Isotope Ratio by HRGC-MS of Citrus junos Tanaka (Yuzu) Essential Oils: m/z 137/136 of Terpene Hydrocarbons

The isotope ratios of monoterpene hydrocarbons in Citrus junos Tanaka (yuzu) essential oils from different origins were determined by ordinary high-resolution gas chromatography-mass spectrometry (HRGC-MS). Both intensities of the molecular mass peaks (m/z 136) and of the isotope peaks (m/z 137) of monoterpene hydrocarbons were measured by single-ion monitoring with an MS analysis. The isotope ratios (m/z 137/136) of the ten monoterpene hydrocarbons commonly contained in citrus essential oils, α-pinene, β-pinene, sabinene, myrcene, α-phellandrene, α-terpinene, limonene, γ-terpinene, β-phellandrene and terpinolene, were determined in yuzu samples of the highest commercial quality from 42 different production districts. Statistical treatment of these data by the t-test and sign test revealed significant differences of the isotope effects in each yuzu sample. It is suggested that this technique will be applicable for evaluating the quality, genuineness and origin of citrus fruits and their products. The isotope fingerprints were also demonstrated in several citrus fruits other than the yuzu samples.     

Isotope ratio analysis by HRGC-MS of monoterpene hydrocarbons from citrus essential oils

The isotope ratio of monoterpene hydrocarbons in citrus essential oils of different origins was measured by ordinary high-resolution gas chromatography-mass spectrometry (HRGC-MS). The isotope ratio (Ir) was determined by the ratio of the isotope peak intensity (m/z 137) to the molecular mass peak intensity (m/z 136) of the monoterpene hydrocarbons. The accuracy of Ir was examined by measuring monoterpene hydrocarbon standards and 13C-labeled compounds. The isotope fingerprints based on the values of monoterpene hydrocarbons from lemon, lime and yuzu essential oils were determined. These citrus essential oils were also discriminated by a principal component analysis of their Ir data. The characteristic vectors showed that alpha-terpinene, beta-pinene and beta-phellandrene were important components for distinguishing between the citrus species. It is suggested that this technique will be applicable to evaluate the quality, genuineness and origin of citrus fruits and their products.     

MS fragment isotope ratio analysis for evaluation of citrus essential oils by HRGC-MS

To evaluate the origin of citrus essential oils, the isotope ratio of fragment peaks on HRGC-MS of the volatile compounds from various citrus oils was measured. The MS fragment ratio was found by the ratio of fragment peak intensity, m+1/m (m/z). This ratio reflects the isotope effect of volatile compounds, that is, it provides information about locality, quality, and species for essential oils. Multivariate analysis based on the MS fragment ratio of monoterpene hydrocarbons clearly distinguished three citrus species, yuzu, lemon, and lime. The carbonyl fractions were also extracted from citrus essential oils by the sodium hydrogensulfite method. The isotope ratio of MS fragments of octanal, nonanal, and decanal was also examined. The results suggest that there was no significant difference in the individual fragment isotope ratios of the three aldehydes.     

Characterization by GC-MS of Vietnamese citrus species and hybrids based on the isotope ratio of monoterpene hydrocarbons

The isotope ratio of monoterpene hydrocarbons was used to characterize the citrus essential oils from different species and hybrids. Citrus cold-pressed peel oils from Vietnam were analyzed for the composition and isotope ratio of monoterpene hydrocarbons by using gas chromatography-mass spectrometry. A profile of citrus essential oils on the basis of their isotope ratio values and levels of monoterpene hydrocarbons was developed for Vietnamese citrus. The molecular isotope ratios were lower than those calculated from natural abundance of 13C and 2H. In addition, the isotope ratio of the base peaks (m/z 94/93) was significantly different among the citrus essential oils from different species and hybrids. The results would be applicable for the characterization of citrus essential oils from different origins.     

Effects of several variable factors on the isotope ratio by HRGC-MS

In the isotope ratio (Ir) analysis using GC-MS, several variable factors in sampling incidental to any food analysis were investigated for yuzu fruit. The Irs of ten monoterpene hydrocarbons in yuzu essential oils from each of six fruiting positions of three trees were measured. The sign test following t-test of all the Ir values demonstrated that there was no significant difference between both sampling years of 2001 and 2002. There was also no significant variation in the Ir values among the three trees and six fruiting positions in the individual two years.     

