NPC Natural Product Communications EDITOR-IN-CHIEF DR. PAWAN K AGRAWAL Natural Product Inc. 7963, Anderson Park Lane, Westerville, Ohio 43081, USA agrawal@naturalproduct.us EDITORS PROFESSOR ALEJANDRO F. BARRERO Department of Organic Chemistry, University of Granada, Campus de Fuente Nueva, s/n, 18071, Granada, Spain afbarre@ugr.es PROFESSOR ALESSANDRA BRACA Dipartimento di Chimica Bioorganicae Biofarmacia, Universita di Pisa, via Bonanno 33, 56126 Pisa, Italy braca@farm.unipi.it PROFESSOR DEAN GUO State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China gda5958@163.com PROFESSOR YOSHIHIRO MIMAKI School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Horinouchi 1432-1, Hachioji, Tokyo 192-0392, Japan mimakiy@ps.toyaku.ac.jp PROFESSOR STEPHEN G. PYNE Department of Chemistry University of Wollongong Wollongong, New South Wales, 2522, Australia spyne@uow.edu.au PROFESSOR MANFRED G. 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All issues are dispatched by airmail throughout the world, excluding the USA and Canada. NPC Natural Product Communications Chemical Compositions and Antimicrobial Activity of the Essential Oils of Piper abbreviatum, P. erecticaule and P. lanatum (Piperaceae) 2014 Vol. 9 No. 12 1795 - 1798 Wan Mohd Nuzul Hakimi Wan Salleha, Farediah Ahmada,*and Khong Heng Yenb a Department of Chemistry, Faculty of Science, Universiti Teknologi Malaysia (UTM), Skudai, 81310 Johor Bahru, Johor, Malaysia b School of Chemistry and Environment Studies, Faculty of Applied Sciences, Universiti Teknologi MARA (UiTM) Sarawak, Jalan Meranek, 94300 Kota Samarahan, Sarawak, Malaysia farediah@kimia.fs.utm.my Received: September 18th, 2014; Accepted: October 24th, 2014 The study was designed to examine the chemical composition and antimicrobial activities of essential oils extracted from the aerial parts of three Piper species: Piper abbreviatum, P. erecticaule and P. lanatum, all from Malaysia. GC and GC/MS analysis showed qualitative and quantitative differences between these oils. GC and GC-MS analysis of P. abbreviatum, P. erecticaule and P. lanatum oils resulted in the identification of 33, 35 and 39 components, representing 70.5%, 63.4% and 78.2% of the components, respectively. The major components of P. abbreviatum oil were spathulenol (11.2%), (E)-nerolidol (8.5%) and -caryophyllene (7.8%), whereas P. erecticaule oil mainly contained -caryophyllene (5.7%) and spathulenol (5.1%). Borneol (7.5%), -caryophyllene (6.6%) and α-amorphene (5.6%) were the most abundant components in P. lanatum oil. Antimicrobial activity was carried out using disc diffusion and broth microdilution method against nine microorganisms. All of the essential oils displayed weak activity towards Gram-positive bacteria with MIC values in the range 250-500 µg/mL. P. erecticaule oil showed the best activity on Aspergillus niger (MIC 31.3 µg/mL), followed by P. lanatum oil (MIC 62.5 µg/mL). This study demonstrated that the essential oils have potential as antimicrobial agents and may be useful in the pharmaceutical and cosmetics industries. Keywords: Essential oil, Antimicrobial, Piper abbreviatum, Piper erecticaule, Piper lanatum, Piperaceae. The genus Piper belongs to the Piperaceae family, comprising five genera and approximately 1400 species distributed throughout the tropical and subtropical regions [1]. Piper species are used in traditional remedies in the Indian Ayurvedic system of medicine and in folklore medicine of Latin America and the West Indies [2]. Piper