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Abstract

The essential oils from aerial parts of young (sample A) and mature leaves (sample B), fruit (sample C), and flowers (sample D) of Zanthoxylum hyemale were obtained by hydrodistillation and analyzed by GC, GC/MS, and chiral phase gas chromatography (CPGC). Thirty-four compounds were identified from the essential oils, representing approximately 90.71, 91.19, 87.33, and 89.08 % of the oils, respectively. The major constituent of the young leaf essential oil was the sesquiterpene trans-nerolidol (51 %), while the main constituent of mature leaf (31 %) and flower oils (22 %) was an as yet unknown humulane-type sesquiterpenoid, which was characterized by spectral techniques (EI-MS and 1D-, 2D-NMR) as 3,7,10,10-tetramethylcycloundeca-3,7-dien-1-ol ( 1) and given the trivial name "hyemalol". In the fruit essential oil, the most abundant components were the monoterpenes beta-pinene (25 %) and alpha-pinene (10 %). The antimicrobial activity of the essential oils and some isolated compounds is also reported.
Introduction
Zanthoxylum hyemale A.St. Hill (Rutaceae), locally called ªcoen-
trilhoº, is a plant native to South America (Southern Brazil, Uru-
guay, Paraguay, and Argentine). In Brazil, Z. hyemale is generally
used in popular medicine as a potent antipyretic, antispasmodic,
astringent, and a tonic agent [1]. In continuation of our chemical
studies on plants of the Rutaceae family [2], [3], [4], [5], this work
reports the composition of essential oils obtained from the
leaves, flowers, and fruit of Zanthoxylum hyemale gathered in
Rio Grande do Sul state (Southern Brazil).
Materials and Methods
General
Optical rotations were measured on a Perkin Elmer-341 digital po-
larimeter. The HR-ESI-MS was recorded on a Finnigan MAT TQS
7000.
1
H- and
13
C -NMR spectra were recorded on a Bruker DPX
400 (400.1/100.6MHz) NMR spectrometer, in CDCl
3
with TMS as
internal standard. TLC were performed on precoated silica gel 60
F
254
plates (Merck) and detection was achieved using UV light
(254 nm) and by spraying with 10% H
2
SO
4
, followed by heating.
Plant material
Zanthoxylum hyemale was collected in the town of Santana do
Livramento, Rio Grande do Sul state, Brazil and identified by
Essential Oil from Zanthoxylum hyemale
EuclØsio Simionatto
1
Carla Porto
1
Ionara I. Dalcol
1
Ubiratan F da Silva
2
Ademir F. Morel
1
Affiliation
1
Departamento de Química, Ncleo de Pesquisa de Produtos Naturais, Universidade Federal de Santa Maria
(UFSM), Santa Maria-RS, Brazil
2
Instituto de Ciencias Exatas e Geocincias, Química, Universidade de Passo Fundo, UPF, Passo Fundo-RS,
Brazil
Correspondence
Prof. Dr. Ademir Farias Morel ´ Departamento de Química ´ Ncleo de Pesquisa de Produtos Naturais ´
Universidade Federal de Santa Maria ´ Campus Camobi ´ CEP 97105±900 ´ Santa Maria RS ´ Brazil ´
E-mail: afmorel@base.ufsm.br
Received October 11, 2004 ´ Accepted January 31, 2005
Bibliography
Planta Med 2005; 71: 759±763 ´ Georg Thieme Verlag KG Stuttgart ´ New York
DOI 10.1055/s-2005-864184 ´ Published online July 29, 2005
ISSN 0032-0943
Abstract
The essential oils from aerial parts of young (sample A) and ma-
ture leaves (sample B), fruit (sample C), and flowers (sample D)
of Zanthoxylum hyemale were obtained by hydrodistillation and
analyzed by GC, GC/MS, and chiral phase gas chromatography
(CPGC). Thirty-four compounds were identified from the essen-
tial oils, representing approximately 90.71, 91.19, 87.33, and
89.08% of the oils, respectively. The major constituent of the
young leaf essential oil was the sesquiterpene trans-nerolidol
(51%), while the main constituent of mature leaf (31%) and flow-
er oils (22%) was an as yet unknown humulane-type sesquiter-
penoid, which was characterized by spectral techniques (EI-MS
and 1D-, 2D-NMR) as 3,7,10,10-tetramethylcycloundeca-3,7-
dien-1-ol (1) and given the trivial name ªhyemalolº. In the fruit
essential oil, the most abundant components were the monoter-
penes
b
-pinene (25%) and
a
-pinene (10%). The antimicrobial ac-
tivity of the essential oils and some isolated compounds is also
reported.
Key words
Zanthoxylum hyemale ´ Rutaceae ´ essential oils ´ hyemalol ´ anti-
microbial activity
Original Paper
759
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Prof. Renato Zµcchia. Leaves and flowers were collected between
July and September, 2002 and fruits were collected in December,
2002 from the wild population of one location at Santana do Liv-
ramento. Voucher specimens (SMDB 5687±5690) have been de-
posited at the Herbarium of the Federal University of Santa Mar-
ia.
Chemical analysis
Fresh young (sample A) and adult leaves (sample B), fruit (sam-
ple C), and flowers (sample D) were subjected to hydrodistilla-
tion for 4 h using a modified Clevenger apparatus followed by ex-
traction of the essential oils from the distillate with diethyl ether.
