全 文 ：广西民族学院学报(自然科学版)
第 10卷第 4 期 JOURNAL OF GUANGXI UNIVERSITY FOR NATIONALITIES Vol.10 No.4
2004年 11 月 (Natural Science Edition) Nov.2004
GC/MS Analysis of Volatile Components of Echinacea Species
YAO Xing-Dong1 , NIE Yuan-Mei1 , Nirmalendu Datta-Gupta2
(1.College of Chemistry and Ecological Engineering , Guangxi University for
Nationalities , Nanning 530006 , China ;2.Department of Physical Sciences ,
South Carolina State University , Orangeburg , SC 29117 , USA)
Abstract:The volatile constituents of three Echinacea species are investigated with GC/MS
analysis of their ethanol extracts.The results show different Echinacea species have different con-
stituents;the volatile parts may also have contribution to the medicinal effect of herb.
Key Words:Echinacea;volatile;GC/MS
中图分类号:O657.63 文献标识码:A 文章编号:1007-0311(2004)04-0078-06
0 INTRODUCTION
Echinacea is one the most widely applied herbs in US.It has been used for immunostimulation and wound healing by
native Americans for a long time.However , we have little knowledge about the constituents of Echinacea and the mechanism
for the immunostimulating effects up to now.Most of researchers paid attentions to Phenolic compounds and the lipophilic
alkamides
[ 1] .Several standardization methods were setup based on the determination of phenolic compounds[ 2～ 3] .The
volatile components of Echinacea , on the other hand , have not been well studied.Mazza[ 4] applied headspace analysis to
investigated three Echinacea species(E.Angustifolia , E.Pallida , and E.Purpurea)and reported 70 volatile components.
Main components include dimethyl sulfide , α-pinene , camphene , β -pinene , limonene , 2-hexenal(cis), 3-hexen-
1-ol acetate.This is the only paper on volatile compounds of Echinacea found inMedline database.Since there are a large
number of volatile compounds in Echinacea plants , it will not be surprise some of them can also contribute to immunostimu-
lation.
In this paper , we investigated ethanol extract of E.Angustifolia , E.Pallida , and E.Purpurea with GC/MS , and
found a lot of components not mentioned before.The main volatile components are 1 , 2-benzenedicarboxylic acid dibutyl
ester;hexanedioic acid dioctyl ester;9 , 12-octadecadienoic acid and phenol , 2 ,4-bis(1 ,1-dimethylethyl)for E.pur-
pura root;germacrene D;1 , 2-benzenedicarboxylic acid dibutyl ester and 9 ,12-octadecadienoic acid for E.Aerial top;
2-butanone , 4-(ethylphenyl);N , N-diethyl-4 ,5-dimethoxy-2-methylbenzamide;N-isobutyl-(2E , 4Z , 8Z ,
10E)-dodecatetraenamide and di(2-ethylhexyl)adipate for E.Pallida root;cyclododecene , tetracyclo[ 4 , 3 ,2 ,0(2 ,9),
78
收稿日期:2004-10-08.
基金项目:美国农业部研究基金资助项目(SCX-420-20-00)
作者简介:姚兴东(1965-),男 , 江苏人 ,广西民族学院化学与生态工程学院副教授 、博士 , 广西民族学院学报编辑部副主任.
主要研究方向:仪器分析 、绿色化学.
DOI :10.16177/j.cnki.gxmzzk.2004.04.020
0(3 ,5)undeca-7 ,10-dien-4-ol , 1-(3-butenyl)cyclobutabenzene , 1[ 7] ,4 , 8-O-methatriene and spiro[ 4 , 5]
decane for E.Angustifolia.1-propanol , 2 ,2-dimetyl;1 ,2-benzenedicarboxylic acid dibutyl ester , 9 , 12-octadeca-
dienoic acid and hexanedioic acid , dioctyl ester for E.Angustifolia leaf.Most of them can not be detected with headspace
analysis technique , possibly due to high boiling point of them.
1 MATERIALS AND METHODS
1.1 Plant Tissue
3 year old plant of E.Angustifolia , E.Pallida , E.Purpurea aerial tops , leaf and root were purchased from Caribon
Ginseng and Starwest Botanical.They are cut and milled before extraction.5 gram of milled plant is extracted with 150 ml
of ethanol for 12h in a Soxhlet apparatus.The extract is evaporated to 10ml in a rotary vacuum evaporator at a temperature
not higher than 50℃ for GC/MS analysis after filtration with 0.45μm nylon membrane.
