synthetic antioxidant, ascorbic acid and BHT, were also determined in parallel experiments as positive control.     

Comparative evaluation of the antibacterial activities of the essential oils of Rosmarinus officinalis L. obtained by hydrodistillation and solvent free microwave extraction methods

Rosmarinus officinalis L. is a perennial herb that belongs to the Lamiaceae family. It is used as a food flavouring agent, and well known medicinally for its powerful antimutagenic, antibacterial and chemopreventive properties. Essential oils were obtained from this plant by hydrodistillation (HD) and solvent free microwave extraction (SFME). GC–MS analyses of the oils revealed the presence of 24 and 21 compounds in the essential oils obtained through HD and SFME, respectively. The total yield of the volatile fractions obtained through HD and SFME was 0.31% and 0.39%, respectively. Higher amounts of oxygenated monoterpenes such as borneol, camphor, terpene-4-ol, linalool, α-terpeneol (28.6%) were present in the oil of SFME in comparison with HD (26.98%). However, HD oil contained more monoterpene hydrocarbons such as α-pinene, camphene, β-pinene, myrcene, α-phellanderene, 1,8-cineole, trans β-ocimene, γ-terpenene, and cis sabinene hydrate (32.95%) than SFME extracted oil (25.77%). The essential oils obtained using the two methods of extraction were active against all the bacteria tested at a concentration of 10 mg ml⁻¹. Minimum inhibitory concentration (MIC) values for all the susceptible bacteria ranged between 0.23 mg ml⁻¹ and 7.5 mg ml^−1.     

Composition of rosemary essential oil (Rosmarinus officinalis) as affected by drying method

The influence of the drying method on volatile compounds of Rosmarinus officinalis was evaluated. The drying methods tested were convective (CD) and vacuum-microwave (VMD), as well as a combination of convective pre-drying and VM finish-drying (CPD–VMFD). Rosemary drying kinetics was described by a simple exponential model for CD and VMD, while VMFD kinetics consisted of two periods: linear until a critical point and exponential beyond that point. Volatile compounds of rosemary samples were extracted by steam-hydrodistillation and analyzed by GC–MS. Thirty-four compounds were tentatively identified, with α-pinene, bornyl acetate, camphene and 1,8-cineole being the major components. The total volatiles concentration of fresh rosemary (135 g kg⁻¹) decreased considerably during both CD (87.2 g kg⁻¹) and VMD (61.9 g kg⁻¹). CPD–VMFD was the best option for drying rosemary because the time required was relatively short (30 min), and the aroma quality was good according to both instrumental (100 g kg⁻¹) and sensory analyses.     

Suppression of LPS-induced inflammatory activities by Rosmarinus officinalis L.

Rosemary (Rosmarinus officinalis L.) has been used in folk medicine to treat headaches, epilepsy, poor circulation, and many other ailments. It was found that rosemary could act as a stimulant and mild analgesic and could reduce inflammation. However, the mechanisms underlying the anti-inflammatory effects of rosemary need more study to be established. Therefore, in this study, the effects of rosemary on the activation of nuclear factor kappa beta (NF-kB) and mitogen-activated protein kinases (MAPKs), the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and the production of nitric oxide (NO), prostaglandin E₂ (PGE₂), and cytokine in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells were investigated. A methanol extract of rosemary and its hexane fraction reduced NO generation with an IC₅₀ of 2.75 and 2.83 μg/ml, respectively. Also, the methanol extract and the hexane fraction inhibited LPS-induced MAPKs and NF-kB activation associated with the inhibition of iNOS or COX-2 expression. LPS-induced production of PGE₂ and tumour necrosis factor-alpha (TNF-α) were blocked by rosemary. Rosemary extract and its hexane fraction are important for the prevention of phosphorylation of MAPKs, thereby blocking NF-kB activation, which in turn leads to decreased expression of iNOS and COX-2, thus preventing inflammation.     

Chemical composition, mineral content and antioxidant activity of Verbena officinalis L.

