全 文 ：Microwave-assisted Extraction and Biological Activity
Determination of Insecticidal Components of Seleng
Wormwood(Artemisia selengensis Turcz．)
CHEN Ze-yu1，LI Shui-qing2* ，LIU Yun2
1． Xishuangbanna Entry-Exit Inspection and Quarantine Bureau，Yunnan Province，Xishuangbanna 666100，China;2． College of Chemical and
Environmental Engineering，Yangtze University，Hubei Province，Jinzhou 434023，China
Abstract ［Objective］The aim was to study the biological activity of insecticidal components of Seleng Wormwood on cabbage butterfly (Pieris ra-
pae)． ［Method］With the assisted condition of microwave heating，ethanol，ethyl ether，water and methylbenzene were selected as solvents to ex-
tract Seleng Wormwood，and the antifeedant，stomach toxicity，contact toxicity and fumigation activities of various solvent extracts against cabbage
butterfly were determined． ［Result］Various solvent extracts all showed certain antifeedant，stomach toxicity，contact toxicity and fumigation activity
against the larvae of cabbage butterfly，and the biological activity of ethanol extract was the strongest． ［Conclusion］The results could provide refer-
ence for development and utilization of botanical pesticides of Seleng Wormwood．
Key words Seleng Wormwood;Cabbage butterfly;Antifeedant;Stomach toxicity;Contact toxicity;Fumigation
Received:October 8，2010 Accepted:November 18，2010
Supported by Doctoral Research Fund of Yangtze University ＆ Educa-
tional Commission of Hubei Province of China(Q200712003)．
* Corresponding author． E-mail:shuiqing2000@163． com
The widespread use of chemical pesticides has caused the
shortcomings difficult to overcome，such as 3R problem(res-
idue，resistance and resurgence)． Thus，the development of
pesticides with biological plausibility which is safe to the envi-
ronment and non-target organisms has become the mainstream
in the development of modern pesticides［1］． Botanical pesti-
cides have several advantages including high efficiency，low
toxicity，low residue and high selectivity，and they are safe to
high animals and the natural enemies［2 －3］，thus becoming the
first choice for the development of reasonable biological
pesticides．
Seleng Wormwood (Artemisia selengensis Turcz．) is a
perennial herb in Asteraceae，which can be used as food and
medicine with good health protection effects． It is commonly
used to cure acute infectious hepatitis with good effects，while it
has no side effects． There are also some reports about the ex-
tracts of Seleng Wormwood for the control of agricultural pests．
LI Bao-tong et al． reported the effect of Seleng Wormwood es-
sential oil on the feeding behavior of cabbage worm larva and
its antifeedant effect on cotton bollworm［4］;XIE Yi-sun et al．
reported that Seleng Wormwood essential oil had strong an-
tifeedant and stomach toxicity on cabbage worm，and it had
weak contact toxicity and no fumigation effect on the pest［5］;
ZHENG Gong-yuan et al． found that the original juice，water
extract and ethanol extract of Seleng Wormwood all had anti-
bacterial effects［6］． Taking microwave-assisted heating as the
research method， the author extracted the biological active
substances of Seleng Wormwood using the solvents with differ-
ent polarities，and tested the biological activity of various sol-
vent extracts against the larvae of cabbage butterfly，this would
provide reference for the development of botanical insecticide．
1 Materials and Methods
1．1 Materials
1． 1． 1 Seleng Wormwood． Seleng Wormwood for the test was
collected from the surroundings of Yangtze University in Jing-
zhou City of Hubei Province．
1． 1． 2 Cabbage butterfly． Young larvae of cabbage butterfly
were collected from the cabbage leaves in field． After indoor
rearing，the 5th instar larvae with the similar size were selected
for test． Indoor rearing conditions were as follows:temperature
(28 ±1)℃，relative humidity (80 ±5)%，photoperiod L∶ D =
16 h∶8 h．
1． 1． 3 Reagents． The reagents for the test were as follows:
alcohol (≥99．5%) ，produced by Anhui Ante Biochemical
Co．，Ltd．;ether (≥99． 5%) ，produced by Tianda Chemical
Reagent Factory in Dongli District of Tianjin City;methyl-
benzene (≥99．5%) ，produced by Wuhan Zhongtian Chemical
Co．，Ltd．;acetone (≥99． 5%) ，produced by Wuhan Zhong-