1.2 GC/MS Analysis
A Hewlett-Packard(HP)model GC6890 gas chromatograph with a Hewlett-Packard 5973 mass-selective detector
were employed to investigate the volatile compounds in the extracts.1μl of extract was injected.Ultrahigh-purity helium
with HP mass spectrometer gas purifier was used as carrier gas.Chromatographic conditions were:Injector:250℃, Detec-
tor 280℃.Column:HP-5MS 30.0 m x 250μm x 0.25μm.Oven temperature was programmed as:Initial oven tempera-
ture 40℃, 2 min.raise temperature from 40℃ to 250℃ at the speed of 5℃/min , hold the final temperature for 11.0
min.For the MS detector , the electron multiplier voltage(EMV)is 1294V.
2 RESULTS AND DISCUSSION
Different E.species have been analyzed with GC/MS and the chromatograms are shown in Figure 1.Different species
show different chromatogram.
(1)3 year old Echinacea purpura root
Table 1 lists the chemicals and their relative percentage in ethanol extract:
Table 1:Components found in 3 year old E.Purpura root ethanol extract
Retention time(min) Area MS result Area %
10.339 a Ethanol , 2-(2-ethoxyethoxy) b
14.374 1041813 2 , 3-dihydro-3 , 5-dihydroxy-6-methyl-4H-py ran-4-one 7%
19.159 309270 4-vinyl-2-methoxy-phenol 2%
21.327 162484 unknown 1%
23.999 a cyclopentanol b
24.097 1669521 phenol , 2 , 4-bis(1 , 1-dimethylethyl) 11%
33.722 4841204 1 , 2-benzenedicarboxylic acid dibutyl ester 32%
36.947 1362269 9 , 12-octadecadienoic acid 9%
37.905 a N-Isobutyl-(2E , 42 , 82 , 10E)-dodecatetraenamide b
39.416 a unknown b
40.073 a unknown b
41.381 5583669 hexanedioic acid dioctyl ester 37%
46.068 a 9 , 12-octadecadienoic acid , 2 , 3-dihydropropyl ester b
53.462 a (23S)-ethylcholest-5-en-3-beta-ol b
a:couldn t measure the area due to low concentration.b:<1%
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2004年第 4期 ○姚兴东 ,聂园梅/不同紫锥花种属中挥发性组分的气相色谱/质谱分析
图 1 不同紫锥花种属乙醇萃取液的 GC/MS 分析图谱
Fig 1 GC/MS chromatogram of different Echinacea species ethanol extracts
The main components include phenol , 2 ,4-bis(1 ,1-dimethylethyl);1 ,2-benzenedicarboxylic acid dibutyl ester;
9 ,12-octadecadienoic acid and hexanedioic acid dioctyl ester.
(2)3 year old Echinacea purpurea aerial top:
Table 2 lists the chemicals and their relative percentage in ethanol extract:
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广西民族学院学报(自然科学版) 2004年 11月 第 10卷
Table 2:Components found in 3 year old E.Purpura Aerial top ethanol extract
Retention time(min) Area MS search result Area %
14.41 1086363 2 , 3-dihydro-3 , 5-dihydroxy-6-methyl-4H-py ran-4-one 3%
16.775 853147 unknown 3%
23.255 4300185 germacrene D 13%
24.354 a delta -cadinene b
25.669 a (+)spathulenol b
27.376 a cycloisolongifol-5-ol b
29.287 a 7-acetyl-2-hydroxy-2-methyl-5-isopropylbicyclo[ 4 , 3 , 0] nonane b
31.332 a 10-oxo-isodam-3-en-15-ol b
31.719 310871 2 , 5-(cyclohexadiene-1 , 4-dione , 2 , 6-bis(1 , 1-dimethylethyl) 1%
33.752 8718702 1 , 2-benzenedicarboxylic acid dibutyl ester 27%
36.497 1444569 phytol or 2-hexadecen-1-ol , 3 , 7 , 11 , 15-tetramethyl- 4%
36.995 11305351 9 , 12-octadecadienoic acid 35%
37.873 2303882 2 , 4-decadienal 7%
38.309 a unknown b
41.331 a di(2-ethylhexyl)adipate b
46.06 2209429 9 , 12-Octadecadienoic acid 2 , 3-dihydroxypropyl ester 7%
50.894 a deptadecane b
53.529 a (23S)-ethylcholest-5-en-3-beta-ol b
a:couldn t measure the area due to low concentration.b:<1%
The main components include 9 , 12-octadecadienoic acid , 1 ,2-benzenedicarboxylic acid dibutyl ester and germa-
crene D.
