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NPC Natural Product Communications

EDITOR-IN-CHIEF

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HONORARY EDITOR

PROFESSOR GERALD BLUNDEN The School of Pharmacy & Biomedical Sciences,

University of Portsmouth, Portsmouth, PO1 2DT U.K.

axuf64@dsl.pipex.com

Essential Oil Composition and Antibacterial Activity of Anthemis mixta and A. tomentosa (Asteraceae) Carmen Formisanoa, Daniela Riganoa, Felice Senatorea, Francesco Maria Raimondob, Antonella Maggioc and Maurizio Brunoc,* aDepartment of Chemistry of Natural Products, University of Naples “Federico II”, Via D. Montesano, 49 - 80131 Naples, Italy

bDipartimento di Biologia Ambientale e Biodiversità, Universita` di Palermo, Via Archirafi 38, Palermo 90123, Italy

cDepartment STEMBIO, Sect. of Organic Chemistry, University of Palermo, Viale delle Scienze, Parco d’Orleans II - 90128 Palermo, Italy maurizio.bruno@unipa.it

Received: July 21st, 2012; Accepted: August 9th, 2012

In the present study the chemical composition of the essential oils from aerial parts and roots of Anthemis mixta L. and A. tomentosa L. was evaluated by GC and GC-MS, and their antibacterial activity tested against ten bacterial species. Hexadecanoic acid (15.2%) was recognized as the main constituent of A. mixta, together with τ-cadinol (6.7%), while in both aerial parts and roots of A. tomentosa nonacosane (21.9% and 20.7%), heptacosane (8.1% and 6.0%), hexadecanoic acid (8.1% and 27.1%) and hexahydrofarnesylacetone (6.8% and 5.5%) prevailed. The oils from aerial parts of both species showed a good activity against Gram-positive bacteria. These results suggest that the plants could be potentially used in food manufacture and cosmetology as preservative agents or in medicine as new antibiotics. Keywords: Asteraceae, Anthemis mixta, Anthemis tomentosa, Essential oil, Antimicrobial activity.

The genus Anthemis L., tribe Anthemideae, subtribe Anthemidinae, is the second largest genus of the Asteraceae family and comprises about 210 species, mainly distributed in western Eurasia, the Mediterranean, and eastern Africa. About 62 species are reported for the European flora [1,2]. Anthemis species are used as herbal tea and for food flavouring and the flowers and aerial parts have well-documented use as antiseptic and healing herbs in folk medicine [1,2]. Extracts, tinctures, tisanes (teas), and salves are widely used as anti-inflammatory, antibacterial, antispasmodic, and sedative agents. Extracts are used to allay pain and irritation, clean wounds and ulcers, and aid prevention as well as therapy for irradiated skin injuries, treatment of cystitis and dental afflictions [3]. Dried flowers of many Anthemis species are used as a replacement for German chamomile and as stimulant, emmenagogue, and carminative infusions [3]. These properties render essential oils from Anthemis ssp. flowers very useful for pharmaceuticals, and food additives, as well as important sources in food, aromatic and cosmetic industries [4,5]. Although Anthemis species have been widely studied as regards the isolation of different metabolites, particularly sesquiterpene lactones [6], their essential oil composition is known only for a limited number of taxa [5]. So, as a continuation of our studies on this interesting genus [7] here we describe the chemical composition and antibacterial activity of essential oils from aerial parts and roots of two species collected in the Mediterranean area, Anthemis mixta L. and A. tomentosa L. Such information is valuable as plant extracts and essential oils containing high biological activities can prove beneficial for maintenance of optimal health and may increase the demand of these bioactive substances by the food, cosmetic and pharmaceutical industries.