After solvent removal, crude oil yields were 0.30%, 045%, 0.25%,
and 0.70% (v/w) for samples A, B, C, and D, respectively.
± Sample A: d
20:
: 0.89 g mL
±1
; n
20
: 1.4701; [
a
]
25
: ±9.4 (c 0.016,
CHCl
3
);
± Sample B: d
20
: 0.86 g mL
±1
; n
20
: 1.5502; [
a
]
25
: ±19.4 (c 0.013,
CHCl
3
);
± Sample C: d
20
: 0.83 g mL
±1
; n
20
: 1.4301; [
a
]
25
: ±14.9 (c 0.012,
CHCl
3
);
± Sample D: d
20
: 0.85 g mL
±1
; n
20
: 1.5301; [
a
]
25
: ±11.6 (c 0.017,
CHCl
3
).
The oils were analyzed by GC and GC-MS. GC analyses were per-
formed using a Varian CP-3800 gas chromatograph with a data
handling system and FID and SE-54 fused-silica column (25
m0.25 mm i.d., film thickness 0.25
m
m). Operating conditions
were as follows: injector and detector temperatures, 220 and
2808C, respectively; carrier gas, H
2
; oven temperature program
from 508C to 2508Cat48C/min. GC-MS analyses were performed
using a Varian model 3800 Saturn system operating in the EI
mode at 70 eV equipped with a CP-SIL cross-linked capillary col-
umn (30 m0.25 mm). The identity of the oil components was
established from their GC retention times, by comparison of their
MS with those reported by Adams [6], by computer matching
with the Wiley 5 mass spectra library [7], and by co-injection
with standards available in our laboratories whenever possible.
Chiral monoterpene constituents (
a
-pinene,
b
-pinene, and limo-
nene) and the sesquiterpene nerolidol of Z. rhoifolium oils were
identified by peak enrichment on enantioselective capillary GC
with two fused capillary columns, 25 m 0.25 mm, coated with
heptakis-(6-O-methyl-2,3-di-O-pentyl)-
b
-cyclodextrin and octa-
kis-(3-O-methyl-2,6-di-O-butyryl)-
g
-cyclodextrin, each diluted
with polysiloxane OV-1701. A Varian-3800 apparatus equipped
with a flame ionization detector (FID) was used with hydrogen
as the carrier gas. All runs were performed with the temperature
program 358C for 15 min, and from 358C to1808Cat38C/min.
Part of the resulting oils of mature (50 mg) and young leaves (50
mg) and fruit (100 mg) were further subjected to preparative TLC
(SiO
2
; hexane-EtOAc, 90:10) and afforded hyemalol (1, 7 mg),
nerolidol (2, 20 mg), and cadinol (3, 10 mg), respectively. The au-
thenticity of nerolidol and cadinol was determined by direct
comparison with authentic samples.
Hyemalol (1): Oil; [
a
]
D
106(c 0.0033, CHCl
3
); EI-MS: m/z = 222
[M]
+
, 205, 180, 149, 135, 109, 95; HR-ESI-MS: m/z =[M +
H]
+
223.2060 (calcd. for: C
15
H
27
O + H: 223.2062);
1
H-NMR
(CDCl
3
, 400.1 MHz):
d
= 4.86 (1H, m, H-4), 4.85 (1H, m, H-8),
3.40 (1H, m, H-1), 2.11±2.25 (2H, m, H-5), 2.06±2.14 (2H, m, H-
6), 2.05±2.13 (2H, m, H-2), 1.91±1.99 (2H, m, H-9), 1.06±1.96
(2H, m, H-11), 1.63 (3H, s, H-12), 1.46 (3H, s, H-13), 1.07 (3H, s,
H-14), 0.90 (3H, s, H-15);
13
C-NMR (CDCl
3
, 100.6 MHz):
d
= 133.6 (C-7), 132.1 (C-3), 126.8 (C-4), 124.6 (C-8), 70.1 (C-1),
49.6 (C-2), 47.0 (C-11), 39.3 (C-6), 39.1 (C-9), 33.5 (C10), 31.2 (C-
15), 27.3 (C-14), 25.5 (C-5).
Determination of absolute configuration of 1 by Horeau's
method
Approximately 6
m
mol of racemic
a
-phenylbutyric anhydride
and 30
m
L of dry pyridine were added to approx. 5
m
mol of 1 and
kept at room temperature for 1 h. After standing with 10
m
Lof
water for 30 min, usual work-up gave a mixture of the enantio-
mers of 2-phenylbutyric acid. A solution of diazomethane in di-
ethyl ether (50
m
L) was added until a permanent yellow color
was attained. The solution was reduced to approximately half of
its volume under an N
2
stream and used for gas chromatography.
Applying Horeau's modified rule for chiral GC [8], the excess en-
antiomer (±)-(R) observed indicated the (R) configuration for C-1
in compound 1.
