Development and validation of HPLC methods for the analysis of phenethylamine and indoloquinazoline alkaloids in Evodia species

The aim of this study was to evaluate the chromatographic performance of a PEG stationary phase, in comparison with those of C18 columns, for the HPLC analysis of phenethylamine ((+/-)-synephrine) and indoloquinazoline (rutaecarpine and evodiamine) alkaloids in methanolic extracts of fruits of Evodia rutaecarpa (Juss.) Benth. and E. rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang (i.e., E. officinalis Dode) (Rutaceae family). The method was validated and showed good linearity, precision, accuracy, sensitivity, and specificity. The highest content of both phenethylamine and indoloquinazoline alkaloids was found in methanolic fruit extracts of E. rutaecarpa, and it was closely related to the degree of maturity. E. officinalis fruits displayed low amounts of both types of alkaloids. Furthermore, an enantioselective HPLC method for the enantioseparation of (+/-)-synephrine from Evodia fruits was applied, by using a protein-based chiral stationary phase with cellobiohydrolase (CBH) as the chiral selector (Chiral-CBH). Isolation of synephrine from Evodia aqueous fruit extracts was carried out by strong cation-exchange SPE. The results of the application of the method to the analysis of Evodia samples showed that (-)-synephrine was the main component while (+)-synephrine was present in low concentration.     

Headspace solid-phase microextraction-gas chromatography--mass spectrometry analysis of the volatile compounds of Evodia species fruits

In this study the investigation of the aroma compounds of dried fruits of Evodia rutaecarpa (Juss.) Benth. and E. rutaecarpa (Juss.) Benth. var. officinalis (Dode) Huang (i.e. E. officinalis Dode) (Rutaceae family) was carried out to identify the odorous target components responsible for the characteristic aroma of these valuable natural products. To avoid the traditional and more time-consuming hydrodistillation, the analyses were carried out by means of headspace solid-phase microextraction (HS-SPME) coupled to gas chromatography-mass spectrometry (GC-MS). The SPME headspace volatiles were collected using a divinylbenzene-carboxen-polydimethylsiloxane (DVB-CAR-PDMS) fiber. The extraction conditions were optimized using a response surface experimental design to analyze the effect of three factors: extraction temperature, equilibrium time and extraction time. The best response was obtained when the extraction temperature was around 80 degrees C, equilibrium time near 25 min and extraction time close to 18 min. Analyses were performed by GC-MS with a 5% diphenyl-95% dimethyl polysiloxane (30 m x 0.25 mm I.D., film thickness 0.25 microm) capillary column using He as the carrier gas and a programmed temperature run. The main components of the HS-SPME samples of E. rutaecarpa (concentration >3.0%) were limonene (33.79%), beta-elemene (10.78%), linalool (8.15%), myrcene (5.83%), valencene (4.73%), beta-caryophyllene (4.62%), linalyl acetate (4.13%) and alpha-terpineol (3.99%). As for E. officinalis, the major compounds were myrcene (32.79%), limonene (18.36%), beta-caryophyllene (9.92%), trans-beta-ocimene (6.04%), linalool (5.88%), beta-elemene (7.85%) and valencene (4.62%).     

Preparative isolation and purification of alkaloids from the Chinese medicinal herb Evodia rutaecarpa (Juss.) Benth by high-speed counter-current chromatography

High-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water system (5:5:7:5, v/v) was applied to the isolation and purification of alkaloids from the Chinese medicinal plant Evodia rutaecarpa (Juss.) Benth. Five kinds of alkaloids were obtained and yielded 28 mg of evodiamine (I), 19 mg of rutaecarpine (II), 21 mg of evocarpine (III), 16mg of 1-methy-2-[(6Z,9Z)]-6,9-pentadecadienyl-4-(1H)-quinolone (IV), 12 mg of 1-methyl-2-dodecyl-4-(1H)-quinolone (V) from 180 mg of crude extract in a one-step separation, with the purity of 98.7%, 98.4%, 96.9%, 98.0%, 97.2%, respectively, as determined by high performance liquid chromatography (HPLC). The structures of these compounds were identified by 1H NMR and 13CNMR.     

