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Aromas from Quebec. I. Composition of the essential oil of the rhizomes of Acorus calamus L



The chemical composition of the essential oil obtained from the rhizomes of Acorus calamus (sweet flag) collected in the Grondines region, Province of Quebec, was determined by GC/FID and GC/MS analyses. Several components were isolated by liquid chromatography and were identified by various NMR experiments such as: H-NMR, C-NMR, HSQC, HMBC, and NOESY. The major compounds were identified as preisocalamenediol, acorenone, shyobunone, and cryptoacorone. The complete stereochemical structure of cryptoacorone was elucidated
Aromas from Quebec. I. Composition of the essential oil of the rhizomes of
Acorus calamus L.
François-Xavier GARNEAU* and Guy COLLIN
Corporation LASEVE, Université du Québec à Chicoutimi (Québec),Saguenay, Canada G7H 2B1
3 Centre de recherche et de développement en horticulture, 400, boul. Gouin, Saint-Jean-sur-Richelieu (Québec),
Canada J3B 3E6
Serge LAVOIE, Hélène GAGNON, Nadia SAVARD and André PICHETTE
Laboratoire LASEVE, Université du Québec à Chicoutimi (Québec), Canada G7H 2B1
* address for correspondence
Correspondence should be addressed to
François-Xavier Garneau
Corporation LASEVE
Université du Québec à Chicoutimi
555 boul. Université
Chicoutimi, Québec, Canada
G7H 2B1
+1-(418)-545-5011 ext. 5071
The chemical composition of the essential oil obtained from the rhizomes of Acorus calamus
(sweet flag) collected in the Grondines region, Province of Quebec, was determined by GC/FID
and GC/MS analyses. Several components were isolated by liquid chromatography and were
identified by various NMR experiments such as: 1H-NMR, 13C-NMR, HSQC, HMBC, and
NOESY. The major compounds were identified as preisocalamendiol, acorenone, shyobunone,
and cryptoacorone. The complete stereochemical structure of cryptoacorone was elucidated.
Key Word Index
Acorus calamus, Araceae, essential oil composition, preisocalamendiol, acorenone, shyobunone,
epiacorone, stereochemistry, cryptoacorone.
The essential oils of different anatomical structures of Acorus calamus L. Araceae, such as the
rhizomes (1) and the leaves (2) were described in several papers. Other papers reported the
chemical composition of the essential oils of plants growing in different countries such as Japan
(3), Bangladesh (4) and Turkey (5). Bélanger et al also published a preliminary study on the oil
composition of plants from Quebec (6). Reviews of the literature appeared elsewhere (7) as well
as a short description of commercial samples (8). In this study, we determined the structure of the
major components using spectroscopic methods. Of particular interest was the structure of
cryptoacorone. Vrkoc et al (9) proposed the stereochemical formula of cryptoacorone based on
the Hudson-Klyne rule, the octant rule and dipole moment measurements. They determined the
configuration of the chiral centers except for the configuration of the methyl group on the
cyclopentanone ring. In this paper, we confirmed these configurations and unambiguously
determined the relative configuration of the methyl group in the five-member ring using various
NMR experiments.
Extraction of essential oil: Rhizomes of A. calamus were collected from plants cultivated in the
Grondines region, Quebec. The rhizomes (40.86 kg) of the plant were subjected to steam
distillation in a 400- litre Stainless Steel Still for 12 h giving 428 ml of a pale yellow oil in a yield
of 1.05% (v/w).
Oil Analysis: GC/FID analyses were carried out on a Hewlett-Packard 5890 gas chromatograph
fitted with both an apolar DB-5 capillary column and a polar Supelcowax 10 column (both 30 m
× 0.25 mm; film thickness 0.25 µm). GC/MS analyses were performed on an HP 5972 mass
spectrometer at 70 eV coupled to an HP 5890 GC equipped with a DB-5 column (same as above).
Temperature program for both GC-FID and GC-MS analyses was 40 °C for 2 min, then 2 °C/min
to 210 °C and held constant for 33 min. Identification of the components was done by
comparison of their retention indices and mass spectra with those of the literature (10, 11, 12).
Quantitative data were obtained electronically from GC-FID area percentages.
Refraction indices and relative density were measured according to standard methods NF ISO
279 and 280. The optical rotation was measured on a Autopol IV polarimeter from Rudolph
Research Analytical. HPLC-MS were carried out on an Agilent 1100 LC-MS system equipped
with a UV-VIS diode array detector (DAD) and an atmospheric pressure chemical ionisation
mass selective detector (APCI-MSD). Analytical HPLC separations were achieved using a
Zorbax Eclipse XDB-C18 5 µm column (4.6×150 mm) at a constant temperature of 20 °C. The
mobile phase was 55 % methanol and 45 % water and was delivered at a flow-rate of 1 mL/min.