Antimicrobial bioassay
The antibacterial activity of samples A ± D was assayed using the
minimal inhibitory concentration (MIC) determination. A collec-
tion of nine microorganisms was used, including three Gram-posi-
tive bacteria: Staphylococcus aureus (ATCC 6538p), Staphylococcus
epidermidis (ATCC 12228), Bacillus subitilis (ATCC 6633); and four
Gram-negative bacteria: Klebsiella pneumoniae (ATCC 10 031),
Eschericchia. coli (ATCC 11103), Pseudomonas aeruginosa (ATCC
27873), Salmonella setubal (ATCC 19196); and two yeasts:
Candida albicans (ATCC 10231), and Saccharomyces cerevisae
(ATCC 2601). Standard microorganism strains were obtained
from American Type Culture Collection (ATCC), and standard anti-
biotics chloramphenicol and nistatine were used in order to con-
trol microbial test sensitivity [9]. The minimal inhibitory concen-
tration (MIC) was determined in 96-well culture plates bya micro-
dilution method using a microorganism suspension with a density
of 10
5
CFU/mL in casein soy broth (CSB) incubated for 24 h at 378C
for bacteria, and Sabouraud broth (SB) incubated for 72 hours at
258C for yeasts.The cultures that did not grow were used to inocu-
late solid medium plates (Muller Hinton agar and Sabouraud agar)
in order to determine the minimal lethal concentration (MLC).
Proper blanks were assayed simultaneously, and samples were
tested in triplicate. Technical data have been described previously
[10], [11], [12].
Bioautographic bioassay
The antimicrobial activity of isolated compounds 1, 2, and 3 was
assayed using the bioautography technique [13], [14], [15] and
the collection of microorganisms described above. Compound
quantities ranging from 50
m
g to 1.56
m
g were applied to pre-
coated TLC plates. Muller Hinton agar and Sabouraud agar media
inoculated with mocroorganisms suspended in saline solution
(10
5
CFU/mL) were distributed over TLC plates. Bacteria and
yeast plates were incubated for 24 h at 378C, and 72 h at 258C,
respectively. Standard antibiotics, chloramphenicol and nista-
tine, were used in order to control microbial test sensitivity [9].
The results were stained with an aqueous solution of 2,3,5-tri-
phenyltetrazolium chloride (TTC, 1 mg/mL). The appearance of
Simionatto E et al. Essential Oil from ¼ Planta Med 2005; 71: 759 ± 763
Original Paper
760
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inhibition zones was used to determine the lowest sample
amount capable of inhibiting microbial growth. Samples were
tested in triplicate.
Results and Discussion
As shown in Table 1, the qualitative and quantitative oil composi-
tions displayed significant differences. Young (sample A) and
mature leaf (sample B), and flower (sample D) oils were rich in
sesquiterpenes, mainly nerolidol in sample A, and an unknown
sesquiterpene, named hyemalol (1), in samples B and D. In the
fruit oil (sample C), monoterpenes
a
-pinene and
b
-pinene predo-
minated. Interestingly, an inversion of the quantities of
a
-pinene
isomers in flower, leaf, and fruit oils was observed. In flower and
leaf oils, the major isomer is (±)-
a
-pinene (85% and 71%, respec-
tively), while in fruit oil, the major isomer is (+)-
a
-pinene (71%).
Hyemalol (1) was obtained as a colorless oil. The molecular for-
mula of C
15
H
16
O for 1 was established by means of HR-ESI-MS,
which gave a pseudomolecular ion peak at m/z = 223.2060 [M +
H]
+
, in combination with
13
C-NMR. The
1
H-NMR spectrum of 1
showed five pairs of methylene hydrogens at
d
= 2.25±2.11
(H
2
±5, m), 2.13±2.05 (H
2
±2, m), 2.14±2.06 (H
2
±6, m), 1.99±
1.91 (H
2
±9, m), and 1.96±1.06 (H
2
±11, m), three methynic hy-
drogens, two of which represented olefinic hydrogens at
d
= 4.86 (H-4, m) and 4.85 (H-8, m), and one oxymethine hydro-
gen at
d
= 3.41 (H-1, m), as well as four methyl groups at
d
= 0.90 (H-14, s), 1.07, (H-15, s), 1.46 (H-13, s), and 1.63 (H-12,
s). The
13
C-NMR spectrum of 1 showed fifteen signals. DEPT
Table 1 Percentual composition of Zanthoxylum hyemale essential oils
Compounds
a
KI
b
KI
c
Samples Identification
ABCD
01 (+)-
a
-Pinene 939 1 016 0.84 ± 7.4 2.85 GC,MS, Co
02 (±)-
a
-Pinene 939 1016 2.06 17.3 2.6 16.15 GC,MS, Co
03 (+)-
b
-Pinene 987 1 124 0.1 ± 0.16 0.3 GC,MS, Co
04 (±)-
b
-Pinene 987 1 124 5.5 9.12 24.9 10.2 GC,MS, Co
05 (+)-Limonene 1 025 1 210 ± 1.50 1.0 3.45 GC,MS, Co
06 (±)-Limonene 1025 1210 ± 1.18 0.89 0.9 GC,MS, Co