Hydrodistillation of A. mixta aerial parts (AmA) and roots (AmR) gave two pale yellow oils. Overall, 66 compounds were identified (56 in AmA and 16 in AmR), representing 90.6% and 90.3% of the total components, respectively. The components are listed in Table 1 according to their retention indices on a HP 5MS column and are classified on the basis of their chemical structures into 10 classes. The two oils presented a different composition and the identity of the most dominant components differed notably. Hexadecanoic acid (15.2%) was recognized as the main constituent of AmA, together with santolina alcohol (11.7%), τ-cadinol (6.7%), spathulenol (6.5%), caryophyllene oxide (3.8%), artemisia alcohol (3.8%), τ-muurolol (3.0%), hexahydrofarnesylacetone (2.7%), mintketone (2.5%), (E)-nerolidol (2.1%), and α-cadinol (2.1%), whereas in AmR the most abundant components were hexadecanoic acid (52.0%), 1,2-benzenedicarboxylic acid bis(2-methylpropyl) ester (12.3%), tetradecanoic acid (4.8%), dodecanoic acid (4.2%), hexahydrofarnesylacetone (3.0%), and hexadecanoic acid methyl ester (2.2%). Generally, AmA consisted mainly of oxygenated sesquiterpenes (32.7%) and fatty acids and esters (23.9%), while AmR was composed almost entirely of acids and esters (78.7%). Monoterpenes, represented especially by irregular monoterpenes (16.5%) and oxygenated monoterpenes (5.9%), contributed 23% for AmA and were absent from AmR. Irregular monoterpenes, santolina alcohol (11.7%), artemisia alcohol (3.8%) and yomogi alcohol (1.0%), were present only in AmA. The occurrence of the irregular monoterpenes, such as cis-chrysanthenyl acetate, artemisyl acetate and yomogi alcohol is a common feature of the tribe Anthemideae [8]. Santolina alcohol is also reported in A. cotula L. from Greece [3] and artemisia alcohol in A. altissima L. grown in Iran [9]. Generally, a significant amount of irregular monoterpenoids was found in A. xylopoda, A. montana, A. carpatica, A. melampodina and A. altissima var. altissima [5].

NPC Natural Product Communications 2012 Vol. 7 No. 10

1379 - 1382

1380 Natural Product Communications Vol. 7 (10) 2012 Formisano et al.

Table 1: Composition of essential oils from of A. mixta aerial parts (AmA) and roots (AmR), and A. tomentosa aerial parts (AtA) and roots (AtR).

COMPONENT Ki a LRI b AmAc AmRd AtAe AtRf Identificationd

Hydrocarbons 1.5 5.4 42.1 34.2 1,2-Dihydro-1,4,4-trimethyl naphtalene 1236 1808 1.5 1, 2 Docosane 2200 2200 0.7 1, 2, 3 Tricosane 2300 2300 1.5 2.6 2.5 2.8 1, 2, 3 Tetracosane 2400 2400 0.9 1, 2, 3 Tetracosane 2400 2400 t 1, 2, 3 Pentacosane 2500 2500 0.8 4.5 3.7 1, 2, 3 Hexacosane 2600 2600 0.4 0.2 1, 2, 3 Heptacosane 2700 2700 0.4 8.1 6.8 1, 2, 3 Octacosane 2800 2800 0.7 1, 2, 3 Nonacosane 2900 2900 21.9 20.7 1, 2 Hentriacontane 3100 3100 2.5 1, 2 Carbonylic compounds 4.4 3.6 8.3 5.9 Phenylacetaldehyde 1045 1663 0.8 1,2,3 (E,E)-2,4-Decadienal 1315 1827 0.3 1, 2 (E)-β-Damascenone 1358 1787 0.7 1, 2 cis-Jasmone 1397 1959 0.1 1, 2 Hexahydrofarnesylacetone 1845 2131 2.7 3.0 6.8 5.9 1, 2 (E,E)-Farnesylacetone 1920 2387 0.5 1, 2 9,12,15-Octadecatrienal 2111 2367 0.8 1, 2 Eicosanal 2221 2571 0.6 1, 2 Monoterpene hydrocarbons 0.6 1.0

p-Cymene 1025 1278 0.6 1.0 1, 2, 3 Limonene 1030 1203 t 1, 2, 3 Sesquiterpene hydrocarbons 4.3 7.2