UV-VIS detector was set at 250 nm with a 100 nm bandwidth. The APCI source of the MS
system worked with a nebulizing nitrogen gas pressure of 40 psi at 10 L/min and at a temperature
of 350 °C. Capillary voltage was set at 4.0 kV with a current of 4 µA and the fragmentor was set
at 70 V. Positive ions in the 100-700 m/z range were registered in the conventional scanning
mode. Preparative HPLC separations were achieved on an Agilent 1100 LC equipped with a
multiple wavelength detector and using a Zorbax ODS-C18 7 µm (21.2 × 250 mm) column. High
Resolution MS analyses were done with electrospray ionisations (ESI) on an Applied
Biosystems/MDS Sciex QSTAR XL QqTOF MS system (serial #T0970304, Toronto, ON,
Canada). Infrared spectra were acquired on KBr using a Perkin-Elmer apparatus. 1H-NMR, 13C-
NMR and 2D NMR spectra were recorded in deuterated chloroform on a Bruker Avance 400
spectrometer (5 mm QNP with Z-gradient probe) operating at 400.13 MHz (1H) or 100.61 MHz
(13C). Chemical shifts are expressed in parts per million (ppm) and were referenced with residual
chloroform (δH = 7.26 and δC = 77.0 ppm). Coupling constants (J) are expressed in hertz and
splitting patterns are designated as follows: s (singlet), d (doublet), t (triplet), q (quardruplet) and
m (multiplet). Purity of the isolated compounds was evaluated with the area of the GC peaks.
Isolation. Compound 1 (20.1 mg, 75.4% pure) was isolated from the essential oil by preparative
TLC using hexane/Et2O 25:1 as eluent. Thereafter, 5.0 g of the essential oil were
chromatographed on silica gel using a gradient of hexane (100%) to hexane/Et2O (10:1). Nine
fractions (F1-F9) were collected and evaporated. 80.0 mg of F8 (1.14 g) was submitted to
preparative HPLC and three compounds were isolated: 2 (TR = 56.5 min, 5.8 mg, 92.5% pure), 3
(TR = 24.7 min, 28.6 mg, 98.0% pure) and 4 (TR = 27.2 min, 20.1 mg, 49.6% pure). All
compounds were submitted to NMR for structure elucidation.
Cryptoacorone [3]: amorphous solid; [α]D = +120.6 (c=0.87, CHCl3); HREIMS m/z 236.1773
[M+] (calcd for C15H24O2, 236.1776); EIMS, see Figure 3; IR (KBr) ν 2961, 2929, 2873, 1734,
1707, 1459, 1169 cm
1; 1H and 13C, see table II.
The pale yellow essential oil had the following physical properties, n: 1.4983,
[α]D = + 36.5 ± 0.1° (net), d20 : 0.964. The chemical composition is shown in Table I. The most
important components are monooxygenated sesquiterpenes (ca. 70 %). Preisocalamendiol [1]
(18 %), acorenone (14.2 %), shyobunone (10.8 %), and cryptoacorone [3] (7.5 %) are the main
products. From the oil relative density, its rotation index value, and the low percentage of β-
asarone, the plant corresponds to a triploid (European) variety (8).
Because many compounds could not be unambiguously identified, some where isolated using a
variety of chromatographic techniques (TLC, CC, HPLC). Thus, preisocalamendiol [1],
isocalamendiol [2], and epiacorone [4] (Fig.1) were identified by 1D and 2D NMR spectral
analyses and by comparison of the MS and NMR spectra published in the literature (13, 14 and
15 respectively). The mass spectrum of isocalamendiol [2] closely resembles the MS published
recently (16), the only exception being the m/z =159 peak. This peak is the base peak in ref. (16)
and counts for less than 5 % in our spectrum.
The molecular formula of compound 3 C15H24O2, an amorphous solid, was determined by HRMS
(m/z 236.1773 obs. 236.1776 calc.) and was supported by 13C and DEPT-135 spectra, which
revealed 4 methyls, 4 methylenes, 4 methines and 3 quaternary carbons, including two carbonyls
at δC 212.8 and 217.6. Since epiacorone [4] was already identified and that its 1H and 13C spectra
were similar to those of compound 3, the latter was suspected to be another acorone stereoisomer.
This was confirmed by the systematic analysis of COSY, TOCSY, HSQC and HMBC spectra
(table II). The relative stereochemistry was determined with the NOESY spectrum. Figure 2
presents the most important NOESY correlations, which clearly show that methyls 15 and 13
reside on the same side as methylene 6. Concerning the stereochemistry of methine 8, no clear
NOESY correlation could be used. However, the S stereochemistry would lead to the same
structure as epiacorone [4], which was already identified. The uncertainty regarding the
stereochemistry of the methyl group at C-4 in Vkroc’s study was cleared up by means of the
NOESY correlation which ascertained the β configuration (9). So the structure of compound 3
was determined to be (1S,4S,5S,8R)-4,8-dimethyl-1-(2-propyl)-spiro[4.5]decane-2,8-dione or
cryptoacorone. The complete NMR data of this compound is reported for the first time. The mass
spectrum of cryptoacorone [3] is shown in Figure 3 and closely resembles that of epiacorone [2].