07 Menth-2-en-1-ol 1 140 1 595 ± ± 0.46 ± GC,MS
08 trans-
b
-Terpineol 1 163 1 586 ± ± 1.15 ± GC, MS
09 neo-Isopulegol 1 168 ± ± ± 0.45 ± GC, MS
10
a
-Terpineol 1 185 1 609 ± ± 0.67 ± GC,MS, Co
11 3-Decanone 1 190 1 580 ± ± 0.35 ± GC,MS
12 Carvacrol ethyl ether 1 204 1 572 ± ± 0,36 ± GC,MS, Co
13 2-Undecanone 1 289 1 623 ± 5.60 2.14 2.90 GC,MS, Co
14
d
-Elemene 1 334 1 483 0.45 0.1 0.24 0.20 GC,MS
15
b
-Elemene 1 414 1 595 2.69 0.74 1.84 1.0 GC,MS
16 E-
b
-Caryophyllene 1 449 1 586 1.05 2.0 1.58 2.36 GC,MS
17
a
-Acoradiene 1463 ± 1.04 ± 0.39 ± GC,MS
18
d
-Himachalene 1 476 1 657 0.13 0.1 2.44 ± GC,MS
19 Humulene 1 480 1 674 9.68 3.63 2.27 3.6 GC,MS
20 Germacrene-D 1 493 1 706 4.74 5.24 3.08 5.8 GC,MS, Co
21
b
-Bisabolene 1 509 1 724 3.55 0.27 0.58 ± GC,MS
22
d
-Cadinene 1 513 1 767 0.64 0.41 0.27 ± GC,MS
23 Bicyclogermagrene 1 520 1 718 1.55 0.22 1.08 6.60 GC,MS
24 Elemol 1 547 ± 0.14 0.1 0.37 ± GC,MS
25 Cadinene 1 551 1 744 0.17 0.34 0.33 2.61 GC,MS
26
b
-Calacoralene 1 563 1 912 ± ± 0.94 ± GC,MS
27 (±)-E-Nerolidol 1 566 2 087 51.5 ± 1.44 0.28 GC,MS, Co
28 Caryophyllene alcohol 1 571 1 954 ± 1.28 3.39 ± GC,MS
29 Caryophyllene oxide 1 580 1 986 ± 0.47 0.65 ± GC,MS, Co
30 epi-
a
-Muurolol 1 640 2 187 0.93 2.90 4.5 0.7 GC,MS
31 epi-
a
-Cadinol 1 642 2 185 0.74 0.85 1.23 2.2 GC,MS
32 Cadinol 1 653 2 145 2.31 5.72 9.58 4.5 GC,MS, NMR
33 Hyemalol 1 664 2 205 0.66 31.8 7.0 22.31 GC,MS, NMR
34 epi-
a
-Bisabolol 1 687 2 235 0.24 0.32 1.47 0.17 GC,MS
Total 90.71 91.19 87.33 89.08
a
Compounds listed in order of elution from a SE-54 column.
b
RI:Kovats Indices determined on an apolar SE-54 column.
c
RI:Kovats Indices determined on a polar PEG-20M column.
Co: peak identifications are based on standard comparison with relative retention time.
Simionatto E et al. Essential Oil from ¼ Planta Med 2005; 71: 759 ± 763
Original Paper
761
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135
o
, HMQC, and HMBC experiments showed the presence of five
methylenes at
d
= 25.1 (C-5), 39.1 (C-9), 39.3 (C-6), 47.0 (C-11),
and 49.6 (C-2), three methines at
d
= 70.1 (C-1), 124.6 (C-7),
and 126.8 (C-4), four methyls at
d
= 15.8 (C-13), 18.3 (C-12),
27.7 (C-14), and 31.2 (C-15), and three non-hydrogenated car-
bons at
d
= 47.0, 132.1, and 133.6. After all hydrogen resonances
had been assigned to those of their directly bonded carbon atoms
via HMQC measurements and after analysis of
1
H-
1
H COSY and
HMBC experiments, it was possible to deduce four molecular
fragments in 1 (a, b, c, and d). Thus, from the COSY spectrum of
1, three spin systems could be deduced. The first spin system
shows coupling between H-1 and H
2
±2, and between H-1 and
H
2
±11 (fragment a). Furthermore, couplings were observed be-
tween H-4 and H
2
±5, which, in turn, coupled with H
2
±6, leading
to the second spin system (fragment b). The third spin system
shows connectivity between H-8 and H-9 (fragment c). Further-
more, HMBC cross-peaks observed between the two geminal
methyl hydrogens and the two methylene carbons, C-9 and C-
11, define the position of C-10, allowing fragment d to be joined
to fragments a and c. The partial fragments a, b, c, and d were
then combined with the aid of the HMBC spectrum to give the
structure of 1, as shown in Fig.1. According to the above data,
the structure of 1 was elucidated as 3,7,10,10-tetramethylcy-
cloundeca-3,7-dien-1-ol, a new natural sesquiterpene derived
from a humulane-skeleton. In addition, the absolute configu-
ration of the secondary alcohol function (C-1) was determined
as (R), based on Horeau's method.
The antimicrobial activity of the oils was evaluated by determin-
ing the minimal inhibitory concentration (MIC). The results are
given in Table 2. The oils were active against all microorganisms
tested, while the best MIC value observed was 0.67 mg/mL for
sample C (flower essential oil) against K. pneumoniae. Because
of the low quantity and the low solubility of the isolated com-
pounds hyemalol (1), nerolidol (2), and cadinol (3), they were
evaluated against the same strains by the bioautography method
in a TLC bioassay. (±)-trans-Nerolidol exhibited antibacterial ac-
tivity against E. coli (1.56
m
g), while cadinol was active against K.
pneumoniae (3.12
m
g) and S. cerevisiae (6.25
m
g). Hyemalol (1)
only showed weak activity against the tested microrganisms.
