全 文 ：天然产物研究与开发 Nat Prod Ｒes Dev 2016，28:547-550
文章编号:1001-6880(2016)4-0547-04
Ｒeceived:October 13，2015 Accepted:January 12，2016
Foundation item:Talent Fund of Anhui agricultural university (yi2010-
03)
* Corresponding author E-mail:yanxu@ ahau． edu． cn
萹蓄乙酸乙酯部位化学成分的抑菌活性研究
李曼曼1，2，刘增辉3，朱 创1，2，凌铁军1，2，李成网3，王莉莉1，2，徐 燕1，2*
1安徽农业大学茶与食品科技学院;2 茶树生物学与资源利用国家重点实验室，合肥 230036;
3安徽省医学科学研究院，合肥 230061
摘 要:萹蓄(Polygonum aviculare L．)是一种常用中药，具有杀虫止痒，治疗黄疸等功效。本文采用打孔法分别
对其乙酸乙酯部位分离得到的十个化合物进行了抑菌活性实验，结果表明，除化合物 1、2、9、10 外，其它化合物
均对不同的菌种呈现出不同的抑菌活性，并且呈一定的剂量效应。本实验为萹蓄及其化学成分作为天然的抑
菌活性物质研究开发提供理论依据。
关键词:萹蓄;抑菌活性;Myricetin-3-O-(3-O-galloyl)-rhamnopyranoside;抑菌圈
中图分类号:Ｒ284． 1 文献标识码:A DOI:10． 16333 / j． 1001-6880． 2016． 4． 014
Antibacterial Activities of Compounds from Ethyl
Acetate Extract of Polygonum aviculare L．
LI Man-man1，2，LIU Zeng-hui3，ZHU Chuang1，2，Ling Tie-jun1，2，
LI Cheng-wang3，WANG Li-li1，2，XU Yan1，2*
1School of Tea ＆ Food Science，Anhui Agricultural University;2State Key Laboratory of Tea Biology
and Utilization，Anhui Agricultural University，Hefei 230036，China;3Anhui Academy of
Medical Science，Hefei 230061，China．
Abstract:Polygonum aviculare L． was used for the treatment of cutaneous pruritus and jaundice． In current study，anti-
bacterial activities of ten compounds isolated from ethyl acetate extract of P． aviculare were tested． The results showed
that these compounds (except for 1，2，9 and 10)showed inhibitory activity against different bacteria with dose-depend-
ent effect． This indicated that P． aviculare and its components can be regarded as a potential source of natural antibacte-
rial agent in the prevention and treatment of microbial infections．
Key words:Polygonum aviculare L．;antibacterial activity;myricetin- 3-O-(3-O-galloyl)-rhamnopyranoside;inhibi-
tion zone
Introduction
Polygonum aviculare L．，belonging to the family of Po-
lygonaceae，is an annual or perennial prostrate herba-
ceous plant with small elliptic lanceolate leaves，and
was widely distributed in the world［1］． P． aviculare con-
tains a variety of chemical active constituents，such as
flavonoids，phenylpropanoids，phenolic acids，amino
acids and carbohydrates (flavonoids are the main
chemical compositions)［2，3］． Some studies proved that
P． aviculare had many functions and was used as diu-
retic，insecticide detoxification and antioxidant，etc［4-7］．
We had reported the antibacterial activities and constit-
uents of ethyl acetate extract of P． aviculare in the pub-
lished paper． The ethyl acetate extract of P． aviculare
(EAE)had obvious antibacterial activities for Esche-
richia coli，enteropathogenic Escherichia coli，Staphyloc-
cocus aureus，Salmonella typhi and Shigella dysenteri-
ae［8］． In this study，the main work was evaluating the
antibacterial activities in vitro of ten compounds puri-
fied from EAE． There were methyl gallate (1) ，stig-
mast-5-en-3-O-β-D-glucopyranoside (daucosterol，2) ，
kaempferol (3) ，quercetin (4) ，gallic acid (5) ，arabi-
nofuranoside (avicularin，6) ，quercetin-3-O-α-L-rham-
nopyrannoside (quercitrin，7) ，myricetin-3-O-α-L(3-
O-galloyl)-rhamnopyranoside (8) ，3，3，4，5． 5，7-
hexahydroxyflavone-3-O-α-L-rhamnopyrannoside (myr-
icitrin，9)and kaempferol-3-O-α-L-rhamnopyrannoside
(juglanin，10)． The results of this study provided theo-
retical foundation for the utilization of P． aviculare in
the health needs．
Materials and Methods
Samples
Ten compounds were isolated from ethyl acetate extract
of P． aviculare (Fig． 1)． The separation and identified
of the compounds were described in the published pa-
per［8］． The plant materials were gathered from Dabie
Mountain (in Jinzhai County，Anhui Province，PＲ Chi-
na． in September 2009 and was identified by Prof．
Shoujin Liu in School of Pharmacy，Anhui University of