We would like to thank Professor Bernd Keller, Mass Spectrometry Laboratory., Dept. of
chemistry, Queen’s University, Kingston, Ontario for the measurement of the exact mass of
1. M. N. Todorova, I. V. Ognyanov and S. Shatar, Chemical composition of essential oil
from Mongolian Acorus calamus L. rhizomes. J. Essent. Oil Res., 7, 191-193 (1995).
2. P. R. Venskutonis and A. Dagilyte, Composition of essential oil of sweet flag (Acorus
calamus L.) leaves at different growing phases. J. Essent. Oil Res., 15, 313-318 (2003).
3. M. Niwa, A. Nishiyama, M. Iguchi and S. Yamamura, Sesquiterpenes from Acorus
calamus L. Bull. Chem. Soc. Japan, 48, 2930-2934 (1975).
4. I. Bonaccorsi, A. Cotroneo, J.U. Chowdhury and M. Yusuf, Studies on essential oils
bearing plants of Bangladesh. Part VII. Composition of the rhizomes oil of Acorus calamus
L.(sweet flag). Essenze, Deriv. Agrum., 67, 392-402 (1997).
5. M. Özcan, A. Akgül and J.-C. Chalchat, Volatile constituents of the esssential oil of
Acorus calamus L. grown in Konya province (Turkey). J. Essent. Oil Res., 14, 366-368
6. A. Bélanger, L. Dextrase, H. Goudmand, F.-X. Garneau and G. Collin, Essential oil
composition of Acorus calamus from Quebec. Riv. Ital. EPPOS, Spec. Num. 15ièmes Journées
Int. Huiles Ess., Digne-les-Bains, France, 529-534 (1997).
7. a) B. Lawrence, Progress in essential oils. Perfum. & Flavor., 11, 52-54 (1986);
b) B. Lawrence, Progress in essential oils. Perfum. & Flavor., 22, 65-67 (March/April 1997).
8. C. M. Bertea, C.M. Azzolin, S. Bossi, G. Doglia and M. E. Maffei. Identification of an
EcoRI restriction site for a rapid and precise determination of
-asarone-free Acorus
calamus cytotypes. Phytochem., 66, 507-514 (2005).
9. J. Vrkoč, J. Jonáš, V. Herout and Šorm, F.; On terpenes. CLVII. Steric structure of
acorone, isoacorone, and cryptoacorone. Collection Czechoslov. Chem. Commun., 29,
539-550 (1964).
10. R.P. Adams, Identification of Essential Oil Components by Gas Chromatography/
Quadrupole Mass Spectrometry. Allured Publishing Corp., Carol Stream, IL (2001).
11. D. H. Hochmuth, MassFinder 3, 2004, Hamburg, Germany (
12. D. Joulain and W. A. König, The atlas of spectral data of sesquiterpene hydrocarbons,
E. B. Verlag Hamburg (1998).
13. C. Zdero, F. Bohlmann, J.C. Solomon, R. M. King and H. Robinson, Ent-Clerodanes
and other constituents from Bolivian Baccharis species. Phytochem., 28, 531-542 (1989).
14. M. Iguchi and A. Nishiyama, Isolation and Structure of Isocalamendiol. Tetrahedron
Letters, 3729 (1969).
15. K. Nawamaki and M. Kuroyanagi, Sesquiterpenoids from Acorus calamus as
germination inhibitors. Phytochem., 43, 1175-1182 (1996).
16. M. Gonny, P. Bradesi and J. Casanova, Identification of the components of the
essential oil from wild Corsican Daucus carota L. using 13C-NMR spectroscopy. Flav. &
Fragr. J., 19, 424-433 (2004).
Table I. Composition of the essential oil of the rhizomes of Acorus calamus L.
Compound Retention index Area
Mode of
DB5 Supelcowax
α-pinene 940 1020 0.1 a,b,c
camphene 953 1065 0.2 a,b,c
β-pinene 978 1107 0.2 a,b,c
myrcene 992 1169 tr. a,b,c
limonene 1032 1194 tr. a,b,c
camphor 1149 1506* 0.1 a,b,c
decanal 1204 1506* 0.1 a,b,c
octyl acetate 1215 1485 0.1 a,b,c
2,3-dimethoxytoluene 1244 1801 0.2 a,b,c
bornyl acetate 1293 1573 0.5 a,b,c
nonyl acetate 1315 1589 tr. a,b,c
α-funebrene 1380 1558 0.2 a, b
β-elemene 1390 1582 0.3 a,b,c
7-epi-α-cedrene 1396 1539 0.1 b,d
(Z)-isoeugenol 1403 2275 0.2 a,b,c
methyl eugenol 1403 2029* tr. a,b,c
β-funebrene 1408* 1571 1.6 b
