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. REINECKE Department of Chemistry, Texas Christian University, Forts Worth, TX 76129, USA m.reinecke@tcu.edu PROFESSOR WILLIAM N. SETZER Department of Chemistry The University of Alabama in Huntsville Huntsville, AL 35809, USA wsetzer@chemistry.uah.edu PROFESSOR YASUHIRO TEZUKA Institute of Natural Medicine Institute of Natural Medicine, University of Toyama, 2630-Sugitani, Toyama 930-0194, Japan tezuka@inm.u-toyama.ac.jp PROFESSOR DAVID E. THURSTON Department of Pharmaceutical and Biological Chemistry, The School of Pharmacy, University of London, 29-39 Brunswick Square, London WC1N 1AX, UK david.thurston@pharmacy.ac.uk HONORARY EDITOR PROFESSOR GERALD BLUNDEN The School of Pharmacy & Biomedical Sciences, University of Portsmouth, Portsmouth, PO1 2DT U.K. axuf64@dsl.pipex.com ADVISORY BOARD Prof. Berhanu M. Abegaz Gaborone, Botswana Prof. Viqar Uddin Ahmad Karachi, Pakistan Prof. Øyvind M. 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All issues are dispatched by airmail throughout the world, excluding the USA and Canada. NPC Natural Product Communications Composition of Essential Oils from Seeds of Abies koreana 2013 Vol. 8 No. 2 227 - 230 Anna Wajs-Bonikowskaa,*, Karol Olejnika, Radosław Bonikowskia and Piotr Banaszczakb a Technical University of Lodz, Faculty of Biotechnology and Food Sciences, Institute of General Food Chemistry, Stefanowskiego 4/10, 90-924 Lodz, Poland b Arboretum - Rogów Forestry Experimental Station, Warsaw University of Life Sciences, 95-063 Rogów, Poland anna.wajs@p.lodz.pl Received: October 26th, 2012; Accepted: December 3rd, 2012 The essential oils from seeds of nine Abies koreana specimens have been studied using GC-MS-FID and NMR methods, leading to the determination of 96 volatiles, which constituted over 99% of the oils. The hydrodistilled oils of fresh, resinous scent were isolated with yields in the range of 3.8-8.5%. The results showed that the essential oil of Korean fir seeds contained 70-95% monoterpenes and 1-20% oxygenated monoterpenes as the dominant groups. The numerous sesquiterpenes, diterpenes and their oxygenated derivatives constituted only 2-8% of the oil. The major component of the seed essential oil was limonene (4172 g/100g); the laevorotary form of this terpene predominated. A. koreana seeds seem to be a rich source of both essential oil and (-)-limonene, whose average enantiomeric excess was above 95%. Keywords: Abies koreana, Fir, Seed, Essential oil, Terpenes, Volatiles, Limonene. The genus Abies (Pinaceae) comprises about 50 species of evergreen conifers, ranged in Europe, North and Central America, Asia, and North Africa, occurring in mountains over most of the range. A. koreana is a fir native to the higher mountains of South Korea. This species was discovered in 1907 on the volcanic island of Cheju (South Korea) and one year later was introduced into France. From a Korean natural forest this fir was introduced into Polish Arboreta in 1929 [1]. Nowadays, A. koreana is cultivated as an ornamental plant in many places, in temperate climates. Korean fir is grown both for its foliage and for its abundant blue cones. The winged ripe seeds are released when the cones disintegrate at maturity about 5-6 months after pollination, in Poland usually in September [1,2]. A. koreana, like other species of Abies, has been used in folk medicine to treat colds, stomachache, indigestion, and vascular and pulmonary diseases [3]. Abies needle oils are used industrially by soap and perfume manufacturers to add pleasant odors to their products [4]. A. koreana needle essential oil has been previously investigated [5]. The volatile composition of intact cones and twigs has also been published [5c-d]. However, to our knowledge, no studies on the volatiles from the seeds of Korean fir have been reported. Therefore, the present investigations were carried out to isolate and identify the compounds of seed essential oils. This paper is the second in a series in which we analysed the fir seed volatiles [6]. The hydrodistilled oil of Korean fir seeds has a characteristic pleasant, balsamic, resinous scent with a fresh lemon note. The yield of essential oils was in the range of 3.2-8.5%. Hydrodistillation of the seeds of