New adduct of abietane-type diterpene from Salvia leriifolia Benth.

A new adduct of abietane-type diterpene, salvialeriicone (1), was isolated from Salvia leriifolia Benth., along with a new chemical entity nor-abietane diterpene, 2-isopropyl-8,8-dimethyl-7,8-dihydrophenanthrene-1,4,5(6H)-trione (2). Their structures were determined using mass spectrometry, and 1D- and 2D-NMR spectroscopy.     

Standardization of officinal herb mixture by gas-liquid chromatography

An antibacterial herb mixture is investigated by gas-liquid chromatography with a mass-selective detector. The mixture consists of Matricaria chamomilla (flores), Bidem tripartite (folia), Calendula officinalis (flores), Salvia officinalis (folia), Eucalyptus globulus (folia), and Glycyrrhiza glabra (radices). The specific substances are described. Terpenoids for Salvia, eucalyptol and ledol for Eucalyptus, and naphtalene derivatives for Calendula are among them.     

Comparison of solid-phase and single-drop microextractions for headspace analysis of herbal essential oils

The analytical microextraction methods of gas chromatography coupled with flame ionisation detector (GC-FID) for determination of selected essential oils in herbs were proposed. Two microextraction methods for the isolation of essential oils from plants such as Lavandula spica L., Melissa officinalis L., Mentha piperita L. and Salvia officinalis L. were used. The methods of solid-phase and single-drop microextractions, were optimised and compared. The obtained LOD values for all studied essential oils were found to be within 2.5–20.5 μg for SDME and 57.0–139.8 μg for SPME method per 100 g of dried sample leaves. The appropriate LOQ values were then 8.4–68.4 μg for SDME and 189.8–466.1 μg for SPME of target analytes per 100 g of dried sample leaves.      

Forensic analysis of <Emphasis Type="Italic">Salvia divinorum</Emphasis> using multivariate statistical procedures. Part I: discrimination from related <Emphasis Type="Italic">Salvia</Emphasis> species

Salvia divinorum is a hallucinogenic herb that is internationally regulated. In this study, salvinorin A, the active compound in S. divinorum, was extracted from S. divinorum plant leaves using a 5-min extraction with dichloromethane. Four additional Salvia species (Salvia officinalis, Salvia guaranitica, Salvia splendens, and Salvia nemorosa) were extracted using this procedure, and all extracts were analyzed by gas chromatography–mass spectrometry. Differentiation of S. divinorum from other Salvia species was successful based on visual assessment of the resulting chromatograms. To provide a more objective comparison, the total ion chromatograms (TICs) were subjected to principal components analysis (PCA). Prior to PCA, the TICs were subjected to a series of data pretreatment procedures to minimize non-chemical sources of variance in the data set. Successful discrimination of S. divinorum from the other four Salvia species was possible based on visual assessment of the PCA scores plot. To provide a numerical assessment of the discrimination, a series of statistical procedures such as Euclidean distance measurement, hierarchical cluster analysis, Student’s t tests, Wilcoxon rank-sum tests, and Pearson product moment correlation were also applied to the PCA scores. The statistical procedures were then compared to determine the advantages and disadvantages for forensic applications.     

Chemical Composition Analysis and Antimicrobial Screening of the Essential Oil of a Rare Plant from Jordan: Ducrosia flabellifolia