α-Longipinene 1345 1483 0.3 1, 2 Cyclosativene 1363 1492 0.4 1, 2 -Copaene 1377 1497 0.8 0.2 1, 2 β-Patchoulene 1378 1527 1.5 1, 2 (E)-Caryophyllene 1418 1612 0.5 0.6 1, 2, 3 γ-Elemene 1438 1652 0.4 1, 2 (E)-β-Farnesene 1452 1672 0.8 1, 2 α-Humulene 1455 1689 1.5 1, 2 β-Selinene 1475 1715 0.3 1, 2 Germacrene D 1477 1726 1.2 1,2 β-Guaiene 1490 1612 0.9 1, 2 Valencene 1492 1741 0.6 1, 2 -Muurolene 1503 1740 1.0 1, 2 β-Bisabolene 1510 1743 0.3 1, 2 δ-Cadinene 1526 1773 0.2 1, 2 Irregular monoterpenes 16.5 Yomogi alcohol 994 1405 1.0 1, 2 Santolina alcohol 1024 1402 11.7 1, 2 Artemisia alcohol 1085 1512 3.8 1, 2 Oxygenated monoterpenes 5.9 3.5 1,8-Cineole 1034 1213 t 0.9 1, 2, 3 cis-p-Ment-2-en-1-ol 1125 1638 0.3 1, 2 α-Campholenal 1128 1487 0.2 1, 2 trans-Verbenol 1152 1683 0.4 1, 2 Borneol 1167 1718 1.6 1, 2, 3 cis-Pinocamphone 1174 1565 0.2 1, 2 Terpinen-4-ol 1176 1611 0.3 1, 2, 3 p-Cymen-8-ol 1185 1856 0.4 1, 2 -Terpineol 1189 1706 0.3 1, 2, 3 Myrtenol 1196 1804 0.2 1, 2 trans-Carveol 1217 1845 0.5 1, 2 Bornyl acetate 1290 1597 2.3 1, 2, 3 trans-Pinocarvyl acetate 1295 1661 0.7 1, 2 Piperitone 1343 1748 0.7 1, 2 Neryl acetate 1358 1734 0.4 1, 2 Oxygenated sesquiterpenes 32.7 2.6 7.5 2.2 cis-α-Copaen-8-ol 1553 2076 4.1 1, 2

1564 2.1 1, 2 Aromadendrene oxide 1568 2402 0.1 1, 2 Spathulenol 1578 2150 6.5 1, 2, 3 Caryophyllene oxide 1579 2208 3.8 t 1, 2, 3 β-Copaen-4--ol 1585 2135 0.7 0.8 1, 2 Viridiflorol 1593 2103 1.7 1, 2 Salvial-4(14)-en-1-one (Mintketone) 1602 2037 2.5 1, 2 nor-Copaanone 1622 2182 0.5 1, 2 Caryophylla-3,8(13)-dien-5α-ol 1632 2371 1.2 1, 2 t-Cadinol 1640 2185 6.7 1, 2 t-Muurolol 1641 2209 3.0 1, 2 Caryophyllenol II 1649 2396 t 1, 2 β-Eudesmol 1650 2258 0.8 2.6 0.7 1, 2

Essential oils of Anthemis mixta and A. tomentosa Natural Product Communications Vol. 7 (10) 2012 1381

α-Cadinol 1652 2255 2.1 t 0.6 1, 2 α-Eudesmol 1654 2206 1.8 0.9 1, 2 Germacrene D-1,10-epoxide 1659 2053 0.7 1, 2 cis-(Z)-α-bisabolene epoxide 1675 0.2 1, 2 β-Costol 1780 2607 0.9 1, 2 Fatty acids and derivatives 23.9 78.7 10.8 49.7