α-cedrene 1408* 1555 0.5 c
decyl acetate 1412 1698 0.1 a, b
β-cedrene 1415 1580* 1.0 a,b,c
α-cedrene, isomer1429 1503 0.1 a, b
(E)-α-bergamotene 1437 1580* 0.8 a,b,c
prezizaene isomer 1444 1610 0.8 a,b,c
(E)-isoeugenol 1444 2334 0.1 a,b,c
prezizaene 1449 1617 1.7 a,b,c
zizaene 1453 0.15 d
acoradiene1458 0.4 a, b
trans-β-farnesene 1463* 1671 1.1 a,b,c
Compound Retention index Area
Mode of
DB5 Supelcowax
(Z)-methyl isoeugenol 1463* 2239 0.6 a.b
acora-3(10),14-diene 1469 1664 0.4 b
β-acoradiene 1476* 1668* 0.4 b
4,5-di-epi-aristolochene 1476* tr. b
α-neocallitropsene 1478 0.1 b
germacrene D 1484* 1701* 0.4 a,b,c
γ-curcumene 1484* 1686 0.2 a,b,c
ar-curcumene 1488 1768 0.6 a,b,c
5-epi-aristolochene 1491 1716 0.5 b
6-epi-shyobunone 1497 1840 3.1 c
hinesene 1499* 0.2 a
bicyclogermacrene 1499* 1722 0.2 a,b,c
α-muurolene 1503 1718 0.2 a,b,c
isogermacrene A 1505 1712 0.2 b,c
cuparene 1507 1801 0.2 a,b,c
shyobunone 1518 1887 13.3 c
δ-cadinene 1525 1749* 0.5 a,b,c
β-sesquiphellandrene 1528 1763 2.1 a,b,c
isoshyobunone 1532 1867 1.3 c
(E)-nerolidol 1566 2042* 1.5 a,b,c
spathulenol 1575* 2118* 0.7 a,b,c
germacren-D-4-ol 1575* 2036 0.1 a,b,c
vulgarone A 1584 2042* 0.3 a
Preisocalamendiol [1] 1600 1999 18.0 e
sesquithuriferol 1608 2115 0.3 a
unknown A1625 2067 1.6
unknown B1628 2016 0.6
unknown C1641* 2042* 2.1
tau-muurolol 1645 2171 0.1 a,b,c
trans-isoelemicin 1650* 2389 tr a.c
Compound Retention index Area
Mode of
DB5 Supelcowax
α-cadinol 1654 2212 0.8 a,b,c
unknown D1659 2118* 1
4-epi-acorenone 1679 2131 0.7 a
acorenone 1688 2158 14.2 a
6α-hydroxygermacra-1(10),4-diene 1703 2304 0.2 b.c
khusiol 1709 2295 0.1 a.c
acora-7(11),9-dien-2-one1723 2239 0.6 b
Isocalamendiol [2] 1741* 2450* 3.1 c,e
unknown E1741* 2357 0.5
calamendiol1746* 2430 0.5 c
eudesma-3,11-dien-2-one1782 2450* 0.1 b
unknown F1792 2450* 0.5
Cryptoacorone [3] 1797 2524 7.5 a,e
epi-acorone [4] 1801 2524 1 a,e
acorone, isomer 1 1807 2524 0.9 a
acorone, isomer 2 1819 2519 2.1 a
unknown G1839 0.3
Total 94.6
*at least two products are co-eluted; ** a) Adams (10), b) MassFinder (11), c) LASEVE data bank. d) Joulain (12),
e) these compounds were isolated and analyzed by various spectroscopic techniques; tentatively identified
Unidentified peaks: A. 220[M]+(9), 192(100), 81(81), 149(76), 41(72), 69(71), 55(62), 107(53), 95(53), 177(51); B.
222[M]+(30), 138(100), 151(67), 111(59), 41(56), 95(51), 55(45), 81(44), 109(41), 69(39), 67(37); C. 222[M]+(28),
138(100), 111(78), 151(68), 95(65), 41(62), 81(55), 109(52), 69(51), 55(51); D. 159(69), 147(100), 121(85), 81(75),
69(75), 41(65), 109(50), 79(48), 91(46), 43(44); E. 236[M]+(6), 193(100), 137(55), 43(51), 41(45), 166(44), 69(40),
95(37), 55(34), 81(33); F. 218[M]+(42), 121(100), 91(78), 108(66), 133(64), 79(62), 41(59), 147(53), 107(51),
93(50); G. 234[M]+(9), 135(100), 82(68), 136(62), 122(42), 121(42), 41(42), 110(41), 164(40), 109(35);
Table II. 1H and 13C NMR data of Cryptoacorone [3].
Position δC (multa)δH (multb)
1 65.7 (d) 1.95 (t, J = 1.6 Hz)
2 217.6 (s) -
3α45.7 (t) 2.34 (m)
3β1.88 (m)
4 40.2 (d) 1.92 (m)
5 49.7 (s) -
6α41.9 (t) 2.45 (dd, J = 15.1, 2.2 Hz)
6β2.21 (dd, J = 15.1, 1.0 Hz)
7 212.8 (s) -
8 44.2 (d) 2.36 (m)
9α30.8 (t) 1.66 (m)
9β2.11 (m)
10α37.5 (t) 2.01 (m)
10β1.7 (m)
11 27.9 (d) 1.69 (m)
12 25.2 (q) 1.17 (d, J = 6.9 Hz)
13 18.4 (q) 0.92 (d, J = 6.7 Hz)
14 15.3 (q) 1.11 (d, J = 6.6 Hz)
15 15.0 (q) 1.07 (d, J = 6.4 Hz)
a Multiplicities were determined by DEPT. b Multiplicities and coupling constants in Hz are in parentheses.