The composition of the essential oil isolated from the fresh and dry leaves of Ducrosia flabellifolia Boiss. (Apiaceae) was determined by gas chromatography and gas chromatography–mass spectrometry using hydrodistillation and solid phase microextraction (SPME). The hydrodistilled oil of the fresh leaves yielded 38 components, accounting for 98.67% of the total oil content, while thirty components were detected from the fresh leaves by solid phase microextraction (94.85%). Fifty-one and 36 components were identified in the hydrodistilled and SPME oils of the dried leaves amounting to 98.78% and 94.52%, respectively. A total of 25 components accounting for 97.24% of the total composition were characterized in the SPME oil of the fresh flowers. Aliphatic compounds predominated in the volatile fractions of the leaves and flowers of both methods with n-decanol, n-decanal, and dodecanal as the main constituents. The α- and ß-pinene were the major monoterpenoids in the oils. The hydrodistilled oil was screened for its antimicrobial and antioxidant activities. The minimal inhibitory concentration of the volatile oil was determined using a microdilution method in 96 well plates against a panel of gram (+), gram (?) bacteria, and fungi. Overnight cultures of reference strains of Candida albicans, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus were used as test microorganisms. The oil exhibited the best activity against C. albicans (MIC 234 µg/mL) and S. aureus (MIC 234 µg/mL) whereas weak activity was detected against E. coli and P. aeruginosa. No antioxidant activity could be detected.     

1‐Methoxyhexane‐3‐thiol,a Powerful Odorant of Clary Sage (Salvia sclarea L.)

The peculiar and highly diffusive odor signal of flowering clary‐sage plants (Salvia sclarea L.) was identified to derive from trace amounts of 1‐methoxyhexane‐3‐thiol ( 1 ) by mass‐spectrometry analysis and confirmed by comparison with synthetic racemic thiol (±)‐ 1 . The enantiomers (S)‐ and (R)‐ 1 were prepared by enantioselective synthesis, and the absolute configuration of (S)‐ 1 was fully corroborated by X‐ray‐diffraction analysis of the crystalline thioester (1′S,1S)‐ 2 . Compound (S)‐ 1 is one of the most powerful odorants known, with a detection threshold of 0.04⋅10⁻³ ng/l air, and is, with its herbaceous‐green, alliaceous, and perspiration profile, key to the fragrance of clary‐sage flowers and of the freshly distilled essential oil. As a consequence of its unique odor, 1 was also suspected to be part of the volatiles of a Ruta species where it was subsequently identified together with its homologue, 1‐methoxyheptane‐3‐thiol ( 3 ), 1‐methoxy‐4‐methylpentane‐3‐thiol ( 4 ), and the known 4‐methoxy‐2‐methylbutane‐2‐thiol ( 5 ). The syntheses of (±)‐ 3 and (±)‐ 4 as well as of the enantiomer (R)‐ 4 are described. In both natural fractions, the ratio (S)‐ 1 /(R)‐ 1 was slightly in favor of the (S)‐enantiomer. Natural 4 has (R)‐configuration.     

The effect of harvest periods on the chemical compositions of essential oils of sage (Salvia aucheri L.) leaves

The essential oils of sage leaves (Salvia aucheri Bentham var. canescens Boiss. & Heldr.), growing wild in South Anatolia, were extracted by hydrodistillation and analysed by GC and GC-MS. The percentage yields of the essential oils from sage leaves harvested at different years were 1.0%, 1.3%, 1.3%, 1.0%, 1.4%, 1.5% and 1.2%, respectively. In this study, 1,8-cineole, camphre, camphene α-pinene and β-pinene were identified as the major components of sage leaves collected at different periods. The main constituents of sage oil collected over the years were 1,8-cineole (35.01-48.06%), camphre (13.58-23.92%), camphene (6.77-8.82%), α-pinene (5.79-8.54%) and β-pinene (4.32-6.28%).     

Comparative essential oil composition of aerial parts of Tanacetum dumosum Boiss. from Southern Zagros,Iran.