Chloramphenicol for bacteria (0.4±0.7
m
g) and nistatine for
yeasts (2.0±2.4
m
g) were used as positive control.
Acknowledgements
This work was supported by FAPERGS (FundacË o de Amparo à
Pesquisa do estado do Rio Grande do Sul), CAPES (FundacË o Co-
ordenacË o de AperfeicË oamento de pessoal de Nível Superior),
and CNPq (Conselho Nacional de Desenvolvimento Cientíifco e
Tecnológico).
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a
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Fig. 1 Fragments derived from the
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H-
1
H-COSY spectrum of 1 (left)
and diagnostic COSY
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H-
1
H(
¨
) and HMBC (-
®
1
Hto
13
C) correla-
tions (right).
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... Ensaios antimicrobianos realizados com óleos essenciais extraídos por hidrodestilação de diferentes partes da planta Z. hyemale apresentaram atividade contra bactérias Grampositivas: S. aureus (ATCC 6538p), S. epidermidis (ATCC 12228), B. subitilis (ATCC 6633); e Gram-negativas: K. pneumoniae (ATCC 10031), E. coli (ATCC 11103), P. aeruginosa (ATCC 27873), S. setubal (ATCC 19196), com o melhor valor da CIM de 0,67 mg/mL para o óleo essencial de flores contra K. pneumoniae. Este mesmo estudo relatou ainda atividade antifúngica contra C. albicans (ATCC 10231) e S. cerevisiae (ATCC 2601) com melhor valor da CIM de 2,5 mg/mL contra S. cerevisiae (Simionatto et al. 2005). ...
... A análise do óleo essencial revelou a presença de limoneno (12), β-cariofileno (16), α-pineno (35), β-mirceno (36), β-elemeno (42), germacreno -D (53), biciclogermacreno (56), δ-cadineno (57), germacreno-A (66), menth-2-en-1-ol (71), epi-α-bisabolol (75), β-bisabolene (78), α-terpineol (79), β-pineno (111), δ-elemeno (117), α-humuleno (119), óxido de cariofileno (129) (277), hiemalol (278) e cadineno (279) (figura 1) (Moura et al. 2002, Simionatto et al. 2005, Chan et al. 2016. ...
Article
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This review covers the documentation of the isolated and biologically active chemical constituents of 20 species of the genus Zanthoxylum Linnaeus distributed all over the world. The species extracts and oils chemical constituents were isolated and characterized by different analytical methods. This review was carried out through the survey of the Master’s Dissertations, Doctoral Theses and mainly published scientific articles about the biological activity and phytochemical characterization of Zanthoxylum species, available at in the CAPES, PubMed, ScienceDirect, Scopus, Scielo and Google Scholar databases from 1985 to 2019. The species presented a diverse chemical composition in which alkaloids, terpenes and coumarins predominate. In addition, those chemical compounds perform different types of biological activity for the plant, such as larvicidal, anti-inflammatory, analgesic, antinociceptive, antioxidant, antibiotic, hepatoprotective, antiplasmodial, cytotoxic, antiproliferative, anthelmintic, antiviral and antifungal.
... Simionatto et al. previously described the isolation of the corresponding non-acetylated structure, i.e., 10-hydroxydihydro-α-humulene, from Zanthoxylium hiemale [50]. The absolute configuration of the secondary alcohol at C-10 was determined as (R) based on Horeau's method. ...
Article
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Australia’s endemic desert shrubs are commonly aromatic, with chemically diverse terpenes and phenylpropanoids in their headspace profiles. Species from the genus Eremophila (Scrophulariaceae ex. Myoporaceae) are the most common, with 215 recognised taxa and many more that have not yet been described, widely spread across the arid parts of the Australian continent. Over the years, our research team has collected multiple specimens as part of a survey to investigate the chemical diversity of the genus and create leads for further scientific enquiry. In the current study, the diversity of volatile compounds is studied using hydrodistilled essential oils and leaf solvent extracts from 30 taxa. Several rare terpenes and iridoids were detected in chemical profiles widely across the genus, and three previously undescribed sesquiterpenes were isolated and are assigned by 2D NMR—E-11(12)-dehydroisodendrolasin, Z-11-hydroxyisodendrolasin and 10-hydroxydihydro-α-humulene acetate. Multiple sampling from Eremophila longifolia, Eremophila arbuscular, Eremophila latrobei, Eremophila deserti, Eremophila sturtii, Eremophila oppositifolia and Eremophila alternifolia coneys that species in Eremophila are highly chemovariable. However, taxa are generally grouped according to the expression of (1) furanosesquiterpenes, (2) iridoids or oxides, (3) mixtures of 1 and 2, (4) phenylpropanoids, (5) non-furanoid terpenes, (6) mixtures of 4 and 5, and less commonly (7) mixtures of 1 and 5. Furthermore, GC–MS analysis of solvent-extracted leaves taken from cultivated specimens conveys that many heavier ‘volatiles’ with lower vapour pressure are not detected in hydrodistilled essential oils and have therefore been neglected in past chemical studies. Hence, our data reiterate that chemical studies of the genus Eremophila will continue to describe new metabolites and that taxon determination has limited predictive value for the chemical composition.