Decanoic acid 1367 2316 1.4 1, 2, 3 Isoamyl phenyl acetate 1490 2016 1.7 1, 2 Dodecanoic acid 1566 2503 4.2 3.7 1, 2, 3 γ-Dodecalactone 1680 2396 1.6 0.5 1, 2 Tetradecanoic acid 1767 2675 2.5 4.8 3.5 1, 2, 3 Benzyl phenylacetate 1802 0.8 1, 2 2-Phenyl ethylbenzoate 1858 2733 0.4 1, 2 Benzyl salicylate 1867 2810 0.7 1, 2, 3 Pentadecanoic acid 1873 2825 1.2 1.1 1, 2, 3 1,2-Benzenedicarboxylic acid bis(2-methylpropyl) ester 1874 2523 12.3 5.0 1, 2 Hexadecanoic acid methyl ester 1925 2208 2.2 1.5 1, 2, 3 Hexadecanoic acid 1957 2932 15.2 52.0 8.2 32.1 1, 2, 3 (Z)-9-Octadecenoic acid 2115 3168 1.6 0.5 1, 2, 3 (Z,Z)-9,12-Octadecadienoic acid 2122 3160 2.6 1.8 1,2,3 Phenolic compounds 10.4 4-Vinylguaiacol 1312 2180 3.6 1,2 Eugenol 1353 2186 1.6 1,2,3 Isoeugenol 1396 2296 5.2 1, 2 Others 0.8 0.3 0.1 Indole 1290 2471 0.3 0.1 1, 2, 3 (E)-Phytol 2135 2625 0.8 1, 2 Total amount of compounds 90.6 90.3 91.1 92.1

a: Linear retention index on a HP 5MS column; b: Linear retention index on a HP Innowax column c: Mass of compounds in mg/100 mg oil; t, trace (<0.1%); d: 1: linear retention index, 2: mass spectrum, 3: co-injection with authentic compound. Hydrodistillation of A. tomentosa aerial parts (AtA) and roots (AtR) gave two pale yellow oils. Overall, 40 compounds were identified (34 in AtA and 16 in AtR), representing 91.1% and 92.1% of the total components, respectively. The main components of the oil of both aerial parts (AtA) and roots (AtR) of A. tomentosa were nonacosane (21.9% and 20.7%), heptacosane (8.1% and 6.0%), hexadecanoic acid (8.1% and 27.1%), hexahydrofarnesylacetone (6.8% and 5.5%), and pentacosane (4.5% and 3.0%). The AtA consisted of hydrocarbons (42.1%), phenolic compounds (10.4%), carbonylic compounds (8.3%), oxygenated sesquiterpenes (7.5%), and sesquiterpene hydrocarbons (7.2%). In addition, the aerial parts of A. tomentosa contain a good amount of isoeugenol (5.2%) and 4-vinylguaiacol (3.6%), which were not present in the roots. A. tomentosa roots are instead rich in fatty acids and esters (49.7%), and hydrocarbons (34.2%), which were not so abundant in A. mixta aerial parts and roots. The oils from A. tomentosa collected in two different localities of Greece have been previously investigated [3]. They were both rich in terpenoids, but their composition differed notably.

The oils from Chamaemelum mixtum (syn. Anthemis mixta) collected in different localities of Corsica and Sardinia [10] and Morocco [11] were also studied. The chemical variability between the oils reported in the papers can be explained by environmental factors. Because Anthemis species are widely used for alimentary purposes and in traditional medicine, the in vitro antibacterial activity was evaluated of the essential oils of A. tomentosa and A. mixta against the bacterial species Bacillus cereus (ATCC 11778), B. subtilis (ATCC 6633), Staphylococcus aureus (ATCC 25923), S. epidermidis (ATCC 12228), Streptococcus faecalis (ATTC 29212), Escherichia coli (ATCC 25922), Proteus mirabilis (ATCC 25933), P. vulgaris (ATCC 13315), Pseudomonas aeruginosa (ATCC 27853), and Salmonella typhi Ty2 (ATCC 19430) by determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) using the broth method, as previously described [12]. Both oils appeared to be not active

