Detection and identification of 1-methylethyl and methyl radicals generated by irradiating tea tree (Melaleuca alternifolia) oil with visible light (436 nm) in the presence of flavin mononucleotide and ferrous ion

Abstract

Here, we determined the electron spin resonance (ESR) spectra of standard reaction mixtures (I) containing 25 μM flavin mononucleotide (FMN), 0.018% tea tree (Melaleuca alternifolia) oil, 1.9 M acetonitrile, 20 mM phosphate buffer (pH 7.4), 0.1 M α-(4-pyridyl-1-oxide)-N-tert-butylnitrone (4-POBN), and 1.0 mM FeSO₄(NH₄)₂SO₄ irradiated with 436 nm visible light (7.8 J/cm²). Prominent ESR signals (α^N = 1.58 mT and α^Hβ = 0.26 mT) were detected, suggesting that free radicals form in the standard reaction. In order to know whether singlet oxygen (¹O₂) is involved in the radical formation or not, ESR measurement was performed for the standard D₂O reaction mixture (I) which contained 25 μM FMN, 0.0036% tea tree oil, 1.9 M acetonitrile-d3, 20 mM phosphate buffer (pH 7.4), 0.1 M 4-POBN and 1.0 mM FeSO₄ in D₂O. The ESR peak height of the standard D₂O reaction increased to 169 ± 24% of the control. Thus, ¹O₂ seems to be involved in the formation of the radicals because D₂O increases the lifetime of singlet oxygen. High-performance liquid chromatography-ESR-mass spectrometry analyses detected 1-methylethyl and methyl radicals in the standard reaction. The radicals appear to form through the reaction of ferrous ion with α-terpinene endoperoxide (ascaridole), which generated from the reaction of α-terpinene with ¹O₂. The 1-methylethyl and methyl radicals may exert a pro-oxidant effect under these conditions.     

植物精油化学成分及其抗菌活性的研究进展

植物精油是一类从植物中萃取的芳香味油状液体,是一类优良的天然抗菌材料。作为抗菌材料,植物精油具有以下优点:具有广谱高效的抗菌活性;具有熏蒸特性、气味芳香;取自天然植物,绿色环保;来源广,提取容易。植物精油因其多种优点,在抗菌领域具有巨大的潜在应用价值。本文从植物精油的分布及化学成分、抗细菌活性和抗真菌活性的研究,以及植物精油化学成分与抗菌活性之间的联系等方面对植物精油的抗菌性能进行评述,以期促进植物精油在抗菌领域的广泛应用,同时给从事植物精油抗菌研究的科研工作者提供参考。     

Components of essential oils extracted from leaves and shoots of abies species in Japan

The essential oils extracted from the leaves and the shoots of five Abies species (Pinaceae) growing in Japan, i.e., A. firma, A. homolepis, A. veitchii, A. mariesii, and A. sachalinensis, were characterized by GC-FID and GC/MS analyses. The yields of the essential oils extracted from A. sachalinensis were the highest among them. A significant amount of α-pinene was contained in the essential oils of all the Abies species examined. In A. homolepis and A. veitchii, significant differences in the content of the essential oils were found depending on whether these were extracted from the leaves or from the shoots. Regarding the enantiomeric ratio of α-pinene, the (+)-enantiomer was predominant in the oil extracted from the leaves of A. firma, while (-)-α-pinene was present in higher amounts in the oils of A. veitchii (leaves and shoots), A. mariesii (leaves and shoots), and A. sachalinensis (shoots). The fact that there may be a quantitative and qualitative difference in the components of the essential oils extracted from the different parts of a plant was investigated by cluster analysis.     

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.     

