Volatile constituents and key odorants in leaves, buds, flowers, and fruits of Laurus nobilis L

The volatiles of fresh leaves, buds, flowers, and fruits from bay (Laurus nolilis L.) were isolated by solvent extraction and analyzed by capillary gas chromatography-mass spectrometry. Their odor quality was characterized by gas chomatography-olfactometry-mass spectrometry (HRGC-O-MS) and aroma extract dilution analysis (AEDA). In fresh bay leaves 1,8-cineole was the major component, together with alpha-terpinyl acetate, sabinene, alpha-pinene, beta-pinene, beta-elemene, alpha-terpineol, linalool, and eugenol. Besides 1,8-cineole and the pinenes, the main components in flowers were alpha-eudesmol, beta-elemene, and beta-caryophyllene, in fruits (E)-beta-ocimene and biclyclogermacrene, and in buds (E)-beta-ocimene and germacrene D. The aliphatic ocimenes and farnesenes were absent in leaves. By using HRGC-O-MS 21 odor compounds were identified in fresh leaves. Application of AEDA revealed (Z)-3-hexenal (fresh green), 1,8-cineole (eucalyptus), linalool (flowery), eugenol (clove), (E)-isoeugenol (flowery), and an unidentified compound (black pepper) with the highest flavor dilution factors. Differences between buds, flowers, fruits, and leaves with regard to the identified odor compounds are presented.     

Insecticidal activity and chemical composition of volatile oils from Hyptis martiusii Benth

The essential oils from leaves and inflorescences of Hyptis martiusii Benth were analyzed by GC-MS. Twenty-six compounds representing 93.2% of the essential oil of leaves were characterized; Delta-3-carene (22.5%), 1,8-cineole (24.27%), beta-caryophyllene (6.15%), and bicyclogermacrene (6.32%) were found as the major components. In the essential oil of inflorescences 27 compounds representing 87.7% of the oil were identified. The major components were Delta-3-carene (13.5%), alpha-pinene (5.78%), beta-caryophyllene (6.59%), viridiflorene (8.25%), and germacrene B (5.21%). The essential oil of leaves and 1,8-cineole showed pronounced insecticidal effect against Aedes aegypti larvae and Bemisia argentifolii, the vectors of dengue fever and white fly fruit plague, respectively.     

Evolution and occurrence of 1,8-cineole (eucalyptol) in Australian wine

A new method has been developed for the quantitation of 1,8-cineole in red and white wines using headspace solid-phase microextraction (SPME) combined with stable isotope dilution analysis (SIDA) and gas chromatography-mass spectrometry (GC-MS). An extensive survey of Australian wines (44 white and 146 red) highlighted that only red wines contained significant amounts of 1,8-cineole (up to 20 μg/L). Hydrolytic studies with limonene and α-terpineol, putative precursors to 1,8-cineole, showed a very low conversion into 1,8-cineole (< 0.6%) over a 2 year period, which does not account for the difference between white and red wines. 1,8-Cineole was chemically stable in model wine solution over 2 years, and absorption from a Shiraz wine by bottle closures was most evident for a synthetic closure only (14% absorption after 1 year). Two commercial ferments at two different locations were monitored daily to investigate the evolution of 1,8-cineole throughout fermentation. Both ferments showed daily increases in 1,8-cineole concentration while in contact with grape solids, but this accumulation ceased immediately after pressing. This observation is consistent with the extraction of 1,8-cineole into the ferment from the solid portions of the grape berries.     