against Gram-negative bacteria (MICs >100 mg/mL), whereas the oils extracted from the aerial parts showed antibacterial activity against the Gram-positive bacteria tested. In particular, B. cereus and S. faecalis were slightly sensitive to the action of AtA (MIC = 100 mg/mL), whereas the oil exerted an appreciable activity against B. subtilis (MIC = 50 mg/mL) and S. epidermidis (MIC = 25 mg/mL). As regards AmA, it was particularly active against S. aureus and S. faecalis (MIC = 25 mg/mL), while it was slightly sensitive to the action of B. cereus (MIC = 50 mg/mL) and B. subtilis (MIC = 100 mg/mL). Only in a few cases the antibacterial activity of essential oils from Anthemis species has been studied [7, 10, 11, 13-15]. Generally, the oils studied are more effective against Gram-positive bacteria than Gram-negative ones; these findings are in accordance with our results. Volatile oils as food preservatives may have great potential use; if they are added to foodstuffs they would cause no loss of organoleptic properties, would retard microbial contamination, and, therefore, reduce the onset of spoilage. In addition, only small quantities would be required for this effect. Experimental

Plant material: A. mixta aerial parts (AmA) and roots (AmR) were collected at full flowering stage at Lami, Lipari, Sicily, in August 2011. A. tomentosa aerial parts (AtA) and roots (AtR) were collected at full flowering stage at Petralona, 5 km north of Skiathos, Greece, in August 2011. Isolation of the essential oil: For the isolation of the essential oils, the air-dried samples (lots of 25 g) were ground in a Waring blender and then subjected to hydrodistillation for 3 h using n-hexane as a solvent, according to the standard procedure previously described [8]. The oils were dried over anhydrous sodium sulfate and stored under N2 at +4°C in the dark until tested and analyzed. The hydrodistillation yielded 0.08% (AmA), 0.03% (AmR), 0.1% (AtA) and 0.03% (AtR), respectively, of yellowish oils.

1382 Natural Product Communications Vol. 7 (10) 2012 Formisano et al.

Gas chromatography-mass spectrometry: Analytical gas chromatography and gas chromatography-mass spectrometry analysis were carried out under the experimental conditions reported earlier [16]. Most constituents were identified by GC by comparison of their retention indices (LRI) with either those of the literature [17-19] or with those of authentic compounds available in our laboratories. The linear retention indices were determined in relation to a

homologous series of n-alkanes (C8-C28) under the same operating conditions. Further identification was achieved by comparison of their MS on both columns, either with those stored in NIST 02 and Wiley 275 libraries or with MS from the literature [18,19] and our home-made library. Acknowledgments – The GC-MS spectra were performed at the "C.S.I.A.S." of the University "Federico II" of Napoli. The assistance of the staff is gratefully appreciated.

References

[1] Bremer K. (1994) Asteraceae, Cladistics & Classification, Timber Press, Portland, Oregon. 435-478 [2] Fernandes R. (1976) Genus Anthemis L., In Flora Europaea, Vol. 4, Tutin TG, Heywood VH, Burges NA (Eds). Cambridge University Press,

Cambridge, UK. 145-149. [3] Saroglou V, Dorizas N, Kypriotakis Z, Skaltsa HD. (2006) Analysis of the essential oil composition of eight Anthemis species from Greece. Journal

of Chromatography A, 1104, 313-322. [4] Baytop T. (1999) Therapy with medicinal plants in Turkey (past and present) (2nd ed.). Nobel Medical Book House, Istanbul, Turkey. [5] Radulovic NS, Blagojevic BK, Palic RM. (2009) Chemotaxonomically important volatiles of the genus Anthemis L. - detailed GC and GC/MS

analyses of Anthemis segetalis Ten. from Montenegro. Journal of the Chinese Chemical Society, 56, 642-652. [6] Çelik S, Rosselli S, Di Maggio AM, Raccuglia RA, Uysal I, Kisiel W, Bruno M. (2005) Sesquiterpene lactones from Anthemis wiedemanniana.