tian Chemical Co．，Ltd． All the reagents used in the test were
analytical pure．
1． 2 Extraction method Four copies of 50 g stem and leaf
powders were accurately weighed，and were dissolved in the
solvents of ethanol，ethyl ether，water and methylbenzene，re-
spectively． The dissolved solutions were subsequently radiated
by microwave (power 400 W)for 5 min． In order to avoid the
evaporation of solvent and thus affect the extraction of active
substances，the samples should be taken out after radiation for
20 s，and carried out radiation again after cooling． After the ra-
Plant Diseases and Pests 2010，1(6) :51 －53，57
diation process was finished，the solutions were depressurized
and filtered，and concentrated by rotary evaporator． Then the
solvents were recovered，and about 1 g sticky material like
cream was obtained． The various sticky materials were diluted
into 5 mg/L solution by acetone and preserved for the test．
1． 3 Bioassay experimental method
1． 3． 1 Determination of antifeedant activity． Small leaf disc
adding method by YU Xiang-yang et al．［7］ was adopted for the
test． The leaves of cabbage were fully cleaned，and hole
puncher was used to punch the leaves into several circular leaf
discs with the diameter of 2 cm． Then the leaf discs were
soaked in the extract solution for 5 s and taken out． After dried，
the leaves were placed in the Petri dish with the diameter of 9
cm which had been covered with absorbent paper． The larvae
of cabbage butterfly that had been hungry for 4 h were inocula-
ted in Petri dishes with 20 heads in each dish． The larvae
soaked with acetone were set as control． Each treatment con-
tained three repeats(the same as follows)． The feeding condi-
tion of leaves were investigated after inoculating pests for 24 h，
and the feeding indexes and antifeedant rates were calculated
according the following grading criteria and formula:zero level:
no feeding traces;Ⅰ level:sporadic feeding;Ⅱ level:obvious
feeding notch;Ⅲ level:feeding area accounting for about 1 /3
of the total area;Ⅳ level:feeding area accounting for about
1 /2 of the total area;Ⅴ level :only a small amount of residue
remaining．
Feeding index(%)=［∑(damage level × total number of
leaf discs in various levels)/(total number of leaf discs × the
highest level × total number of pests in the test) ］×100
Antifeedant rate(%)=［(feeding index in control － feeding
index in treatment)/ feeding index in control］× 100
1．3． 2 Determination of stomach toxicity． The extracts from
different solvents were evenly coated on the one side of cab-
bage leaf disc with the diameter of 2 cm using tapered brush，
and the leaves were air dried． Another side of leaf disc was
covered on solution membrane using double-sided adhesive． 20
leaf discs were placed in the Petri dish，and each leaf dish was
inoculated with 20 heads of larvae． The mortality rate of the lar-
vae was investigated after 12，24，36 and 48 h，and the correc-
ted mortality was calculated．
Mortality rate(%)=(the number of dead pests in the test /
total number of pests in the test)× 100
Corrected mortality rate(%)=［(mortality rate in treatment
－mortality rate in control)/(100 －mortality rate in control)］×100
1． 3． 3 Determination of contact toxicity． Dripping method［8］
was adopted to determine the contact toxicity． The extracts of
different solvents were dripped on the pronotum of larvae by
macro dropper with 1 μl in each larva． The larvae after dripping
were placed in the Petri dish which had been covered with ab-
sorbent paper with 20 heads in each dish． The mortality condi-
tion of the larvae was checked after 12，24，36 and 48 h，and fi-
nally mortality rate and corrected mortality rate were calculated．
1． 3． 4 Determination of fumigation activity． Proper amount of
leaves were placed in 250 ml flask，and they were inoculated
with 20 heads of test pests． The internal mouth of the flask was
pasted with 8 cm ×1 cm filter paper，then the filter paper was
dripped with 2 ml acetone extract of Seleng Wormwood，finally
rubber stopper was quickly plugged． The flask was placed un-
der room temperature，and the mortality condition of test pests
was investigated after 24 h．
2 Results and Analysis