... 蛇麻烷型倍半萜(蛇麻烯),又称葎草烷型倍半萜,作为单环倍半萜的一种,是一个十一元碳环 和 4 个角甲基组成的天然萜类化合物群 [1] 。蛇麻烷型倍半萜生物合成需要的最基本结构单元是异戊烯 基焦磷酸 (3-甲基丁-3-烯-1-基焦磷酸酯, IPP) 和二甲基烯丙基焦磷酸 (3-甲基丁-2-烯-1-基焦磷酸酯, DMAPP)。生物体中 IPP 和 DMAPP 来源有 2 种,一种为甲羟戊酸 (MEV) 途径,另一种为脱氧木酮 糖磷酸酯途径,其中原核生物的 IPP 和 DMAPP 多来源于脱氧木酮糖磷酸酯途径 [2][3] 。蛇麻烷型倍半 萜由 2 分子 IPP 和 1 分子 DMAPP 在倍半萜合酶的催化下形成法尼基焦磷酸 (farnesyl diphosphate, FPP), 然后 FPP 又在倍半萜环化酶如 α-蛇麻烯合酶的作用下形成十一元环的蛇麻烷型倍半萜类成分 [4][5][6] ,其生物合成见图 1。 收稿日期:2018-06-08 基 金 项 目 : 国 家 自 然 科 学 基 金 项 目 ( 81774001, 81473426 ) ; 江 西 民 族 传 统 药 现 代 科 技 与 产 业 发 展 2011 协 同 创 新 基 金 项 目 (JXXT2018003) 作者简介:焦顺刚,硕士研究生, E-mail: Shungangjiao@126.com 网络首发时间: 网络首发地址: 中国中药杂志 图 1 蛇麻烷型倍半萜类骨架的生物合成途径示意图 Fig. 1 Biosynthesis pathway of humulane 蛇麻烷型倍半萜在植物和微生物中均有报道,广泛分布于姜科 Zingiberaceae、菊科 Compositae、 伞 形 科 Apiaceae 中 , 在 芸 香 科 Rutaceae 、 萝 藦 科 Asclepiadaceae 、 樟 科 Lauraceae 、 荨 麻 科 Urticaceae、桃金娘科 Myrtaceae、藤黄科 Guttiferae、唇形科 Labiatae、锦葵科 Malvaceae、木犀科 Oleceae、及岩蕨科个别植物种属也有报道,此外,真菌和苔藓中亦有分布。 蛇麻烷型倍半萜具有抗肿瘤、抗炎、抗菌、抗病毒等药理活性,广泛应用于医药、食品、保健品 等领域 [7][8][9][10][11][12] 。鉴于其独特的化学结构及显著的药理活性,蛇麻烷型倍半萜相关研究持续不断。本文对 其化学结构和药理作用进行分类整理,以期为此类成分的进一步研究提供参考,并为相关的构效关系 研究提供依据。 2 蛇麻烷型倍半萜的化学研究 蛇麻烷型倍半萜以蛇麻烯及其衍生物为代表,是自然界广泛分布的双环倍半萜前体之一。第一个 蛇 麻 烯 衍 生 物 α-humulene (1) 早 在 1895 年 就 从 植 物 中 分 离 得 到 , 其 完 全 氢 化 后 得 到 蛇 麻 烷 (humulane)。20 世纪 60 年代,α-humulene 的异构体 β-humulene (2) 和 γ-humulene (3) 相继从植物 Agonis abnormis 和植物 Abies magnifica 中分离得到,随后蛇麻烯的酯类、酮类、醇类、环氧化物和其 他衍生物也陆续被发现 [13] [17][18][19][20][21][22][23][24][25] 。双键无环氧化的 α-humulene 的结构差别主 要为取代基不同及 Δ 9,10 位双键是否被氢化。在此 25 个化合物中,于 C-8, C-9, C-10 位有取代的化合 Fig. 2 Structures of α-humulenes without epoxidation 2.1.2 双键环氧化 α-humulene 双键环氧化 α-humulene 指结构中 Δ 2,3 ,Δ 6,7 或 Δ 9,10 双键氧化成环氧基团的一类结构。有些化合物 同时存在 2 或 3 个环氧基,其中环氧基在 Δ 2,3 和 Δ 6,7 位出现较多,而于 Δ 9,10 位较少。目前已从植物和 真菌中报道了 2 个 Δ 2,3 位环氧基成分(29,30)、12 个 Δ 6,7 位环氧基成分(31~42)、2 个 Δ 2,3 ,Δ 6,7 上均有环氧基的成分(43, 45)、1 个 Δ 2,3 ,Δ 9,10 均有环氧基的成分(44)共 17 个该类化合物。该类 结构变化还体现在 C-9 位双键是否被还原等,其中有 12 个化合物(29~34, 37~42)的 C-9 位双键被 还原氢化。此外,十一元环上不同种类、不同数量以及不同位置的取代基也增加了结构新颖度。 值得一提的是,化合物 29-33 在结构上可以看成 C-9 位羟基与对羟基肉桂酸的酯化产物,其中 29 与 30,32 与 33 因 Δ 6,7 和 Δ 2,3 的构型不同而分别构成顺反异构体。2014 年,陈子明等从真菌 Antrodiella albocinnamomea 中分离得到 antrodol A(34),其 C-11 位甲基重排到 C-10 位上,并与同 时分离得到的 antrodol C(45)表现出对蛋白酪氨酸激酶中等抑制作用 [26] 。6,7-Epoxy-2,9-humuladiene (35) 还具有对蚜虫 L. decemlineata 拒食活性 [29] 。化合物 29-45 的结构见图 3,来源见表 1。 ...