individual woods was performed in triplicate; the results (calculated as an average yield with standard deviation value) are presented in Table 1. It is noticeable that the yield of essential oils from the seeds of trees no. 2-9 are at a similar level, while the yield from tree no. 1 was 37% higher than the lowest yield (tree no. 4). The yield differences may be caused by different degrees of seed ripeness or different sun exposure of individual trees. However, the eight Korean firs tested were of the same age (woods no. 1-8) and grown in the same area. The average essential oil yield was 4.9%, which was much higher than those of the needles (0.9-1.0%) and cones (0.42%) reported in the literature [5-8]. It is noticeable that the yields from seeds of A. koreana, growing in Poland are not much lower than those of hydrodistilled oils from the seeds of other fir species: Abies maroccana (5.3%, north Morocco), [7] A. nordmanniana (6.0%, Georgia) [8], and A. alba (7.4%, south Poland) [6]. In total, 96 compounds were identified, of which, to the best of our knowledge, 51 are new records for Korean fir. The identified compounds presented in Table 1 constituted 99.1-99.9% of the 9 examined oils; the volatiles new for A. koreana seeds are marked with asterisks. The hydrodistilled oils contained 15 monoterpene hydrocarbons, which constituted 70.4-94.9% of the seed oils. The main compound was limonene, which formed 39.8% to 69.6% of the total. This was also one of the main components of A. koreana cone oils, but occurred at a lower concentration (25.6%) than in the examined seeds [5c]. The other major monoterpenes in the tested essential oils were: α-pinene (7.3-38.6%), camphene (0.7-12.6%), -pinene (1.87.6%) and -myrcene (1.3-1.9%). They were also predominant in the oils of Korean fir cones, needles and twigs [5]. In order to provide the real concentration of the main oil constituents, we carried out a methodology based on the true quantitation. This experiment (listed in Table 1) showed that the amount of limonene (41.0-71.7 g/100 g essential oil) corresponded to almost 40%-70%. The concentrations (g/100 g) of other monoterpenes were for α- and -pinene 7.4-39.4 g/100 g and 1.8-7.8 g/100 g, respectively. The monoterpene hydrocarbons identified in the nine individual trees of A. koreana were qualitatively the same, but differed in the percentages of their major components. The second identified group was oxygenated monoterpenes. These 17 compounds constituted 1-20% of the oils. Predominant was bornyl acetate, the structure of which, as well as that of geranyl acetate, were confirmed by NMR spectroscopy; the amount of bornyl acetate varied strongly between the specimens from 0.4% to 18.5%. The quantitative determination of this ester was carried out through the calibration curve, and the concentration (Table 1) was 0.6-28.5 g/100 g. This compound was also a predominant component of needle oil from the Korean fir grown in Poland (30.2%) [7] and in Korea on Jeju Island (30.4%) [5b]. Borneol 228 Natural Product Communications Vol. 8 (2) 2013 Wajs-Bonikowska et al. Table 1: The composition of essential oils from seeds of 9 specimens of Abies koreana No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Tree no. Compound Santene Tricyclene α-Thujene α-Pinene [%] α-Pinene [g/100g] Camphene Sabinene β-Pinene [%] -Pinene [g/100g] β-Myrcene δ-2-Carene α-Phellandrene δ-3-Carene p-Cymene Limonene [%] Limonene [g/100g] -Terpinene Terpinolene α-Fenchol Limonene oxide trans-Pinocarveol cis-Verbenol -Terpineol Borneol Myrtenal α-Terpineol Myrtenol Verbenone trans-Carveol Fenchyl acetate cis-Carveol Bornyl acetate [%] Bornyl acetate [g/100g] α-Terpinyl acetate Neryl acetate α-Cubebene Geranyl acetate -Ylangene α-Copaene -Elemene -Longipinene Longifolene α-Cedrene (E)- -Caryophyllene -Elemene -Copaene cis-Muurola-3,5-diene Cadina-3,5-diene α-Humulene cis-Muurola-4(15),5-diene α-Acoradiene (Z,Z)-α-Farnesene ar-Curcumene -Muurolene Germacrene D -Selinene -Humulene 4-epi-Cubebol α-Selinene α-Muurolene -Bisabolene -Cadinene cis/trans-Calamenene δ-Cadinene -Bisabolene ω-Cadinene α-Cadinene -Elemol -Calacorene (E)-Nerolidol Germacrene-1(10),5-dien-4-ol Caryophyllene oxide Globulol Longiborneol Cedrol 10-epi- -Eudesmol Selin-6-en-4-ol -Eudesmol 1-epi-Cubenol τ-Cadinol τ -Muurolol trans-10-Hydroxycalamenene 1(10)-Spirovetiven-7 -ol -Eudesmol cis-10-Hydroxycalamenene α-Cadinol α-Eudesmol Intermedeol Tetradecan-1-ol α-Bisabolol RIexp. 