全 文 ：天然产物研究与开发 Nat Prod Res Dev 2012，24:590-593
文章编号:1001-6880(2012)05-0590-04
Received October 24，2011 Accepted March 5，2012
Foundation Item:The National Natural Science Foundation of China
(81160391 and 20862005) ;The Natural Science Foundation of Hain-
an Province (211015)．
* Corresponding author Tel:86-898-65889422;E-mail:sxp628 @ 126．
com
海南暗罗叶挥发油化学成分及其抗菌活性
李小宝1，2，陈光英1，2，宋小平1，2* ，韩长日1，2，邵泰明1，2
1海南师范大学省部共建-热带药用植物化学教育部重点实验室;2 海南师范大学化学与化工学院，海口 571158
摘 要:本文采用水蒸气蒸馏萃取法提取海南暗罗叶挥发油化学成分，利用气相色谱-质谱(GC-MS)从中分离鉴
定出 40 种化学成分，占挥发油总量的 67． 03%。其中主要成分为 1-甲基-5-亚甲基-8-(1-甲基乙基)-1，6-二烯环
十烷(18． 305%) ，丁子香烯(6． 256%)和 γ-榄香烯(6． 211%)。所得的挥发油抑菌试验表明其对大肠杆菌和枯
草芽孢杆菌表现出显著的抑制活性。
关键词:海南暗罗;挥发油;气相色谱-质谱技术;抗菌活性
中图分类号:R284. 2 文献标识码:A
Composition and Antimicrobial Activities of Essential Oil of
Leaves of Polyalthia laui Merr．
LI Xiao-bao1，2，CHEN Guang-ying1，2，SONG Xiao-ping1，2* ，HAN Chang-ri1，2，SHAO Tai-ming1，2
1Key Laboratory of Tropical Medicinal Plant Chemistry of Ministry of Education;Hainan Normal University;
2College of chemistry and chemical engineering，Hainan Normal University;Hainan 571158，China
Abstract:The essential oil obtained by water-steam distillation extraction of the leaves of Polyalthia laui was analyzed
by gas chromatography-mass spectrometry (GC-MS)． Forty compounds were characterized accounting for 67． 03% in the
oil． The major components were 1-methyl-5-methylene-8-(1-methylethyl)-1，6-Cyclodecadiene (18． 305%) ，Caryo-
phyllene (6． 256%)and γ-Elemene (6． 211%)． The bacteriostatic test showed that the essential oil had remarkable in-
hibitory effects for Escherichia coli and Bacillus subtilis．
Key words:Polyalthia laui Merr;essential oil;GC-MS;antimicrobial activity
Introduction
Genus Polyalthia (Annonaceae)comprises of about 70
species［1］，among which only 7 are indigenous in Hain-
an province． including P． cerasoides (Roxb．)Benth，
P． consanguinea Merr，P． laui Merr，P． nemoralis A．
DC，P． plagioneura Diels，P． rumphii (Bl． ex Hensch．)
Merr and P． suberosa (Roxb．)Thw． Plants of the ge-
nus is known for its folkloric applications against a
number of ailments such as hepatitis，lymphadenitis，
pneumonia and so on［2］． Some plants of this genus are
also used in folk medicines as abortifacien，febrifuge，
antimalarial and anti-fungal agents［1-3］． In our previous
study against human cancer cell lines of P． laui，its eth-
anol extract showed that it possessed some inhibitory
activities towards four human tumor cells，including
BEL-7402，K562，SPCA-1 and SGC-7409［4］． However，
there was no report about chemical constituents and an-
timicrobial activity of the oil from P． laui． In this pa-
per，we describe the chemical composition and antimi-
crobial activity of the volatile oils from the leaves of P．
laui extracted by water-steam distillation to provide evi-
dences for further development and application of the
Chinese traditional herbal medicine．
Materials and Methods
Plant material
The leaves of P． laui were collected from Bawangling
mountain in Hainan Province，P． R． China in May
2011，and identified as P． laui by vice-professor Qion-
gxin Zhong from the College of Life Science in Hainan
Normal University． A voucher specimen has been pre-
DOI:10.16333/j.1001-6880.2012.05.027
served in the Key Laboratory of Tropical Medicinal
Plant Chemistry of Ministry of Education，Hainan Nor-
mal University．
Microbial strains and media
The essential oils from P． laui against 3 strains of bac-
teria (Escherichia coli，Bacillus subtilis，and Staphylo-
coccus aureus)． All microbial cultures were maintained
on nutrient agar slants and were regular subculture． The
purity of cultures was checked regularly by Gram-stai-
ning and standard microbiological methods．
Essential oil extraction
The fresh leaves (495 g)of P． laui were water distilled