2．1 Antifeedant activity of various solvent extracts of Se-
leng Wormwood against the larvae of cabbage butterfly
The antifeedant activity of various solvent extracts of Seleng
Wormwood against the larvae of cabbage butterfly was deter-
mined，and the results were shown in Table1．
Table 1 The antifeedant activities of various solvent extracts
against the larvae of cabbage butterfly
Treatment Feeding index∥% Antifeedant rate∥%
Ethanol extract 2． 32 ±0． 08 34． 10 ±0． 70 a
Ethyl ether extract 2． 38 ±0． 09 32． 45 ±0． 53 b
Water extract 2． 40 ±0． 10 31． 75 ±0． 28 bc
Methylbenzene extract 2． 42 ±0． 11 31． 04 ±0． 30 c
Control 3． 52 ±0． 15 －
Note:The same letter in the same column represented that the differ-
ence was not significant after Duncan new multiple range test(P
＞0． 05)．
As shown in Table 1，various solvent extracts of Seleng
Wormwood had certain antifeedant activity on the 5th instar lar-
vae of cabbage butterfly after treating for 24 h，and the an-
tifeedant rate reached over 30%． The antifeedant activity of
ethanol extract was the strongest，and the antifeedant activity
of methylbenzene extract was the weakest．
2． 2 Stomach toxicity of various solvent extracts of Se-
leng Wormwood against the larvae of cabbage butterfly
The stomach toxicity of various solvent extracts of Seleng
Wormwood against the larvae of cabbage butterfly was deter-
mined，and the results were shown in Table 2．
Table 2 The stomach toxicity of various solvent extracts against the larvae of cabbage butterfly
Treatment
Corrected mortality rate∥%
12 h 24 h 36 h 48 h
Ethanol extract 26． 75 ±1． 79 a 31． 62 ±1． 48 a 38． 83 ±2． 67 a 42． 37 ±2． 79 a
Ethyl ether extract 13． 37 ±0． 90 b 19． 49 ±3． 86 b 25． 12 ±2． 10 b 33． 23 ±3． 18 b
Water extract 6． 69 ±0． 45 c 14． 70 ±1． 18 b 16． 74 ±1． 40 c 14． 95 ±4． 35 d
Methylbenzene extract 13． 37 ±0． 90 b 19． 49 ±3． 88 b 22． 09 ±3． 52 b 24． 09 ±3． 72 c
Note:The same letter in the same column represented that the difference was not significant after Duncan new multiple range test(P ＞0． 05)．
25 Plant Diseases and Pests 2010
As shown in Table 2，various solvent extracts of Seleng
Wormwood had certain stomach toxicity on the larvae of cab-
bage butterfly，and the stomach toxicity of ethanol extract was
the strongest． The stomach toxicity of ethanol extract reached
42．37% after treating for 48 h;followed by ether extraction and
methylbenzene extract;and the stomach toxicity of water ex-
tract was the weakest of only 14． 95%．
2． 3 Contact toxicity of various solvent extracts of Seleng
Wormwood against the larvae of cabbage butterfly The
contact toxicity of various solvent extracts of Seleng Wormwood
against the larvae of cabbage butterfly was determined，and the
results were shown in Table 3．
Table 3 The contact toxicity of various solvent extracts against the larvae of cabbage butterfly
Treatment
Corrected mortality rate∥%
12 h 24 h 36 h 48 h
Ethanol extract 11． 97 ±0． 70 a 27． 64 ±3． 24 a 32． 24 ±1． 88 a 33． 93 ±4． 26 a
Ethyl ether extract 5． 90 ±0． 35 b 17． 08 ±4． 02 b 24． 92 ±1． 80 ab 25． 26 ±5． 39 ab
Water extract 5． 90 ±0． 35 b 10． 56 ±3． 37 b 14． 57 ±5． 88 c 16． 58 ±6． 54 b
Methylbenzene extract 5． 90 ±0． 35 b 14． 86 ±3． 39 b 19． 92 ±3． 21 bc 22． 69 ±2． 52 b
Note:The same letter in the same column represented that the difference was not significant after Duncan new multiple range test(P ＞0． 05)．
As shown in Table 3，the contact toxicity of various solvent
extracts of Seleng Wormwood against the larvae of cabbage
butterfly gradually increased with the prolongation of treating
time． In contrast，the contact toxicity of ethanol extract was the
strongest．
2． 4 Fumigation activity of various solvent extracts of Se-
leng Wormwood against the larvae of cabbage butterfly
The fumigation activity of various solvent extracts of Seleng
Wormwood against the larvae of cabbage butterfly was deter-
mined，and the results were shown in Table 4．
Table 4 The fumigation activity of various solvent extracts
against the larvae of cabbage butterfly
Treatment Mortality rate∥% Corrected mortality rate∥%
Ethanol extract 36． 67 ±7． 64 26． 92 ±0． 76 a
Ethanol extract 26． 67 ±7． 64 15． 26 ±1． 73 b
Water extract 20． 00 ±8． 66 7． 59 ±2． 61 c
Methylbenzene extract 23． 33 ±10． 41 11． 66 ±1． 48 b