881 922 925 935 946 968 974 984 995 990 1008 1014 1033 1053 1082 1102 1120 1128 1132 1137 1154 1175 1177 1183 1186 1202 1210 1215 1273 1336 1343 1353 1361 1379 1382 1392 1407 1415 1421 1426 1433 1434 1448 1449 1459 1464 1466 1468 1473 1477 1483 1487 1491 1494 1498 1500 1506 1515 1521 1522 1526 1533 1539 1542 1548 1551 1572 1575 1581 1598 1602 1607 1615 1619 1629 1636 1637 1638 1642 1649 1647 1648 1653 1657 1664 1676 1 2 3 4 tr 0.9 tr 17.3 17.6 9.1 tr 3.5 3.6 1.5 tr 0.2 0.8 tr 51.8 53.4 tr 0.2 tr tr 0.1 0.1 0.6 0.7 tr tr tr tr tr tr tr 9.4 14.4 0.1 tr tr 0.1 tr tr 0.1 tr tr tr 0.1 0.1 tr tr tr 0.1 tr tr tr tr 0.1 tr 0.3 0.1 tr 0.2 0.1 0.1 0.2 tr 0.4 tr tr tr tr tr 0.1 tr tr tr tr tr tr 0.1 tr tr tr tr tr tr tr tr tr tr 1.3 tr tr tr tr tr 38.6 39.4 0.4 tr 7.6 7.8 1.8 tr tr 0.8 tr 43.5 44.8 tr 0.1 0.2 tr 0.1 0.1 0.1 tr tr tr tr 0.1 tr tr tr 0.5 0.8 tr tr tr 0.4 tr tr tr tr 0.3 tr 0.2 0.2 tr tr tr 0.1 tr tr tr tr tr tr 0.4 0.2 tr 0.3 0.1 0.1 tr tr tr tr tr tr tr tr tr tr tr tr tr tr tr 0.1 tr tr tr tr tr tr 0.1 tr tr tr 2.6 tr tr tr 0.4 tr 9.5 9.7 5.3 tr 1.8 1.8 1.7 tr 0.3 0.5 tr 67.3 69.3 tr 0.1 0.1 tr 0.1 0.2 0.3 0.3 tr tr tr tr tr 0.1 tr 6.5 9.9 tr tr tr tr tr tr 0.1 tr 0.1 tr 0.2 tr tr tr tr 0.1 tr tr tr tr 0.1 tr 0.3 0.2 tr 0.3 0.2 0.1 0.3 tr 0.5 tr tr tr tr tr tr tr tr tr tr tr tr 0.1 tr tr 0.1 tr tr tr 0.1 tr tr tr 2.4 tr tr tr tr tr 20.0 20.4 0.7 tr 4.4 4.5 1.6 tr 0.3 0.1 tr 64.7 66.6 tr tr 0.1 tr 0.1 0.2 0.1 0.1 tr tr tr tr tr tr tr 1.1 1.7 tr tr tr tr tr tr tr tr tr tr 0.1 0.1 tr tr tr 0.1 tr tr tr tr tr tr 0.4 0.2 tr 0.3 0.1 tr 0.1 tr 0.2 tr tr tr tr tr tr tr tr tr tr tr tr 0.2 tr tr tr tr tr tr tr tr tr tr 3.9 tr tr 5 Content [%] tr 0.3 tr 11.3 11.5 2.9 tr 2.5 2.6 1.8 tr 0.2 0.1 tr 69.6 71.7 tr 0.1 tr tr 0.1 0.1 0.1 0.2 tr tr tr tr tr tr tr 3.3 5.0 tr tr tr tr tr tr tr tr tr tr 0.9 0.2 tr tr tr 0.5 tr tr tr tr 0.1 tr 0.3 0.2 tr 0.3 0.2 tr 0.3 tr 0.5 tr tr tr tr tr tr tr tr tr tr tr tr 0.1 tr tr 0.1 tr tr tr 0.1 tr tr tr 2.8 tr tr 6 7 8 9 Identif. method tr 0.6 tr 17.3 17.6 7.9 tr 3.2 3.3 1.7 tr tr 0.4 tr 49.3 50.8 tr 0.1 0.1 tr 0.1 0.1 0.4 0.5 tr tr tr 0.1 tr tr tr 10.1 15.5 tr tr tr 0.1 tr tr 0.2 tr 0.1 tr 0.2 0.1 tr tr tr 0.1 tr tr tr tr 0.1 0.1 0.4 0.2 tr 0.4 0.2 0.1 0.3 tr 0.5 tr tr tr tr tr 0.1 tr tr tr tr tr tr 0.2 tr tr 0.1 tr tr tr 0.1 tr tr tr 3.7 tr tr tr 1 tr 15.1 15.4 12.6 tr 2.6 2.7 1.4 tr tr 0.9 tr 39.8 41.0 tr 0.2 tr tr 0.2 0.3 0.6 0.9 tr tr 0.1 0.1 tr 0.1 tr 15.7 24.0 0.1 tr tr tr tr tr 0.1 tr tr tr 0.2 0.1 tr tr tr 0.1 tr tr tr tr 0.1 0.1 0.4 0.2 tr 0.4 0.2 0.1 0.2 tr 0.4 tr tr tr tr tr 0.2 tr tr tr tr tr tr 0.2 tr tr 0.1 tr tr tr 0.1 tr tr tr 4.5 tr tr tr 0.2 tr 7.3 7.4 4.7 tr 2 2.0 1.3 tr 0.2 0.5 tr 54.0 55.6 tr 0.2 0.1 tr 0.1 0.1 0.3 0.7 tr tr tr tr tr tr tr 18.6 28.5 tr tr tr 0.5 tr tr 0.1 tr tr tr 0.2 0.1 tr tr tr 0.1 tr tr tr tr 0.1 0.1 0.4 0.1 tr 0.3 0.1 0.1 0.1 tr 0.2 tr tr tr tr tr 0.1 tr tr tr tr tr tr 0.2 tr tr tr tr tr tr 0.1 tr tr tr 5.2 tr 0.3 tr tr tr 17.5 17.9 0.2 tr 5.3 5.4 1.9 tr 0.1 1 tr 68.6 70.7 tr 0.1 0.1 0.1 0.1 0.2 tr tr tr tr tr 0.1 tr tr tr 0.4 0.6 tr tr tr 0.4 tr 0.1 tr tr tr tr 0.1 0.1 tr tr tr tr tr tr tr tr 0.1 tr tr 0.1 tr tr 0.1 tr 0.3 tr 0.4 tr tr tr tr tr tr tr tr tr tr tr tr tr tr tr 0.1 tr tr tr 0.1 tr tr tr 0.8 0.1 0.2 RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS ¹H,RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS ¹H,RI,MS RI,MS RI,MS RI,MS ¹³C,¹H,RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS ¹H,RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS ¹³C,¹H,RI,MS RI,MS ¹³C,¹H,RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS ¹³C,¹H,RI,MS RI,MS RI,MS ¹H,RI,MS RI,MS RI,MS ¹H,RI,MS RI,MS RI,MS ¹H,RI, MS RI,MS RI,MS Abies koreana seed essential oil Natural Product Communications Vol. 8 (2) 2013 229 87 * Eudesm-7(11)-en-4α-ol 88 Manoyl oxide 89 * Isopimara-8,15-diene 90 * Abietatriene 91 * Abieta-7,13-diene 92 * (8α,12Z)-Abienol 93 * Abieta-8(14),13(15)-diene 94 * trans-Totarol 95 * Dehydroabietal 96 * Abietal Monoterpene hydrocarbons Oxygenated monoterpenes Sesquiterpene hydrocarbons Oxygenated sesquiterpenes Diterpene hydrocarbons