Eudesmane-type sesquiterpenoids from the aerial parts of Artemisia lavandulaefolia and their anti-pancreatic cancer activities.

Five undescribed eudesmane sesquiterpenoids, artemilavanins A-E, and one undescribed rearranged eudesmane sesquiterpenoid, artemilavanin F, were isolated from the 95% ethanol extract of the aerial parts of Artemisia lavandulaefolia DC., along with ten known compounds. The structures and configurations of undescribed compounds were mainly elucidated by spectroscopic analyses and single-crystal X-ray diffraction analysis. Among all isolated compounds, artemilavanin F exhibited inhibitory activity on PANC-1 pancreatic cancer cells with IC50 of 9.69 ± 2.39 µM. Artemilavanin F inhibited PANC-1 cell proliferation by induction of G2/M cell cycle arrest and apoptosis mediated by downregulation of cyclin-dependent kinases and accumulation of reactive oxygen species. Moreover, artemilavanin F inhibited the colony formation, cell migration and sphere formation of PANC-1 cells, indicating the suppression of stem-cell-like phenotype of PANC-1 cells. Further results confirmed that the expression of cancer stem cell markers such as Bmi1, CD44, CD133 were inhibited by artemilavanin F. Downregulation of epithelial-mesenchymal transition (EMT) markers such as N-cadherin and Oct-4 indicated the potential of artemilavanin F in prevention of metastasis.

Two new eudesmane sesquiterpene glucosides from the aerial parts of Artemisia vulgaris.

Using combined chromatographic methods, two new sesquiterpene glucosides, vulgarosides A (1) and B (2), and two known analogs ainsliaside E (3) and pumilaside A (4) were isolated from the aerial parts of Artemisia vulgaris. Their chemical structures were established by spectroscopic methods, including one and two-dimensional nuclear magnetic resonance (1 D and 2 D-NMR) spectroscopy and high-resolution electrospray ionization mass spectrometry (HR-ESI-MS). In addition, their cytotoxicity on five human cancer cell lines, including KB (epidermoid carcinoma), HepG2 (hepatocarcinoma), MCF7 (breast carcinoma), SK-Mel-2 (melanoma), and LNCaP (prostate cancer) was also evaluated by the SRB assay. However, none of the tested eudesmane sesquiterpene glycosides showed significant cytotoxicity (IC50>100 µM).

Polyhydroxylated eudesmane sesquiterpenoids and sesquiterpenoid glucoside from the flower buds of Tussilago farfara.

Chemical fractionation of the n-BuOH partition, which was generated from the EtOH extract of the flower buds of Tussilago farfara, afforded a series of polar constituents including four new sesquiterpenoids (1-4), one new sesquiterpenoid glucoside (5) and one known analogue (6) of the eudesmane type, as well as five known quinic acid derivatives (7-11). Structures of the new compounds were unambiguously characterized by detailed spectroscopic analyses, with their absolute configurations being established by X-ray crystallography, electronic circular dichroism (ECD) calculation and induced ECD experiments. The inhibitory effect of all the isolates against LPS-induced NO production in murine RAW264.7 macrophages was evaluated, with isochlorogenic acid A (7) showing significant inhibitory activity.

[New eudesmane sesquiterpenoids from Atractylodis Macrocephalae Rhizoma and their inhibitory activities against SREBPs].

Three sesquiterpenoids were isolated and purified from the 95% ethanol extract of Atractylodis Macrocephalae Rhizoma by column chromatography on silica gel, Sephadex LH-20, ODS, and high-performance liquid chromatography(HPLC). Their chemical structures were identified on the basis of spectroscopic analysis and physiochemical properties as(7Z)-8ß,13-diacetoxy-eudesma-4(15),7(11)-diene(1), 7-oxo-7,8-secoeudesma-4(15),11-dien-8-oic acid(2), and guai-10(14)-en-11-ol(3). Compounds 1 and 2 are new compounds and compound 3 was obtained from Compositae family for the first time. Compounds 1, 2, and 3 showed weak inhibitory activities against sterol regulatory element-binding proteins(SREBPs).