for 6 h using simultaneous distillation extraction． The
extracts were dried with anhydrous sodium sulfate for
two days then concentrated to dryness in a rotary evap-
orator，Essential oil was obtained by the standard ex-
tracting method described in the Chinese Pharmacopoe-
ia and kept in vials at 4 ℃ until GC-MS analysis．
GC-MS analysis
The essential oils were analyzed by GC /MS on a
HP7890A GC coupled to a 5975C MS (Agilent Tech-
nologies，USA )equipped with a HP-5MS silica capil-
lary column (30 m × 0． 25 mm I D ，0． 25 μm film
thickness)． Carrier gas was helium with a flow rate of 1
mL /min． The injector (the spilt flow ration was 50∶ 1)
was set to 280 ℃ ． The GC temperature program was 30
℃，10 ℃ /min to 70 ℃，held for 2 min． The mass spec-
tra in the electron impact mode were recorded at 70 eV
with a scan range from 35-450 amu． The components in
the essential oils of fresh materials were identified by
comparison of their mass spectra with those of NIST08．
L library data．
Antimicrobial activity assay
The antimicrobial activities of the essential oils from P．
laui by the disc diffusion method［5］． In brief，a meas-
ured amount of the test samples was dissolved in defi-
nite volumes of solvent to give solutions of known con-
centrations (20，40，60 mg /mL)． Then，sterile filter pa-
per discs were impregnated with known amounts of the
tested the essential oils and dried． These discs were
placed on plates containing agar media seeded with the
test organisms． The plates were incubated at 37 oC for
24 h and the antimicrobial activities were measured by
the zones of inhibition expressed in mm． Test samples
showing zones of inhibition with a diameter of 5． 36 mm
or more were considered active．
Results and Discussion
Chemical constituents of the essential oils
Essential oils from the fresh leaves (495 g)of P． laui
with water distilled were analyzed by GC /MS． The i-
dentified constituents and the irrelative percentages
were shown in Table 2 and the chromatogram were
showed in Fig1．
Fig. 1 The total ion chromatogram of essential l oil from
P． laui
Table 1 Analysis of the Chemical Composition of Essential Oil from P． laui by GC-MS Method
No． tR(min) Compound Formula
Molecular
weight
Relative
content (%)
Similarity
(%)
1 7． 299 Z-3-Hexen-1-ol C6H12O 100． 089 0． 239 91
2 14． 546 3，7-dimethyl -1，6-octadien-3-ol C10H18O 154． 136 0． 039 94
3 21． 470 4-ethenyl-4-methyl-3-(1-methylethenyl)-1-(1-methylethyl)-Cyclohexene
C15H16O4 204． 188 5． 502 98
4 21． 762 α-Cubebene C15H24 204． 188 0． 084 99
5 22． 355 γ-langene C15H24 204． 188 0． 542 99
6 22． 477 Copaene C15H24 204． 188 0． 786 98
195Vol. 24 LI Xiao-bao，et al:Composition and Antimicrobial Activities of Essential Oil of Leaves of Polyalthia laui Merr．
7 22． 720
［1S-(1α，3aα，3bβ，6aβ，6bα) ］-
decahydro-3a-methyl-6-methylene-1-
(1-methylethyl)-Cyclobuta［1，2，3，4］dicyclopentene
C15H24 204． 188 0． 443 95
8 22． 901 1-ethenyl-1-methyl-2，4-bis(1-methylethenyl)-Cyclohexane C15H24 204． 188 1． 364 91
9 23． 371 ［1aR-(1aα，4α，4aβ，7bα) ］-1a，2，3，4，4a，5，6，7b-octahydro-1，1，4，7-tetramethyl-1H-Cycloprop［e］azulene
C15H24 204． 188 0． 937 99
10 23． 666 Caryophyllene C15H24 204． 188 6． 256 99
11 23． 863
［3aS-(3aα，3bβ，4β，7α，7aS* ) ］-octahydro-7-methyl-3-
methylene-4-(1-methylethyl)-1-
H-Cyclopenta［1，3］cyclopropa1，2］benzene
C15H24 204． 188 0． 323 96
12 24． 000 2，6-dimethyl-6-(4-methyl-3-pentenyl)-Bicyclo［3． 1． 1］hept-2-ene
C15H24 204． 188 0． 807 98
13 24． 510 Z，Z，Z-1，4，7，-Cycloundecatriene，1，5，9，9-tetramethyl C15H24 204． 188 2． 562 98
14 24． 677
［1aR-(1aα，4aβ，7α，7aβ，7bα) ］-
decahydro-1，1，7-trimethyl-4-methylene-
1-H-Cycloprop［e］azulene
C15H24 204． 188 0． 305 99
15 24． 812 2-isopropyl-5-methyl-9-methylene-Bicyclo［4． 4． 0］dec-1-ene C15H24 204． 188 0． 195 93
16 25． 270 ［s-(E，E) ］-1-methyl-5-methylene-8-