The essential oils of leaves and flowers of Tanacetum dumosum Boiss., an endemic medicinal shrub, were extracted by using hydrodistillation method and analysed using GC and GC–MS. A total of 43 and 44 compounds were identified in the essential oils from the leaves and flowers of T. dumosum, respectively. The major chemical constituents of leaves oil were borneol (27.9%), bornyl acetate (18.4%), 1,8-cineol (17.5%), α-terpineol (5.3%), cis-chrysanthenyl acetate (3.3%), camphene (2.7%) and terpinene-4-ol (1.9%), while the main components of the flower oil were isobornyl-2-methyl butanoate (41.1%), trans-linalyl oxide acetate (11.9%), 1,8-cineole (7.7%), thymol (4.2%), linalool (3.9%), camphor (2.9%), isobornyl propanoate (2.9%), α-terpineol (2.1%) and caryophyllene oxide (2.0%). Major qualitative and quantitative variations for some main chemical compounds among different aerial parts of T. dumosum were identified. High contents of borneol, bornyl acetate, 1,8-cineol and linalool in the leaves and flowers of T. dumosum show its potential for use in the food and perfumery industry.     

Comparison of the components of the essential oils from leaves and fruits of Grammosciadium platycarpum

The composition of the hydrodistilled essential oils obtained from dried leaves and fruits of Grammosciadium platycarpum Boiss. & Hausskn. were determined by GC and GC/MS. Twenty-five compounds (87.0%) and sixteen constituents (96.2%) were identified in the leaf and fruit oils, respectively. Linalool (26.1 and 53.9%), (E,E)-α-farnesene (24.1 and 20.4% ) and (Z)-β-santalol (10.6 and 10.9%) were the major components in the leaf and fruit oils. Published in Khimiya Prirodnykh Soedinenii, No. 6, pp. 559–560, November–December, 2006.     

The Changes in Bioactive Compounds and Antioxidant Activity of Chia (Salvia hispanica L.) Herb under Storage and Different Drying Conditions: A Comparison with Other Species of Sage

Studies on herb chia (Salvia hispanica L.) are very limited. Therefore, the aim of this study was to assess how different drying methods and periods of storage affect the bioactive properties of the herb Salvia hispanica and to compare it with other species of sage (Salvia officinalis L. and Salvia sclarea L.). In fresh herbs, directly after drying (freeze-drying, natural drying, and drying at 30, 40, and 50 °C), and after storage (3, 6, and 12 months), the following analyses were performed: content of total carotenoids and total polyphenols, polyphenol profile (including 25 compounds), and antioxidant activity. Additionally, the basic chemical compositions of the herbs were analyzed. To the best of our knowledge, the content of total carotenoids and the quantitative polyphenol profile in Salvia hispanica and Salvia sclarea were evaluated for the first time. The obtained results showed that the barely investigated herb Salvia hispanica is rich in polyphenolic compounds and shows high antioxidant activity. In all the tested species, rosmarinic acid was the most abundant polyphenolic compound. The use of different drying methods allowed us to determine that freeze-drying was the most effective for preserving polyphenols and carotenoids. Long-term storage up to 12 months resulted in a gradual reduction in antioxidant activity and in the content of polyphenols and carotenoids.     

Volatile composition of Brassica oleracea L. var. costata DC leaves using solid‐phase microextraction and gas chromatography/ion trap mass spectrometry

Volatile and semi‐volatile components of internal and external leaves of Brassica oleracea L. var. costata DC, grown under different fertilization regimens, were determined by headspace solid‐phase microextraction (HS‐SPME) combined with gas chromatography/ion trap mass spectrometry (GC/ITMS). Forty‐one volatiles and non‐volatile components were formally identified and thirty others were tentatively identified. Qualitative and quantitative differences were noticed between internal and external leaves. In general, internal leaves exhibited more aldehydes and sulfur volatile compounds than external ones, and less ketone, terpenes and norisoprenoid compounds. The fertilization regimens influenced considerably the volatile profile. Fertilizations with higher levels of sulfur produced Brassica leaves with more sulfur volatiles. In opposition, N and S fertilization led to leaves with lower levels of norisoprenoids and terpenes. Copyright © 2009 John Wiley & Sons, Ltd.     