Simultaneous determination of asarinin, ß-eudesmol, and wogonin in rats using ultraperformance liquid chromatography-tandem mass spectrometry and its application to pharmacokinetic studies following administration of standards and Gumiganghwal-tang.

Asarinin, ß-eudesmol, and wogonin have common antiangiogenic activities and have the potential for use in chemotherapy. Besides, they are multivalent substances that are combined in various herbal medicines. The purpose of this study was to develop a method for simultaneous analysis of asarinin, ß-eudesmol, and wogonin, which are representative pharmacological components of Asarum heterotropoides, Atractylodes lancea, and Scutellaria baicalensis, respectively, in rat biosamples using ultraperformance liquid chromatography-tandem mass spectrometry. The three components were separated using 5 mm aqueous ammonium acetate containing 0.1% formic acid and acetonitrile as a mobile phase, equipped with a KINETEX core-shell C18 column. The analysis was quantitated on a triple-quadrupole mass-spectrometer employing electrospray ionization, and operated in the multiple reaction monitoring mode. The chromatograms showed high resolution, sensitivity, and selectivity with no interference with plasma, urine, and feces constituents. The developed analytical method satisfied international guidance criteria and could be successfully applied to the pharmacokinetic (PK) studies evaluating oral bioavailability of asarinin, ß-eudesmol, and wogonin after oral and intravenous administration and their urinary and fecal excretion ratios after oral administration to rats. Furthermore, the analysis was extended to PK studies following oral administration of Gumiganghwal-tang. This study was the first simultaneous analysis of the aforesaid three constituents in rat plasma, urine, and feces that also determined their PK parameters.

Microscopic Characteristic and Chemical Composition Analysis of Three Medicinal Plants and Surface Frosts.

The accumulation of chemical constituents of some medicinal plants, such as Paeonia ostii T. Hong et J. X. Zhang, Houpoëa officinalis (Rehder and E. H. Wilson) N. H. Xia and C. Y. Wu. and Atractylodes lancea (Thunb.) DC, can precipitate on the surface and form frosts after natural or artificial intervention. The characteristics of these three medicinal plants and their frosts were analyzed by light microscope, polarizing microscope, stereomicroscope, and metalloscope. The results of ordinary Raman of P. ostii and H. officinalis showed that the frosts of P. ostii matched paeonol, while that of H. officinalis matched magnolol and honokiol. In P. ostii and its frost, 19 peaks were identified by UPLC-Q/TOF-MS, and the main component was paeonol. Eleven components were identified in H. officinalis and its frosts, and the main components were magnolol and honokiol. A. lancea and its frosts were analyzed by gas chromatography-mass spectrometry (GC-MS), 21 were identified, and its main components were hinesol and ß-eudesmol. These three medicinal plants accumulate compounds and precipitate frosts on the surface. The results show that the components of the frosts provide a basis for quality evaluation and research on similar medicinal plants, and reveals the scientific connotation of "taking the medicinal materials' precipitated frosts as the best" of P. ostii, H. officinalis, and A. lancea, to some extent.

Chemical investigations of male and female leaf extracts from Schinus molle L.

The pepper-tree Schinus molle is an evergreen ornamental plant with various and diversified list of medical uses. In this article we analysed the chemical composition of male and female leaves of this plant during the off-flowering and flowering seasons. The leaf extracts were obtained by using a sequential extraction with solvents of different polarities and the chemical composition was investigated by GC-MS. The results showed a total of twenty-three components, in which elemol is the most abundant constituent followed by bicyclogermacrene, γ-eudesmol, α-eudesmol, ß-eudesmol and isocalamendiol. The petroleum ether and diethyl ether extracts from male and female flowering and off-flowering leaves consisted of sesquiterpene hydrocarbons as a major constituent followed by monoterpene hydrocarbons, while the acetone extracts showed a different composition. The obtained results show differences in the chemical composition between male and female and flowering and not flowering.