Oxygenated diterpenes Nonterpenes 1677 2007 2027 2059 2100 2134 2166 2239 2252 2275 0.1 tr tr tr tr tr tr tr tr tr 85.3 11.1 1.9 1.6 0 0 tr 99.9 0.1 tr tr tr tr tr tr tr tr tr 92.8 1.5 1.9 2.9 0 0 tr 99.1 tr tr tr tr tr tr tr tr tr tr 86.9 7.6 2.5 2.7 0 0 tr 99.7 tr tr tr tr tr tr tr 0.1 tr tr 91.8 1.7 1.6 4.1 0 0.1 tr 99.3 tr tr 0.1 tr 0.1 tr tr 0.1 tr tr 88.8 3.8 3.5 3.1 0.2 0.1 tr 99.5 tr tr 0.1 tr 0.1 tr tr tr tr tr 80.5 11.5 3.0 4.2 0.2 0 tr 99.4 tr tr tr tr 0.1 tr tr tr tr tr 73.6 18.1 2.6 5.1 0.1 0 tr 99.5 tr tr tr tr 0.1 0.1 tr 0.1 tr 0.1 70.4 20.4 2.0 5.9 0.1 0.3 tr 99.1 tr tr tr tr 0.1 tr tr 0.1 tr 0.2 94.7 1.4 1.3 1.2 0.1 0.3 tr 99.1 RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS RI,MS Total identified Yield of essential oil [%] 8.52 4.34 3.88 3.19 5.22 3.80 5.53 4.93 (standard deviation) (0.41) (0.52) (0.53) (1.14) (0.03) (0.26) (0.51) (0.34) *- new for A. koreana seed volatiles; tr - <0.05%; RIexp. - experimental retention indices calculated on non-polar column; (tr-0.9%), -terpineol (tr-0.6%), cis-verbenol (0.1-0.3%) and transpinocarveol (0.1-0.2%) are also dominant volatiles among the seed monoterpenoids, but they occur at levels lower than 1% in the oils of individual woods. Surprisingly, borneol was identified previously as the main volatile in the needle oil (27.9%) of fir grown in Korea, but at Mt Dukyu, Muju [5a]. The enantiometric distribution of four major monoterpene hydrocarbons of the seed essential oils: limonene, α- and -pinene and camphene (Table 2) was determined on a Chirasil-Dex CB fused silica chiral column. It was found that the laevorotary isomer was dominant for all examined monoterpenes. What is most interesting is that the almost enantiomerically pure (-)-limonene was found in A. koreana seed oil. The enantiomeric excess of (-)limonene was above 97%. Such a high concentration of the laevorotary form of this terpene is also characteristic of A. koreana needles [7] and other fir seeds, e.g: A. alba seeds [6] and A. maroccana seeds [7]. The relatively high enantiomeric excess of the (–) form was also noticed for α-pinene (an average 84%). The ratio of the laevorotary to dextrorotary form of -pinene and camphene was 76%: 24% and 90%: 10%, respectively. The excess of laevorotary enantiomers of the above monoterpenes was also observed previously in Korean fir needles [5d]. Table 2: Enantiomer ratio and enantiomeric excess (ee) of the main A. koreana seed monoterpenes. Tree no. 1 Compound (-)-Limonene 96.6 (+)-Limonene 3.4 ee 93.2 (-)-α-Pinene 94.1 (+)-α-Pinene 5.9 ee 88.2 (-)- -Pinene 66.7 (+)- -Pinene 33.3 ee 33.4 (-)-Camphene 90.1 (+)-Camphene 9.9 ee 80.2 SD – Standard deviation 2 3 4 96.6 3.4 93.2 84.8 15.2 69.6 90.9 9.1 81.8 75.0 25.0 50.0 97.3 2.7 94.6 95.7 4.3 91.4 75.0 25.0 50.0 92.6 7.4 85.2 97.5 2.5 95.0 97.6 2.4 95.2 66.7 33.3 33.4 83.3 16.7 66.6 5 6 7 Content [%] 99.5 96.4 97.1 0.5 3.6 2.9 99.0 92.8 94.2 95.0 87.8 89.2 5.0 12.2 10.8 90.0 75.6 78.4 66.7 62.5 68.8 33.3 37.5 31.2 33.4 25.0 37.6 90.3 92.3 91.4 9.7 7.7 8.6 80.6 84.6 82.8 8 9 Mean ± SD 97.2 2.8 94.4 87.9 12.1 75.8 94.5 5.5 89.0 93.3 6.7 86.6 97.9 2.1 95.8 94.5 5.5 89.0 91.7 8.3 83.4 100.0 0.0 100.0 97.3 ± 0.9 2.7 ± 0.9 94.7 ± 1.9 91.8 ± 4.5 8.2 ± 4.5 83.7 ± 8.9 75.9 ± 12.8 24.1 ± 12.8 51.9 ± 25.5 89.8 ± 7.0 10.2 ± 7.0 79.6 ± 14.0 Although sesquiterpene hydrocarbons and their oxygenated derivatives are the most numerous group in the seed oils, consisting of 31 and 24 compounds, respectively, they constituted only 1.32.6% and 1.2-5.9% of the seed oils, respectively. Intermedeol (0.85.2%) and -caryophyllene (0.1-0.9%) were the major sesquiterpenes; the great majority of the others were found only in traces. However, structures of seven terpenes, isolated by flashchromatography, were additionally confirmed by NMR spectroscopy (Table 1). What is interesting is that intermedeol, identified as the main sesquiterpenoid in the seeds, prevailed also in needles and twigs of Korean fir [5c]. The identified compounds, with many different sesquiterpene skeletons, generally had muurolane, cadinane, eudesmane, elemane, cubebane, bisabolane, caryophyllane, and germacrane skeletons; the most numerous had muurolane (9 compounds) and cadinane (7 volatiles) skeletons. However, farnesenes, curcumenes, acoradienes and