Biochemical Systematic and Ecology, 33, 952-956. [7] Conforti F, Menichini F, Formisano C, Rigano D, Senatore F, Bruno M, Rosselli S, Çelik S. (2012) Anthemis wiedemanniana essential oil prevents

LPS-induced production of NO in RAW 264.7 macrophages and exerts antiproliferative and antibacterial activity in vitro. Natural Product Research, 26, 1594-1601.

[8] Epstein WW, Poulter CD. (1973) A survey of some irregular monoterpenes and their biogenetic analogies to presqualene alcohol. Phytochemistry, 12, 737-747.

[9] Javidnia K, Miri R, Kamalinejad M, Sarkarzadeh H, Jamalian A. (2004) Chemical composition of the essential oils of Anthemis altissima L. grown in Iran. Flavour and Fragrance Journal, 19, 213-216.

[10] Darriet F, Bendahou M, Costa J, Muselli A. (2012) Chemical compositions of the essential oils of the aerial parts of Chamaemelum mixtum (L.) Alloni. Journal of Agricultural Food and Chemistry, 60, 1494-1502.

[11] Satrani B, Ghanmi M, Farah A, Aafi A, Fougrach H, Bourkhiss B, Bousta D, Talbi M. (2007) Composition chimique et activité antimicrobienne de l′huile essentielle de Cladanthus mixtus. Bulletin de la Société de Pharmacie de Bordeaux, 146, 85-89.

[12] Formisano C, Mignola E, Rigano D, Senatore F, Apostolides AN, Bruno M, Rosselli S. (2009) Constituents of leaves and flowers essential oils of Helichrysum pallasii (Spreng.) Ledeb. growing wild in Lebanon. Journal of Medicinal Food, 12, 203–207.

[13] El Hanbali F, Mellouki F, Akssira M, Boira H, Blazquez MA. (2007) Composition and antimicrobial activity of essential oil of Anthemis tenuisecta Ball. Journal of Essential Oil-Bearing Plants, 10, 499-503.

[14] Albay CG, Albay M, Yayli N, Yildirim N, Coskuncelebi K, Yayli N. (2009) Essential oil analysis and antimicrobial activities of Anthemis marschalliana ssp. pectinata and Anthemis cretica ssp. argaea from Turkey. Asian Journal of Chemistry, 21, 1425-1431.

[15] Nasrin S, Azadeh M, Mahdi V, Mahtab S, Zahra Z, Zahra S, Neda A, Vali-O-Allah M, Mahnaz K. Chemical composition and antimicrobial activity of the essential oil of Anthemis altissima L. var. altissima. Natural Product Research, in press.

[16] Canzoneri M, Bruno M, Rosselli S, Russo A, Cardile V, Formisano C, Rigano D, Senatore F. (2011) Chemical composition and biological activity of Salvia verbenaca essential oil. Natural Product Communications, 6, 1023-1026.

[17] Jennings W, Shibamoto T. (1980) Qualitative Analysis of Flavour and Fragrance Volatiles by Glass Capillary Gas Chromatography. Academic Press, New York.

[18] Davies NW. (1990) Gas chromatographic retention indices of monoterpenes and sesquiterpenes on methyl silicone and Carbowax 20M phases. Journal of Chromatography A, 503, 1-24.

[19] Adams RP. (2007) Identification of Essential Oil Components by Gas Chromatography/Mass Spectrometry. Allured Publishing Coro., Carol Stream, IL.