Chemical composition, seasonal variation, and biosynthetic considerations of essential oils from Baccharis microdonta and B. elaeagnoides (Asteraceae)

The chemical composition and seasonal variation throughout one year of the essential oils from leaves of Baccharis microdonta and B. elaeagnoides, collected in Campos do Jordão, SP, were investigated. The composition of the latter species has been described for the first time. By GC (RI) and GC/MS analysis, 43 compounds were identified, and a predominance of oxygenated sesquiterpene derivatives was found in both species. The main components of the B. microdonta oils were elemol ( 31 ; 11.7–30.6%), spathulenol ( 34 ; 4.7–9.1%), β‐caryophyllene ( 19 ; 3.7–6.2%), and germacrene D ( 24 ; 2.9–12.2%), and those of the B. elaeagnoides oils were 34 (10.1–21.5%), viridiflorol ( 35 ; 3.6–18.4%), 24 (0.9–13.8%), and 19 (3.5–9.4%). The identified compounds were grouped according to their respective C‐skeletons, and the percentages of occurrence of the C‐skeletons in the essential oils of leaves collected in the four seasons allowed identifying the preferential accumulation of different types of C‐skeletons as well as the seasonal variation of the biosynthetic routes over the studied period.     

Variability of the needle essential oils of Pinus peuce from different populations in Montenegro and Serbia

The essential-oil composition of Pinus peuce Griseb. is reported at the population level. Macedonian pine is endemic high-mountain Balkan pine relict of an anthropogenically reduced area, with large morphological diversity and insufficiently clear taxonomic position. In the pine-needle terpene profile of two populations from Montenegro and one from Serbia, 78 compounds were detected, 56 of which are identified (Table 3). The dominant constituents were alpha-pinene (36.5%) and germacrene D (11.4%). The following 20 additional components were found to be present in medium-to-high amounts (0.5-10%): camphene (8.5%), bornyl acetate (6.8%), beta-pinene (6.8%), beta-caryophyllene (5.2%), beta-phellandrene (4.7%), terpinen-4-ol acetate (1.6%), (E)-hex-2-enal (1.5%), alpha-muurolene (1.2%), beta-gurjunene (1.1%), beta-myrcene (1.0%), alpha-terpinyl acetate (0.9%), alpha-phellandrene (0.8%), delta-cadinene (0.8%), alpha-humulene (0.8%), sabinene (0.7%), aromadendrene (0.6%), alpha-thujene (0.6%), gamma-muurolene (0.6%), gamma-cadinene (0.6%), alpha-terpinolene (0.5%), and one unknown component (0.5%). The similarity of the populations and the within-population variability were visualized by principle-component analysis (PCA) and genetic analysis of selected terpenes in 90 tree samples. Our study suggests a closer connection between populations II and III compared to population I. Based on the profile of the main terpene components, the studied populations are more similar to populations from Kosovo and Greece than to the population from Mt. Mokra (Montenegro) and the population in France.     

Isolation and identification of a plant growth promotive substance from mixture of essential plant oils

The PCS (commercial products by Field Science Co, Japan, used for air fresheners) was analyzed for the presence of bioactive constituents and their role as root growth promoters. Chromatographic separation of the methanolic solution of PCS resulted in the isolation of an promoting active substance, which was identified using GC-mass spectrometry and NMR spectroscopy as 1,2-propanediol (CH₃CH(OH)CH₂OH). Lettuce seedling growth bioassay as test plant revealed that 1,2-propanediol can act as potent root growth promoter; enhancing the growth of lettuce seedling radicle at a concentration 0.01 ppm. The concentration of 1,2-propanediol in PCS mixture was estimated as 4 g/l. These studies suggest that 1,2-propanediol might play an important role in the plant growth promoting activity of PCS.     

Chemotaxonomic significance of the essential oils of 18 Ferula species (Apiaceae) from Iran

To evaluate the chemotaxonomic significance of the essential oils of 23 populations of 18 Iranian Ferula species, the chemical composition of the oils was investigated by GC/FID and GC/MS. Altogether, 84 constituents, representing 81.3-99.7% of the total composition of the oils, have been identified. The composition of six species of the genus, i.e., F. oopoda, F. foetida, F. behboudiana, F. diversivittata, F. galbaniflua, and F. hezarlalehzarica, has been reported for the first time. The main constituents identified were α-terpinyl acetate (73.3%), 2,3,4-trimethylthiophene (2; 49.0%), sabinene (75.3%), verbenone (5; 69.4%), β-pinene (59.0-66.3%), and (Z)-β-ocimene (41.7%). Cluster analysis (CA) of the percentage content of the essential oil components of the Ferula species resulted in the characterization of four groups, i.e., taxa containing either i) monoterpene hydrocarbons, ii) oxygenated monoterpenes, iii) organosulfur compounds, or iv) monoterpene, sesquiterpene, and aliphatic hydrocarbons as the principal classes of compounds. Based on the results obtained, the chemical independence of F. hirtella from F. szowitsiana and of F. galbaniflua from F. gummosa at the specific level was concluded and their positions as distinct species were confirmed. The chemotaxonomic relationships among the representatives of the genus Ferula have been discussed in detail.     