Essential oil of Daucus glaber Forssk

The composition of the essential oil of the fruits, leaves and stems of Daucus glaber Forssk has been studied by GC/MS. It was found that, the essential oil of the fruits consists of monoterpene hydrocarbons (limonene and sylvestrene are the majors) and phenylpropanoids (elemicin is the major). Sylvestrene has never been reported before in the essential oil of any Daucus species. The study of the essential oil of the leaves revealed the presence of monoterpene hydrocarbons; limonene and gamma-terpinene are the majors and a small amount of sylvestrene. The essential oil of stems consists of monoterpene hydrocarbons (gamma-terpinene is the major), terpene alcohols (mainly 4-terpineol) and phenylpropanoids (myristicin and elemicin are the majors). It is interesting that, the essential oil of the fruits is free from any oxygenated terpenes while that of the stems is free from limonene and sylvestrene which are present in the essential oil of the fruits and leaves in fairly large amounts The essential oil of the fruits, leaves and stems shows broad antimicrobial activities against both gram positive and gram negative bacteria. In addition, the volatile oil of the stem, particularly, show activities against Candida albicans (yeast). Also, the prepared oils have variable cytotoxic activities with LC50 21.52, 36.01 and 42.34 microg/ml, respectively.     

Composition of the essential oils in various organs at different developmental stages of Ammi visnaga (L.) Lam. from Tunisia

The composition of the essential oils isolated by hydrodistillation from various organs at different development stages of Ammi visnaga (L.) Lam. growing in Tunisia was determined by GC/MS analysis. In particular, the oil profiles of the leaves, stems, flower buds, roots, umbels, and fruits have been examined during the whole life cycle. The oil from the flowering aerial parts was characterized by a high content of isoamyl 2-methylbutanoate. After flowering and during desiccation and fructification, the umbels and fruits expressed a high content of linalool. The oils, extracted from the roots collected in the vegetatif, buds floral, and floral stages, were rich in monoterpene aldehydes, oxygenated monoterpenes, and monoterpene hydrocarbons. The highest level of non-terpene hydrocarbons was found at the flower-bud stage, represented by 61.3% of nonane. Among the monoterpenes, sabinene (12.5%) and β-pinene (8.5%) were identified in the flower buds.     

Toxicity of citrus essential oils against Ceratitis capitata (Diptera: Tephritidae) larvae

Citrus peel essential oils are considered to constitute the most important resistance factor of citrus fruits against fruit flies. Essential oils were obtained from three sweet orange varieties, one bitter orange and one lemon variety. Yield, chemical composition and toxicity against neonates of the Mediterranean fruit fly were determined. Based on chemical analysis, the toxicity of commercially purchased major and minor components (monoterpenes and sesquiterpenes) of essential oils was determined. In addition, fractions were prepared to evaluate the role of minor components in the toxicity of crude essential oils. Limonene was by far the most abundant ingredient (96.2–97.4%) in all sweet orange varieties and in bitter orange, while the concentration of limonene was much lower in lemon essential oils (74.3%). Orange and bitter orange essential oils were more toxic than lemon essential oils. The toxicity of orange and bitter orange essential oils was similar to that of their major component limonene. In tests of commercially purchased chemicals, the oxygenated components of essential oils were more toxic than hydrocarbons but their low concentration in citrus essential oils could not affect the toxic activity of essential oils. The presence of α-pinene and β-pinene seems to account for the lower toxicity of lemon essential oils in relation to other citrus essential oils. The importance of understanding the toxicity of essential oils in relation to their composition and their role regarding the resistance of citrus fruits to Ceratitis capitata infestation is discussed.     