Two New Indole Alkaloids from Evodia rutaecarpa

The tyuit of Evodia rutaecarpa (Juss.) Benth is a Chinese traditional drug (Wu-Chu-Yu).The components of Wu-Chu-Yu have been studied by several groups, including indolealkaloids,quinolone alkaloids. limonoids and other kinds]. Further chemical investigationofthis drug led us to isolate two new indole alkaloids 1 and 2. This is the first time toisolate indolinone alkaloids from this plant.Wuchuyuamide I (l ), was isolated as colorless needles, m,p. 261 -262<> C (CHCI,MeOH), [Q].=# 0(c 0…     

Evodiagenine and dievodiamine, two new indole alkaloids from Evodia rutaecarpa

<正>Two new indole alkaloids,evodiagenine 1 and dievodiamine 2 were isolated from the fruits of Evodia rutaecarpa(Juss.) Benth. The structure of compounds 1 and 2 were elucidated by comprehensive spectroscopic analysis and compound 1 was confirmed by X-ray crystallographic analysis.     

Essential oil composition of aerial parts of Cyclospermum leptophyllum (Pers.) Sprague ex Britton and P. Wilson

The essential oil of Cyclospermum leptophyllum (Pers.) Sprague ex Britton and P. Wilson syn. Apium leptophyllum (Pers.) F. Muell. Ex Benth., family Apiaceae (Umbellifereae), was analysed by GC and GC-MS. The essential oil showed the presence of thymohydroquinone dimethyl ether (50.7%), thymol methyl ether (11.2%), γ-terpinene (10.4%), p-cymene (9.5%) and carvacrol methyl ether (5.9%), along with trans-ocimene, β-pinene, germacrene D, cumin aldehyde, α-humulene, β-myrcene, sabinene, limonene, β-phellandrene, caryophyllene oxide, α-pinene and α-thujene as minor constituents. To the best of our knowledge, this is the first study of the essential oil composition of C. leptophyllum collected from India.     

Chemical investigation of the essential oil from berries and needles of common juniper (Juniperus communis L.) growing wild in Estonia

The essential oils obtained by simultaneous distillation and extraction (SDE) from the fresh and dried needles and dried berries of Juniperus communis L. of Estonian origin were subjected to GC-FID and GC-MS analyses. The yields of the oils ranged between 0.2% and 0.6% from juniper berries and between 0.5% and 1.0% from needles (dried weight). A total of 87 compounds were identified, representing over 95% of the oil. The major compounds in the needle oil were monoterpenes α-pinene (33.3-45.6%), sabinene (0.2-15.4%), limonene (2.8-4.6%) and sesquiterpenes (E)-β-caryophyllene (0.8-10.3%), α-humulene (0.8-6.2%) and germacrene D (3.0-7.8%). The juniper berry oil was rich in α-pinene (53.6-62.3%), β-myrcene (6.5-6.9%) and germacrene D (4.5-6.1%). The main oxygenated terpenoids found in the needle oil were germacrene D-4-ol (0.4-4.0%) and α-cadinol (to 2.7%). The oil from fresh needles contained high amounts of (E)-2-hexenal (3.7-11.7%).     

Evollionines A–C,Three New Alkaloids Isolated from the Fruits of Evodia rutaecarpa

A phytochemical study of the CHCl₃ extract of the dried unripe fruits of Evodia rutaecarpa (Juss. ) Benth. (Rutaceae) resulted in the isolation of three new alkaloids, evollionines A–C ( 1 – 3 , resp.), together with two known compounds, evodianinine and wuzhuyumide I. The structures of the new compounds were elucidated on the basis of detailed spectroscopic evidence and confirmed in the case of compound 1 by single‐crystal X‐ray analysis.     