spirovetivenes were identified as isomer pairs in many of the examined essential oils, but at concentrations lower than 0.05%. Diterpene hydrocarbons and their oxygenated derivatives (9 compounds) were identified in quantities varying from tr-0.5%, with abietal, trans-totarol, abieta-7,13-diene and isopimara-8,15diene being the main diterpenes in some of the examined tree oil specimens. Eight of them were identified for the first time in A. koreana. In summary, the qualitative composition of all oils was exactly the same, but quantitative differences between them were observed; for many volatiles these differences were very significant, irrespective of the population examined. For example, tree no. 1-8 grown in the neighborhood and introduced to the Arboretum from a Korean natural forest from the same seed packet. It is worth noting that when the amount of monoterpene hydrocarbons in the seed oils decreased the concentration of their oxygenated derivatives at the same time increased (Figure 2). This relationship was observed between the main compounds, like limonene and bornyl acetate, as well as α-pinene, camphene and borneol, while the amount of sesquiterpene hydrocarbons and their oxygenated derivatives did not change as strongly (2-8%) as the monoterpene group did. Concerning the previous publication in which the composition of essential oils from needles of 20 A. koreana specimens (the same population as ours) were examined, the qualitative composition was very similar, while especially the content of the main volatiles like camphene (2.4-24.5%), limonene (1.9-37.3%), bornyl acetate (10.742.8%) and intermedeol (0.1-25.6%) differed significantly [5d]. As a result of chemical polymorphism of an aroma plant, huge differences can be observed in the composition of essential oils, even among one population. These differences may arise from the fact that biosynthesis of terpenoids is controlled by genetic factors. This process could also be a result of ontogenetic variability and could be influenced by many environmental factors. These correlations have been previously very well documented [9]. Moreover, like other conifer tissues, seed tissues appear to rely on the reserves of essential oils to protect against the attack by insects and fungal pathogens. The differences in the quantitative composition of volatiles of the examined individual woods may be caused by insect attack or different environmental conditions [10]. In summary, the seeds of A. koreana constitute a rich source of essential oil (up to 8.5%) that may be potentially interesting for the perfume industry. The essential oil contains the laevorotary enantiomer of limonene in almost pure form and can be used to isolate this monoterpene hydrocarbon. 230 Natural Product Communications Vol. 8 (2) 2013 Experimental Plant material: Seeds were collected from 9 specimens of Abies koreana E.H. Wilson growing in central Poland, identified by the director of Rogów Arboretum. The material (seeds with cones) was hand-picked in September 2009. The samples were stored in tight plastic bags in a freezer (-24°C) until needed. Before hydrodistillation, the seeds were separated from cones and ground in a mill. Eight trees, about 30-years old (assayed as 1-8), were grown in Rogów Arboretum, in the same (about 200m2) area, while one tree, about 15-years old, was grown in a Łódź suburb. The voucher specimen of plant material was deposited in the Institute of General Food Chemistry, Lodz, Poland. Isolation of essential oil: The fir seeds were crushed using a simple mill. The essential oils were isolated from the crushed seeds by hydrodistillation for 4 h using a Clevenger-type apparatus. Hydrodistillation of the seeds was performed 3 times in parallel. However, the mass of seeds of tree no. 8 was too small to calculate the yield of essential oil. Isolation of essential oil components: To isolate the volatiles, the seed oil (20.8 g) – mixture of hydrodistilled oils from all tested trees, was fractionally distilled under reduced pressure using a 20 cm Vigreux column. Eight fractions (1.1 – 1.8) were obtained containing only monoterpenes: mainly limonene, and α- and pinene. From fraction 1.6 (4.2 g) limonene (89%) was identified by 1 H NMR spectroscopy. The residue from