Isolation and structural elucidation of some glycosides from the rhizomes of smaller galanga (Alpinia officinarum Hance)

Glycosidically bound compounds were isolated from the methanol extract of fresh rhizomes of smaller galanga (Alpinia officinarum Hance). Nine glycosides (1-9) were finally obtained by reversed-phase HPLC and their structures were elucidated by MS and NMR analyses. They were the three known glycosides, (1R,3S,4S)-trans-3-hydroxy-1,8-cineole beta-D-glucopyranoside (1), benzyl beta-D-glucopyranoside (3), and 1-O-beta-D-glucopyranosyl-4-allylbenzene (chavicol beta-D-glucopyranoside, 4); and the six novel glycosides, 3-methyl-but-2-en-1-yl beta-D-glucopyranoside (2), 1-hydroxy-2-O-beta-D-glucopyranosyl-4-allylbenzene (5), 1-O-beta-D-glucopyranosyl-2-hydroxy-4-allylbenzene (demethyleugenol beta-D-glucopyranoside, 6), 1-O-(6-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl)-2-hydroxy-4-allylbenzene (demethyleugenol beta-rutinoside, 7), 1-O-(6-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl)-4-allylbenzene (chavicol beta-rutinoside, 8), and 1,2-di-O-beta-D-glucopyranosyl-4-allylbenzene (9). Compounds 2-9 were detected for the first time as constituents of galanga rhizomes.     

Vineyard and fermentation studies to elucidate the origin of 1,8-cineole in Australian red wine

Preliminary investigations revealed that the proximity of Eucalyptus trees to grapevines can directly influence the concentration of the aroma compound 1,8-cineole present in the corresponding red wines. For two different vineyards, the closer the grapevines were to the trees, the greater was the amount of 1,8-cineole in the wines elaborated from those grapes. This led us to carry out further studies to quantify the levels of 1,8-cineole found in grape berries, leaves, and stems at set distances from Eucalyptus trees over multiple vintages. Generally, the highest concentration of 1,8-cineole was found in the grapevine leaves, followed by grape stems and then grapes. In each sample type, we observed greater concentrations of 1,8-cineole in samples closer to the trees. Various fermentation treatments carried out with Shiraz grapes showed that matter other than grapes (MOG, e.g., Eucalyptus or grape leaves) could contribute significant amounts of 1,8-cineole to the finished wines. These studies confirmed that vineyard position and winemaking conditions can determine the 1,8-cineole concentration in red wine. The fermentation study also showed for the first time that the concentration of rotundone in red wine can be strongly influenced by grapevine leaves and stems in the ferment.     

Chemotypical variation of tansy (Tanacetum vulgare L.) from 40 different locations in Norway

Between 2001 and 2002, plant collections from wild populations of Norwegian tansy (Tanacetum vulgare L.) were studied with a focus on essential oil (EO) yield and composition in order to characterize the chemotypical EO variability. Tansy collections of 40 different locations from North, Mid-, and South Norway were transplanted to the Apelsvoll Research Centre Div. Kise in 2000 and grown for 2 years before the aerial parts (leaves and flower buds) were harvested in June 2002. The EO from individual plants was isolated from dried plant material by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS) on a DB5 column at the Plant Biocenter. The EO yield ranged between 0.35 and 1.90% (v/w) (average: 0.81%); the most abundant thujone plants were especially rich in EO volatiles (0.95%). On the basis of GC-MS data, seven chemotypes could be identified as follows: A, alpha-thujone (two individuals); B, beta-thujone (22); C, camphor (six); D, chrysanthenyl acetate/chrysanthenol (three); E, chrysanthenone (two); F, artemisia ketone/artemisia alcohol (three); and G, 1,8-cineole (two). The thujone chemotype was dominated by beta-thujone (81%) associated with alpha-thujone, but tansy plants rich in alpha-thujone were also detected (61%). The chemotypical classification of Norwegian tansy genotypes was underscored by preliminary studies from 2001, indicating the genetic uniformity and biochemical stability of the domesticated plants.     

Investigation of bound aroma constituents of yellow-fleshed nectarines (Prunus persica L. Cv. Springbright). changes in bound aroma profile during maturation