Allelopathic effect of Schinus molle essential oils on wheat germination

Gas chromatography-flame ionisation detection (GC-FID) and gas chromatography–mass spectrometry (GC–MS) analyses of the essential oils of leaves and fruits of the ornamental Shinus molle L. were reported and their allelopathic effect on wheat (Triticum aestivum L.) was evaluated. Qualitative and quantitative differences between fruit and leaf oils were observed. Both oils were rich in monoterpene hydrocarbons and the major constituents were limonene and β-phellendrene (35.9–65.4%), α-phellendrene (24.3–20.1%), myrcene (12.8–7.7%) and α-pinene (5.9–1.7%) for fruits and leaves, respectively. Both essential oils showed a dose-dependent allelopathic activity on wheat germination and radicle elongation with leaf oil being the more phytotoxic.     

Characterization of gamma-terpinene synthase from Citrus unshiu (Satsuma mandarin)

Gamma-terpinene is a monoterpene and a major component of essential oils made from citrus fruits and shows strong antioxidant activity in various assay systems. Plant gamma-terpinene synthase is a member of the monoterpene cyclase family, which produces a specific monoterpene through cyclization of geranyl diphosphate (GPP), but the monoterpene cyclases have not been fully characterized. It is necessary to prepare large amounts of gamma-terpinene synthase from Citrus unshiu (Satsuma mandarin) for the characterization, on this purpose we expressed the protein in Escherichia coli (E. coli) cells. As most monoterpene synthases have plastid-targeting signals, a gene lacking these signals was prepared and functionally expressed in E. coli cells harboring extra copies of the argU gene. The purified enzyme was incubated with GPP and the main product was confirmed to be gamma-terpinene by GC/MS.     

Chemical Composition of Essential Oils and Aromatic Waters from Different Italian Anthemis maritima Populations

The chemical composition of the essential oils and aromatic waters isolated from six Italian Anthemis maritima populations was determined by GC‐FID and GC/MS analyses. In total, 122 and 100 chemical compounds were identified in the essential oils and the aromatic waters, respectively. The main compound classes represented in the oils were monoterpene hydrocarbons, oxygenated monoterpenes, sesquiterpene hydrocarbons, oxygenated sesquiterpenes, and terpene esters. Multivariate chemometric techniques such as cluster analysis (CA) and principal coordinate analysis (PCO) were used to classify the samples according to the geographical origin. Statistical analysis allowed the attribution of the analyzed populations to different chemotype groups.     

Chemical composition and bioactivity of Boswellia serrata Roxb. essential oil in relation to geographical variation

Oleo–gum–resin of Boswellia serrata Roxb. (Burseraceae) was collected from Shivpuri forest situated in northwestern district of Madhya Pradesh, India. The commercial samples were purchased from Mandsaur and Neemuch districts (the major herbal markets of Madhya Pradesh) for comparative study of their essential oils. The average essential oil contents in the commercial Neemuch samples (11.1%) were much better as compared to the wild collection from Shivpuri (6.1%) as well as commercial collection from Mandsaur (6.9%). The commercial samples contained higher percentage of monoterpene hydrocarbons (81.9–88.1%) including α-thujene (61.4–69.8%) as the major compound. The wild habitat contained higher percentage of oxygenated monoterpenoids/benzenoids (15.7%) and sesquiterpenes (19.2%) including α-terpineol (7.8%), terpinyl isobutyrate (5.1%), and eudesmol (11.5%). Further, the antimicrobial activities of these collections showed remarkable variation among the essential oils. On the other hand, wild habitat contained improved percentage of semi-polar compounds (34.9%) which might be responsible for enhanced antibacterial activity. Similarly, the winter collection from Neemuch (BS-N2) showed significant antioxidant activity, whereas the wild habitat did not show any antioxidant activity. The higher percentage of monoterpene hydrocarbons in commercial samples, especially δ-3-carene, α-pinene, and α-thujene, might be responsible for enhanced antifungal and antioxidant activities. The Pearson correlation values justify the bioactivity relationship of the major compounds with different microbial strains. It also justified the interaction among the major individual compounds.     