fractional distillation (2.3 g) was next flash-chromatographed (FC) on silica gel 60 (0.0400.063 mm, Merck) with n-hexane and increasing amounts of diethyl ether. The separation was monitored by TLC and GC-MS. Twenty fractions (2.1 – 2.20) were obtained and analysed by GC-MS. Structures of 7 volatiles from the following fractions were confirmed using NMR spectroscopy. Frs. 2.4 (31 mg): -bisabolene (47%); 2.9 (300 mg): bornyl acetate (85%); 2.11 (42 mg): geranyl acetate (68%); 2.13 (18 mg): (E)- -caryophyllene oxide (42%); 2.15 (19 mg): 1-epi-cubenol (20%); 2.16 (66 mg): (E)-nerolidol (48%); 2.17 (431 mg): intermedeol (87%). The repeated FC of sample 2.2 (326 mg) gave a fraction containing cis- and trans-10hydroxycalamenene; their structures were confirmed additionally by 1 H NMR spectroscopy. Wajs-Bonikowska et al. Main component quantification: Quantification of the main components of the essential oil was carried out using a calibration curve. Calibration curves were prepared for limonene α-pinene, pinene and bornyl acetate by diluting each standard in n-heptane, at 4 different concentrations, each concentration being injected in triplicate. The application of this analytical procedure allowed the accurate determination of the concentration (g/100 g) of all tested components to be made. The linear regression equation of the examined compounds had the coefficient of determination (R2) of 0.999. Chromatographic analysis: Essential oils were analysed by GCMS-FID. The analyses were performed on a Trace GC Ultra column coupled with a DSQII mass spectrometer (Thermo Electron). A simultaneous GC-FID and MS analysis was performed using a MS-FID splitter (SGE Analytical Science). Mass range was 33-550 amu, ion source-heating: 200°C, ionization energy: 70eV. Operating conditions: capillary column Rtx-1 MS (60 m × 0.25 mm i.d., film thickness 0.25 m), temperature program: 50 (3 min) - 300°C (30 min) at 4°C/min. Injector and detector temperatures were 280°C and 300°C, respectively. Carrier gas was helium (constant pressure: 300 kPa). Resolution of (-) and (+) enantiomers of limonene, α- and -pinene, and camphene was performed on a Chirasil-Dex CB column having the following dimensions: 30 m × 0.25 mm i.d., 0.25 m df. Temperature program: 50 (3 min)-220°C (30 min) at 4°C/min. Injector and detector temperatures were 240°C and 250°C, respectively. Carrier gas was nitrogen at the flow rate of 1.0 mL/min. Identification of compounds: The identification of compounds was based on a comparison of their MS with computer mass library NIST 08, Wiley Registry of Mass Spectral Data 8th Edn, and MassFinder 4.1, along with the relative retention indices (RI, nonpolar column). The identification was also based on a comparison of 1H- and 13C-NMR spectra with literature data and a home-made NMR data base. The identification of limonene, camphene, and αand -pinene enantiomers was based on (-)- and (+)-standards of these terpenes. References [1] [2] [3] [4] [5] [6] [7] [8] [9] [10] Bugala W. (1983) Trees and bushes, 3rd edn. PWRiL, Warsaw, 80-82. Chojnowska E. (2000) Conifers in the garden. Multico Oficyna Wydawnicza, Warsaw, 171-172, 201-202. Yesilda E, Honda G, Sezik E, Tabata M, Fujita T, Tanaka T, Takeda Y, Takaishi Y. (1995) Traditional medicine in Turkey; V: Folk medicine in the inner Taurus Mountains. Journal of Ethnopharmacology, 46, 133-152. Kim YG. (1997) Studies on the extractive of Abies koreana. Mokchae Konghak, 25, 1-9. (a) Jeong SI, Lim JP, Jeon H. (2007) Chemical composition and antibacterial activities of the essential oil from Abies koreana. Phytotherapy Research, 21, 1246-1250; (b) Yoon K, Weon-Jong Y, Sang-Suk K, Tae-Heon O, Nam Ho L, Chang-Gu H. (2009) Abies koreana essential oil inhibits drug-resistant skin pathogen growth and LPS-induced inflammatory effects of murine macrophage. Lipids: ProQuest Agricultural Journal, 44, 471-476; (c) Baran Sz, Stephan von Reuss H, König WA, Kalemba D. (2007) Composition of the essential oil of Abies koreana Wils. Flavour and Fragrance Journal, 22, 78-83; (d) Baran Sz, (2006) Biodiversity of essential oils in Abies. Ph.D. Thesis, Technical University of Lodz, Poland. Wajs A, Urbańska J, Zaleśkiewicz E, Bonikowski R. (2010) Composition of essential oil from seeds and cones of Abies