Article
Humulane-type sesquiterpenoids, widely distributed in plants and microbes, include three types: α-humulene, β-humulene, and γ-humulene. Up to now, 98 humulane-type sesquiterpenoids have been reported, which possessed anti-tumor, anti-inflammatory, antibacterial, and antiviral activities. Herein, this paper describes their chemical constituents and pharmacological activities, hoping to bring benefits for further research and lay a foundation for investigating the structure-activity relationships.
... Some of these metabolites functions independently or in synergy to exert the pharmacological effect (Evans, 2002). For instance Zanthoxylum tingoassuiba and Z. hyemale essential oils have also been reported for their antimicrobial activity (Detoni et al., 2009;Simionatto et al., 2005) and Z. leprieurii and Z. xanthoxyloides for their antioxidant activity (Dongmo et al., 2008). The antimalarial, artemisinin, was isolated from Artemisia annua, and the anticancer alkaloid drugs, vinblastine and vincristine, were isolated from Catharanthus roseus (Evans, 2002). ...
... [11], on the other hand, tend to be dominated by (-)-limonene. Z. hyemale leaves and fruits have nearly racemic mixtures of limonene [12], while Z. schinifolium fruit has the (-)-enantiomer predominating (84%) [13]. ...
Article
The bark essential oils of Zanthoxylum clava-herculis and Ptelea trifoliata (Rutaceae) were obtained by hydrodistillation and analyzed by both gas chromatography as well as chiral gas chromatography coupled with mass spectrometry. Z. clava-herculis bark oil was dominated by sabinene [47.0%, 95% (–)-sabinene], limonene [18.7%, 99% (+)-limonene], and terpinen-4-ol [12.9%, 75% (–)-terpinen-4-ol]. The major components in P. trifoliata bark oil were limonene [15.2%, 99% (–)-limonene], sabinene [6.9%, 79% (–)-sabinene], and β-phellandrene [6.2%, 87% (–)-β-phellandrene].
... It is a family of pronounced morphological variety. Research has demonstrated that Rutaceae is rich and diverse source of specialized metabolites, such as essential oils ( Craveiro et al., 1979;Ferronatto et al., 2012;Simionatto et al., 2005), limonoids ( Dreyer et al., 1972;Nebo et al., 2015), flavonoids ( Itoigawa et al., 1994;Moraes et al., 2003), coumarins ( Gunatilaka et al., 1994;Kassim et al., 2013), lignans ( Kassim et al., 2013;Parhoodeh et al., 2011), and alkaloids ( de Moura et al., 2002de Moura et al., , 1997Tarus et al., 2005), have been shown possess medicinal properties such as antitumor ( Barret and Sauvaire, 1992;Kem enyBeke et al., 2006;Messmer et al., 1972;Panzer et al., 2001;Tillequin, 2007), antimicrobial ( Cabral et al., 2015;Navarro and Delgado, 1999;Nissanka et al., 2001;Odebiyi and Sofowora, 1979), anti-inflammatory ( Lenfeld et al., 1981), and antiplasmodial ( Basco et al., 1994;Wansi et al., 2009) activities. Considering the wide variety of metabolites, the family is considered one of the most versatile in terms of natural products. ...
Article
Extraction and characterization of natural products from the bark of the trunk of Helietta apiculata Benth (Rutaceae) afforded nine alkaloids, eight furoquinoline and one quinolone, limonine, three cinnamic acid derivatives, three neolignans, tetracosanoic acid, six coumarins, of which apiculin A and apiculin B (neolignans), and tanizin (coumarin) are previously undescribed compounds. The structures of all compounds were determined by spectroscopic methods, and the crystal structures of two of the newly undescribed compounds, apiculin A and apiculin B, were determined by X-ray analysis. Extracts and pure compounds isolated from Helietta apiculata showed promising antimicrobial activities.
Article
New copaene-type and nerolidol-type sesquiterpenoids, 7-hydroxymustakone (1) and 15-hydroxynerolidol (2), and a 15-norlabdane diterpenoid, kaempcandiol (3), together with four known compounds (4–7) were isolated from the chloroform extract of Kaempferia candida roots and rhizomes. The structures of the new compounds 1–3 were elucidated based on 1D and 2D NMR and HRESIMS spectroscopic analyses. The extract of the K. candida roots and rhizomes and all isolated compounds 1–7 possessed HIV-1 viral protein R (Vpr) inhibitory activities on the TREx-HeLa-Vpr cell line at a 5 μM concentration, without detectable cytotoxicity.