species have been investigated as a source of new natural products with potential antioxidant [3], antimicrobial [4], antifungal [5], anti-inflammatory [6], antileishmanial [7] and insecticidal activities [8]. In recent years, several reports have been published regarding the composition and the biological activities of the essential oils of Piper species. These studies have highlighted the existence of marked chemical differences among oils extracted from different species or varieties. The chemical diversity observed in these oils can influence their biological activity, which is generally a function of three factors: genetics, physiological conditions and environment [9]. P. abbreviatum, which grows in Indonesia and the Philippines, is a branching climber hugging trees with pendent lateral branches. The stems are fissured longitudinally, rooting and articulated. The leaves are simple, spiral and exstipulate. In the Philippines, a paste of its leaves is used externally to treat splenomegaly [10]. P. erecticaule commonly known as ‘lada hutan’, is a shrubby, woody herb. The leaves are rather thin, and chartaceous, the underside of which is glaucous [11]. P. lanatum is locally known as ‘chabai hutan’ or ‘akar kalong’ [12]. The medicinal properties of this plant have not been studied. As part of an exhaustive research of the composition of the essential oils of the aromatic and medicinal plants from Malaysia, we report herein the results of the microbial properties of the oils of three Piper species, P. abbreviatum, P. erecticaule and P. lanatum, for which no data have been previously reported. The hydrodistillation of aerial parts of P. abbreviatum, P. erecticaule and P. lanatum gave pale yellow oils with a pungent smell in mean yields of 0.22%, 0.18% and 0.25% (w/w), respectively. The essential oil components were identified on the basis of their RI values and by comparison of their mass spectra with those reported in the literature [13]. The chemical compositions of the essential oils are presented in Table 1. Thirty three components were identified from the essential oil of P. abbreviatum, representing 70.5% of the total oil, among which spathulenol (11.2%), (E)-nerolidol (8.5%), -caryophyllene (7.8%) and ar-curcumene (5.8%) were the major components. From the essential oil of P. erecticaule, thirty five components were identified (63.4%) of which -caryophyllene (5.7%), spathulenol (5.1%), -cadinene (3.8%) and α-amorphene (3.8%) were the major components. The essential oil of P. lanatum yielded thirty nine components, which represented 78.2% of the total oil, with borneol (7.5%), caryophyllene oxide (6.6%) and α-amorphene (5.6%) identified as the most abundant. Jantan et al. [14] have reported that the essential oil of P. lanatum contains chavibetol (42.7% of the oil) as the major component. However, this component was not detected in this current oil. The differences in the composition of the essential oils may be due to variations in environmental parameters, such as irradiance, climate, nutrients, soil water availability, or to seasonal adaptations. It is well known that medicinal plant materials derived from the same species can show significant differences in quality when collected at different sites, owing to the influence of soil, climate, and other factors. These differences may also relate to physical appearance or to variations in their constituents, the biosynthesis of which may be affected by extrinsic environmental conditions, including ecological and geographical variables [15]. In all cases, in this study, the most abundant group components were the sesquiterpene hydrocarbons (41.9-53.9%), followed by the oxygenated sesquiterpenes (6.6–25.2%). A large number of studies have reported that the essential oils of Piper species are among the most potent essential oils with regard to their antimicrobial properties which have been confirmed in this study [16-21]. 