Glycosidically bound volatile constituents of yellow-fleshed clingstone nectarines (cv. Springbright) were identified and quantified at three stages of maturity. Glycoconjugates were isolated by LC on a C(18) reversed phase column with methanol elution followed by hydrolysis with a commercial pectinase enzyme. Forty-five bound aglycons were identified for the first time in yellow-fleshed nectarine. Thirty were terpene derivatives, and the most abundant ones were (E)- and (Z)-furan linalool oxides, linalool, alpha-terpineol, (E)-pyran linalool oxide, 3,7-dimethylocta-1,5-diene-3,7-diol, linalool hydrate, 8-hydroxy-6,7-dihydrolinalool, (E)- and (Z)-8-hydroxylinalools, and (E)- and (Z)-8-hydroxygeraniols. The group of C(13) norisoprenoids included 3-hydroxy-beta-damascone, 3-hydroxy-7,8-dihydro-beta-ionone, 3-oxo-alpha-ionol, 3-hydroxy-7,8-dihydro-beta-ionol, 3-hydroxy-beta-ionone, 3-oxo-7,8-dihydro-alpha-ionol, 3-hydroxy-5,6-epoxy-beta-ionone, 3-oxo-retro-alpha-ionol (isomers I and II), 3-hydroxy-7,8-dehydro-beta-ionol, 4,5-dihydrovomifoliol, and vomifoliol. Generally, levels of bound compounds, in particular monoterpenols and C(13) norisoprenoids, increased significantly with maturation. delta-Decalactone was the only lactone found in the enzymatic hydrolysate of yellow-fleshed nectarine, but its level was much lower than that of its free form.     

Near infrared spectroscopy for cost effective screening of foliar oil characteristics in a Melaleuca cajuputi breeding population

The identification of Melaleuca cajuputi leaf samples (trees) that demonstrate enhanced oil characteristics using near infrared (NIR) spectroscopy is described. Leaf samples from an unthinned M. cajuputi seedling seed orchard in Indonesia were collected and air-dried, and their 1,8-cineole content and oil concentrations were determined. NIR spectra of the leaves were obtained, and calibrations for 1,8-cineole content and oil concentration were developed using spectra that had been selected using spectral features; that is, no knowledge of 1,8-cineole content or oil concentration was used to select the calibration samples. The calibrations were used to predict the 1,8-cineole content and oil concentration of the remaining samples. It was demonstrated that NIR spectroscopy could be used to identify leaf samples that had high 1,8-cineole contents and oil concentrations. The technique has the potential to greatly reduce the time involved in ranking large numbers of samples for these attributes, as is a requirement in tree breeding programs to enhance oil production.     

Screening of chemical composition and antifungal and antioxidant activities of the essential oils from three Turkish artemisia species

The compositions of essential oils isolated from the aerial parts of Artemisia absinthium, Artemisia santonicum, and Artemisia spicigera by hydrodistillation were analyzed by GC-MS, and a total of 204 components were identified. The major components of these essential oils were camphor (34.9-1.4%), 1,8-cineole (9.5-1.5%), chamazulene (17.8-nd%), nuciferol propionate (5.1-nd%), nuciferol butanoate (8.2-nd%), caryophyllene oxide (4.3-1.7%), borneol (5.1-0.6%), alpha-terpineol (4.1-1.6%), spathulenol (3.7-1.3%), cubenol (4.2-0.1%), beta-eudesmol (7.2-0.6%), and terpinen-4-ol (3.5-1.2%). The antifungal activities of these essential oils were tested against 11 plant fungi and were compared with that of a commercial antifungal reagent, benomyl. The results showed that all of the oils have potent inhibitory effects at very broad spectrum against all of the tested fungi. Pure camphor and 1,8-cineole, which are the major components of the oils, were also tested for antifungal activity against the same fungal species. Unlike essential oils, these pure compounds were able to show antifungal activity against only some of the fungal species. In addition, the antioxidant and DPPH radical scavenging activities of the essential oils, camphor, and 1,8-cineole were determined in vitro. All of the studied essential oils showed antioxidant activity, but camphor and 1,8-cineole did not.     

Volatile compounds released by enzymatic hydrolysis of glycoconjugates of leaves and grape berries from Vitis vinifera Muscat of Alexandria and Shiraz cultivars.