Mosquitocidal and repellent activities of essential oils against Culex pipiens L.

Plants are a prospective source of novel natural insect repellents and botanical insecticides. This study was conducted to investigate the chemical composition of the essential oils of three plants growing in Saudi Arabia, namely Ducrosia anethifolia, Achillea fragrantissima, and Teucrium polium; and to evaluate their potential mosquitocidal and repellent activities against adult female Culex pipiens L. The main components of the three oils were found to be decanal (28.9%) and chrysanthenyl acetate (10.04%), (D. anethifolia); sabinyl acetate (35.79) and artemesia ketone (18.28%) (A. fragrantissima); α‐cadinol (49.53%) and δ‐cadinene (10.23%) (T. polium). The oil of A. fragrantissima was the most toxic (LC₅₀ = 0.11 μL/L air) followed by D. anethifolia and T. polium with LC₅₀ values of 5.22 and 25.98 μL/L air, respectively. T. polium oil was the most repellent (292 min at 2 μL/cm²), followed by D. anethifolia and A. fragrantissima. The results indicate that the essential oils have a potential fumigant insecticidal and repellent activities for mosquito control.     

Chemical composition of the essential oils of ripe berries of Juniperus oxycedrus L., growing wild in Kosovo

The main objective of this work was to study the essential oil composition of ripe Juniperus oxycedrus L. berries and its natural variation among wild populations in Kosovo. Essential oil was analysed using GC-FID and GC–MS. Plant materials were collected from five locations in Kosovo in August and September of 2011. In total, twenty-seven compounds were identified in the essential oils. The main components were β-myrcene (45.5–56.9%), α-pinene (10.2–36.6%), dl-limonene (3.6–13.8%) and germacrene D (1.7–8.7%). Of the total identified compounds, monoterpenes constituted the highest percentage of all components (70.24–88.22%), followed by oxygenated sesquiterpenes (4.9–11.4%), sesquiterpenes (3.5–11.0%), oxygenated monoterpenes (0.2–2.7%) and oxygenated diterpenes (0.0–1.7). Hierarchical Cluster Analysis (HCA) and Principal Component Analyses (PCA) were used to identify any geographical variations in essential oil composition. Statistical analysis suggests that the clustering of populations is not related to their geographic location, but rather seemed to be linked to local selective forces acting on chemotype diversity.     

Using essential oils to overcome bacterial biofilm formation and their antimicrobial resistance

The increase of resistant bacteria puts a huge pressure on the antimicrobials in current use. Antimicrobial resistance (AMR) results from antibiotic misuse and abuse over many years and is a global financial burden. New polices must be developed for the use of antimicrobials and to continue research efforts to mitigate AMR. It is essential to target the most harmful bacteria and concentrate on their mechanisms of resistance to develop successful antimicrobials. Essential oils (EOs) are occur naturally in plants and have long been used as antimicrobials, but most have not been researched. This review explores EOs as alternative antimicrobials, investigating their ability to decrease or inhibit biofilm formation, and assess their ability to contribute to AMR control. Low concentrations of EOs can inhibit Gram-positive and Gram-negative pathogenic bacteria. Some EOs have demonstrated strong anti-biofilm activities. If EOs are successful against biofilm formation, particularly in bacteria developing AMR, they could be incorporated into new antimicrobials. Therefore, there is a need to investigate these EOs’ potential, particularly for surface disinfection, and against bacteria from food, clinical and non-clinical environments.     

杉木心材精油抑菌活性及其化学成分研究

通过水蒸气蒸馏法提取杉木心材精油,并进行柱层析分离、气-质联用分析和抑菌活性试验,比较分析了精油含量、化学组成和抑菌活性成分.结果表明,杉木心材精油含量为1.794～2.076(w/w);气-质联用分析共分离出47个色谱峰,鉴定出27个化合物(占精油总量的99%),其中主要成分为柏木脑(76.27%);杉木心材精油对大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌、伤寒沙门氏菌等均有较明显的抑制作用;柏木脑是杉木精油的主要抑菌活性成分.     