Chemical composition and α-amylase inhibitory activity of the essential oil from Sabina chinensis cv. Kaizuca leaves.

Sabina chinensis cv. Kaizuca (SCK) is a variant of S. chinensis L. The essential oil from its leaves exhibited α-amylase inhibitory activity in vitro and the IC50 value was 187.08 ± 0.56 µg/mL. Nineteen compounds were identified from this essential oil by gas chromatography-mass spectrometry (GC-MS) analysis. The major compounds identified were bornyl acetate (42.6%), elemol (20.5%), ß-myrcene (13.7%) and ß-linalool (4.0%). In order to study the reason of the α-amylase inhibitory activity of this essential oil, the identified compounds were docked with α-amylase by molecular docking individually. Among these compounds, γ-eudesmol exhibited the lowest binding energy (-6.73 kcal/mol), followed by α-copaen-11-ol (-6.66 kcal/mol), cubedol (-6.39 kcal/mol) and α-acorenol (-6.12 kcal/mol). The results indicated that these compounds were the active ingredients responsible for the α-amylase inhibitory activity of essential oil from SCK.

Silver perchlorate in the mobile phase for rapid separation and determination of a pair of positional isomers in Inula racemosa Hook.f. with RP-HPLC.

Alantolactone and isoalantolactone isolated from many species of plants are a pair of positional isomers of CC bond. Previously, alantolactone and isoalantolactone have been proved to be good lead compounds for future anticancer agent development. Similarity of their molecular structures increases the separation difficulty for these two isomers on a conventional C18 column. Silver perchlorate (AgClO4) as mobile phase additives with RP-HPLC for improving the separation was developed for rapid determination of the positional isomers in Inula racemosa Hook.f. The effects of the concentration of silver perchlorate on the separation of the analytes were investigated. The composition of acetonitrile and water containing 5.0% silver perchlorate in a 65:35 (v/v) ratio was used as mobile phase, in which they were well separated within a short period of time on the C18 column. The method was successfully applied to determine them in an extract of Inula racemosa Hook.f. root. Silver perchlorate in mobile phase can efficiently improve the separation of the positional isomers and could be applied to rapidly determinate their content in this plant.

Simultaneous determination of chlorogenic acid, caffeic acid, alantolactone and isoalantolactone in Inula helenium by HPLC.

A rapid and sensitive high-performance liquid chromatographic (HPLC) method was developed for the simultaneous separation and determination of chlorogenic acid, caffeic acid, alantolactone and isoalantolactone in Inula helenium. The HPLC separation was performed on an Elite Hypersil C18 column (200 × 4.6 mm i.d., 5 µm particle size) with a gradient elution of solvent A (acetonitrile) and solvent B (0.1% phosphoric acid in water) at a flow rate of 1.0 mL/min. Detection was monitored at 225 nm. The recovery of chlorogenic acid ranged from 95.6 to 107.7%, the recovery of caffeic acid ranged from 95.4 to 104.2%, the recovery of alantolactone ranged from 95.8 to 100.8% and the recovery of isoalantolactone ranged from 96.5 to 102.3%. The retention times for chlorogenic acid, caffeic acid, alantolactone and isoalantolactone were 5.2, 7.1, 25.6 and 26.6 min with the limits of detection of 0.069, 0.021, 0.039 and 0.051 µg/mL, respectively. Relative standard deviation for the intra-day and inter-day was ≤2.5%. The validated method is reliable for the routine control of these four compounds in I. helenium.

Volatile composition and sensory properties of Indian herbal medicine-Pavonia odorata-used in Ayurveda.