Labdane Diterpenoids from Salvia tingitana Etl. Synergize with Clindamycin against Methicillin-Resistant Staphylococcus aureus

Quorum-sensing (QS) is a regulatory mechanism in bacterial communication, important for pathogenesis control. The search for small molecules active as quorum-sensing inhibitors (QSI) that can synergize with antibiotics is considered a good strategy to counteract the problem of antibiotic resistance. Here the antimicrobial labdane diterpenoids sclareol (1) and manool (2) extracted from Salvia tingitana were considered as potential QSI against methicillin-resistant Staphylococcus aureus. Only sclareol showed synergistic activity with clindamycin. The quantification of these compounds by LC–MS analysis in the organs and in the calli of S. tingitana showed that sclareol is most abundant in the flower spikes and is produced by calli, while manool is the major labdane of the roots, and is abundant also in the leaves. Other metabolites of the roots were abietane diterpenoids, common in Salvia species, and pentacyclic triterpenoids, bearing a γ-lactone moiety, previously undescribed in Salvia. Docking simulations suggested that 1 and 2 bind to key residues, involved in direct interactions with DNA. They may prevent accessory gene regulator A (AgrA) binding to DNA or AgrA activation upon phosphorylation, to suppress virulence factor expression. The antimicrobial activity of these two compounds probably achieves preventing upregulation of the accessory gene regulator (agr)-regulated genes.     

Analysis of Volatile Organic Compounds from Dendranthema indicum var. aromaticum by Headspace Gas Chromatography-Mass Spectrometry and Accurate Mass Measurement

The volatile organic compounds from flowers, leaves, and stems of Dendranthema indicum var. aromaticum, obtained through a static headspace technique, were analyzed by gas chromatography-mass spectrometry (GC-MS) and accurate mass measurement. The qualitative approach, comprising accurate mass measurement, retention index, and mass spectral search, was utilized to identify compounds. A total of 162 components were identified, representing 97.55–98.72% of the volatiles of individual samples. The principal chemical components in flowers were bornyl acetate (15.40%), α-phellandrene (14.18%), p-cymene (9.64%), camphor (9.54%), β-linalool (8.61%), and α-thujone (7.06%). In leaves, the main components were p-cymene (20.42%), bornyl acetate (20.41%), α-phellandrene (13.67%), and β-linalool (5.46%). As for stems, trans-β-farnesene (17.95%), germacrene D (12.89%), β-phellandrene (12.70%), β-caryophyllene (10.18%), and bicyclogermacrene (8.01%) were the dominant volatile compounds. Comparative studies on the volatiles from various species of genus Dendranthema indicated that Dendranthema indicum var. aromaticum contains significantly more aroma compounds than its morphologically similar species.     

Fatty Acid Composition of Seed Oils of Twelve Salvia Species Growing in Turkey

Seed oils of 12 Salvia species collected from different regions in Turkey (S. Albimaculata Hedge & Hub.-Mor., S. Candidisima Vahl., S. Cedronella Boiss., S. Cryptantha Montbret & Aucher ex Bentham, S. Forskahlei L., S. Fruticosa Miller (Sin. S. Triloba L. Fil), S. Halophila Hedge, S. Hypargeia Fisch. & Mey., S. Sclarea L., S. Tomentosa Miller, S. Tchihatcheffii (Fisch. & Mey.) Boiss., S. Virgata Jacq.) were obtained by Soxhlet apparatus using hexane. The oil yields were found to be between 2.0% and 20.9%. Fatty acids in the oils were converted to methyl esters and determined by GC/MS in methyl ester form. The main fatty acid components of S. halophila, S. Hypargeia, and S. Sclarea are unsaturated oleic, linoleic, and linolenic acids. In others except S. Candidissima, the dominant acids are oleic, linoleic and palmitic acids.     