Identifying the characteristic secondary ions of lignin polymer using ToF-SIMS

The chemical structure of lignin, a complex, irregular polymer of phenylpropane units that occurs in plant cell walls, was investigated using time-of-flight secondary ion mass spectrometry (ToF-SIMS). The positive ToF-SIMS spectra of lignin isolated from pine and beech wood showed prominent secondary ions possessing guaiacyl (at m/z 137 and 151) or syringyl (at m/z 167 and 181) rings, which are the basic building units of lignin polymer. This shows that ToF-SIMS is a useful tool for lignin structural analysis. The peaks at m/z 137 and 167 were assigned as the C6-C1 ion, and the peaks at m/z 151 and 181 may be double-component, the C6-C1 ion and the C6-C2 ion. We confirmed the characteristic guaiacyl ions using a synthetic lignin model compound, dehydrogenation polymer (DHP), which was formed by polymerizing of unlabeled and deuterium-labeled coniferyl alcohols. The formation mechanism of the main secondary ions was deduced by labeling specific positions of coniferyl alcohols with a stable isotope to study the relationship between chemical structure and secondary ion formation in ToF-SIMS.     

Salvia verbenaca L. essential oil: Variation of yield and composition according to collection site and phenophase

The effect of the collection sites and phenophase on yield and chemical composition of Salvia verbenaca essential oils was evaluated. The essential oil constituents were assessed by gas chromatography (GC) and gas chromatography–mass spectrometry (GC–MS). The highest essential oil yields were observed for samples of the higher semi-arid bioclimate and at the flowering period. Eighty-five volatile constituents were identified and their percentages varied significantly (p < 0.05) depending on the collection site and the phenological stage. According to the plants origin, essential oils were dominated by monoterpene hydrocarbons, oxygenated monoterpenes and sesquiterpene hydrocarbons. The monoterpene hydrocarbons (31.9%) predominate at the flowering stage whereas oxygenated sesquiterpenes (27.5%) at the early fruiting stage. The sesquiterpene hydrocarbons (28.2%) was the most represented chemical class at late fruiting. On the basis of GC-MS data, the major identified volatile constituents were viridiflorol (3.4–17.7%), α-pinene (0.7–15.9%), β-caryophyllene (1.0–15.3%) and p-cymene (1.3–14.2%). S. verbenaca contains a diversity of bioactive constituents which shows large variations as affected by the collection sites and phenophase.     

Geographic variation in the essential oil monoterpenes of Liriodendron tulipifera L

The monoterpene component of Liriodendron tulipifera L. (Magnoliaceae) essential oils was quantitatively analysed in branch samples collected from an Ohio provenance test of 11 geographic seed sources representing the natural range of the species. Mean monoterpene composition of the essential oil is described and significant differences in the monoterpene composition among geographic sources are discussed.     

A Comparative GC/MS Analysis of Citrus Essential Oils: Unveiling the Potential Benefits of Herb-Drug Interactions in Preventing Paracetamol-Induced Hepatotoxicity

Our study aimed to test the potential of Citrus oils in protecting against paracetamol (PAR)-induced hepatotoxicity. The essential oils of Pineapple sweet orange (OO), Murcott mandarin (MO), Red grapefruit (GO), and Oval kumquat (KO) were investigated using gas chromatography coupled with mass spectrometry (GC/MS). Twenty-seven compounds were identified, with monoterpene hydrocarbons being abundant class. d-Limonene had the highest percentage (92.98 %, 92.82 %, 89.75 %, and 94.46 % in OO, MO, GO, and KO, respectively). Hierarchical cluster analysis (HCA) and principal components analysis (PCA) revealed that octanal, linalool, germacrene D, and d-limonene were the principal discriminatory metabolites that segregated the samples into three distinct clusters. In vitro antioxidant capacities were ranged from 1.2–12.27, 1.79–5.91, and 235.05–585.28 μM Trolox eq/mg oil for 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic (ABTS), ferric-reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC), respectively. In vivo, citrus oils exhibited a significant reduction in alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and nitric oxide (NO). Additionally, there was an increase in glutathione reductase (GSH), and the liver architecture was nearly normal. Molecular docking revealed that d-limonene exhibited a good inhibitory interaction with cytochrome P450 (CYP450) isoforms 1A2, 3A4, and 2E1, with binding energies of −6.17, −4.51, and −5.61 kcal/mol, respectively.     