Article
Abstract Campomanesia aurea O. Berg is a native plant of the biome Pampa, which belongs to the family Myrtaceae. The present study aimed to evaluate the antimicrobial and antibiofilme activity of the essential oil of Campomanesia aurea (EOCA) against Listeria monocytogenes ATCC 19114, Staphylococcus aureus ATCC 25923, Salmonella enteritidis ATCC 13076 and Pseudomonas aeruginosa ATCC 27853. An analysis of the chemical composition of the EOCA was realized through gas chromatography coupled to mass spectrometry (GC-MS). The analysis for in vitro evaluation of the antimicrobial and antibiofilm activity was realized by determination of the minimal inhibitory concentration (MIC) and that of the antibiofilm through utilization of 96-well plates with crystal violet, respectively. The action of standard (E)–nerolidol (major compound of the EOCA) was also tested. GC-MS analysis revealed the presence of the sesquiterpene (E)-nerolidol (56.04%) as the main compound in the EOCA. Antimicrobial activity of the EOCA against L. monocytogenes (MIC 5.0 mg mL−1) and S. aureus (MIC 0.7 mg mL−1) was observed. Inhibition of biofilm formation against L. monocytogenes, S. aureus and S. enteritidis could be observed for EOCA and (E)-nerolidol. The results demonstrate that the EOCA was efficient against inhibition of biofilm formation for most of the tested pathogens.
Article
A terpene cyclase from Streptomyces pristinaespiralis was characterized as the synthase for (+)-(2S,3S,9R)-pristinol. The structure of this sesquiterpene alcohol, which has a new carbon skeleton, was established by NMR spectroscopy and single-wavelength anomalous-dispersion X-ray crystallography. Extensive isotopic labelling experiments were performed to distinguish between various possible cyclization mechanisms of the terpene cyclase and to decipher the EI-MS fragmentation mechanism for pristinol.
Article
Eine Terpencyclase aus Streptomyces pristinaespiralis wurde als (+)-(2S,3S,9R)-Pristinol-Synthase charakterisiert. Die Struktur dieses Sesquiterpenalkohols mit einem neuen Gerüst wurde durch NMR-Spektroskopie und monochromatische anomale Röntgendispersion aufgeklärt. Isotopenmarkierungsexperimente ermöglichten es, zwischen verschiedenen Cyclisierungsmechanismen der Terpencyclase zu unterscheiden und den EI-MS-Fragmentierungsmechanismus des Pristinols zu studieren.
Article
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A simple bioautographic agar overlay assay using Candida albicans as the indicator organism for the detection and activity-guided fractionation of antifungal compounds by thin layer chromatography has been developed. Inhibition of fungal growth was assessed by the detection of dehydrogenase activity with thiazolyl blue (methylthiazolyltetrazolium chloride; MTT). A series of clinically used antimycotic agents were tested in order to determine the sensitivity of the assay. The compatibility of the agar overlay technique with chemically modified silica gel (Diol and RP-18) plates and with various organic solvents was evaluated. The methodology is also applicable to the search for antibacterial compounds, as shown with Bacillus subtilis as a test organism.
Article
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A novel benzophenanthridine alkaloid, named zanthoxyline, was isolated from the bark of Zanthoxylum rhoifolium, together with the known compounds, dihydronitidine, 6-oxynitidine and skimmianine. The structure of the new compound was elucidated on the basis of spectroscopic investigations and elemental analysis.
Article
The reaction of optically active secondary alcohols with excess of racemic 2-phenylbutyric acid anhydride in pyridine proceeds at different rates to the diastereoisomeric esters (kinetic partial resolution). According to Horeau the (unknown) absolute configuration of the alcohol can be derived from the optical rotation of the remaining excess of 2-phenylbutyric acid in the reaction mixture. Measuring the optical rotation may be very difficult in cases of small absolute rotation values and may be inaccurate due to the necessity to completely remove all chiral impurities. The application of Horeau's method is greatly facilitated by gas chromatographic determination of the enantiomeric ratio of the remaining 2-phenylbutyric acid after methylation using a short capillary column with heptakis(2,6-di-O-methyl-3-O-pentyl)-β-cyclodextrin as a chiral stationary phase. Baseline resolution of the enantiomers is achieved after approximately 10 min of retention time. Due to the high selectivity of capillary gas chromatography the probability of impurities in the mixture to interfere with the measurement of the enantiomeric ratio is extremely low. © 1994 Wiley-Liss, Inc.
Book
The book is in two parts: first part Essential Oil includes compositae; labiatae; verbenaceae; oleaceae; umbelliferae; myrtaceae; euphorbiaceae; rutaceae; geraniaceae; rosaceae; lauraceae; myristicaceae; anonaceae; santalaceae; moraceae; piperaceae; zingiberaceae; araceae; gramineae; and cupressaceae written in English and Japanese. Part two includes essential oil; gas chromatography, and mass spectrometry written in Japanese. (DP)
Article
A simple bioautographic techniqud according to WELTZIEN', and modified by DEISHUI JZEN~ for detection of fungitoxic substances has been in use for many years in this laborattiry. Chromatograms on Whatman No. 3MM paper are develpped with propanol-water (85: 15) and after drying are sprayed with a conidial suspension of Glomerella cingzclata. After incubation, clearly visible inhibition zones indicate the preserice of. fungitoxic compounds. Chromatography thus permits not only the detection sf fungitoxic substances per se, but also makes the study of the conversion reactions and of decomposition of such compounds possible.
Article
A simple bioassay for the direct detection of antibacterial compounds on tlc plates has been developed. A series of natural products and different stationary phases were tested in order to establish the utility of the assay for the isolation of antibacterial compounds from higher plants.