Natural Product Communications Vol. 7 (10) 2012 Published online (www.naturalproduct.us)

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Antifungal and Antibacterial Activity of the Essential oil of Chamaecyparis lawsoniana from Spain Jesús Palá-Paúl, Jaime Usano-Alemany, Elena Granda and Ana-Cristina Soria 1383

Chemical Composition and Biological Activities of Essential Oil from Hyptis crenata Growing in the Brazilian Cerrado Ivana Maria Póvoa Violante, Walmir Silva Garcez, Carolina da Silva Barbosa and Fernanda Rodrigues Garcez 1387

Larvicidal Activity against Aedes aegypti of Essential Oils from Northeast Brazil Patrícia L. Lavor, Gilvandete M. P. Santiago, Roberto W. da Silva Gois, Leôncio M. de Sousa, Gabrieli da P. Bezerra, Nirla R. Romero, Ângela M. C. Arriaga, Telma L. G. Lemos, Péricles B. Alves and Paulo C. S. Gomes 1391

Composition and Antipseudomonal Effect of Essential Oils Isolated from Different Lavender Species Anna Végh, Tímea Bencsik, Péter Molnár, Andrea Böszörményi, Éva Lemberkovics, Krisztina Kovács, Béla Kocsis and Györgyi Horváth 1393

Essential Oil of Croton argyrophylloides: Toxicological Aspects and Vasorelaxant Activity in Rats Aldair de França-Neto, Ana Carolina Cardoso-Teixeira, Thiago Coutinho Medeiros, Maria do Socorro Quinto-Farias, Celia Maria de Souza Sampaio, Andrelina Noronha Coelho-de-Souza, Saad Lahlou and José Henrique Leal-Cardoso 1397

Antimicrobial Activity of Blended Essential Oil Preparation Sarin Tadtong, Supatcha Suppawat, Anchalee Tintawee, Phanida Saramas, Suchada Jareonvong and Tapanee Hongratanaworakit 1401

Natural Product Communications 2012

Volume 7, Number 10

Contents

Original Paper Page

HPLC Analysis and Cytotoxic Activity of Vernonia cinerea Mom Khay, Phirom Toeng, Valérie Mahiou-Leddet, Fathi Mabrouki, Kim Sothea, Evelyne Ollivier, Riad Elias and Sok-Siya Bun 1259

Antiplasmodial Activity of the Ethnobotanical Plant Cassia fistula Mary H. Grace, Carmen Lategan, Rocky Graziose, Peter J. Smith, Ilya Raskin and Mary Ann Lila 1263

Trichilone, a New C21 Steroid from Trichilia connaroides Qiang Zhang, Yu Zhang, Hong-Ping He, Ying-Tong Di and Xiao-Jiang Hao 1267

Three Novel Immunosuppressive Steroidal Glycosides from the Stems of Stephanotis mucronata Xiao-yu Li, Shu-ling Zong, Feng-yang Chen, Shi-fang Xu and Yi-ping Ye 1269

Optimization of Carbon Source for Hairy Root Growth and Withaferin A and Withanone Production in Withania somnifera Ganeshan Sivanandhan, Manoharan Rajesh, Muthukrishnan Arun, Murugaraj Jeyaraj, Gnanajothi Kapil Dev, Markandan Manickavasagam, Natesan Selvaraj and Andy Ganapathi 1271

Quantitative Analysis of Substituted N,N-Dimethyl-tryptamines in the Presence of Natural Type XII Alkaloids Bojidarka Ivanova and Michael Spiteller 1273

HPLC Analysis of Stemokerrine and Oxystemokerrine in the Thai Medicinal Plant Stemona kerrii Sumet Kongkiatpaiboon, Vichien Keeratinijakal and Wandee Gritsanapan 1277

Identification of Pavinane Alkaloids in the Genera Argemone and Eschscholzia by GC-MS Lucie Cahlíková, Radim Kučera, Anna Hošťálková, Jiří Klimeš and Lubomír Opletal 1279

Effect of Piperine on the Pharmacokinetics and Pharmacodynamics of Glimepiride in Normal and Streptozotocin - induced Diabetic Rats Ciddi Veeresham, Samala Sujatha and Thatipamula Sandya Rani 1283