Analysis of the volatile essential oils of Murraya koenigii and Pandanus latifolius

Using well-established techniques, samples were obtained of the volatile essential oils of the two types of curry leaf, Murraya koenigii and Pandanus latifolius. Both contained mainly terpenes, and M. koenigii produced less than 4% of other components with eight monoterpene hydrocarbons (ca 16%) and 17 sesquiterpene hydrocarbons (ca 80%) being obtained. The most important constituents of M. koenigii are β-caryophyllene, β-gurjunene, β-elemene, β-phellandrene and β-thujene. The volatile essential oil of P. latifolius also contained mainly sesquiterpene hydrocarbons (6–42%) but the only monoterpene was linalool (ca 6%). Nearly 2000 times the total quantity of aroma volatiles was produced by M. koenigii compared with P. latifolius, and this partly explains the observed stronger flavour potency of the former.     

Chemical composition and intraspecific variability of the essential oils of five populations of Hypericum triquetrifolium Turra growing in North Tunisia

The chemical composition of the essential oils of five populations of Hypericum triquetrifolium Turra from Tunisia and their intraspecific variability were analyzed in detail by GC/MS. One hundred seventy-four compounds were identified, representing averages of 87.9 to 98.7% of the oil composition. The components are represented here by homologous series of monoterpene hydrocarbons, oxygenated monoterpenes, sesquiterpenes hydrocarbons, oxygenated sesquiterpenes, non-terpenic hydrocarbons, and others. Sesquiterpene hydrocarbons were the most abundant chemical compounds. Multivariate chemometric techniques, such as cluster analysis (CA) and principal-component analysis (PCA), were used to characterize the samples according to the geographical origin. By statistical analysis, the analyzed populations were classified into four chemotype groups.     

Chemical composition and biological activities of essential oils extracted from Korean endemic citrus species

The aim of this study was to analyze the chemical composition of 14 kinds of citrus oils and to test their biological activities. Citrus essential oils were obtained by steam distillation from immature fruits collected from Jeju Island and were analyzed using gas chromatograph (GC)- flame ionization detectors (FID) and GC-MS. Limonene (55.4% to 91.7%), myrcene (2.1% to 32.1%), alpha-pinene (0.6% to 1.6%) and linalool (0.4% to 6.9%) were the major components in most citrus species. To evaluate in vitro antibacterial activity, all essential oils were tested against Propionibacterium acnes and Staphylococcus epidermidis. Nine out of fourteen citrus oils exhibited antibacterial activity against P. acnes, but not against S. epidermidis. The effects of the citrus oils on DPPH radical scavenging, superoxide radical anion scavenging, nitric oxide radical, and cytotoxicity were also assessed. Three essential citrus oils, Joadeung, Dongjunggyul, and Bujiwha, exhibited potent inhibitory effects on nitric oxide production. Two essential oils, Dongjunggyul and Joadeung, showed potent free radical scavenging activities in the DPPH assay. For future applications in cosmetic products, we also performed MTT assays in a human dermal fibroblast cell line. The majority of the essential oils showed no cytotoxicity. The results indicate that citrus essential oils can be useful natural agents for cosmetic application.     

Enantioselective GC–MS analysis of volatile components from rosemary (Rosmarinus officinalis L.) essential oils and hydrosols

Essential oils and hydrosols were extracted from rosemary harvested in different seasons, and the chemical compositions of volatile components in the two fractions were analyzed by gas chromatography–mass spectrometry (GC–MS). Enantiomers of some volatile components were also analyzed by enantioselective GC–MS. Classification of aroma components based on chemical groups revealed that essential oils contained high levels of monoterpene hydrocarbons but hydrosols did not. Furthermore, the enantiomeric ratios within some volatile components were different from each other; for example, only the (S)-form was observed for limonene and the (R)-form was dominant for verbenone. These indicate the importance of determining the enantiomer composition of volatile components for investigating the physiological and psychological effects on humans. Overall, enantiomeric ratios were determined by volatile components, with no difference between essential oils and hydrosols or between seasons.