Glycoconjugates from Muscat of Alexandria and Shiraz leaves and grape berries were isolated by adsorption on Amberlite XAD-2 resin, and enzymatically released aglycons were analyzed by GC-FID and GC-MS. About 120 aglycons were fully or tentatively identified. Compositional differences were observed between leaves and berries of the two varieties in five aglycon chemical groups: C6 alcohols, aliphatic alcohols, monoterpenes, shikimates, and C(13)-norisoprenoids, which were much more abundant in the leaves than in the berries. The differences observed for C(13)-norisoprenoids were in agreement with their hypothetical independent biosynthesis in leaves and berries. Thus, 3-hydroxy-beta-damascone, an important norisoprenoid aglycon of grape berries, was not detected in leaves, whereas its oxidized derivative, 3-oxo-alpha-damascone, was absent in berries. Compositional differences were also observed between Muscat and Shiraz leaves. 3-Oxo-alpha-ionol was not detected in Shiraz leaves, and its retro derivatives were less abundant than in Muscat of Alexandria leaves. Conversely, in Shiraz leaves the levels of 7,8-dihydroionone derivatives, such as megastigman-3,9-diol and 3-oxo-7,8-dihydro-alpha-ionol, were higher than in Muscat of Alexandria leaves.     

Seasonal and phenological variations of the essential oil from the narrow endemic species Artemisia molinieri and its biological activities

The volatile components of the aerial parts of Artemisia molinieri, an endemic wormwood of southern France, were analyzed by GC and GC-MS. Among the 69 compounds identified, major components were ascaridole (19-76%), alpha-terpinene (traces-36%), p-cymene (1-17%), 1,8-cineole (0.3-8%), and germacrene D (0.6-15%). Quantitative variations have been characterized following the season, the phenological cycle, and the aging of the plants. Bioassays have been performed on a sample of essential oil, which has shown a strong inhibition of the growth of both tested yeasts (Candida albicans and Saccharomyces cerevisiae var. chevalieri) and minor activity on both tested Gram-negative bacteria (Escherichia coli and Enterococcus hirae). The oils have shown interesting antioxidant activities on the basis of alpha-tocopherol as reference compound, up to 400-1200%.     

Chemical composition of volatiles in Sardinian myrtle (Myrtus communis L.) alcoholic extracts and essential oils

The chemical composition of the volatile fraction of myrtle (Myrtus communis L.) alcoholic extracts and essential oils from leaves and berries collected in different places in Sardinia (Italy) was studied. A simple and rapid liquid-liquid extraction method was used to isolate volatile compounds from myrtle alcoholic extracts followed by GC and GC-MS analysis allowing the detection of 24 compounds. The volatile fraction was characterized by the terpenes fraction corresponding to that of the essential oils and by a fatty acid ethyl esters fraction. The variation during extraction of the volatile fraction in alcoholic extracts of berries and leaves was evaluated. Essential oils were obtained by hydrodistillation, and the yields were on average 0.52 +/- 0.03% (v/w dried weight) and 0.02 +/- 0.00% for leaves and berries, respectively. The essential oils were analyzed by GC and GC-MS, and a total of 27 components were detected, accounting for 90.6-98.7% of the total essential oil composition. Strong chemical variability depending on the origin of the samples was observed. The major compounds in the essential oils were alpha-pinene (30.0 and 28.5%), 1,8-cineole (28.8 and 15.3%), and limonene (17.5 and 24.1%) in leaves and berries, respectively, and were characterized by the lack of myrtenyl acetate.     

Volatile metabolites from Salvia fruticosa as antifungal agents in soilborne pathogens

The volatile metabolites of Salvia fruticosa plants, growing wild in 15 localities scattered across Greece, were analyzed by means of GC and GC-MS. The essential oil content ranged from 0.69 to 4.68%, and the results of the analyses showed a noticeable variation in the amounts of the five main components [1,8-cineole, alpha-thujone, beta-thujone, camphor, and (E)-caryophyllene]. The antifungal activities of the essential oils from two localities, belonging in two different groups of cluster and principal component analysis, and their main components (1,8-cineole and camphor) were evaluated in vitro against five phytopathogenic fungi. Both oils were slightly effective against Fusarium oxysporum f. sp. dianthi and Fusarium proliferatum, whereas against Rhizoctonia solani, Sclerotinia sclerotiorum, and Fusarium solani f. sp. cucurbitae the oils exhibited high antifungal activities.     