Aqueous and hydroalcoholic extracts from Verbena officinalis L. were obtained and characterised. The analysis by HPLC-DAD and LC-MS allowed the detection and identification of three iridoids, fifteen flavonoids and four phenolic acid derivatives. Four flavonoids, scutellarein 7-diglucuronide (9), scutellarein 7-glucuronide (13), pedalitin 6-galactoside (15) and scutellarein 7-glucoside (19) are reported for the first time from this plant. In addition, three new flavonoids have been isolated: scutellarein 7-O-(2-O-feruloyl)-diglucuronide (5), pedalitin 6-O-diglucuronide (6) and pedalitin 6-O-(2-O-feruloyl)-diglucuronide (13). To our knowledge, these flavonoids have not been reported as natural products. Both extracts showed significant antioxidant activity using three in vitro model systems and the results have been correlated with total phenolic and total flavonoid contents. The results have allowed establishing an important relation structure-activity and significant correlations have also been found between the mineral content and the flavonoids present in both extracts.     

Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component

Buddleia officinalis Maxim, commonly used as rice dye for festivals, was extracted with ethanol using microwave-assisted extraction and Soxhlet extraction. The antioxidant activities of microwave-assisted extract of B. officialis (MEB) and Soxhlet extract of B. officianils (SEB) at the optimum extraction conditions were evaluated and compared with synthetic antioxidant butylated hydroxytoluene (BHT) employing DPPH free radical assay, ABTS assay, total antioxidant activity and reducing power. MEB and SEB had stronger antioxidant activities than BHT in all assays except reducing power, and the effects decreased as follows: MEB > SEB > BHT. The total phenolic contents of MEB and SEB reached 113.56 mg/g and 100.94 mg/g dry weight of extract, respectively, expressed as pyrocatechol equivalents, while the total flavonoids contents were 75.33 mg/g and 62.56 mg/g dry weight of extract, respectively, expressed as catechin equivalents (P < 0.05). Higher phenolic and flavonoids compounds may be major contributors to their higher antioxidant activities. Following activity-oriented separation, luteolin was isolated as an active principle, which exhibited excellent free radical scavenging activities with DPPH IC₅₀ 3.09 μg/ml and ABTS IC₅₀ 2.20 μg/ml.     

Antibacterial activity of the essential oils of Salvia officinalis L. and Salvia triloba L. cultivated in South Brazil

The essential oils of Salvia officinalis and Salvia triloba cultivated in South Brazil were analyzed by GC–MS. The major constituents of the oil of S. officinalis were α-thujone, 1,8-cineole, camphor, borneol and β-pinene, whereas those of S. triloba were α-thujone, 1,8-cineole, camphor, and β-caryophyllene. The essential oils of both species exhibited remarkable bacteriostatic and bactericidal activities against Bacillus cereus, Bacillus megatherium, Bacillus subtilis, Aeromonas hydrophila, Aeromonas sobria, and Klebsiella oxytoca. Moreover, the essential oil of S. triloba efficiently inhibited the growth of Staphylococcus aureus. S. aureus and A. hydrophila growth were drastically reduced even in the presence of 0.05 mg/ml of the essential oil of S. triloba.     

Chemical composition and in vitro antioxidative activity of a lemon balm (Melissa officinalis L.) extract

The leaf material of lemon balm (Melissa officinalis L.) was extracted with 450 ml/l aqueous ethanol by medium pressure liquid-solid extraction. The total phenolic content of the extract was estimated as gallic acid equivalents by Folin-Ciocalteu reagent method and a qualitative-quantitative compositional analysis was carried out using high performance liquid chromatography coupled with photodiode array detection. The lemon balm extract contained hydroxycinnamic acid derivatives and flavonoids with caffeic acid, m-coumaric acid, eriodictyol-7-O-glucoside, naringin, hesperidin, rosmarinic acid, naringenin, hesperetin being identified based on their chromatographic behaviour and spectral characteristics. The extract was also investigated for potential in vitro antioxidant properties in iron(III) reduction, iron(II) chelation, 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulphonate), superoxide anion and nitric oxide free-radical scavenging, and inhibition of β-carotene-linoleic acid bleaching assays. The extract demonstrated antioxidant activity in all the assays. However, it was not as potent as the positive controls except in the β-carotene-linoleic acid bleaching assay, where its activity was superior to that of gallic and caffeic acids and statistically indistinguishable from quercetin and BHA. The exceptionally high antioxidant activity and the fact that this assay is of biological relevance warrants further investigation of lemon balm extract in ex vivo and in vivo models of oxidative stress.     