11 13
(5) Acorone
(4) Epiacorone(3) Cryptoacorone
(1) Preisocalamendiol (2) Isocalamendiol
Figure 1. Structures of isolated sesquiterpenes and acorone.
Figure 2: Important NOESY correlations of cryptoacorone [3].
40 60 80 100 120 140 160 180 200 220 240
Scan 7450 (67.046 min): NSAC01.D
109 123
Figure 3. Mass spectrum of cryptoacorone [3].
... The results are quite different from the previous reports. For example, preisocalamenediol (18.0%), acorenone (14.2%), shyobunone (13.3%) and cryptoacorone (7.5%) were major compounds of the essential oil of A. calamus collected from Quebec, Canada (Garneau, et al., 2008) [6] . Preisocalamendiol (17.3%), isoshyobunone (13.0%), 1,4-(trans)-1,7-(trans)acorenone (10.5%), camphor (5.9%) 2,6-diepishyobunone (2.6%) and β-gurjunene (2.5%) were the main components of the essential oil of A. calamus roots obtained from Turkey. ...
... The results are quite different from the previous reports. For example, preisocalamenediol (18.0%), acorenone (14.2%), shyobunone (13.3%) and cryptoacorone (7.5%) were major compounds of the essential oil of A. calamus collected from Quebec, Canada (Garneau, et al., 2008) [6] . Preisocalamendiol (17.3%), isoshyobunone (13.0%), 1,4-(trans)-1,7-(trans)acorenone (10.5%), camphor (5.9%) 2,6-diepishyobunone (2.6%) and β-gurjunene (2.5%) were the main components of the essential oil of A. calamus roots obtained from Turkey. ...
Full-text available
Sweet flag (Acorus calamus) is mentioned in Ayurveda and belongs to the genus Acorus L. of the family Acoraceae and is widely distributed temperate to sub temperate regions. It is commonly used in traditional medicinal systems of Asian and European countries to treat appetite loss, diarrhoea, digestive disorders, bronchitis indigestion, chest pain, nervous disorders. This review focuses on the therapeutic potential of essential oils, phenyl propanoids, sesquiterpenes and monoterpenes as well as xanthone, glycosides, flavones, lignans, steroids and inorganic constituents.
... The fruits of Grewia asiatica gave 2,10dimethyl-6-methylene dodecan-1-oic acid (5) and cerotic acid (6). The leaves of Ocimum sanctum yielded a carotenol (7) and kaur-5,15(17)-dien-7β-olyl vanillate (8). This work has enhanced understanding about the phytoconstituents of these plants. ...
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The rhizomes of Acorus calamus L. (Acoraceae), aerial parts of Digera muricata (L.) Mart. (Amaranthaceae), fruits of Grewia asiatica L. (Malvaceae) and leaves of Ocimum sanctum L. (Lamiaceae) are used to treat various diseases. This study was planned to isolate phytoconstituents from these plant materials and to characterize their structures. The air-dried powders of the herbal drugs (1.0 kg each) were exhaustively extracted with methanol individually and the concentrated each extract was adsorbed on silica gel separately for preparation of slurries. Each dried slurry was subjected to silica gel column packed in petroleum ether. The columns were eluted with organic solvents in order of increasing polarity to isolate the compounds. The rhizomes of A. calamus afforded stearyl oleate (1) and eudesman-11-ol-8β, 13-olide (2). The aerial parts of D. muricata furnished phenolic glucosides identified as 3-isopropanoic acid phenyl 1-O-α-D-glucopyranoside (3) and resorcinyl 1-O-β-D- glucopyranosyl-(6′→1′′)-O- β-D-glucopyranoside (4). An acyclic sesquiterpenic acid 2,10-dimethyl-6-methylene dodecan-1-oic acid (5) and cerotic acid (6) were isolated from the fruits of Grewia asiatica. The leaves of O. sanctum gave a carotenol carot-4,6,8,10,12,14,2′(17′), 6′(8′), 10′(19′),14′(20′)-decaene-1′-ol (ocimum xanthin, 7) and a diterpenic ester kaur-5,15(17)-dien-7β-olyl vanillate (kaurdienoyl vanillate, 8). The structures of these phytoconstituents have been established on the basis of spectral data analysis and chemical reactions.
... (Xin Chao Liu et al., 2013). Garneau et al. (2008) reported preisocalamendiol (18.0%), acorenone (14.2%), shyobunone (13.3%) and cryptoacorone (7.5%) were major compounds of the essential oil of A. calamus collected from Quebec, Canada. Preisocalamendiol (17.3%), isoshyobunone (13.0%), 1,4-(trans)-1,7-(trans)-acorenone (10.5%), camphor (5.9%) 2,6-diepishyobunone (2.6%) and β-gurjunene (2.5%) were the main components of the essential oil of A. calamus roots obtained from Turkey. ...