Traditional Chinese Medicine．
Fig. 1 Chemical structures of compounds from ethyl acetate extract of P． aviculare
Ｒeagents
Berberine hydrochloride was purchased from Nanjing
Baijingyu Pharmaceutical Co．，Ltd． China and norfloxa-
cin was purchased from Anhui Sanjinwansen Pharma-
ceutical Co．，Ltd． China． All other chemicals and rea-
gents used in this study were of analytical grade and
supplied by Tianjin Bodi Chemicals Co．，Ltd． (Tian-
jin，China)．
Microorganisms
The tested microorganisms included Escherichia coli，
enteropathogenic Escherichia coli，Staphyloccocus au-
reus，Salmonella typhi and Shigella dysenteriae． All of
them were obtained from Key Laboratory of Tea Bio-
chemistry ＆ Biotechnology (LTBB) ，Anhui Agricultur-
al University． All bacterial strains were respectively
cultivated in fluid nutrient medium (Beef extract，5 g /
L;Peptone，10 g /L;NaCl 5 g /L;pH 7． 4) ，and incuba-
ted at 37 ℃ for 24 h． After incubated，1 mL bacterial
suspension was added in the 100 mL new fluid nutrient
medium，and incubated at 37 ℃ for 12 h． The new bac-
terial suspension was ready for further processing．
Antibacterial assays
The hole plate diffusion method was used to investigate
the antimicrobial activities of the purified compounds．
They were diluted by DMSO to different gradient con-
centrations (200 μg /mL，400 μg /mL and 800 μg /mL
for each compound)． Berberine hydrochloride (25． 6
mg /mL)and norfloxacin (25 μg /mL)diluted by DM-
SO as standard antimicrobials were used for compari-
son．
For the determination of antibacterial activity，300 μL
bacterial suspension were inoculated onto 15 cm diame-
ter plates with 45 mL nutrient agar medium (Beef ex-
tract，5 g /L;Peptone，10 g /L;NaCl 5 g /L;Agar，20 g;
845 Nat Prod Ｒes Dev Vol. 28
pH 7． 4)． 50 μL of testing solution was injected in the
hole of 6 mm size prepared using sterile steel tuber．
These plates were incubated at 37 ℃ for 24 h． Antimi-
crobial activity of test solution was determined by meas-
urement of inhibition zone against the reagent (DMSO)
blank． All the tests were performed in triplicate．
Data analysis
Data were analyzed by SPSS (Version 11． 0 for Win-
dows，SPSS Inc．，Chicago，IL)and expressed as mean
± SD of triplicate determinations．
Ｒesults and Discussion
Table 1 illustrated the antibacterial activities of ten
Table 1 The inhibition zone of compounds for testing microorganisms
Concentration
(μg /mL)
Diameter of inhibition zone (mm)
Escherichia
coli
Enteropathogenic
Escherichia coli
Staphyloccocus
aureus
Salmonella
typhi
Shigella
dysenteriae
DMSO － 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Berberine Hydrochloride 25． 6 × 103 10． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Norfloxacin 25 24． 2 ± 0． 1 ＊＊ 32． 2 ± 0． 1 ＊＊ 33． 2 ± 0． 2 ＊＊ 32． 2 ± 0． 3 33． 9 ± 0． 1 ＊＊
Methyl gallate 200 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Daucosterol 200 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Kaempferol 200 9． 9 ± 0． 1 ＊＊ 6． 0 ± 0． 0 10． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0
400 10． 1 ± 0． 1＊＊ 11． 9 ± 0． 2 ＊＊ 15． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 9． 9 ± 0． 2＊＊
800 11． 1 ± 0． 1 ＊＊ 12． 1 ± 0． 1 ＊＊ 13． 0 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0
Quercetin 200 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 8． 2 ± 0． 2 ＊＊ 10． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 10． 2 ± 0． 1 ＊＊ 11． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0
Gallic acid 200 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 9． 3 ± 0． 2＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Avicularin 200 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 10． 94 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 11． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Quercitrin 200 6． 0 ± 0． 0 6． 0 ± 0． 0 10． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 6． 0 ± 0． 0 13． 1 ± 0． 2 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 12． 1 ± 0． 1 ＊＊ 15． 1 ± 0． 2 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0