Antianoxic action and active constituents of evodiae fructus

In order to develop new drugs from natural products, constituents of natural medicines were examined for their effectiveness in the KCN-induced anoxia model in mice. Methanol extract from a Chinese medicine, evodia (fruits of Evodia rutaecarpa Benth. or E. officinalis Dode), had a significant effect in the KCN-induced anoxia model in mice and therefore the active constituents were further examined. The results indicated that the antianoxic action of evodia was found in the fraction containing evodiamine. Further analysis of the active constituent indicated that evodiamine and rutaecarpine, indole-alkaloids found in large amounts in the Chinese medicine evodia, were mainly responsible for the antianoxic action.     

Seasonal variations of volatile constituents of Hemizygia bracteosa (Benth.) Briq. aerial parts from Benin

Essential oils from fresh aerial parts of Hemizygia bracteosa (Benth.) Briq. were extracted by steam distillation. The oil yield from plants collected during the hot season (February) and during the cold season (August) were 0.12 ± 0.01% and 0.25 ± 0.02%, respectively. GC/FID and GC/MS analyses allowed us to identify a total of 65 compounds, representing 97% of the hydrodistillate. The main components of the oil from the hot period were (E)-β-farnesene (64 ± 0.04%), β-elemene (7.4 ± 0.05%), trans-nerolidol (6.2 ± 0.04%), and α-muurolene (2.7 ± 0.03%). The essential oil from the cold season was characterized by the presence, as major compounds, of (E)-β-farnesene (67 ± 0.04%) along with β-caryophyllene (3.6 ± 0.06%), β-elemene (3.3 ± 0.05%), 7-epi-α-selinene (3.1 ± 0.01%) and p-cymene (2.5 ± 0.04%). This is the first report of these components in the essential oil of Hemizygia bracteosa (Benth.) Briq.     

Chemical composition and antimicrobial activities of the essential oil of (Tunisian) Chrysanthemum trifurcatum (Desf.) Batt. and Trab. flowerheads

The chemical composition of essential oil isolated from the flowerheads of Chrysanthemum trifurcatum (Desf.) Batt. and Trab. var. macrocephalum (viv.) Beg. (Asteraceae) by hydrodistillation was analysed by GC and GC/MS. A total of 56 compounds representing 97.48% of the oil were identified: limonene (20.89%), γ-terpinene (19.13%), 1,8-cineole (10.64%), β-pinene (8.77%), α-pinene (5.32%), 2-hexenal (4.85%), 4-terpenyl acetate (3.42%), β-myrcene (2.31%), germacrene-B (2.01%), β-spathulenol (1.62%), longifolene (1.39%), α-cadinol (1.39%), α-thujene (1.23%) and β-bourbobene (1.06%) were found to be the major components. Essential oil of flowerheads of C. trifurcatum was tested for antibacterial activity against eight strains, using a microdilution method, and for cytotoxicity and antiviral activity against Herpes simplex virus type 1 using a neutral red incorporation method. The oil showed a great potential of antibacterial effect against Staphylococcus epidermidis and Bacillus subtilis, in the inhibition range of 64–66% and IC₅₀ ranging from 62.5 to 125 μg/ml. On Vero cells, the CC₅₀ of the oil was 735.9 μg/ml and it did not exhibit a significant antiviral activity.     

A new caffeoylgluconic acid derivative from the nearly ripe fruits of Evodia rutaecarpa

A new caffeoylgluconic acid derivative, trans-caffeoyl-6-O-d-gluconic acid methyl ester (1), together with two known compounds named trans-caffeoyl-6-O-d-glucono-γ-lactone (2) and trans-caffeoyl-6-O-d-gluconic acid (3), was isolated from the nearly ripe fruits of Evodia rutaecarpa (Juss.) Benth.. These compounds were isolated by various separation methods associated with the UPLC-Q-TOF-MS technique. Their structures were elucidated on the basis of extensive spectroscopic methods.     