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The present investigations entitled “Evaluation of accessions and standardization of cultivation practices of Acorus calamus Linn.” were carried out in the experimental field as well as in laboratory of the Department of Forest Products, College of Forestry, Dr Y S Parmar University of Horticulture and Forestry, Nauni, Solan (HP) during 2015-16 & 2016-17. In the first experiment “Morphological studies of A. calamus under natural conditions” Acorus calamus Linn. was found to be perennial, semi-aquatic, monocot, aromatic herb with creeping rhizomes. Rhizomes were cylindrical or somewhat compressed, creeping, horizontal, greenish white on the outside, pinkish white and spongy inside with distinct nodes and internodes. Leaves were linear, smooth, erect, distichously alternate, ensiform, base equitant, moderate yellow green in colour and having a single prominent mid vein with margin almost entire. In second experiment “Evaluation of different accessions of A. calamus (Bach)” accession C4 (Nikyar) gave maximum fresh rhizome weight (31.29 g/plant), dry rhizome weight (15.33 g/plant), estimated fresh rhizome yield (17.37 q/ha), estimated dry rhizome yield (8.51 q/ha), essential oil content in fresh rhizome (1.07 %) and estimated essential oil yield (18.57 kg/ha) among all the ten accessions evaluated. No inter-accession difference in leaf shape, colour and rhizome colour were noticed amongst the ten accessions evaluated. In third experiment “Effect of different node cuttings on growth and yield of A.calamus” three node cutting (N3) of rhizome gave maximum fresh rhizome weight (28.13 g/plant), dry rhizome weight (13.78 g/plant), estimated fresh rhizome yield (31.25 q/ha), estimated dry rhizome yield (15.32 q/ha) and estimated essential oil yield (16.80 kg/ha) whereas higher benefit cost ratio (1.80) was obtained from one node cutting of rhizome which was followed by two node cutting of rhizome (0.93). In fourth experiment “Effect of different planting time and harvesting schedules on growth and yield of A.calamus” crop planted in July and harvested after 2nd growing season resulted in maximum fresh rhizome weight (33.85 g/plant), dry rhizome weight (15.91 g/plant), estimated fresh rhizome yield (37.61 q/ha), estimated dry rhizome yield (17.68 q/ha) and estimated essential oil yield (26.50 kg/ha). Higher benefit cost ratio (2.14) was obtained when crop was planted in July and harvested after 2nd growing season which was followed by planted in March and harvested after 2nd growing season (1.81). In fifth experiment “Management of A. calamus under different plant spacings, organic manures and fertilizers” Treatment combination of 5t Vermicompost + NPK (100:60:40) with 20x20 cm spacing resulted in maximum estimated fresh rhizome yield (63.25 q/ha), estimated dry rhizome yield (31.63 q/ha) and estimated essential oil yield (30.24 kg/ha) whereas higher benefit cost ratio (2.32) was observed when plants were planted at 20x20 cm spacing and NPK (100:60:40 Kg/ha) was applied which was followed by 30x20 cm spacing and NPK (100:60:40) Kg/ha (1.84).
... (m) 9 1, 5, 6, 9, 11, 14, 15 1a, 14 11 34. (Garneau et al., 2008;Nawamaki and Kuroyanagi, 1996). Thus, we focused toward an epimer of acorone whose 1 H and 13 C assignments were established with accurate studying of HSQC, HMBC and COSY spectra. ...
The MAP sector (Medicinal and Aromatic Plants) is a booming sector. The demand for essential oil is constantly increasing, and in addition to the historical purchasers such perfumers, there is an increasing demand from the general public, especially since the advent of the "organic" trend. Their fields of application are multiple, from aromatherapy to cosmetics or perfumery or agri-food. In this context, this work had two main objectives. Initially, the study of the essential oils of several coniferous species introduced in Corsica (Larix decidua, Pseudotsuga menziesii, Pinus ponderosa, Sequoiadendron giganteum, Cryptomeria japonica and Calocedrus decurrens), which could contribute to a broader range of the essential oils produced on the island. In the second phase, we focused on the essential oils of various endemic species to Madagascar (Melicope belahe, Vepris unifoliolata, Cinnamosma madagascariensis and Elionurus tristis), which are widely used in traditional medicine. Analysis of all essential oils was carried out by combination of GC(RI), GC-MS and 13C NMR. The first part of this thesis concerns the analysis of the oils of six coniferous species introduced in Corsica. Among them, the oils of L. decidua, P. menziesii, P. ponderosa, S. giganteum and C. japonica are dominated by a high content of monoterpenes. Those of L. decidua, P. menziesii and C. japonica also contain a few sesqui- and diterpenes at appreciable contents, while those of P. ponderosa and S. giganteum are rich in phenylpropanoids. On the other hand, C. decurrens oils are more complex and required several fractionation steps to identify four unknown molecules in our databases: pin-2-en-8-ol, pin-2-en-8-yl acetate, methyl pin-2-en-8-oate, and pin-2-en-8-al. The last two are new molecules. The second part was devoted to the analysis of the essential oils of endemic Madagascar species. The leaves essential oil of M. belahe led to the identification of 56 constituents, majors of which are: α-pinene (42.6%), linalool (6.2%), (E)-β-caryophyllene (5.2%) and β-elemene (4.4%). V. unifoliolata exhibited a bark essential oil with high amounts of thermosensitive molecules: isofuranodiene (43.9%), germacrone (17.1%), curzerene (9.6%) and germacrene B (5.3%). 13C NMR allowed the identification and quantification of these compounds. In addition to β-pinene (49.9%) and α-pinene (19.5%) as major constituents, the bark oil of C. madagascariensis contains cyclolocopacamphene (2.0%), an epimer of cyclosativene. We report for the first time its 13C NMR chemical shifts. The essential oils of E. tristis are characterized by great chemical complexity. We have identified several oxygenated sesquiterpenes that are rarely encountered and four unknown compounds: (i) 7-epi-khusian-2-ol and 4,8-di-epi-acorone (respectively two new epimers of khusian-2-ol and acorone); (ii) 2-epi-ziza-5-en-2-ol; and (iii) antsorenone, which is so far the one and only molecule with the new terpenic skeleton that we have named antsorane. We suggest a biosynthesis pathway for this molecule, which seems to be directly linked to that of the zizaane skeleton.