In Silico Prediction of the Cosmetic Whitening Effects of Naturally Occurring Lead Compounds Pedro Fong and Henry H. Y. Tong 1287

Profiling Flavonoid Cytotoxicity in Human Breast Cancer Cell Lines: Determination of Structure-Function Relationships Sina Yadegarynia, Anh Pham, Alex Ng, Duong Nguyen, Tetiana Lialiutska, Anthony Bortolazzo, Valentin Sivryuk, Martina Bremer and J. Brandon White 1295

Two New Flavonoids from the Seeds of Derris scandens Rachakunta Munikishore, Aluru Rammohan, Adivireddy Padmaja, Duvvuru Gunasekar, Alexandre Deville and Bernard Bodo 1305

Dihydroflavonol and Flavonol Derivatives from Macaranga recurvata Mulyadi Tanjung, Euis H. Hakim, Elfahmi, Jalifah Latip and Yana M. Syah 1309

Flavonoids Bearing an O-arabinofuranosyl-(1→3)-rhamnoside Moiety from Cladocolea micrantha: Inhibitory Effect on Human Melanoma Cells Anderson C. Guimarães, Alvicler Magalhães, Marcos J. Nakamura, Antonio C. Siani, Christina Barja-Fidalgo and André L. F. Sampaio 1311

Effect of Seasonality on Chemical Composition and Antibacterial and Anticandida Activities of Argentine Propolis. Design of a Topical Formulation María Inés Isla, Yanina Dantur, Ana Salas, Carolina Danert, Catiana Zampini, Myriam Arias, Roxana Ordóñez, Luis Maldonado, Enrique Bedascarrasbure and María Inés Nieva Moreno 1315

Larvicidal Activity of Isoflavonoids from Muellera frutescens Extracts against Aedes aegypti Charlotte Nirma, Alice M S. Rodrigues, Charlie Basset, Lionel Chevolot, Romain Girod, Christian Moretti, Didier Stien, Isabelle Dusfour and Véronique Eparvier 1319

Phenolic Compounds in Five Epilobium Species Collected from Estonia Indrek Remmel, Lauri Vares, Lauri Toom, Vallo Matto and Ain Raal 1323

A New Prenylated Acetophenone from the Root Bark of Derris indica Naushad Edayadulla and Penugonda Ramesh 1325

Phytotoxic Furanocoumarins from the Shoots of Semenovia transiliensis Shobha Sondhia, Stephen O. Duke, Solomon Green III, Nadezhda G. Gemejiyeva, Leonid K. Mamonov and Charles L Cantrell 1327

Chemoenzymatic Synthesis of Two New Halogenated Coumarin Glycosides as Potential Antifungal Agents Liangbin Zhou, Ling Liu, Tian Tian, Bailin Xue and Rongmin Yu 1331

Two New Naphthoquinone Derivatives from the Stem Bark of Callicarpa maingayi Sumayah Mohammed Asiri, Khozirah Shaari, Faridah Abas, Nabil Ali Al-Mekhlafi and Nordin H. Lajis 1333

Novel Sorbicillin Analogues from the Marine Fungus Trichoderma sp. Associated with the Seastar Acanthaster planci Wen-Jian Lan, Yang Zhao, Zhong-Liang Xie, Li-Zhen Liang, Wei-Yan Shao, Long-Ping Zhu, De-Po Yang, Xiao-Feng Zhu and Hou-Jin Li 1337

Cytotoxic Bioactivity of some Phenylpropanoic Acid Derivatives Guillermo F. Reta, Carlos E. Tonn, Carla Ríos-Luci, Leticia G. León, Eduardo Pérez-Roth, José M. Padrón, and Osvaldo J. Donadel 1341

Anti-Helicobacter pylori Activities of Natural Isopentenyloxycinnamyl Derivatives from Boronia pinnata Cécile Ribeiro da Silva, Valérie Michel, Salvatore Genovese, Marie-Christine Prévost, Francesco Epifano and Eliette Touati 1347

Continued inside backcover