The essential oils of the aerial parts of two Hypericum taxa (Hypericum triquetrifolium and Hypericum aviculariifolium subsp. depilatum var. depilatum (Clusiaceae)) from Turkey

In this study, essential oil compositions of two Hypericum L. taxa (Hypericum triquetrifolium Turra. and Hypericum aviculariifolium Jaub. et Spach subsp. depilatum var. depilatum (Freyn et Bornm.) Robson) naturally grown in Turkey were determined using gas chromatography and gas chromatography-mass spectrometry systems. The essential oils were obtained by hydrodistillation from the aerial parts of plants. A total of 45 compounds were identified in the essential oils of H. triquetrifolium; 1-hexanal (18.8%), 3-methylnonane (12.5%) and α-pinene (12.3%). In this study, 41 components were identified in H. aviculariifolium subsp. depilatum var. depilatum oil; α-pinene (52.1%), germacrene D (8.5%) and β-pinene (3.6%) were the predominant constituents. The essential oil analysis showed that monoterpene concentrations were higher than that of the sesquiterpenes in both oils. The results were discussed in the meaning of the usefulness of these plants and their chemicals as natural products and potential usage in chemotaxonomy.     

Volatile constituents of Amomum maximum Roxb and Amomum microcarpum C. F. Liang & D. Fang: two Zingiberaceae grown in Vietnam

The chemical composition of essential oils obtained from the hydrodistillation of different parts of Amomum maximum Roxb and Amomum muricarpum C. F. Liang & D. Fang (Zingiberaceae) grown in Vietnam are reported. The analysis was performed by means of gas chromatography–flame ionisation detectoorand gas chromatography coupled with mass spectrometry. The major compounds identified in the oils of A. maximum were β-pinene (20.4–40.8%), α-pinene (6.8–15.0%), β-elemene (2.5–12.8%) and β-caryophyllene (2.3–10.3%). Moreover, β-phellandrene (11.6%) was present in the root oil. The main compound identified in all the oil samples of A. muricarpum was α-pinene (24.1–54.7%) and β-pinene (9.2–25.9%). In addition, limonene (7.4%) and δ-3-carene (9.4%) were present in the leaves and stem oils, respectively. However, while β-phellandrene (8.3%) could be seen prominent in the root oil, the fruits contained significant amount of zingiberene (6.3%). The largest amount of τ-muurolol (13.0%) was found in the flower oil.     

Analysis of Essential Oil from Stems of Schisandra Chinensis（Turc z. ） Baill

The essential oil of Schisandra Chinensis(Turc z. ) Baill was extracted and analyzed. The essential oilwas obtained in a 1.8% (mL/100 g material) yield. Forty eight compounds were isolated and characterizedfrom the oil. The results show that the content(20. 42%) of β-pinene of naturally essential oil is the highestamong the components of the oil, which has laid a foundation for the luther development and application ofthe stems of Schisandra Chinensis(Turc z. ) Baill.     

Antibacterial activity and anticancer activity of Rosmarinus officinalis L. essential oil compared to that of its main components

In this study, Rosmarinus officinalis L. essential oil and three of its main components 1,8-cineole (27.23%), α-pinene (19.43%) and β-pinene (6.71%) were evaluated for their in vitro antibacterial activities and toxicology properties. R. officinalis L. essential oil possessed similar antibacterial activities to α-pinene, and a little bit better than β-pinene, while 1,8-cineole possessed the lowest antibacterial activities. R. officinalis L. essential oil exhibited the strongest cytotoxicity towards three human cancer cells. Its inhibition concentration 50% (IC??) values on SK-OV-3, HO-8910 and Bel-7402 were 0.025‰, 0.076‰ and 0.13‰ (v/v), respectively. The cytotoxicity of all the test samples on SK-OV-3 was significantly stronger than on HO-8910 and Bel-7402. In general, R. officinalis L. essential oil showed greater activity than its components in both antibacterial and anticancer test systems, and the activities were mostly related to their concentrations.     

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.     

