3146
J. Agric. Food Chem. 2006, 54, 3146−3150
Characterization of Volatile Constituents of Haplopappus greenei and Studies on the Antifungal Activity against Phytopathogens BETUL DEMIRCI,*,† KEMAL HUSNU CAN BASER,† NURHAYAT TABANCA,‡ DAVID E. WEDGE‡
AND
Department of Pharmacognosy, Faculty of Pharmacy, Anadolu University, 26470 Eskisehir, Turkey, and Natural Products Utilization Research Unit, Agricultural Research Service, U.S. Department of Agriculture, The University of Mississippi, University, Mississippi 38677
Essential oil of Haplopappus greenei A. Gray was obtained by hydrodistillation of aerial parts, which were subsequently analyzed by gas chromatography and gas chromatography-mass spectrometry. Major components were identified as carvacrol (8.7%), β-pinene (7.6%), trans-pinocarveol (6.2%), and caryophyllene oxide (5.8%), respectively. In total, 104 components representing 84.9% of the investigated essential oil were characterized. Furthermore, the essential oil was evaluated for antimalarial, antimicrobial, and antifungal activities. However, only antifungal activity was observed against the strawberry anthracnose-causing fungal plant pathogens Colletotrichum acutatum, Colletotrichum fragariae, and Colletotrichum gloeosporioides using the direct overlay bioautography assay. Major essential oil components were also evaluated for antifungal activity; the carvacrol standard demonstrated nonselective activity against the three Colletotrichum species and the other compounds were inactive. KEYWORDS: Haplopappus greenei; Asteraceae; essential oil; GC-MS; carvacrol; β-pinene; strawberry pathogenic fungi; biological activity; antifungal activity
INTRODUCTION
The genus Haplopappus Cass, which belongs to the family of Asteraceae, is characterized by perennial herbs or shrubs with yellow flowers and is distributed in North and South America. Some Haplopappus species are used in Chilean folk medicine as choleretics and cholagogues (1, 2). Furthermore, the exudates of the various Haplopappus species are used to treat infected wounds and gastrointestinal infections, to promote wound healing, and as digestive stimulants (3). Haplopappus greenei A. Gray [) Ericameria greenei (A. Gray) G.L. Nesom, H. bloomeri (Gray) var. greenei, Macronema greenei (Gray)] is commonly known as “Greene’s goldenweed”. This species produced a significant amount of resinous exudates from the twigs and leaves (1, 2). Previous phytochemical investigations of Haplopappus species have revealed the presence of triterpenes (4, 5), diterpenes (3, 6-14), flavonoids (5, 15-20), coumarins (1, 12, 16, 20, 21), monoterpenes (22, 23), and sesquiterpenes (22-26). In the published literature only a few volatile compounds from Haplopappus species have been reported (23-25). For example, R-pinene, β-phellandrene, and phellandral were found as major
compounds in the steam-distilled oil of the leaves and stems of H. laricifolius (24). Isocomene, modhelphene, 1,2,3,4-tetrahydro-1,1,5,6-tetramethylnaphthalene, β-caryophyllene, caryophyllene oxide, limonene, borneol, bornyl acetate, and carvone were reported in the H. heterophyllus volatile oil (25). Urzua et al. described the chemical composition of the resinous exudates from H. foliosus and H. uncinatus and identified a number of monoterpenes, sesquiterpenes, hydrocarbons, and phenyl propanoids (23). Urzua et al. recently reported the antibacterial diterpenoids of the resinous exudate (3). Several groups investigating the biological activities of Haplopappus species have reported antioxidant, antimicrobial, and antibacterial activities (2, 3, 13, 21, 27, 28). The aim of this study was to evaluate the antimalarial, antimicrobial, and antifungal activities of H. greenei essential oil and its major constituents for activity against various plant and human pathogenic microorganisms. Furthermore, to the best of our knowledge we are reporting for the first time the volatile constituents of H. greenei characterized by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). MATERIALS AND METHODS
* Corresponding author tel: +90 222 335 05 80-3717; fax: +90 222 335 07 50; e-mail:
[email protected] or
[email protected]. † Anadolu University. ‡ U.S. Department of Agriculture.
General. Pure essential oil compounds (β-pinene, carvacrol, transpinocarveol, and caryophyllene oxide) (>95%, Aldrich-Sigma, St. Louis, MO) and fungicide technical grade standards benomyl, cyprodi-
10.1021/jf053222v CCC: $33.50 © 2006 American Chemical Society Published on Web 03/23/2006
Antifungal Haplopappus greenei Essential Oil
J. Agric. Food Chem., Vol. 54, No. 8, 2006
3147
Table 1. Composition of the Essential Oil of H. greenei no.
compound
RRIa
%b
ID methodc
no.