Optimization of the Extraction Conditions and Simultaneous Quantification by RP-LC of Six Alkaloids in Evodiae Fructus

A reversed phase liquid chromatographic (LC) method coupled with DAD (250 nm) has been developed and validated for simultaneous quantification of six alkaloids, dehydroevodiamine (1), wuzhuyuamide-I (2), 5-hydroxyrutaecarpine (3), 14-formyldihydrorutaecarpine (4), evodiamine (5) and rutaecarpine (6), in 12 batches evodiae fructus [the dried, unripe fruits of Evodia rutaecarpa (Juss.) Benth. or E. rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang, E. rutaecarpa (Juss.) Benth. var. bodinieri (Dode) Huang] as a traditional Chinese medicine. The method was carried out by a C₁₈ column (250 × 4.6 mm) with a gradient mobile phase of methanol, acetonitrile, and phosphoric acid–triethylamine–buffer solution. The contents of 1–6 in the evodiae fructus could easily be determined within 70 min. The experimental results were satisfactory for the intra-day and inter-day precision and accuracy of the method for simultaneous determination. The linear calibration ranges of 1–6 were 40–1,000, 20–500, 1–100, 10–500, 40–1,000 and 80–1,000 μg mL⁻¹. The recoveries of 1–6 were 97.43–103.73% with RSDs from 0.21 to 1.99%. The limits of detection for 1–6 were 2.0, 2.0, 0.1, 1.0, 5.0 and 5.0 μg mL⁻¹, and the limits of quantification were 6.6, 6.6, 0.3, 3.3, 16.5 and 16.5 μg mL⁻¹. The method was successfully applied to the quantification of six alkaloids in the evodiae fructus.     

Forensic analysis of <Emphasis Type="Italic">Salvia divinorum</Emphasis> using multivariate statistical procedures. Part II: association of adulterated samples to <Emphasis Type="Italic">S. divinorum</Emphasis>

Salvia divinorum is a plant material that is of forensic interest due to the hallucinogenic nature of the active ingredient, salvinorin A. In this study, S. divinorum was extracted and spiked onto four different plant materials (S. divinorum, Salvia officinalis, Cannabis sativa, and Nicotiana tabacum) to simulate an adulterated sample that might be encountered in a forensic laboratory. The adulterated samples were extracted and analyzed by gas chromatography–mass spectrometry, and the resulting total ion chromatograms were subjected to a series of pretreatment procedures that were used to minimize non-chemical sources of variance in the data set. The data were then analyzed using principal components analysis (PCA) to investigate association of the adulterated extracts to unadulterated S. divinorum. While association was possible based on visual assessment of the PCA scores plot, additional procedures including Euclidean distance measurement, hierarchical cluster analysis, Student’s t tests, Wilcoxon rank-sum tests, and Pearson product moment correlation were also applied to the PCA scores to provide a statistical evaluation of the association observed. The advantages and limitations of each statistical procedure in a forensic context were compared and are presented herein.     

Comparison of different extraction methods for the determination of α‐ and β‐thujone in sage (Salvia officinalis L.) herbal tea

Salvia officinalis L. (sage) is an important industrial plant used both for food and pharmaceutical purposes. The terpene fraction of this plant is responsible for many of its therapeutic and culinary properties. We used different extraction methods Tenax TA® purge and trap, headspace (HS) solid‐phase microextraction, HS sorptive extraction, and stir bar sorptive extraction to analyze the terpene fraction extracted from sage tea by GC–MS. Twenty compounds were identified, including α‐, β‐thujone, and several other oxygenated monoterpenes (1,8‐cineole, linalool, camphor, boneol, and bornyl acetate) and oxygenated sesquiterpenes (caryophyllene oxide, viridiflorol, humulene epoxide I, II, and III). Tenax TA® and HS sorptive extraction extracted a lower number of identified compounds, whereas HS solid‐phase microextraction allowed the complete extraction of volatiles with particular reference to α‐ and β‐thujone. The importance of the determination of thujones content in sage herbal tea is also discussed.     

LC Determination of Peroxynitrite Scavenging Activity of Phenols from Salvia spp.