1796 Natural Product Communications Vol. 9 (12) 2014 Salleh et al. Table 2: Antimicrobial activity of three Piper speciesa. Table 1: Constituents identified from the essential oils of three Piper species. No Components 1 α-Pinene 2 Camphene 3 -3-Carene 4 p-Cymene 5 1,8-Cineole 6 -Terpinene 7 Camphor 8 iso-Borneol 9 Borneol 10 Verbenone 11 Geraniol 12 Bornyl acetate 13 α-Cubebene 14 Cyclosativene 15 α-Ylangene 16 α-Copaene 17 -Patchoulene 18 -Panasinsene 19 iso-Ledene 20 -Bourbonene 21 -Elemene 22 α-Gurjunene 23 Longifolene 24 α-Bergamotene 25 -Caryophyllene 26 Ledene 27 -Gurjunene 28 -Elemene 29 -Selinene 30 Aromadendrene 31 α-Humulene 32 allo-Aromadendrene 33 α-Caryophyllene 34 -Cadinene -Gurjunene 35 36 -Muurolene 37 ar-Curcumene 38 epi-Bicyclosesquiphellandrene 39 α-Amorphene 40 Germacrene D 41 -Selinene 42 -Guaiene 43 –Selinene 44 α-Zingiberene 45 Cadina-1,4-diene 46 Valencene 47 α-Selinene 48 Epizonarene 49 α-Muurolene 50 -Cadinene 51 -Cadinene 52 -Sesquiphellandrene 53 cis-Calamenene 54 α-Calacorene 55 Germacrene B 56 (E)-Nerolidol 57 Spathulenol 58 Caryophyllene oxide 59 t-Cadinol 60 -Eudesmol 61 t-Muurolol 62 Cadalene 63 Phytol 64 n-Hexadecanoic acid Group components Monoterpene hydrocarbons Oxygenated monoterpenes Sesquiterpene hydrocarbons Oxygenated sesquiterpenes Others Identified components (%) KI a 930 945 1008 1020 1026 1055 1142 1152 1165 1204 1249 1287 1345 1369 1372 1374 1379 1381 1382 1387 1389 1405 1407 1412 1417 1419 1431 1434 1438 1440 1452 1458 1472 1473 1475 1478 1479 1482 1483 1485 1490 1492 1492 1493 1495 1496 1498 1501 1502 1513 1520 1521 1528 1545 1560 1562 1578 1582 1630 1632 1644 1675 1942 1959 PA 0.3 0.8 0.6 0.7 0.7 1.5 0.5 0.4 1.0 7.8 1.50 2.0 0.9 1.3 0.8 5.8 1.1 0.4 1.1 0.8 0.4 1.7 1.0 2.6 1.2 1.1 0.4 3.4 8.5 11.2 5.5 2.4 1.0 Percentage (%) PE 0.5 0.7 1.7 3.6 3.0 0.3 0.7 0.7 2.8 5.7 0.6 0.8 0.9 2.3 0.6 0.8 3.8 0.8 3.0 2.7 3.8 0.6 0.8 0.8 3.1 0.4 1.9 0.4 0.8 0.9 3.1 3.4 0.8 5.1 1.5 - PL 0.8 2.3 0.3 0.4 0.3 0.4 0.4 2.5 7.5 0.7 0.4 5.0 0.5 0.4 2.2 1.0 1.9 1.8 0.8 0.4 2.2 1.3 4.1 1.6 5.6 0.6 1.2 1.9 1.5 1.6 1.1 1.3 3.2 4.1 3.7 6.6 0.7 3.5 2.4 - 41.9 25.2 3.4 70.5 0.5 2.4 53.9 6.6 63.4 4.9 11.1 47.7 14.5 78.2 a Retention indices on Ultra-1 capillary column; PA – P. abbreviatum; PE – P. erecticaule; PL – P. lanatum. These essential oils were tested in both disc-diffusion and broth micro-dilution assays against a panel of microorganisms including six bacterial strains and three fungal. The results obtained along with the activity (Minimum Inhibitory Concentration) for the standard antibiotics are presented in Table 2. The results of the antimicrobial activity showed that the oils of P. erecticaule and P. lanatum exhibited the best activity towards Samples/ Microbes Bacillus cereus Staphylococcus aureus Enterococcus faecalis Pseudomonas putida Escherichia coli Klebsiella pneumoniae Candida glabrata Aspergillus niger Saccharomyces cerevisiae DD MIC DD MIC DD MIC DD MIC DD MIC DD MIC PA 8.2±0.3 250 8.0±0.2 250 8.5±0.2 250 6.5±0.2 >1000 7.2±0.3 500 6.5±0.2 >1000 PE 7.8±0.2 500 6.8±0.3 1000 7.2±0.2 500 6.5±0.2 