compound
RRIa
%b
ID methodc
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
R-pinene β-pinene sabinene myrcene R-terpinene limonene 1,8-cineole β-phellandrene (Z)-β-ocimene γ-terpinene (E)-β-ocimene p-cymene terpinolene artemisia ketone perillen γ-campholene aldehyde R,p-dimethylstyrene 7β-(H)-silphiperfol-5-ene R-cubebene trans-sabinene hydrate (Z)-3-hexenyl 2-methylbutyrate cyclosativene R-campholene aldehyde (E)-theaspirane camphor dihydroachillene linalool cis-sabinene hydrate trans-p-menth-2-en-1-ol pinocarvone nopinone terpinen-4-ol β-caryophyllene hotrienol cis-p-menth-2-en-1-ol myrtenal pulegone trans-pinocarveol trans-verbenol cryptone R-terpineol trans-sabinol verbenone thujol p-mentha-1,5-dien-8-ol valencene phellandral carvone cis-piperitol δ-cadinene γ-cadinene ar-curcumene cumin aldehyde myrtenol
1032 1118 1132 1174 1188 1203 1213 1218 1246 1255 1266 1280 1290 1358 1429 1439 1452 1454 1466 1474 1482 1492 1499 1516 1532 1547 1553 1556 1571 1586 1601 1611 1612 1616 1638 1648 1662 1670 1683 1690 1706 1720 1725 1729 1738 1740 1744 1751 1758 1773 1776 1786 1802 1804
0.3 7.6 1.3 0.2 0.3 0.8 1.0 trd 0.1 0.8 0.1 0.5 0.3 0.1 0.1 0.1 0.1 0.1 0.3 0.1 tr tr 0.5 0.1 0.4 0.2 0.4 0.1 0.2 4.2 0.7 2.0 0.4 0.7 0.2 4.9 0.2 6.2 0.9 0.8 0.9 0.3 0.5 0.1 0.7 0.2 0.6 0.3 0.1 0.4 0.1 0.6 0.6 4.1
GC-MS GC-MS GC-MS GC-MS GC-MS GC-MS GC-MS MS MS GC-MS MS GC-MS GC-MS MS MS MS MS MS MS GC-MS MS MS MS MS GC-MS MS GC-MS MS MS GC-MS MS GC-MS GC-MS MS GC-MS MS GC-MS GC-MS MS MS GC-MS GC-MS GC-MS MS MS MS MS GC-MS MS MS MS MS GC-MS MS
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 87 88 89 90
perilla aldehyde p-mentha-1,5-dien-7-ol trans-carveol p-cymen-8-ol (E)-geranyl acetone epi-cubebol R-calacorene cubebol (E)-β-ionone γ-calacorene isocaryophyllene oxide caryophyllene oxide perilla alcohol presilphiperfolan-9R-ol salvial-4(14)-en-1-one (E)-nerolidol humulene epoxide II p-mentha-1,4-dien-7-ol caryophylla-2(12),6(13)-dien-5-one 1-epi-cubenol hexyl benzoate viridiflorol cumin alcohol hexahydrofarnesyl acetone spathulenol T-cadinol nonanoic acid thymol ar-turmerol carvacrol 1-methylethyl hexadecanoatee R-cadinol epi-R-bisabolol selin-11-en-4R-ol decanoic acid caryophylla-2(12),6(13)-dien-5β-ol () caryophylladienol I) caryophylla-2(12),6(13)-dien-5R-ol () caryophylladienol II) eudesma-4(15),7-dien-4β-ol caryophylla-2(12),6-dien-5R-ol () caryophyllenol I) caryophylla-2(12),6-dien-5β-ol () caryophyllenol II) kaur-16-ene pentacosane dodecanoic acid phytol benzyl benzoate tetradecanoic acid heptacosane pentadecanoic acid nonacosane hexadecanoic acid
1807 1814 1845 1864 1868 1900 1941 1957 1958 1984 2001 2008 2029 2030 2037 2050 2071 2073 2074 2088 2095 2104 2113 2131 2144 2187 2192 2198 2214 2239 2251 2255 2256 2273 2298 2316
0.1 0.3 0.4 0.1 0.1 0.3 0.3 0.2 0.2 0.1 0.2 5.8 0.6 0.2 0.4 0.1 0.7 0.2 0.3 0.2 0.3 0.1 0.3 1.2 3.4 1.0 0.2 0.4 0.2 8.7 0.2 1.0 0.6 0.5 0.3 0.5
GC-MS MS GC-MS GC-MS MS MS MS MS MS MS GC-MS GC-MS GC-MS MS MS GC-MS MS MS MS MS MS MS GC-MS MS MS MS GC-MS GC-MS MS GC-MS MS MS MS MS GC-MS MS
2324
1.1
MS
2369 2389
1.1 0.7
MS MS
2392
1.7
MS
2438 2500 2503 2622 2655 2670 2700 2822 2900 2931
0.1 0.5 0.4 0.1 0.2 0.8 0.2 0.2 0.5 0.8
MS GC-MS GC-MS MS MS GC-MS GC-MS GC-MS GC-MS GC-MS
monoterpene hydrocarbons oxygenated monoterpenes sesquiterpene hydrocarbons oxygenated sesquiterpenes others total
91 92 93 94 95 96 97 98 99 100 101 102 103 104
12.3 41.5 3.0 20.9 7.2 84.9
a RRI, relative retention indices calculated against n-alkanes on the HP Innowax column. b Percent calculated from flame ionization detector (FID) data. c Method of identification: GC, identification based on retention times of genuine compounds on the HP Innowax column; MS, tentatively identified on the basis of computer matching of the mass spectra of peaks with the Wiley (29) and MassFinder (30) libraries. d tr, trace (