(3)3 year old Echinacea pallida root
Table 3 lists the chemicals and their relative percentage in ethanol extract:
Table 3:Components found in 3 year old E.Pallida root ethanol extract
Retention time(min) Area MS result Area %
14.424 1438310 2 , 3-dihydro-3 , 5-dihydroxy-6-4H-py ran-4-one 2%
27.802 a unknown b
32.58 1551059 2-exo-(1-oxacyclohex-2-yloxy)bicyclo[ 2 , 2 , 1] hepta-5-ene 2%
33.704 4784650 1 , 2-benzendicarboxylic acid dibutyl ester 6%
34.078 12502341 2-butanone , 4-[ ethylphenyl] 15%
35.62 1957252 unknown 2%
35.989 8245848 N-Isobutylundeca-2[ E] -ene-8 , 10-diynamide 10%
36.947 4081638 linoleic acid(9 , 12-octadecadienoic acid) 5%
37.162 11656262 N , N-diethyl-4 , 5-dimethoxy-2-methylbenzamide 14%
37.911 21826449 N-Isobutyl-[ 2E , 4Z , 8Z , 10E] -dodecatetraenamide 26%
39.502 10780025 unknown 13%
40.036 2144993 2-mino-5-(4-amino-2-methoxyphenyl)tropone 3%
41.314 4289708 di-(2-ethylhexyl)ester of adipic acid b
42.542 a cyclododecyne b
43.126 a hexadecanoic acid , 2 , 3-dihydroxypropy l ester b
53.457 a (23S)-ethylcholest-5-en-3-beta-ol b
a:couldn t measure the area due to low concentration.b:<1%
The main components are N-Isobutylundeca-2[ E] -ene-8 ,10-diynamide , N ,N-diethyl-4 ,5-dimethoxy-2
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2004年第 4期 ○姚兴东 ,聂园梅/不同紫锥花种属中挥发性组分的气相色谱/质谱分析
-methylbenzamide , 1 ,2-benzendicarboxylic acid dibutyl ester and a unknown compound.
(4)3 year old Echinacea Angustifolia
Table 4 lists the chemicals and their relative percentage in ethanol extract:
Table 4:Components found in 3 year old E.Angustifolia ethanol extract
Retention time(min) Area MS search result Area %
12.839 472114 4-Ethyl-2-methy lhexa-2 , 3-diene b
14.528 3288578 2 , 3-dihydro-3 , 5-dihydroxy-6-methyl-4H-py ran-4-one 1%
15.99 226337 unknown b
17.261 598248 2-furancarboxaldehyde b
20.768 225575 Unknown b
23.041 36682304 spiro[ 4 , 5] decane or 9-octadecyne 12%
23.495 a 1-pentadecene b
27.967 185736909 Cyclododecene 59%
28.477 55190075 tetracyclo[ 4 , 3 , 2 , 0(2 , 9), 0(3 , 5)undeca-7 , 10-dien-4-ol 17%
29.066 1628356 1-dodecen-3-yne 1%
29.669 16415220 1[ 7] , 4 , 8-O-menthatriene 5%
31.535 2136583 4-methyl-5-nonanone 1%
31.855 1142299 Unknown b
32.605 10336096 1-(3-butenyl)cyclobutabenzene 3%
41.339 2834263 hexanedioic acid dioctyl ester 1%
a:couldn t measure the area due to low concentration.b:<1%
The main components include cyclododecent;tetracyclo[ 4 ,3 ,2 ,0(2 , 9), 0(3 , 5)undeca-7 , 10-dien-4-ol and
spiro[ 4 ,5] decane.