1000 6.4±0.2 >1000 6.8±0.4 1000 PL 8.2±0.2 250 8.5±0.2 250 7.8±0.3 250 7.2±0.2 500 7.2±0.2 500 6.5±0.2 1000 DD MIC DD MIC DD MIC 6.4±0.2 >1000 7.0±0.2 >1000 6.5±0.2 >1000 6.4±0.2 >1000 9.0±0.2 31.3 6.5±0.2 >1000 6.8±0.3 >1000 9.2±0.2 62.5 6.9±0.2 >1000 SS 17.8±0.2 7.8 17.5±0.1 7.8 17.3±0.2 7.8 17.4±0.1 7.8 16.0±0.2 7.8 17.2±0.2 7.8 NYS 15.8±0.2 7.8 16.5±0.1 7.8 16.8±0.1 7.8 a Data represent mean±standard deviation of three independent experiments; DD – disc diffusion; MIC – Mininum inhibitory concentration (µg/mL); SS – streptomycin sulfate; NYS – nystatin; ND – not determined; PA – P. abbreviatum; PE – P. erecticaule; PL – Piper lanatum. Aspergillus niger with MIC values of 31.3 µg/mL and 62.5 µg/mL, respectively. Previous studies on the single compound showed that 1,8-cineole, myrcene, α-pinene, -pinene and camphor are the most frequently found components in different essential oils, and were associated with antifungal activity. However, these components were absent in the currently investigated oils. It is therefore evident that the antifungal activity of the essential oil does not depend solely on these compounds, and it is reasonable to assume that it results from synergic activity of all the components present in the oil [22]. The essential oils of P. abbreviatum and P. lanatum showed weak activity towards all Gram-positive bacteria (Bacillus cereus, Staphylococcus aureus, and Enterococcus faecalis) with MIC values of 250 µg/mL. The essential oils showed even weaker activity against Gram-negative bacteria with MIC values in the range 500-1000 µg/mL. These results are consistent with previous reports in the literature indicating that Gram-positive bacteria are more susceptible to essential oils than Gram-negative bacteria [23]. Among volatile constituents, phenolics (thymol, carvacrol, eugenol) and oxygenated monoterpenes (α-terpineol, terpinen-4-ol, linalool) have been reported to possess not only strong antimicrobial effects, but also a wide spectrum of activity. The lack of activity against these microbial strains might be due to the high content of monoterpene hydrocarbons in the essential oils. The low antimicrobial activity of hydrocarbons has been attributed to their low hydrogen bound capacity and water solubility [24]. Experimental Plant materials: Samples of P. abbreviatum (UiTMKS01), P. erecticaule (UiTMKS02) and P. lanatum (UiTMKS03) were collected from Kuching, Sarawak, Malaysia, in March 2012. This species were identified by Mrs. Mohizar Mohamad from the Forest Research Centre, Kuching, Sarawak and the voucher specimens were deposited at the Natural Products Research & Development Centre (NPRDC), UiTM Sarawak. Solvents and chemicals: Analytical grade methanol, ethanol and dimethylsulfoxide (DMSO), HPLC grade chloroform, magnesium sulphate, nutrient agar (NA), nutrient broth (NB), sobouraud dextrose agar (SDA), and sobouraud dextrose broth (SDB) were purchased from Merck (Germany). Stretopmycin sulfate, and nystatin were purchased from Oxoid (Italy). All tested microorganism were purchased from Mutiara Scientific (Malaysia). Essential oil of three Piper species Extraction of essential oils: The whole plant part of P. abbreviatum, P. erecticaule and P. lanatum were subjected to hydrodistillation in an all glass Dean-stark apparatus for 8 h. The oils were dried over anhydrous MgSO4 and stored at 4–6 °C. The oils yield (w/w) was calculated based on their fresh weight. Gas chromatography (GC): GC analysis was performed on a Hewlett Packard 