(1-methylethyl)-1，6-Cyclodecadiene
C15H24 204． 188 18． 305 97
17 25． 794 6-ethenyl-6-methyl-1-(1-methylethyl)-3-
(1-methylethylidene)-(S)-Cyclohexene
C15H24 204． 188 1． 082 93
18 25． 993 1，2，4a，5，6，8a-hexahydro-4，7-dimethyl-1-
(1-methylethyl)-Naphthalene
C15H24 204． 188 1． 001 97
19 26． 203 1，2，3，5，6，8a-hexahydro-4，7-dimethyl-1-
(1-methylethyl)-(1S-cis)-Naphthalene
C15H24 204． 188 2． 600 98
20 26． 363 ［1S-(1α，7α，8aα) ］-1，2，3，5，6，7，8，8a-octahydro-1，8a-dimethyl-7-(1-methylethenyl)-Naphthalene
C15H24 204． 188 0． 243 93
21 26． 553 ［1S-(1α，4aβ，8aα) ］-1，2，4a，5，6，8a-hexahydro-4，7-dimethyl-1-(1-methylethyl)-Naphthalene
C15H24 204． 188 0． 374 94
22 26． 839 Cyclohexanemethanol C7H14O 222． 198 0． 769 91
23 27． 047 Cyclohexane C15H24 204． 188 3． 842 91
24 27． 571 ［1aR-(1aα，4aα，7β，7aβ，7bα) ］-decahydro-1，1，7-trimethyl-4-methylene-1H-Cycloprop［e］azulen-7-ol
C15H24O 220． 183 4． 28296
25 27． 689 Caryophyllene oxide C15H24O 220． 183 3． 319 93
26 28． 096 Naphthalene C15H24 204． 188 0． 968 92
27 28． 482 Ledol C15H26O 222． 198 1． 490 90
28 29． 001 tau-Muurolol C15H26O 222． 198 1． 194 96
29 29． 097 1，2，3，4，4a，7，8，8a-octahydro-1，6-dimethyl-(1-methylethyl)-，［1S-(1α，4α，4aβ，8aβ) ］-1-Naphthalenol
C15H26O 222． 198 1． 300 96
30 29． 294 α-Cadinol C15H26O 222． 198 3． 246 99
31 29． 562 ［1S-(1α，7α，8aα) ］-1，2，3，5，6，7，8，8a-octahydro-1，8a-dimethyl-7-(1-methylethenyl)-Naphthalene
C15H24 204． 188 0． 830 92
32 30． 215 ［2R-(2α，4aα．，8aβ) ］-1，2，3，4，4a，5，6，8a-octahydro-4a，8-dimethyl-2-(1-methylethenyl)-Naphthalene
C15H24 204． 188 0． 145 93
33 30． 453 Isocaryophillene C15H24 204． 188 0． 230 90
34 30． 694 7R，8R-8-Hydroxy-4-isopropylidene-7-methylbicyclo［5． 3． 1］undec-1-ene
C15H24O 220． 183 0． 560 90
35 31． 968 1，4-dimethyl-7-(1-methylethyl)-Azulene C15H18 198． 141 0． 266 99
36 32． 210 1，2，3，4，5，6-Hexahydro-1，1，5，5-tetramethyl-2，4a-methanonaphthalen-7(4a-H)-one
C15H22O 218． 167 0． 207 90
37 34． 196 cis，cis，cis-7，10，13-Hexadecatrienal C16H26O 234． 198 0． 079 96
38 35． 235 n-Hexadecanoic acid C16H32O2 256． 24 0． 131 99
295 Nat Prod Res Dev Vol. 24
39 37． 196 Phytol C20H40O 296． 308 0． 044 91
40 37． 680 mono(2-ethylhexyl)ester1-2-Benzenedicarboxylic acid C16H22O4 278． 152 0． 143 91
The oil constituents are classified into sesquiterpene
hydrocarbons (44． 2%) ，oxygenated sesquite -rpenes
(8． 2%)and others． Among 40 identified constituents，
representing 67． 03% of the oil，1-methyl-5-methylene-
8-(1-methylethyl)-1，6-cyclodecadiene (18． 305%) ，
caryophyllene (6． 256%) ，γ-elemene (6． 211%)were
the most abundant． The oil was characterized by the
high percentage of sesquiterpene hydrocarbons and oxy-
genated sesquiterpenes． The antimicrobial activity of
the essential oil suggested this plant had the potential
use as an important antimicrobial source．
Antimicrobial activity
All experiments were carried out in triplicate． The anti-
bacterial activity was assessed by evaluating the pres-
ence of inhibition zone values． Results of inhibition
zone (Table 1)showed that the essential oil of P． laui
had different antibacterial activities against the three
bacteria tested． The bacteriostatic test showed that the
essential oil had remarkable inhibitory efffects on E． co-
li and B． subtilis．
Table 1 The bacterial inhibition zone of different concentrations (x ± s，mm，n = 3)
Bacteria
Diameter (mm)
20 mg /mL 40 mg /mL 60 mg /mL Benzylpenicillinsodium (1 mg /mL) Ethanol
E． coli 15． 06 ± 1． 12 17． 44 ± 1． 54 22． 24 ± 2． 07 14． 32 ± 0． 70 －
B． subtilis 17． 02 ± 1． 47 17． 84 ± 1． 59 26． 18 ± 2． 07 12． 91 ± 0． 65 －
S． aureus 13． 56 ± 1． 16 14． 72 ± 1． 23 18． 13 ± 1． 98 40． 29 ± 0． 50 －
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395Vol. 24 LI Xiao-bao，et al:Composition and Antimicrobial Activities of Essential Oil of Leaves of Polyalthia laui Merr．