Antioxidant abietane diterpenoids from Salvia barrelieri

The root extract of endemic Algerian Salvia species Salvia barrelieri Ettling and its diterpenoids were investigated for potential antioxidant activity. From its acetone extract, a new natural abietane diterpenoid 7-oxoroyleanone-12-methyl ether (1) and six known diterpenoids 7α-acetoxyroyleanone-12-methyl ether (2), royleanone (3), horminone (4), 7-acetylhorminone (5), cryptojaponol (6) and inuroyleanol (7) were isolated, and their structures were elucidated by spectroscopic means. Among the diterpenoids, the new diterpenoid 7-oxoroyleanone-12-methyl ether (1) showed highest superoxide anion scavenging activity while inuroyleanol (7) showed both the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity and inhibition of lipid peroxidation in β-carotene–linoleic acid system. These findings indicate that S. barrelieri extract as well as isolated abietane diterpenes, particularly inuroyleanol are promising antioxidants which can be used as food additives.     

Phenolic profiles of cultivated, in vitro cultured and commercial samples of Melissa officinalis L. infusions

Melissa officinalis L. (lemon balm) is normally consumed as an infusion and presents therapeutic properties, such as sedative, carminative and antispasmodic, also being included in some pharmaceutical preparations. The phenolic profiles of different samples of lemon balm, prepared as infusions, were evaluated by HPLC-DAD-ESI/MS. The profiles were compared in order to understand the differences between cultivated, in vitro cultured and commercial (bags and granulated) samples. All the samples showed a similar phenolic profile, presenting differences only in the quantities found of each compound. Rosmarinic acid was the most abundant compound, being higher in commercial samples, especially in tea bag sample (55.68 mg/g of infusion) and lower in in vitro cultured sample (15.46 mg/g). Moreover, dimers, trimers and tetramers of caffeic acid were identified and quantified for the first time in lemon balm. Only one flavonoid, luteolin-3′-O-glucuronide was found in all the samples, ranging from 8.43 mg/g in commercial granulate sample to 1.22 mg/g in in vitro cultured sample. Overall, cultivated and in vitro cultured samples presented the lowest amounts of phenolic compounds (59.59 and 30.21 mg/g, respectively); otherwise, commercial samples showed the highest contents (109.24 mg/g for tea bag and 101.03 mg/g for granulate sample). The present study shows that infusion of lemon balm can be a source of phenolic compounds, known for their bioactive effects.     

Changes in valerenic acids content of valerian root (Valeriana officinalis L. s.l.) during long-term storage

Dried valerian (Valeriana officinalis) root powder was stored at 5, 14 and 30 °C under low, moderate and high humidity conditions for 6 months, and the level of the valerenic acids monitored on a monthly basis. From an initial moisture content of 5 g/kg, the moisture level during storage decreased in root powder stored at low humidity and increased during storage at high humidity, with the effect greatest at 30 °C.     

Flavonoid pattern of sage (Salvia officinalis L.) unifloral honey

The aim of the present paper was to determine the flavonoids in monofloral sage (Salvia officinalis L.) honey which is characteristic and specific for the area of Croatian coast and islands. For that purpose 38 sage honey samples from two production seasons were analysed. After specific pollen content determination, and analyses of selected physicochemical parameters which confirmed that samples are in compliance with national and international regulations and can be regarded as unifloral sage honeys, flavonoid fraction was isolated and analysed using RP-HPLC/DAD method. The HPLC analysis showed that all examined sage honey samples contain quercetin (3,3′,4′,5,7-pentahydroxyflavone), luteolin (3′,4′,5,7-tetrahydroxyflavone), kaempferol (3,4′,5,7-tetrahydroxyflavone), apigenin (4′,5,7-trihydroxyflavone), chrysin (5,7-dihydroxyflavone) and galangin (3,5,7-trihydroxyflavone), as well as p-coumaric (trans-4-hydroxycinnamic acid) and caffeic acid (3,4-dihydroxycinnamic acid). Total amount of identified flavonoids varied from 109.4 μg/100 g of honey to 589.9 μg/100 g of honey, with the average of 288.5 μg/100 g of honey. All analysed honey samples showed common and specific flavonoid profile which could be the basis for differentiating sage from other monofloral honeys.