(5)Echinacea Anustifolia leaf
Table 5 lists the chemicals and their relative percentage in ethanol extract:
Table 5:Components found in E.Angustifolia leaf ethanol extract
Retention time(min) Area MS search result Area %
14.435 1294421 2 , 3-dihydro-3 , 5-dihydroxy-6-methyl-4H-py ran-4-one 8%
16.707 719249 hexanol , 5-methyl 4%
23.519 1744304 1-propanol , 2 , 2-dimethyl 11%
27.818 1075356 1[ S] , 4[ S] -dihydroxy-p-menth-2-one 7%
33.764 3964307 1 , 2-benzenedicarboxylic acid dibutyl ester + hexadecanoic acid 24%
36.423 661039 Phytol 4%
36.958 4326370 9.12-octadecadienoic acid 27%
37.873 a Unknown b
41.349 2498412 hexanedioic acid , dioctyl ester 15%
46.085 a Heptadecane b
49.721 a stigmasta-5 , 22-dien-3-ol b
50.882 a Nonacosane b
53.364 a (23S)-ethylcholest-5-en-3-beta-ol b
a:couldn t measure the area due to low concentration.b:<1%
The main components include 9 ,12-octadecadienoic acid;1 ,2-benzenedicarboxylic acid dibutyl ester;hexanedioic
acid dioctyl ester;1-propanol , 2 ,2-dimethyl and 2 ,3-dihydro-3 ,5-dihydroxy-6-methyl-4H-pyran-4-one.
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广西民族学院学报(自然科学版) 2004年 11月 第 10卷
Most of the compounds in Mazza s paper can not be found here.Compare the retention of germacrene D , it s clear
Mazza s method is good for low boiling point compounds.The method employed here is good for analysis of compounds with
high boiling point , which can t be detected with Mazza s method.
The main volatile components are 1 ,2-benzenedicarboxylic acid dibutyl ester;hexanedioic acid dioctyl ester;9 ,12-
octadecadienoic acid and phenol , 2 ,4-bis(1 ,1-dimethylethyl)for E.purpura root;germacrene D;1 ,2-benzenedicar-
boxylic acid dibutyl ester and 9 ,12-octadecadienoic acid for E.Aerial top;2-butanone , 4-(ethylphenyl);N , N-di-
ethyl-4 , 5-dimethoxy -2-methylbenzamide;N-isobutyl-(2E , 4Z , 8Z , 10E)-dodecatetraenamide and di(2-
ethylhexyl)adipate for E.Pallida root;cyclododecene , tetracyclo[ 4 ,3 ,2 ,0(2 ,9), 0(3 ,5)undeca-7 ,10-dien-4-ol ,
1-(3-butenyl)cyclobutabenzene , 1[ 7] ,4 ,8-O-methatriene and spiro[ 4 ,5] decane for E.Angustifolia.1-propanol ,
2 ,2-dimetyl;1 ,2-benzenedicarboxylic acid dibutyl ester , 9 ,12-octadecadienoic acid and hexanedioic acid , dioctyl es-
ter for E.Angustifolia leaf.Investigation to these compounds may help us understanding the biological effect of E.Plants on
immunostimulation.For example , germacrene D has been found in some original oils of herbs , which show positive effect on
immunostimulation.It s reasonable the compound has contribution to the medicinal effect of herb.Detail studies on these
compounds are needed.
REFERENCES
[ 1] Bauer , R.Chemistry , analysis and immunological investigations of Echinacea phytopharmaceut icals.In Immunomodulatory Agents from plants[M] .Birkhauser
Verlag:Basel;1999 , 41-88.
[ 2] The Institute for Nutraceutical Advancement HPLC method:http://www.nsf-ina.org/methods/ echinaceaset.html .
[ 3] Perry NB , Burgess EJ , Glennie VL.Echinacea Standardization:AnalyticalMethods for Phenolic Compounds and Typical Levels inMedicinal species [ J] .J.A-
gric.Food Chem.2001, 49:1702-1706.
[ 4]Mazza G , Cottrell T.Volatile Components of Roots , Stems , Leaves , and Flowers of Echinacea Species[ J] .J.Agric.Food Chem.1999 , 47:3081-3085.
[责任编辑 黄招扬] [责任校对 黄祖宾]
不同紫锥花种属中挥发性组分的
气相色谱/质谱分析
姚兴东1 , 聂园梅1 ,Nirmalendu Datta-Gupta2
(1.广西民族学院 化学与生态工程学院 ,广西 南宁 530006;
2.Department of Physical Sciences , South Carolina State University , Orangeburg , SC 29117)
摘 要:通过气相色谱/质谱联用技术分析了三种常见紫锥花种属 E.Angustifolia , E.Pallida 和 E.Pur-
purea中挥发性组分.从结果分析 , 这些挥发性组分对于其药效也有部分贡献.
关键词:紫锥花;挥发性组分;气相色谱/质谱分析
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2004年第 4期 ○姚兴东 ,聂园梅/不同紫锥花种属中挥发性组分的气相色谱/质谱分析