... These differences in the composition of the oil may be caused by climatic and seasonal factors, sampling times, and plant material storing conditions (Chen et al., 2015). On the other hand, the diploid sample from Canada had no β-asarone and contained high percentage of preisocalamendiol (18.0%), acorenone (14.2%), shyobunone (13.3%), and acorone derivative (7.5%) (Garneau et al., 2008). ...
Sweet flag rhizome (Acorus calamus L., Araceae) exhibits various biological activities such as sedative, anticonvulsant, immunosuppressant, antidiabetic, and anti-inflammatory. This plant is protected in Serbia, but due to its high market requirements, the cultivation of this species could be the final solution. The content and composition of essential oils from 24 cultivated samples and five samples of sweet flag from natural populations, as well as the influence of nitrogen fertilization on the composition of essential oils were studied. Essential oils were analysed using GC and GC/MS. Statistical analysis included the ANOVA and PCA analysis. The total content of essential oil was not significantly different between the natural (0.80–1.10%) and cultivated samples (1.15–1.30%). In all essential oils main compounds were β-asarone (4.82–17.98%), cadinadienol (10.14–13.95%), camphor (3.55–13.20%), and acorenone (9.68–12.64%). The cultivation of sweet flag showed significant (p < 0.05) impact on the composition of essential oil and the content of β-asarone (4.82–17.07%, mean 11.95±4.76 vs. 17.51–17.98%, mean 17.71 ± 2.23). The PCA analysis also separated the essential oils from natural and cultivated conditions based on the content of β-asarone. On the other hand, the amount of the fertilizer did not show any effect (p > 0.05) on the composition of investigated samples. The process of cultivation increased the level of active, but potentially genotoxic and carcinogenic compound β-asarone. It would be significant to determine the content of β-asarone routinely in the sweet flag rhizome, especially in the cultivated plants.
... The two signals at 209.7 ppm and 215.5 ppm indicated the presence of two ketone groups. The set of 13 C NMR values of 78 were similar to those of acorone, 1,8di-epi-acorone (also named epi-acorone), and 1-epi-acorone (commonly named cryptoacorone) except for some carbons (Garneau et al., 2008;Nawamaki and Kuroyanagi, 1996). Thus, we focused our attention toward an epimer of acorone for which the 1 H and 13 C assignments (continued on next page) G.P. Garcia, et al. ...
Essential oils (EOs) obtained from aerial parts and roots of Elionurus tristis were investigated by GC, GC-MS, pc-GC and NMR. Both aerial parts and roots EOs contained common molecules such as α-pinene, camphene, trans-α-bergamotene and calarene. Moreover, we identified several unusual sesquiterpenes and four undescribed compounds, 7-epi-khusian-2-ol, 4,8-di-epi-acorone, 2-epi-ziza-5-en-2-ol and antsorenone. The last one exhibits an undescribed natural sesquiterpene skeleton. All undescribed compounds were isolated and fully characterized by MS, 1D and 2D-NMR. Furthermore, the formation pathway of the Antsorane skeleton is discussed.
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Preliminary phytochemical investigations were performed on the fruit essential oil and antioxidant-rich methanolic extracts of the fruits and roots of Ferula drudeana, the putative Anatolian ecotype of the Silphion plant, to corroborate its medicinal plant potential and identify its unique characteristics amongst other Ferula species. The essential oil from the fruits of the endemic species Ferula drudeana collected from Aksaray was analyzed by GC and GC/MS. The main components of the oil were determined as shyobunone (44.2%) and 6-epishyobunone (12.6%). The essential oil of the fruits and various solvent extracts of the fruits and roots of F. drudeana were evaluated for their antibacterial and anticandidal activity using microbroth dilution methods. The essential oil of the fruits, methanol, and methylene chloride extracts of the fruits and roots showed weak to moderate inhibitory activity against all tested microorganisms with MIC values of 78–2000 μg/mL. However, the petroleum ether extract of the roots showed remarkable inhibitory activity against Candida krusei and Candida utilis with MIC values of 19.5 and 9.75 μg/mL, respectively. Furthermore, all the samples were tested for their antioxidant activities using DPPH• TLC spot testing, online HPLC–ABTS screening, and DPPH/ABTS radical scavenging activity assessment assays. Methanolic extracts of the fruits and roots showed strong antioxidant activity in both systems.