6890 series II A gas chromatograph equipped with an Ultra-1 column (100% polymethylsiloxanes) (25 m long, 0.33 μm thickness and 0.20 mm inner diameter). Helium was used as a carrier gas at flow rate of 0.7 mL/min. Injector and detector temperature were set at 250 °C and 280 °C, respectively. Oven temperature was kept at 50 °C, then gradually raised to 280 °C at 5 °C/min and finally held isothermally for 15 min. Diluted samples (1/100 in diethyl ether, v/v) of 1.0 µL were injected manually (split ratio 50:1). The injection was repeated three times and the peak area percentages were reported as means ±SD of triplicates. Calculation of peak area percentage was carried out by using the GC HP Chemstation Software (Agilent Technologies). Gas chromatography-mass spectrometry (GC-MS): GC-MS chromatograms were recorded using a Hewlett Packard Model 5890A gas chromatograph and a Hewlett Packard Model 5989A mass spectrometer. The GC was equipped with Ultra-1 column (25 m long, 0.33 μm thickness and 0.20 mm inner diameter). Helium was used as a carrier gas at a flow rate of 1 mL/min. Injector temperature was 250 °C. Oven temperature was programmed from 50 °C (5 min hold) at 10 °C/min to 250 °C and finally held isothermally for 15 min. For GC-MS detection, an electron ionization system, with ionization energy of 70 eV was used. A scan rate of 0.5 s (cycle time: 0.2 s) was applied, covering a mass range from 50-400 amu. Identification of components: The constituents of the oils were identified by comparison of their mass spectra with reference spectra in the computer library (Wiley) and also by comparing their retention indices, with those of authentic compounds or data in the literature [13]. The quantitative data were obtained electronically from FID area percentage without the use of correction factor. Antimicrobial activity - Microbial strains: Antimicrobial activity of the oils was tested against the Gram-positive bacteria, Bacillus cereus (ATCC11778), Staphylococcus aureus (ATCC29737) and Enterococcus faecalis (ATCC19433): the Gram-negative bacteria, Escherichia coli (ATCC10536), Pseudomonas putida (ATCC49128), and Klebsiella pneumoniae (ATCC13883). Three fungi were also used, namely Aspergillus niger (ATCC16888), Candida glabrata (ATCC2001) and Saccharomyces cerevisiae (ATCC7754). The strains were grown on nutrient agar for the Natural Product Communications Vol. 9 (12) 2014 1797 bacteria and sabouraud dextrose agar for fungal/yeast. For the activity tests, nutrient broth for bacteria and sabouraud dextrose broth for fungal/yeast strains were used. Disc diffusion: Antimicrobial tests were carried out by the disc diffusion method as previously described [25]. The essential oils were dissolved in DMSO (4 mg/mL). Antimicrobial tests were carried out by the disc diffusion method using 400 μL of suspension containing 108 CFU/mL of bacteria and 106 CFU/mL of fungi, spread on the nutrient agar (NA) and sabouraud dextrose agar (SDA) mediums, respectively. The disc (6 mm diameter) impregnated with 10 μL of the essential oil and DMSO (negative control) was placed on the inoculated agar, which was incubated for either 24 h at 37 °C (bacteria) or 48 h at 30 ºC (fungi). Streptomycin sulfate (10 μg/mL) and nystatin (100 IU) were used as the positive controls for bacteria and fungi, respectively. Clear inhibition zones around the discs indicated the presence of antimicrobial activity. All tests and analyses were carried out in triplicate. Minimum inhibitory concentration (MIC): The MIC was determined by the broth micro-dilution method as previously described using 96-well microplates [26]. The inoculate of the microbial strains was prepared from 24 h broth cultures and suspensions were adjusted to 0.5 McFarland standard turbidity. Essential oils was dissolved in DMSO (1 mg/mL) to obtain 1000 μg/mL stock solution. A number of wells were reserved in each plate for positive and negative controls. Sterile broth (100 μL) was added to the well from row B to H. The stock solutions of samples (100 μL) were added to the wells in rows A and B. Then, the mixture of samples and sterile broth (100 μL) in row B was transferred to each well in order to obtain a twofold serial dilution of the stock samples (concentration of 1000-7.8 μg/mL). The inoculum (100 μL) was added to each well. The final volume in each well was 200 μL. Streptomycin sulfate for bacteria and nystatin for fungi were used as positive controls. Plates were incubated at 37 °C for 24 h. Microbial growth was indicated by the presence of turbidity and a pellet at the bottom of the well. 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Gulluce M, Sahin F, Sokmen M, Ozer H, Daferera D, Sokmen A, Polissiou M, Adiguzel A, Ozkan H. (2007) Antimicrobial and antioxidant properties of the essential oils and methanol extract from Mentha longifolia L. ssp. longifolia. Food Chemistry, 103, 1449-1456. Natural Product Communications Vol. 9 (12) 2014 Published online (www.naturalproduct.us) Reactive Oxygen Species Scavenging Activity of Jixueteng Evaluated by Electron Spin Resonance (ESR) and Photon Emission Toshizo Toyama, Satoko Wada-Takahashi, Maomi Takamichi, Kiyoko Watanabe, Ayaka Yoshida, Fumihiko Yoshino, Chihiro Miyamoto, Yojiro Maehata, Shuta Sugiyama, Shun-suke Takahashi, Kazuo Todoki, Masaichi-Chang-il Lee and Nobushiro Hamada Selective COX-2 Inhibitory Properties of Dihydrostilbenes from Liquorice Leaves–In Vitro Assays and Structure/Activity Relationship Study Domenico Trombetta, Salvatore V. 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Setzer Chemical Composition and Biological Activities of the Essential Oils from Two Pereskia Species Grown in Brazil Lucéia Fátima Souza, Ingrid Bergman Inchausti de Barros, Emilia Mancini, Laura De Martino, Elia Scandolera and Vincenzo De Feo 1755 1761 1765 1769 1773 1777 1781 1783 1787 1791 1795 1799 1805 Acconts/Reviews Stemona Alkaloids: Biosynthesis, Classification, and Biogenetic Relationships Feng-Peng Wang and Qiao-Hong Chen 1809 Natural Product Communications 2014 Volume 9, Number 12 Contents Original Paper Page Biobased Lactams as Novel Arthropod Repellents Kamlesh R. Chauhan, Hemant Khanna, Nagendra Babu Bathini, Thanh C. Le and John Grieco Two New Aromadendrane Sesquiterpenes from the Stem Bark of Alafia multiflora Alembert T. Tchinda, David E. Tsala, Nnanga Nga, Ewa Cieckiewicz, Robert Kiss, Joseph D. Connolly and Michel Frédérich Isolation and Structural Characterization of a New Minor Diterpene Glycoside from Stevia rebaudiana Venkata Sai Prakash Chaturvedula and Julian Zamora ent-Kaurane Diterpenes from Annona glabra and Their Cytotoxic Activities Hoang Le Tuan Anh, Nguyen Thi Thu Hien, Dan Thi Thuy Hang, Tran Minh Ha, Nguyen Xuan Nhiem, Truong Thi Thu Hien, Vu Kim Thu, Do Thi Thao, Chau Van Minh and Phan Van Kiem Chemical Constituents of Tilia taquetii Leaves and their Inhibition of MMP-1 Expression and Elastase Activities Su Yeong Kim, Jung Eun Kim, Hee Jung Bu, Chang-Gu Hyun and Nam Ho Lee Novel C-ring