The chemical composition of volatile oil obtained from aerial parts of Pavonia odorata were investigated using gas chromatography-mass spectrometry (GC-MS). Its aroma-active compounds were identified using gas chromatography-olfactometry (GC-O) and aroma extraction dilution analysis (AEDA). In order to determine the relative contribution of each compound to the aroma of P. odorata, relative flavour activity (RFA) was calculated. The hydrodistillation of P. odorata afforded yellowish oil and the yield was 0.009% (w/w) with a spicy, sweet, and green odour. Eighty-five compounds were identified in the oil by GC-MS; the major constituents of the volatile oil were ageratochromene (11.95%), palmitic acid (9.95%), hexahydrofarnesyl acetone (5.96%), ß-eudesmol (4.53%) and ß-caryophyllene oxide (3.08%). The most characteristic aroma compounds in the volatile oil were identified for ß-caryophyllene oxide (FD-factor = 128, spicy), (E)-pinocarveol (FD-factor = 64, sweet), 3-butylpyridine (FD-factor = 64, spicy), and 2-nonanone (FD-factor = 32, green) by GC-MS, GC-O and AEDA. It seems that these compounds are responsible for the spicy, sweet and green odour of the aerial parts of P. odorata. The antioxidant activity of the volatile oil was also investigated by the oxygen radical absorbance capacity (ORAC) assay using fluorescein (FL) as the fluorescent probe. The ORAC value of the oil was 594.2 ± 25.9 µM TE/g. The results indicated that the volatile oil from the aerial parts of P. odorata could be considered as a natural antioxidant effect agent.

Apolar Laurus nobilis leaf extracts induce cytotoxicity and apoptosis towards three nervous system cell lines.

In the course of a bioactivity screening of Mediterranean plants, the assessment of neuroprotective properties of Laurus nobilis L. was of interest. Dried leaves were extracted by sonication using CHCl3 as solvent. The CHCl3 parental extract (CHCl3-pe) was fractionated to yield CHCl3 (LnC-1), EtOAc (LnC-2), MeOH (LnC-3) fractions. Each fraction underwent an extensive screening towards human neuroblastoma (SK-N-BE(2)-C, and SH-SY5Y) and rat glioma (C6) cell lines. MTT and SRB cytotoxicity tests were performed. The effect on the plasma membrane integrity was evaluated by assessment of LDH release. The caspase-3 activation enzyme and DNA fragmentation were also evaluated. The oxidant/antioxidant ability of all the extracts were evaluated using different methods. Furthermore, a metabolite profiling of the investigated extracts was carried out by GC-EI-MS. CHCl3-pe contained terpenes, allylphenols, and α-tocopherol. Dehydrocostus lactone was the main constituent. As result of the fractionation technique, the LnC-1 extract was mainly composed of α-tocopherol, whereas the LnC-2 fraction was enriched in guaiane and eudesmane terpenes. The most cytotoxic LnC-2 fraction induced apoptosis; it was ineffective in preventing in vitro free radicals production. Overall, the experimental results support a possible role of LnC-2 preparation as a chemopreventive agent for neuronal cells or other cells of the CNS.

Five new eudesmane-type sesquiterpenoid lactones biotransformed from atractylenolide I by rat hepatic microsomes.

The work presented here is the first study performed on the biotransformation and/or metabolism of atractylenolide I (1) as a valuable anti-inflammatory and chemopreventive agent, using liver microsomes from rats pre-treated with sodium phenobarbital. Two known eudesmane-type sesquiterpenoid lactones, namely 1ß-acetoxyatractylenolide I (2) and 1ß-hydroxy-atractylenolide I (3), and five new ones, namely 3ß-hydroxy-atractylenolide I (4), 1ß,13-dihydroxy-atractylenolide I (5), 1ß,2α-dihydroxy-atractylenolide I (6), 1ß,3α-dihydroxy-atractylenolide I (7), and 1ß,3ß-dihydroxy-atractylenolide I (8) were obtained. Their chemical structures were unambiguously established by both 1D and 2D NMR as well as mass spectroscopic techniques. The result indicated that the parent prototype compound 1 could be specifically oxidized at C-1, C-2 and C-3 of A-ring, suggesting that the oxidizable of 1 may contribute to its in vivo anti-inflammatory and chemopreventive effects. And the result also provided valuable information for further investigation of relationship among metabolic activation and liver microsomal cytochrome P450 enzyme isoforms.

Aroma evaluation of gamazumi (Viburnum dilatatum) by aroma extract dilution analysis and odour activity value.