The negative side effects of synthetic pesticides have drawn attention to the need for environmentally friendly agents to control arthropod pests. To identify promising candidates as botanical pesticides, we investigated the acaricidal and insecticidal activities of 44 plant-derived essential oils (EOs) against Tetranychus urticae Koch and Myzus persicae Sulzer. Among the tested EOs, Tasmannia lanceolata (Poir.) A.C.Sm. (Tasmanian pepper) essential oil (TPEO) exhibited strong acaricidal and insecticidal activity. Mortality rates of 100% and 71.4% against T. urticae and M. persicae, respectively, were observed with TPEO at a concentration of 2 mg/ml. Polygodial was determined to be the primary active component after bioassay-guided isolation of TPEO using silica gel open-column chromatography, gas chromatography, and gas chromatography–mass spectrometry. Polygodial demonstrated acaricidal activity against T. urticae with mortality rates of 100%, 100%, 61.9%, and 61.6% at concentrations of 1, 0.5, 0.25, and 0.125 mg/ml, respectively. Insecticidal activity against M. persicae was also evident, with mortality rates of 88.5%, 85.0%, 46.7%, and 43.3% at respective concentrations of 1, 0.5, 0.25, and 0.125 mg/ml. Insecticidal and acaricidal activities of TPEO were greater than those of Eungjinssag, a commercially available organic agricultural material for controlling mites and aphids in the Republic of Korea. These findings suggest that TPEO is a promising candidate for mites and aphids control.
The comminution and distillation time are two important factors, which affect the essential oil yield and/or composition of aromatic plants significantly. To find out the suitable comminution process and optimum distillation time for isolation of essential oil from Acorus calamus L., an experiment was conducted. The comminuted rhizomes (sliced and powdered) were hydrodistilled for different durations and their yields and compositions were compared. Distillation of uncomminuted rhizome for 8 hours gave 2.37% of essential oil. However, in comminuted rhizomes, yield varied from 2.27 to 6.60% under different distillation durations. The resulting essential oils were analyzed using GC-FID and GC-MS. Twenty-seven constituents, representing 94.7–97.2% of the total oil compositions were identified. Major constituents of the oils were (Z)-asarone (84.8–91.2%), and (E)-asarone (2.5–5.2%). The comminution and distillation time showed noteworthy effect on the essential oil yield; however, they showed no major change on the essential oil composition of A. calamus. It was concluded that powdering of dried rhizomes and effecting distillation for 4 hours are appropriate parameters for extracting maximum essential oil from A. calamus at laboratory scale.
Five new sesquiterpenes were isolated from Acorus calamus L., in addition to calameone (or calamendiol), the structure of which was revised, and their structures were established. Furthermore, chemical co-relation among these sesquiterpenes was carried out. In particular, the thermal isomerization of shyobunone (I), an elemene-type sesquiterpene, to preisocalamendiol (VI), a germacrone-type compound, should be noted.
Investigation of seven Bolivian Baccharis species afforded in addition to known compounds 17 ent-clerodane, 10 ent-labdane, two bisabolene, a germacrane, a cadinane, a guaiane and a p-coumaric acid derivative. In one case the ent-labdane was linked with a bisabolene derivative via a malonate group. The structures were elucidated mainly by high field NMR techniques.
The rhizome oil of Acorus calamus L. was analyzed by GC and GC/MS and 30 compounds were identified, the main ones were shyobunones (17.3%) and acorenone (14.4%).
In the search for germination inhibitors from plant sources, the methanol extract of Acori rhizoma (Acorus calamus) was shown to inhibit germination of lettuce seeds. From Acori rhizoma, eight new sesquiterpenes were isolated together with some known sesquiterpenes. These sesquiterpenoids have cadinane, acorane and eudesmane skeletons and their structures were elucidated from the spectral evidence. Some of the compounds showed potent anti-germination activity. Copyright (C) 1996 Elsevier Science Ltd
The essential oil composition of Acorus calamus (sweet flag) leaves collected in Lithuania at different growing phases was examined by GC and GC/MS. Rhizome oils of A. calamus, having been more thoroughly investigated, were used for comparison purposes. The content of the oil in dried sweet flag rhizomes was 1.20± 0.12% and in the leaves, depending on the vegetation phase, was from 0.56–1.01%. Ninety-one constituents were positively or tentatively identified in the oils—66 in the leaves and 55 in the rhizomes. Possible formation of calacorene hydrates is suggested for the first time on the basis of mass spectral data. δ-Asarone [(Z)-asarone] was the major constituent in the leaves (27.4–45.5%), whereas acorenone was dominant in the rhizomes (20.86%) followed by isocalamendiol (12.75%). A higher content of some aliphatic and oxygenated monoterpenes was found in oils of the leaves at their earliest growth phase (May), while the β-asarone content was at its lowest level.