Flavor release and perception in hard candy: influence of flavor compound-compound interactions

The influence of flavor compound-compound interactions on flavor release properties and flavor perception in hard candy was investigated. Hard candies made with two different modes of binary flavor delivery, (1) L-menthol and 1,8-cineole added as a mixture and (2) L-menthol and 1,8-cineole added separate from one another, were analyzed via breath analysis and sensory time-intensity testing. Single-flavor candy containing only L-menthol or 1,8-cineole was also investigated via breath analysis for comparison. The release rates of both L-menthol and 1,8-cineole in the breath were more rapid and at a higher concentration when the compounds were added to hard candy separate from one another in comparison to their addition as a mixture (conventional protocol). Additionally, the time-intensity study indicated a significantly increased flavor intensity (measured as overall cooling) for hard candy made with separate addition of these flavor compounds. In conclusion, the flavor properties of hard candy can be controlled, at least in part, by flavor compound-compound interactions and may be altered by the method of flavor delivery.     

Characterization of the bound volatile extract from baby kiwi (Actinidia arguta)

The glycosidically bound volatile fraction of baby kiwi ( Actinidia arguta ) was studied. Glycosidic precursors were isolated from juice by adsorption onto an Amberlite XAD-2 column. After enzymatic hydrolysis with Rapidase AR2000, the released aglycones were analyzed by GC-MS. Alcohols, terpenoids, and benzenoids were the most abundant compound classes. Aromatic compounds and norisoprenoids showed the highest concentrations. Major compounds were 2,5-dimethyl-4-hydroxy-3(2H)-furanone (Furaneol), benzyl alcohol, 3-hydroxy-β-damascone, hexanal, and (Z)-3-hexen-1-ol. Precursors of aroma compounds including benzoic acid, cinnamic acid, and coniferyl alcohol were also found. Eugenol, raspberry ketone, and 4-vinylguaiacol were identified for the first time in the fruit of an Actinidia species. The high concentration of 2,5-dimethyl-4-hydroxy-3(2H)-furanone in bound form (95.36 μg/kg) is particularly interesting and justifies further investigation.     

Essential oils from Mediterranean lamiaceae as weed germination inhibitors

The essential oils obtained from rosemary (Rosmarinus officinalis L.), thyme (Thymus vulgaris L.), and savory (Satureja montana L.) and the four monoterpenes that are their major constituents have been analyzed by GC and GC-MS and tested for their allelopathic properties on the seeds of three different annual weeds (Chenopodium album, Portulaca oleracea, and Echinochloa crus-galli) and three crops (Raphanus sativus, Capsicum annuum, and Lactuca sativa), with the aim to evaluate in vitro their potential as germination inhibitors. The essential oil composition varied with the species, thymol being the main constituent (44%) of thyme and carvacrol (57%) that of savory oil. Differences in essential oil composition were observed within two different rosemary ecotypes, type A, with alpha-pinene (37%) and 1,8-cineole (23%), and type B, characterized by a 2-fold content of 1,8-cineole (47%). This latest essential oil inhibited completely the germination of weeds while concurrently displaying little effect on pepper. The other two oils showed less selective action. S. montana essential oil, with 57% carvacrol, is the most active compound, completely inhibiting germination both of crops and weeds. Borneol, one of the main constituents of the oil of rosemary type B, showed an activity comparable to that of the whole oil. Crop and weed seeds treated with 1,8-cineole showed germination values that were not significantly different from controls, even if a slowing of the germination process expressed in terms of a significant increase in mean germination time was observed. Monoterpene compounds also present in the essential oils mainly represented the volatile fraction released from the crops and their residues into the soil.     