INTRODUCTION: Viburnum dilatatum (gamazumi) is widely distributed in Japan and China. Recently, juice from V. dilatatum fruits has been manufactured in Japan. Concerning the aroma of V. dilatatum, phenethyl alcohol, 3Z-hexenol and l-linalool have been identified in the essential oil from the flowers of V. dilatatum, however, there are no detailed reports on the aroma of V. dilatatum elucidated using sensory evaluation. OBJECTIVE: To clarify odourants contributing to the characteristic aroma, the aroma extract dilution analysis (AEDA) method was performed through gas chromatography olfactometry (GC-O) analysis. METHODOLOGY: The aroma-active compounds were identified by GC-O and AEDA, and in order to determine the relative contribution of each compound to the aroma of V. dilatatum, odour activity value (OAV) has been used. RESULTS: The hydrodistillation of the leaf and branch of V. dilatatum afforded pale yellowish oils, with yields of 0.008 and 0.015% (w/w). The main components of the leaf oil were 3Z-hexenal (12.7%) and linalool (10.8%). In branch oil, palmitic acid (18.3%) and linoleic acid (8.2%) were identified. With regard to aroma components, 24 and 14 compounds were identified in the leaf and branch oils respectively, by GC-O analysis. CONCLUSION: On the basis of AEDA, OAVs and sensory evaluations, nonanal is estimated as the main aroma compound of leaf and branch oil, as the other aroma compounds, C6 compounds and 2-pentyl furan make green odour; linalool, eugenol and ß-ionone play important role in the sweet odour of leaf oil. In branch oil, cis-furanlinalool oxide and eugenol make sweet odour, and ß-eudesmol contributes to woody odour.

Chemical composition and antioxidant activity of the essential oil of aerial parts of Petasites albus from Iran: a good natural source of euparin.

The chemical composition of the essential oil of Petasites albus (known as 'Baba Adam' in Iran) was investigated by capillary gas chromatography and gas chromatography-mass spectrometry for the first time. Twenty components were identified, accounting for 99.7% of the oil composition. The major compounds were euparin (73%), α-eudesmol (13.2%) and ß-selinene (4.5%). Euparin, the main component of the essential oil, was isolated and characterised by spectroscopic techniques. The antioxidant activities of the essential oil and euparin were evaluated by using the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay and are shown to exhibit a moderate antioxidant activity.

New sesquiterpene coumarins from the roots of Ferula flabelliloba.

Three new sesquiterpene coumarins, namely, farnesiferone B (1), flabellilobin A (2) and flabellilobin B (3), together with nine known compounds, ligupersin A, 7-epi-gamma-eudesmol, persicasulfide A, conferdione, umbelliprenin, conferone, feselol, lehmferin and farnesiferol B were isolated from the roots of Ferula flabelliloba Rech. f. & Aell. (Apiaceae). The structures of these compounds were elucidated by various 1- and 2-D NMR techniques as well as HREIMS.

[Simultaneous determination of atractylone, hinesol, beta-eudesmol, atrctylodin in Atractylodes lancea and hierarchical cluster analysis].

OBJECTIVE: To develop a GC method for simultaneous determination of 4 compounds (atractylone, hinesol, beta-eudesmol and atractylodin) in Atractylodes lancea. METHOD: A HP-1 capillary column (0.25 mm x 30 m, 0.25 microm) was used. The detector was FID:Inlet temperature was 250 degrees C. The detector temperature was 250 degrees C. The column temperature was set at 145 degrees C and held for 25 min after injection, then programmed at 10 degrees C x min(-1) to 250 degrees C and held for 10 min at the temperature. The carrying gas was nitrogen, split ratio was 40:1. Injection volume was 2 microL, Cluster analysis was performed by SPSS13.0 software. RESULT: The linear ranges for atractylone, hinesol, beta-eudesmol and atractylodin were 0.0122. 32 (r = .9998), 0.008-1.68 (r = 0.9998), 0.009-1.76 (r = 0.9999), 0.016-3.20 g x L(-1) (r = 0.9997), respectively. The average recoveries (n = 3) of atractylone, hinesol, beta-eudesmol and atractylodin were 98.0%-99.0%, 97.7%-99.4%, 98.4%-99.2%, 97.8%-99.7%, respectively. The samples analyzed were divided into two classes. CONCLUSION: This method is simple, specific, repeatable and stable. It can be applied for the simultaneous determination of 4 compounds (atractylone, hinesol, beta-eudesmol and atractylodin) in A. lancea, which will provide the basis for the quality control of A. lancea. The contents of 4 active compounds were significantly different between geo-authentic and non-authentic producing areas.