Volatile constituents of Origanum vulgare L., 'thymol' chemotype: variability in North India during plant ontogeny

Essential oils derived from six different phenophases, namely early vegetative stage, late vegetative stage, early flowering stage, full flowering stage (FFS), late flowering stage and seed shattering stage of Origanum vulgare L. grown in Kumaon region of Uttarakhand, India were investigated by GC and GC-MS. A total of 38 constituents, representing 97.4-99.7% of the total oil composition, were identified. Major components of oils were thymol (40.9-63.4%), p-cymene, (5.1-25.9%), γ-terpinene (1.4-20.1%), bicyclogermacrene (0.2-6.1%), terpinen-4-ol (3.5-5.9%), α-pinene (1.6-3.1%), 1-octen-3-ol (1.4-2.7%), α-terpinene (1.0-2.2%), carvacrol (<0.1-2.1%), β-caryophyllene (0.5-2.0%) and β-myrcene (1.2-1.9%). Thymol, terpinen-4-ol, 3-octanol, α-pinene, β-pinene, 1,8-cineole, α-cubebene and (E)-β-ocimene were observed to be higher during FFS. The study showed that plant stage had a significant effect on the essential oil content and composition of O. vulgare grown in the hilly tracks of Northern India.     

Essential Oils of New Lippia alba Genotypes Analyzed by Flow-Modulated Comprehensive Two-Dimensional Gas Chromatography (GC×GC) and Chemometric Analysis

Lippia alba (Mill.) N. E. Br. (Verbenaceae) is an aromatic shrub whose essential oils have stood out as a promising source for application in several industrial fields. In this study, the essential oils chemical characterization of eight new L. alba genotypes was performed. The selected materials were collected from the Active Germplasm Bank of the Agronomic Institute and the essential oils were extracted by hydrodistillation. Flow-modulated comprehensive two-dimensional gas chromatography coupled to mass spectrometry (GC×GC-MS) was employed for chemical characterization and evaluation of possible co-eluted compounds. In addition, the chemical analyses were submitted to multivariate statistical analyses. From this investigation, 73 metabolites were identified in the essential oils of the genotypes, from which α-pinene, β-myrcene, 1,8-cineole, linalool, neral, geranial, and caryophyllene oxide were the most abundant compounds among the accessions. This is the first report disclosing α-pinene in higher amounts in L. alba (19.69%). In addition, sabinene, trans-verbenol, myrtenol, (E)-caryophyllene, α-guaiene, germacrene D, and α-bulnesene were also found in relevant quantities in some of the genotypes, and myrtenal and myrtenol could be well separated through the second dimension. Such results contributed to the understanding of the chemical composition of those new genotypes, being important to drive a future industrial applicability and studies in genetic breeding.     

α-Pinene- and β-myrcene-rich volatile fruit oil of Cupressus arizonica Greene from northwest Iran

Cupressus arizonica Greene is an aromatic evergreen coniferous plant with great importance in urban horticulture and in the pharmaceutical and fragrance industries. The hydrodistilled volatile fruit oil of cultivated C. arizonica from northwest Iran was analysed by GC/MS. Forty-three components were identified, accounting for 96.4% of the total oil. Monoterpenoids (91.9%) dominated the identified components of the essential oil, followed by a lesser portion of sesquiterpenoids (4.2%). Monoterpene hydrocarbons (87.9%) were the principal subclass of components, with α-pinene (54.3%), β-myrcene (11.1%), δ-3-carene (6.5%) and limonene (6%) as main constituents. β-Pinene (4%), terpinolene (2.8%) and camphene (1.1%) were the other monoterpenoids present in notable amounts. α-Terpineol (1.4%) was the only representative of the oxygen-containing monoterpenoids. Sesquiterpenoids had a minor share in the volatile oil's composition. Hydrocarbonic compounds (91.1%) had a higher share compared to the oxygenated components (5%). Comparison of the essential oil profile of C. arizonica Greene plants cultivated in Iran showed remarkable quantitative but slight qualitative differences with previous reports from other parts of the world. In summary, the chemical and percentage composition of the studied oil from cultivated C. arizonica Greene from northwest Iran was characterised by a high occurrence of α-pinene and β-myrcene, and is thus competent for use in related industries and as a favourite ornamental aromatic tree.