alba. Natural Product Communication, 5, 1291-1294 Bazdi B, Oller Lopez JL, Cuerva JM, Oltra J. (2006) Composition of the essential oil from the seeds of Abies maroccana. Journal of Essential Oil Research, 18, 160-161. Sagareishvili TG. (1999) Composition of the essential oil of Abies nordmanniana, Chemistry of Natural Compounds, 35, 586 (a) Biodiversity Convention signed in Rio de Janeiro 5.06.1992 (Journal of Laws of the Republic of Poland 2002 no. 184, position 1532); (b) Hunt RS, von Rudloff E. (1974) Chemosystematic studies in the genus Abies, part I. Canadian Journal of Botany, 52, 477-487; (c) von Rudloff E. (1976) Chemosystematic studies in the genus Abies, part II. Canadian Journal of Botany, 54, 1926-1931; (d) von Rudloff E, Hunt RS. (1977) Chemosystematic studies in the genus Abies, part III. Canadian Journal of Botany, 55, 3087-3092. Person M. (2003) Chemodiversity and functions of monoterpene hydrocarbons in conifers. Ph.D. Thesis, Royal Institute of Technology (KTH), Stockholm. Natural Product Communications Vol. 8 (2) 2013 Published online (www.naturalproduct.us) Essential Oil of Flowers of Anaphalis contorta, an Aromatic and Medicinal Plant from India Rajesh K. Joshi Composition of Essential Oils from Seeds of Abies koreana Anna Wajs-Bonikowska, Karol Olejnik, Radosław Bonikowski and Piotr Banaszczak Chemical Composition of Hypericum richeri subsp. grisebachii Essential Oil from Croatia Igor Jerković, Maja Marasović, Zvonimir Marijanović, Kroata Hazler Pilepić, Željan Maleš and Mladen Miloš Chemical Composition of the Essential Oils from Two Vietnamese Asarum Species: A. glabrum and A. cordifolium Tran Huy Thai, Ophélie Bazzali, Tran Minh Hoi, Nguyen Anh Tuan, Félix Tomi, Joseph Casanova and Ange Bighelli Essential Oils from Five Species of Annonaceae from Vietnam Tran D. Thang, Do N. Dai, Tran M. Hoi and Isiaka A. Ogunwande Essential Oils from the Leaves of Six Medicinal Plants of Nigeria Isiaka A. Ogunwande, Nudewhenu O. Avoseh, Guido Flamini, Alimot-Sadiat O. Hassan, AbdulRazaq O. Ogunmoye, Akindele O. Ogunsanwo, Kamorudeen O. Yusuf, Atuonwu O. Kelechi, Zainab A. Tiamiyu and Godgift O. Tabowei Comparative Study on In Vitro Activities of Citral, Limonene and Essential Oils from Lippia citriodora and L. alba on Yellow Fever Virus Luz Angela Gómez, Elena Stashenko and Raquel Elvira Ocazionez TLC-Bioautographic Evaluation of In Vitro Anti-tyrosinase and Anti-cholinesterase Potentials of Sandalwood Oil Biswapriya B. Misra and Satyahari Dey Composition, Mosquito Larvicidal, Biting Deterrent and Antifungal Activity of Essential Oils of Different Plant Parts of Cupressus arizonica var. glabra (‘Carolina Sapphire’) Abbas Ali, Nurhayat Tabanca, Betul Demirci, K. Husnu Can Baser, Jane Ellis, Sandra Gray, Brett R. Lackey, Christine Murphy, Ikhlas A. Khan and David E. Wedge Composition of Cassia fistula Oil and its Antifungal Activity by Disrupting Ergosterol Biosynthesis Md. Irshad, Aijaz Ahmad, Md. Zafaryab, Farah Ahmad, Nikhat Manzoor, Man Singh and M. Moshahid A. Rizvi Chemical Composition and Biological Activity of the Essential Oil of Amomum biflorum Chakkrapat Singtothong, Michel J. Gagnon and Jean Legault Chemical Composition and Antibacterial Activity of Essential Oils from Myrcia alagoensis (Myrtaceae) Aline do N. Silva, Ana Paula T. Uetanabaro and Angélica M. Lucchese Composition, in-vitro Anticancer, and Antimicrobial Activities of the Leaf Essential Oil of Machilus mushaensis from Taiwan Yu-Chang Su, and Chen-Lung Ho Chemical Constituents and Cytotoxic Evaluation of Essential Oils from Leaves of Porcelia macrocarpa (Annonaceae) Erica Biolcati P. da Silva, Alisson L. Matsuo, Carlos R. Figueiredo, Mariana H. Chaves, Patricia Sartorelli and João Henrique G. Lago 225 227 231 235 239 243 249 253 257 261 265 269 273 277 Book Review Medicinal Plants of China, Korea, and Japan: Bioresources for Tomorrow’s Drugs and Cosmetics by Chritophe Wiart, PharmD, PhD Stephen Pyne 281 Natural Product Communications 2013 Volume 8, Number 2 Contents Original Paper Page Drimendiol, A Drimane Sesquiterpene with Quorum Sensing Inhibition Activity Cristian Paz, Gerardo Cárcamo, Mario Silva, José Becerra, Homero Urrutia and Katherine Sossa Microbial Transformation of Curcumol by Aspergillus niger Chen Li-Xia, Zhang