The aim of this study was to adopt pyrogallol red bleaching test for LC-DAD capable of characterizing chemical constituents and peroxynitrite (ONOO⁻) scavenging activity in parallel for a large number of plant extracts. The hypothesis is that upon reaction with ONOO⁻, the peak areas of compounds in complex mixtures with potential radical scavenging activity in the LC chromatograms will be significantly reduced or disappeared. We validated this approach with a model mixture of 17 phenolic compounds, which mimics a general alcoholic extract of Salvia species. The LC separation conditions were optimized for baseline separation of phenolic reference compounds and for selective detection of the test substrate pyrogallol red. The results indicated that the depletion kinetic of compounds in the model mixture correlates moderately with the individual scavenging activities. Moreover, catechol group dependent relationship between the structure–scavenging activity relationships confirmed the LC based assay parameters. The final demonstration of the assay was by methanolic extract of Salvia miltiorrhiza Bunge, which showed outstanding ONOO⁻ scavenging activity. 13 min total analysis time per sample and excellent resolution allow the method to be applied for chemical fingerprinting coupled with rapid screening for natural antioxidants derived from alcoholic extracts of Salvia species.

     

Determination of Oleanolic, Betulinic and Ursolic Acid in Lamiaceae and Mass Spectral Fragmentation of Their Trimethylsilylated Derivatives

Phytochemical investigations of different species of Lamiaceae family (rosemary-Rosmarinus officinalis L., sage-Salvia officinalis L., winter savory-Satureja montana L., clary sage-Salvia sclarea L. and sticky sage-Salvia glutinosa L.), using gas chromatography and mass spectrometry (GC-MS) were performed. The studies were focused on oleanolic, betulinic and ursolic acid. Since oleanolic and ursolic acid are position isomers with very similar structures, the difficulties in their separation and identification have been reported by several authors. However, both compounds can be well distinguished by order of elution during gas chromatography and by intensities of the fragment ion signals in their mass spectra, where the retro-Diels-Alder reaction was primarily observed. A combination of ultrasonic extraction, solid phase extraction, size exclusion chromatography, trimethylsilylation, and GC-MS resulted in simultaneous separation, identification and quantification of the mentioned compounds. The compounds have been identified by retention time and comparison of mass spectra with those of pure standards. The mass spectral fragmentation behavior of all three derivatised acids was investigated. The obtained characteristic fragment patterns are discussed in the presented work. Good linearity over the concentration range 1–50 mg L⁻¹ for all three compounds was confirmed. The correlation coefficients (r ² ) were in the range of 0.9980–0.9983. Quantitative analyses of different Lamiaceae extracts showed that the oleanolic acid content ranged from 0.09 to 0.9% dry weight, content of betulinic acid ranged from traces to 0.6%, and that of ursolic acid varied from 0.09 to 1.6% dry weight.     

Headspace Sorptive Extraction (HSSE) in the Headspace Analysis of Aromatic and Medicinal Plants

A new sampling technique, Headspace Sorptive Extraction (HSSE), is here applied for the first time to the headspace sampling of medicinal and aromatic plants. The analyte partition coefficient between HSSE‐PDMS stir bar and sample headspace (K₁), the concentration factor (CF), the reproducibility, and the minimum recoverable amount were determined by analyzing standard solution of high volatility C₅–C₇ compounds with different polarities and structures (cyclohexane, propyl acetate, hexanal, 1‐hexen‐3‐ol, isoamyl acetate, and 2‐heptanol). Four aromatic and medicinal plants, viz. rosemary (Rosmarinus officinalis L.), sage (Salvia officinalis L.), thyme (Thymus vulgaris L.), and valerian (Valeriana officinalis L.) were analyzed by HSSE‐GC with PDMS stir bars, and their concentration capacity was compared with those of S‐HS and HS‐SPME with different fibers. HSSE showed very high concentration capability with both standard and real sample components.