Insecticidal effect and chemical composition of the volatile oil from Bergenia ligulata

The chemical composition of the volatile oil from roots of Bergenia ligulalta was analyzed by GC-MS. A total of 97 compounds were identified. (+)-(6S)-Parasorbic acid (1) (47.45%), isovaleric acid (6.25%), 1,8-cineole (4.24%), (Z)-asarone (3.50%), and terpinen-4-ol (2.96%) were the most prominent constituents. (+)-(6S)-Parasorbic acid (1) was isolated and characterized by spectroscopic data. This is the first report of the existence of (+)-(6S)-parasorbic acid in the saxifrage family. The volatile oil and the isolated compound were tested against Drosophila melanogaster . The results obtained showed that the volatile oil from roots could be considered as natural insecticidal effect agents.     

Main agronomic-productive characteristics of two ecotypes of Rosmarinus officinalis L. and chemical composition of their essential oils

The productive potential of two different ecotypes of Rosmarinus officinalis (Cevoli and Lunigiana) cultivated in the littoral area near Pisa (northern Tuscany, Italy) and the differences in the yield and composition of the essential oils of leaves, flowers, and stems obtained from different positions of the plants were used to characterize the two ecotypes. The Cevoli ecotype plant produced the highest yield of dry matter (221 g plant-1) in comparison to the Lunigiana ecotype (72 g plant-1). There were significant differences in dry matter production of different organs of both ecotypes. The essential oil contents of Cevoli and Lunigiana ecotypes were similar. In contrast, the oil contents of the different plant parts showed marked differences. The apical part of the plant and the leaves gave the highest essential oil yields. The major difference between the oils of the two ecotypes consisted in the 1,8-cineole contents (6.6 and 37.9% in Cevoli and Lunigiana, respectively). The Cevoli ecotype was determined to be the most suitable for essential oil extraction because it was characterized by a preponderance of flowers and leaves in the apical portion. The Cevoli ecotype could be classifited as an alpha-pinene chemotype, whereas Lunigiana is a 1,8-cineole chemotype.     

Antimicrobial and bactericidal activities of esters of 2-endo-hydroxy-1,8-cineole as new aroma chemicals

Fourteen kinds of alkyl esters of 2-endo-hydroxy-1,8-cineole were synthesized, with yields of 57.8-98.0%. Each ester had a characteristic and unique odor. Especially, the tert-butyl acetate of 2-endo-hydroxy-1,8-cineole had the most interesting odor of all the synthetic esters. The antimicrobial and bactericidal activities of these synthetic esters against test bacteria (Staphylococcus aureus, Escherichia coli, and Pseudomonas fluorescens) were examined using the broth dilution method. As a result, the tert-butyl acetate of 2-endo-hydroxy-1,8-cineole showed the highest antimicrobial and bactericidal activities against all kinds of the test bacteria.     

Volatile chemicals identified in extracts from leaves of Japanese mugwort (Artemisia princeps pamp.)

Extracts from leaves of Japanese mugwort (Artemisia princeps Pamp.) were obtained using two methods: steam distillation under reduced pressure followed by dichloromethane extraction (DRP) and simultaneous purging and extraction (SPSE). A total of 192 volatile chemicals were identified in the extracts obtained by both methods using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). They included 47 monoterpenoids (oxygenated monoterpenes), 26 aromatic compounds, 19 aliphatic esters, 18 aliphatic alcohols, 17 monoterpenes (hydrocarbon monoterpenes), 17 sesquiterpenes (hydrocarbon sesquiterpenes), 13 sesquiterpenoids (oxygenated sesquiterpenes), 12 aliphatic aldehydes, 8 aliphatic hydrocarbons, 7 aliphatic ketones, and 9 miscellaneous compounds. The major volatile constituents of the extract by DRP were borneol (10.27 ppm), alpha-thujone (3.49 ppm), artemisia alcohol (2.17 ppm), verbenone (1.85 ppm), yomogi alcohol (1.50 ppm), and germacren-4-ol (1.43 ppm). The major volatile constituents of the extract by SPSE were 1,8-cineole (8.12 ppm), artemisia acetate (4.22 ppm), alpha-thujone (3.20 ppm), beta-caryophyllene (2.39 ppm), bornyl acetate (2.05 ppm), borneol (1.80 ppm), and trans-beta-farnesene (1. 78 ppm).