HPLC determination and NMR structural elucidation of sesquiterpene lactones in Inula helenium.

A high performance liquid chromatography (HPLC) method was developed for the simultaneous quantification of three bioactive sesquiterpene lactones in Inula helenium L., namely igalane (1), isoalantolactone (2) and alantolactone (3). The HPLC separation was performed on an Agilent Zorbax XDB-C(18) column (250mmx4.6mm, 5microm) with a mobile phase consisting of 55% acetonitrile and 45% water, at a flow rate of 1.0ml/min, and detected at 210nm. All three regression equations indicated good linear relationships (r(2)>or=0.9994) between the peak area of each compound and its respective concentration. The HPLC assay was reproducible with overall intra- and inter-day variation of less than 2.85%. The recoveries, measured at three concentrations, varied from 96.60 to 104.43%. This assay was successfully applied to the determination of three bioactive sesquiterpene lactones in eleven samples. The results demonstrated that the assay developed herein was rapid, accurate, reliable and could be readily utilized as a quality control method for I. helenium. In addition, two minor isomers in I. helenium were isolated by semi-preparative HPLC. Their structures were elucidated on the basis of NMR analysis.

Montanoa tomentosa glandular trichomes containing kaurenoic acids chemical profile and distribution.

Montanoa tomentosa has been used in traditional medicine in Mexico to treat diverse female health disorders; it is particularly useful in inducing childbirth. Microscopic analysis of leaf surfaces of M. tomentosa revealed the presence of glandular trichomes. The chemical profile and distribution of glandular trichomes from different developmental stages of M. tomentosa leaves were investigated. Two diterpenic acids, kaurenoic and grandiflorenic were detected in glandular trichomes through the glandular microsampling technique and GC/MS analysis. In the glandular trichomes of the leaves also up to twenty-six volatile terpenes were identified, where beta-eudesmol and valencene were the most abundant terpenes.

New eudesmane derivatives and other sesquiterpenes from the epigeal parts of Dittrichia graveolens.

In bioassay-guided searches for novel bioactive natural products from higher plants of the Egyptian flora, two new eudesmane sesquiterpene derivatives, 3alpha-hydroxyilicic acid methyl ester (1) and 2alpha-hydroxy-4-epi-ilicic acid (2), together with 11 known sesquiterpenes were isolated from bioactive fractions of the active epigeal parts extracts of Dittrichia graveolens (L.) GREUTER (Asteraceae) growing in the coastal regions of northwestern Egypt. Four other known sesquiterpene lactones with different carbon skeletons, named 2alpha-hydroxy-2R-xanthalongin (8), 4-epi-isoinuviscolide (9), 8-epi-helenalin (10), and bigelovin (11), were also isolated for the first time from the same source. The stereochemical structures of the isolated compounds were elucidated on the basis of physical and spectroscopic methods including UV, IR, 1H-, 13C-NMR, distortionless enhancement by polarization transfer, 2D NMR, 1H-1H correlation spectroscopy, 1H-13C heteronuclear single-quantum coherence, 1H-13C heteronuclear multiple-bond connectivity, 1H-1H nuclear Overhauser effect spectroscopy experiments, and high-resolution mass spectrometry, as well as some chemical transformations. The antimicrobial, antiinflammatory, and antipyretic activities of D. graveolens extracts and chromatographic fractions were carried out and the various bioactivities of our findings are discussed.