Hui, Zhao Qian, Yin Shi-Yu, Zhang Zhong, Li Tian-Xian and Qiu Feng Chemopreventive Effect of Sarcophine-diol on NOR-1-Induced TPA-Promoted Skin Carcinogenesis in Female HOS:HR-1 Mice Pawel T. Szymanski, Safwat A. Ahmed, Sherief Khalifa, Harukuni Tokuda, Eiichiro Ichiishi, Akira Iida, Nobutaka Suzuki and Hesham Fahmy Carmichaeline A: A New C20-diterpenoid Alkaloid from Aconitum carmichaeli Shu-hua Li, Jun-ruXiong, Yuan-qin Zhang, Qing-xiang Xiang and Feng-zheng Chen Steroidal Saponins from Dracaena marginata Abdelmalek Rezgui, Anne-Claire Mitaine-Offer, David Pertuit, Tomofumi Miyamoto, Chiaki Tanaka, Stéphanie Delemasure, Patrick Dutartre and Marie-Aleth Lacaille-Dubois Zephgrabetaine: A New Betaine-type Amaryllidaceae Alkaloid from Zephyranthes grandiflora Deepali Katoch, Dharmesh Kumar, Upendra Sharma, Neeraj Kumar, Yogendra S. Padwad, Brij Lal and Bikram Singh Antioxidant and Anti-inflammatory Compounds in the Popular Landscape Plant Berberis thunbergii var. atropurpurea Chuan-Rui Zhang, Robert E. Schutzki and Muraleedharan G. Nair Two New Amides from Streptomyces michiganensis Jinghua Xue, Liangxiong Xu, Zi-Hua Jiang and Xiaoyi Wei Determination of Bioactive Compounds in the Juice of Pummelo (Citrus grandis Osbeck) Marina Russo, Ivana Bonaccorsi, Germana Torre, Antonella Cotroneo, Paola Dugo and Luigi Mondello Antiplasmodial Activity of Compounds from the Surface Exudates of Senecio roseiflorus Leonidah Omosa Kerubo, Jacob Ogweno Midiwo, Solomon Derese, Moses K. Langat, Hosea M. Akala, Norman C. Waters, Martin Peter and Matthias Heydenreich Anti-inflammatory, Antioxidant and Cytotoxicity Activities of Methanolic Extract and Prenylated Flavanones Isolated from Leaves of Eysehardtia platycarpa Valeri Domínguez-Villegas, Vanessa Domínguez-Villegas, María Luisa García, Ana Calpena, Beatriz Clares-Naveros and María Luisa Garduño-Ramírez Phenolic Glycosides from Lindera obtusiloba and their Anti-allergic Inflammatory Activities Hyun Gyu Choi, Hwa Dong Lee, Sang Hyun Kim, MinKyun Na, Jeong Ah Kim and Seung Ho Lee Antiproliferative Effects of an Analog of Curcumin in Hep-2 cells: A Comparative Study with Curcumin Mohankumar Kumaravel, Pajaniradje Sankar, Periyasamy Latha, Chellakan S Benson and Rajagopalan Rukkumani Antiproliferative Activity of epi-Cercosporin in Human Solid Tumor Cell Lines Ángel Trigos, César Espinoza, Maricela Martínez, Olivia Márquez, Leticia G. León, José M. Padrón, Manuel Norte and José J. Fernández New Anthraquinone Derivatives from Geosmithia lavendula Lourin G. Malak, Daoud W. Bishay, Afaf M. Abdel-Baky, Ahmed M. Moharram, Stephen J. Cutler and Samir A. Ross Pancreatic Lipase Inhibitory Activity of Cassiamin A, a Bianthraquinone from Cassia siamea Dilip Kumar, Aniket Karmase, Sneha Jagtap, Ruchi Shekhar and Kamlesh K. Bhutani Antifeedant Activity of Spin-Labeled Derivatives of Deoxypodophyllotoxin against Brontispa longissima Larvae Gang Feng, Jing Zhang, Liu Yang, Ying-Qian Liu, Zhi-Wei Zhang, Xuan Tian, Qi-An Jin and Zheng-Qiang Peng Determination of Organic Acids in Salicornia herbacea by Solid-phase Extraction Combined with Liquid Chromatography Dandan Han, Minglei Tian, Dong Wha Park and Kyung Ho Row Hypoglycemic Effect of Bumelia sartorum Polyphenolic Rich Extracts Halliny S. Ruela, Katia C. C. Sabino, Ivana C. R. Leal, Ana M. Landeira-Fernandez, Michelle R. A. de Almeida, Talita S. M. Rocha and Ricardo M. Kuster Protoanemonin Content Variation between Clematis spp.: Leaf, Stem and Root Fangming Jin, Christian Narkowicz and Glenn A Jacobson Methanolic Extract of Nigella sativa Seed Inhibits SiHa Human Cervical Cancer Cell Proliferation Through Apoptosis Tarique N. Hasan, Gowhar Shafi, Naveed A Syed, Muhammad A Alfawaz, Mohammed A. Alsaif, Anjana Munshi, Kai Y. Lei and Ali A. Alshatwi Glucosinolate Biosynthesis in Hairy Root Cultures of Broccoli (Brassica oleracea var. italica) Sun-Ju Kim, Woo Tae Park, Md. Romij Uddin, Yeon Bok Kim, Sang-Yong Nam, Kwang Hyun Jho and Sang Un Park Characterization of Volatile Components of Zingiber roseum Essential Oil Using Capillary GC on Modified Cyclodextrins VPPalayam S Pragadheesh, Anju Yadav, Manju Singh and Chandan S Chanotiya Continued inside backcover 147 149 153 155 157 161 165 169 171 175 177 181 183 187 191 195 199 203 207 211 213 217 221