Control 13． 33 ±5． 61 －
Note:The same letter in the same column represented that the differ-
ence was not significant after Duncan new multiple range test(P
＞0． 05)．
As shown in Table 4，the fumigation activity of various sol-
vent extracts of Seleng Wormwood against the larvae of cab-
bage butterfly was not strong． In contrast，the fumigation activi-
ty of ethanol extract was relatively stronger．
3 Conclusion
The research on botanical pesticides has received more
attention due to the requirements of sustainable development
of agriculture，which has made great progress，and many
compounds with antifeedant and insecticidal activity have been
found． For example，YANG Zheng-min et al． isolated and
identified the pesticide active ingredient in the fruit of Celastrus
angulatus Max．［9］;WU Wen-jun et al． studied the toxic com-
ponents of C． angulatus Max． against insects［10］;and TAN
Wei-hong et al． studied the chemical composition of aza-
dirachtin and its biological activity［11］．
Taking microwave-assisted heating as the research meth-
od，the author extracted the biological active substances of
Seleng Wormwood by solvents with different polarities，and
tested the biological activity of various solvent extracts against
the larva of cabbage butterfly． The results showed that all the
extracts of different solvents had certain antifeedant，stomach
toxicity，contact toxicity and fumigation activity against the lar-
vae of cabbage butterfly，in which the biological activity of eth-
anol extract was the strongest．
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35CHEN Ze-yu et al． Microwave-assisted Extraction and Biological Activity Determination of Insecticidal Components of Seleng Wormwood(Artemisia selengensis Turcz．)
when different extraction methods were adopted． Because the
extraction methods were different，the dissolution degree of ac-
tive substances was also different，thereby affecting the anti-
fungal effect．
(5)Extraction unit is one of the key links of modernization
of antifungal medicinal plants，affecting the final quality of me-
dicinal preparation and cost，and the modernization level of
pharmaceutical industry［9 －10］． There are varieties of traditional
Chinese medicine extraction methods，we should comprehensive-
ly consider from the aspects of efficacy，pharmacology，technolo-
gy，engineering，economics，environmental protection and recy-
cling． The operation process of the experimental study was sim-
ple，the price of equipment was equitable，and the antifungal
effect was significant，which has certain application prospect．
(6)Ethanol and acetone reagent had some antifungal
effect，control must be designed reasonably to exclude effects．
(7)It was essential to consider from production cost and
make plant medicinal extraction better apply in production prac-
tice，and find the aqueous extraction of medicinal plant with
high antifungal effect，the comprehensive effect of Coptis root in
this aspect can not be ignored，which should be further studied．
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8 种植物源农药的提取工艺及其抑制真菌作用研究
高 磊1，谢宇琪2，王兴华1* ，李 颖1
(1．山西大学生命科学学院，山西太原 030006;2．太原市环境保护局，山西太原 030002)
摘要 ［目的］研究黄连、丁香等 8种药用植物抑菌成分的最佳提取工艺及对 2 种植物病原菌的抑菌作用。［方法］采用水提、乙醇提、丙酮提 3
种提取方法提取各种药用植物有效成分，采用药物平板稀释法测定药用植物提取物的最低抑菌浓度(MIC)和抑菌率。［结果］抑制 2种植物病
原菌的最优药用植物及提取方法分别为:抑制地黄枯萎病菌采用丁香乙醇提和丙酮提，肉桂乙醇提，此时 MIC达到 4 mg /ml，抑制肉苁蓉致病菌
采用丁香乙醇提和丙酮提，肉桂丙酮提，此时 MIC达到 4 mg /ml。［结论］8种药用植物的乙醇和丙酮提取物对供试菌种的抑制作用明显优于水
提物。
关键词 植物源农药;提取工艺;最低抑菌浓度;抑菌率
基金项目 山西省科技厅项目(041020)。
作者简介 高磊(1986 －)，女，山西运城人，硕士研究生，研究方向:应用微生物。* 通讯作者，副教授，硕士生导师，从事微生物代谢研究。
收稿日期 2010-09-19 修回日期
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2010-10-26
(Continued from page 53)
蒌蒿杀虫成分的微波辅助提取及生物活性测定
陈泽宇1，李水清2* ，刘 云2 (1．西双版纳出入境检验检疫局，云南西双版纳 666100;2．长江大学化学与环境工程学院，湖北荆州 434023)
摘要 ［目的］研究蒌蒿杀虫成分对菜粉蝶的生物学活性。［方法］在微波辅助加热条件下，以乙醇、乙醚、水、甲苯为溶剂对蒌蒿进行浸提，测定
各溶剂提取物对菜粉蝶幼虫的拒食、胃毒、触杀和熏蒸活性。［结果］各溶剂提取物对菜粉蝶幼虫均表现出一定的拒食、胃毒、触杀和熏蒸活性，
其中，以乙醇提取物的生物活性最强。［结论］该研究结果可为开发利用蒌蒿植物源杀虫剂提供参考。
关键词 蒌蒿;菜粉蝶;拒食;胃毒;触杀;熏蒸
基金项目 长江大学博士科研基金;湖北省教育厅科研项目(Q200712003)。
作者简介 陈泽宇(1970 － )，男，云南版纳人，硕士，从事有机化学分析测试工作。* 通讯作者，博士，教授。
收稿日期 2010-10-08 修回日期 2010-11-18
75GAO Lei et al． Extraction Technology of Eight Botanical Pesticides and Their Antifungal Activities on Fungus