Nano essential oils against the red palm weevil,Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae)

The red palm weevil, Rhynchophorus ferrugineus, is the biggest pest of various palm species. The aim of this study was to evaluate the impact of the essential oils of purslane, mustard and castor (bulk and nano) on R. ferrugineus and to evaluate the oviposition deterrent effect to prevent new infestations. Prepared concentrations of essential oils (bulk and nano) were tested for their impact on R. ferrugineus larvae, pupae and adults. The most effective oil was purslane oil (bulk and nano), followed by mustard and the least effective was castor. The percent mortality of larvae was 75.2, 45.3 and 17.9% (bulk phase) and 92.5, 84.4 and 65.5% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The percent mortality of adults was 67.2, 40.0 and 15.6% (bulk phase) and 83.5, 72.9 and 60.0% (nano phase) in purslane, mustard and castor, respectively, when treated with the high concentration. The number of eggs laid decreased with increasing concentration of the tested oils. Purslane essential oil showed good impact on R. ferrugineus larvae and adults followed by mustard and castor oils as bulk and as nano.     

植物精油对大菱鲆弧菌的体外和体内抗菌活性

【目的】研究天然植物精油对大菱鲆弧菌的体外和体内抗菌活性。【方法】采用纸片扩散法和微量肉汤稀释法对14种植物精油或其组分的体外抑菌活性进行检测;通过细菌形态透射电镜观察、胞内乳酸脱氢酶及核酸释放研究山苍子精油对大菱鲆弧菌的膜损伤作用;采用大菱鲆人工攻毒感染实验研究山苍子精油的体内抗菌作用。【结果】14种植物精油或其组分对大菱鲆弧菌具有不同程度的抑制效果,其中肉桂醛的抗菌活性最强,最低抑菌浓度为0.25μL/m L;百里香酚、丁香酚、柠檬醛和山苍子的抗菌活性次之,最低抑菌浓度为0.5μL/m L;山苍子精油可破坏大菱鲆弧菌的细胞膜,并导致胞内蛋白酶和核酸外泄;经200μL/L山苍子精油浸浴后,大菱鲆攻毒后死亡率由对照组50%降至0。【结论】富含芳香醛、芳香酚和柠檬醛的植物精油对大菱鲆弧菌具有良好抗菌活性,有望替代抗生素用于大菱鲆弧菌病的防治。     

Geographical and environment-related variations of essential oils in isolated populations of Thymus richardii Pers. in the Mediterranean basin

Composition of essential oils of different populations of Thymus richardii grex of six localities from Bosnia-Herzegovina (Konjic, Borci), Spain (Majorca, Ibiza, Valencia) and Italy (Marettimo, Sicily) were determined by GC/FID and GC/MS. The main constituents in most of the samples were aromatic monoterpenes corresponding to non-phenolic cyclic compounds (p-cymene, γ-terpinene). The highest monoterpene concentrations were found in the Bosnian samples (70%), and the lowest in samples from the Balearic Islands (<30%; Ibiza and Majorca). Sesquiterpenes were the major component (average > 50%) in samples from Majorca with β-bisabolene (>40%) being the principal constituent. Discriminant analysis (LDA) shows the differentiation of two chemotypes: A (phenol chemotype), with p-cymene and γ-terpinene as characteristic compounds and B, with β-bisabolene and carvacrol, as major and significative compounds. The occurrence of the chemotypes was related to summer positive precipitation and to deep of soils.     

Leaf essential oils and their application in systematics of Juniperus excelsa complex in Iran

The composition of the leaf essential oils of ten populations of Juniperus excelsa complex in Iran are reported and compared. Hydrodistillation yielded in 0.08%–3.78% dry weight (v/w) clear oils among populations studied. The oils are mostly dominated by α- Pinene, Limonene and β-Myrcene. The simplest oil was found in Qushchi (J. polycarpos var. polycarpos) with 25 components and the most complex oil was found in Lushan1 (J. polycarpos var. polycarpos) with 92 components. Quantitative data is presented for 164 components were found. The southern populations, Fasa and Khabr, show differences in composition of essential oils compared with other populations so that Borneol and isopinocamphone are diagnostic for them. These populations have been identified as Juniperus seravschanica in southeast of Iran and hybrid samples between J. polycarpos and J. seravschanica in southwest of Iran whereas other populations, as is confirmed in this study, are J. polycarpos. It is confirmed by essential oils data that there is no J. excelsa in Iran.     