Myricetin-3-O-(3-O-galloyl )-
rhamnopyranoside 200 6． 0 ± 0． 0 12． 2 ± 0． 2
＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 10． 1 ± 0． 1 ＊＊ 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Myricitrin 200 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Juglanin 200 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
400 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
800 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0 6． 0 ± 0． 0
Note:The diameter of inhibition zone ＞ 6． 0 mm indicated that the substance had antibacterial activity，＊＊ P ＜ 0． 01 means significantly different Vs blank
Control (DMSO)．
945Vol. 28 LI Man-man，et al:Antibacterial Activities of Compounds from Ethyl Acetate Extract of Polygonum aviculare L．
compounds． Methyl gallate，daucosterol，myricitrin and
juglanin did not show any antibacterial activity in this
study． The other compounds showed different anti-bac-
terial activities and displayed significantly different Vs
blank Control (＊＊P ＜ 0． 01)．
Kaempferol can restrain the growth of Escherichia coli
(at 200，400 μg /mL and 800 μg /mL) ，enteropatho-
genic Escherichia coli (at 400 μg /mL and 800 μg /
mL) ，Staphyloccocus aureus (at 200，400 μg /mL and
800 μg /mL) and Shigella dysenteriae (at 400 μg /
mL)． But it did not have any effect for Salmonella
typhi． Quercetin appeared its antibacterial activity
merely at 400 μg /mL and 800 μg /mL against entero-
pathogenic Escherichia coli and Staphyloccocus aureus．
Gallic acid (800 μg /mL)and avicularin (400 μg /mL
and 800 μg /mL)had antibacterial activities only for
enteropathogenic Escherichia coli and had hardly any
effect for other bacterium． Quercitrin (200 μg /mL，400
μg /mL and 800 μg /mL) showed good suppression
effect for Staphyloccocus aureus，and it merely inhibited
the growth of enteropathogenic Escherichia coli at 800
μg /mL． Myricetin-3-O-(3-O-galloyl)-rhamnopyrano-
side was not showing any inhibition for all bacteria ex-
cept enteropathogenic Escherichia coli． The inhibition
zone of myricetin-3-O-(3-O-galloyl)-rhamnopyrano-
side against enteropathogenic Escherichia coli． was 12． 2
mm (200 μg /mL)and 10． 1 mm (400 μg /mL)． But
the activity was not obvious at 800 μg /mL．
The best inhibition performance for Escherichia coli，
enteropathogenic Escherichia coli and Staphyloccocus
aureus，were kaempferol at 800 μg /mL． Myricetin-3-
O-(3-O-galloyl)-rhamnopyranoside can inhibit the
growth of Enteropathogenic Escherichia coli 12． 2 mm (
the diameter of inhibition zone )at 200 μg /mL while
kaempferol 12． 1 mm at 800 μg /mL． Quercitrin can in-
hibit the growth of Staphyloccocus aureus 15． 1 mm as
kaempferol，but its concentration (800 μg /mL)was
higher than kaempferol (400 μg /mL)． These com-
pounds cannot suppress the growth of Salmonella typhi
in our experiments． Through the survey of antibacterial
activities of these compounds，the roles of them in anti-
bacterial test in vitro were confirmed． This study had
improved the possibility of the use of P． aviculare in an-
timicrobial drug and food additive development for hu-
man application．
Acknowledgement
The authors are grateful to the natural product team of
Key Laboratory of Tea Biochemistry ＆ Biotechnology
(LTBB)providing the generous help． We also thank
the Anhui Agricultural University for financial support．
We all thank Professor Guanhu Bao for providing the
help of article amending．
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