Analogs of Ursolic acid: Synthesis and Cytotoxic Evaluation Uppuluri V. Mallavadhani, Banita Pattnaik, Nitasha Suri and Ajit K. Saxena Phenolic Constituents of Erigeron floribundus (Asteraceae), a Cameroonian Medicinal Plant Chiara Berto, Filippo Maggi, Prosper C. Biapa Nya, Anna Pettena, Irene Boschiero and Stefano Dall'Acqua Nortriterpene Saponins from Akebia trifoliata Keiichi Matsuzaki, Kayo Murano, Yurika Endo and Susumu Kitanaka C-24 Stereochemistry of Marine Sterols: (22E)-25,28-Dimethyl- stigmasta-5,22,28-trien-3-ol and 25,28-Dimethylstigmasta-5,28-dien-3-ol Rie Nojo, Shizue Echigo, Noriyuki Hara and Yoshinori Fujimoto Antibacterial Compounds from Glycosmis puberula Twigs Cholpisut Tantapakul, Tawanun Sripisut, Wisanu Maneerat, Thunwadee Ritthiwigrom and Surat Laphookhieo Antimicrobial Activity of Extracts and Isoquinoline Alkaloids of Selected Papaveraceae Plants Lubomír Opletal, Miroslav Ločárek, Adéla Fraňková, Jakub Chlebek, Jakub Šmíd, Anna Hošťálková, Marcela Šafratová, Daniela Hulcová, Pavel Klouček, Miroslav Rozkot and Lucie Cahlíková New Unusual Alkaloids from the Ascidian Eudistoma vannamei Antônia Torres Ávila Pimenta, Paula Christine Jimenez, Letícia Veras Costa-Lotufo, Raimundo Braz-Filho and Mary Anne Sousa Lima Synthesis and Biological Evaluation of Febrifugine Analogues Huong Doan Thi Mai, Giang Vo Thanh, Van Hieu Tran, Van Nam Vu, Van Loi Vu, Cong Vinh Le, Thuy Linh Nguyen, Thi Dao Phi, Bich Ngan Truong, Van Minh Chau and Van Cuong Pham Flavonoids from Twigs of Millettia pubinervis Zhi Na, Qi-Shi Song and Hua-Bin Hu Effect of Osajin and Pomiferin on Antidiabetic Effects from Normal and Streptozotocin-induced Diabetic Rats Hyung-In Moon Two New Homoisoflavonoids from the Bulbs of Bessera elegans Yukiko Matsuo, Risa Kurihara, Nana Akagi and Yoshihiro Mimaki Isolation of Phenolics from Rhizophora mangle by Combined Counter-current Chromatography and Gel-Filtration Fernanda das Neves Costa, Marcos Daniel da Silva, Ricardo Moreira Borges and Gilda Guimarães Leitão Angular-type Furocoumarins from the Roots of Angelica atropurpurea and their Inhibitory Activity on the NFAT Signal Transduction Pathway Azumi Nagasawa, Mitsuyoshi Sakasai, Daishi Sakaguchi, Shigeru Moriwaki, Yoshinori Nishizawa and Takashi Kitahara Isoprenylated Xanthone and Benzophenone Constituents of the Pericarp of Garcinia planchonii Duong Hoang Trinh, Ly Dieu Ha, Phuong Thu Tran and Lien-Hoa Dieu Nguyen Stemofurans X-Y from the Roots of Stemona Species from Laos Dang Ngoc Quang, Vong Anatha Khamko, Nguyen Thi Trang, Lam Thi Hai Yen and Pham Huu Dien A New Stilbenoid Compound from the Lianas of Gnetum microcarpum Nik Fatini Nik Azmin, Norizan Ahmat, Yana M. Syah, Nik Khairunissa’ Nik Abdullah Zawawi and Mohd Izwan Mohamad Yusof Phenolic Acids Profile, Antioxidant and Antibacterial Activity of Chamomile, Common Yarrow and Immortelle (Asteraceae) Ivana Generalić Mekinić, Danijela Skroza, Ivica Ljubenkov, Luka Krstulović, Sonja Smole Možina and Višnja Katalinić Activity-guided Fractionation of Ipomea fistulosa Leaves for Pro-inflammatory Cytokines and Nitric Oxide Inhibitory Constituents Neeraj K. Patel, Ramandeep and Kamlesh K. Bhutani Development and Validation of a High-Performance Liquid Chromatographic Method for the Simultaneous Quantification of Marker Constituents in Cheonwangbosimdan Chang-Seob Seo and Hyeun-Kyoo Shin Continued inside backcover 1671 1673 1677 1681 1683 1687 1691 1695 1699 1705 1709 1713 1717 1721 1723 1725 1729 1733 1737 1741 1743 1745 1749 1751