Chemical composition and antimicrobial activity of the essential oils of Anthospermum emirnense and Anthospermum perrieri (Rubiaceae)

The essential oils of Anthospermum emirnense Baker and Anthospermum perrieri Homolle ex Puff, obtained by hydrodistillation in 0.03 and 0.02% yield, respectively, were analyzed by GC/MS. In both cases, the major constituents consisted of sesquiterpene hydrocarbons and oxygenated sesquiterpenes. The two species showed an important qualitative similarity, with 40 compounds common to A. emirnense and A. perrieri, including β-elemene, trans-β-caryophyllene, caryophyllene oxide, and τ-cadinol, which were major components in both cases. When tested for antimicrobial activity, both essential oils showed similar profiles and exhibited interesting minimal-inhibitory-concentration (MIC) values towards Bacillus subtilis, Chryseobacterium indologenes, Flavimonas oryzihabitans, and Yersinia enterocolitica.     

Effect of increasing doses of a microencapsulated blend of essential oils on performance of lactating primiparous dairy cows

This study determined impacts of a commercial blend of microencapsulated essential oils (EO), fed at increasing dose levels to high yielding primiparous dairy cows, on milk yield and composition. Eight pregnant Holstein heifers, selected to have their parturition within a period of 30 d, were kept in tie stalls equipped for individual feeding, and with free access to water, from the 7th month of gestation. At an average of 40 d postpartum, cows were assigned to one of four dietary EO levels in a replicated 4 × 4 Latin square design. The experimental diets (i.e., corn silage, fescue hay and a compound feed, 6.5, 4.4 and 8.7 kg DM/d respectively) differed only in the addition level of a microencapsulated EO supplement (RumaXol Feed) mixed at different levels into the compound feed (fed at 1 kg/cow day) in order to provide 0, 0.32, 0.64 and 0.96 g/d of the EO mixture. The experiment had four experimental periods of 21 d, with the last 5 d used for collection of faeces and urine, for milk yield and composition recording and measurement of cow biometric data. The dietary EO supplementation had no effect on dry matter (DM) intake, water consumptions or faecal DM, while urine density was (P=0.01) lower at the intermediate EO dosages. Digestibility and biometric measures of cows were not modified by EO feeding. The protein content of milk tended (P=0.06) to be higher at the intermediate EO dosages, as did (P=0.05) the milk energy concentration. However milk and milk component yields were not affected by EO feeding level.     

Comparative larvicidal activity of essential oils from three medicinal plants against Aedes aegypti L

This work aimed at comparing larvicidal activity of essential oils extracted from the dried leaves of Alpinia speciosa, Cymbopogon citratus, and Rosmarinus officinalis against Ae. aegypti larvae. The larvae were observed for 4 h and at 24 h according to a completely randomized design with three replications and the following concentrations [μl/ml]: 0.25, 0.5, 1.0, 1.5, 2.0, 2.5, and controls were distilled water, and commercial standard citral, camphor, eucalyptol, α‐pinene, and β‐myrcene. The essential oil of C. citratus had the lowest LC₅₀ (0.28) and LC₉₀ (0.56) values, followed by that of A. speciosa (0.94 and 1.2, resp.) and of R. officinalis (1.18 and 1.67, resp.), and only the commercial standards citral and α‐pinene were larvicidal.     

Antimicrobial and antioxidative activity of extracts and essential oils of Myrtus communis L.

Since synthetic antimicrobial agents and food additives can cause a number of adverse effects, there is a growing interest from consumers in ingredients from natural sources. Medicinal plants, such as Myrtus communis L. are a source of new compounds which can be used in both the food industry and for medical purposes, primarily as antimicrobial agents. In this review, the characteristics of myrtle essential oils and extracts are summarized, with particular attention to their chemical composition, biological activities and potential applications.