全 文 ：天然产物研究与开发 Nat Prod Ｒes Dev 2015，27:251-254
文章编号:1001-6880(2015)2-0251-04
Ｒeceived:September 20，2014 Accepted:December 31，2014
Foundation Item:This work was financially supported by Natural Sci-
ence Foundation of China (NSFC，No． 21162023)and Bingtuan Inter-
national Cooperation Projects (No． 2011BC003)
* Corresponding author Tel:86-993-2057272;E-mail:yhb tea@ 163． com
叉毛蓬化学成分的体外抗菌及抗氧化活性研究
王 莹，孙 文，张静晗，徐丽丽，杨红兵*
石河子大学化学化工学院，石河子 832003
摘 要:从叉毛蓬全株首次分离出 4-羟基苯乙酮(1)、紫丁香酸(2)、金圣草黄素(3)、香草酸(4)、4-羟基-3-甲基
苯乙醇(5)、N-［2-(3，4-二羟基苯基)-2-羟基乙基］-3-(4-甲氧基苯基)丙-2-稀酰胺(6)和异鼠李素-3-O-芸香糖苷
(7)7 个化合物。通过 MTT法测定这 7 个化合物的体外抗菌活性，结果表明多数化合物有较强的抗菌活性，其
中化合物 2 对枯草芽孢杆菌，化合物 5 对大肠杆菌，化合物 7 对番茄疮痂病菌和番茄早疫病菌的抑制作用均强
于阳性对照硫酸链霉素对同种菌的抑制。用 DPPH和 FＲAP两种方法测定了化合物的抗氧化活性，结果表明在
DPPH方法中化合物 6 的抗氧化活性最好，IC50值为 0． 2452 mg /mL，在 FＲAP方法中化合物 5 有最好的抗氧化活
性，FＲAP值为 9． 402 mmol /g，强于阳性对照抗坏血酸。
关键词:叉毛蓬;抗菌;抗氧化;MTT;DPPH;FＲAP
中图分类号:O629． 9 文献标识码:A DOI:10． 16333 / j． 1001-6880． 2015． 02． 011
Antibacterial and Antioxidant Properties of Compounds
Extracted from Petrosimonia sibirica L．
WANG Ying，SUN Wen，ZHANG Jing-han，XU Li-li，YANG Hong-bing*
College of Chemistry and Chemical Engineering，Shihezi University，Shihezi 832003，China
Abstract:4-Hydroxyacetophenone (1)，syringic acid (2) ，chrysoeriol (3) ，vanillic acid (4) ，4-hydroxy-3-methoxy ben-
zene ethanol (5) ，N-［2-(3，4-dihydroxyphenyl)-2-hydroxyethyl］-3-(4-methoxyphenyl)prop-2-enamide (6)and isorh-
amnetin-3-O-rutinoside (7)were isolated from Petrosimonia sibirica L． for the first time． In vitro antibacterial activity of
these compounds was evaluated by MTT assay． The results revealed that most of these compounds exhibited significant
activity． The antimicrobial activities of compound 2 against Bacillus subtilis，of compound 5 against Escherichia coli，and
of compound 7 against Xanthomonas vesicatoria and Alternaria solani，were found to be stronger than those of the positive
control，streptomycin sulfate． The antioxidant potential was evaluated by DPPH and FＲAP methods． The results demon-
strated that compound 6 was the most effective in DPPH method，with IC50 value of 0． 2452 mg /mL． Compound 5 was rel-
atively active and showed potent antioxidant effect compared to standard ascorbic acid in FＲAP assay，with FＲAP value
of 9． 402 mmol /g．
Key words:Petrosimonia sibirica L．;antibacterial;antioxidant;MTT;DPPH;FＲAP
Introduction
Petrosimonia sibirica L． (Genus Petrosimonia，Family
Chenopodiaceae)is an annual herbaceous plant，which
grows mostly in arid and saline areas［1］． In China，it is
found only in Xinjiang［2］． The harsh environment im-
parts some specific biological features to it;therefore，in
future it could be used as an important source of thera-
peutic agent exhibiting significant antibacterial activity．
Currently，it is used in desert area as sand fixing plant
providing substantial forage yields［3］． The research on
it is limited only to its geographical distribution and ec-
ological study［4，5］，and its chemical composition and bi-
ological activity have not been reported yet．
The results of our preliminary experiments showed that
the crude extracts exhibited commendable antibacterial
and antioxidant properties［6，7］． Therefore，to better il-
lustrate the biological activity of this plant，extensive
research was carried out to study its chemical composi-
tion and activity． In this study，compounds extracted
from Petrosimonia sibirica L． were evaluated for anti-
bacterial and antioxidant activities with the objective of
obtaining more effective antibacterial agent and an ef-
fective potential source of natural antioxidant that might
help in preventing various oxidative stresses．
Materials and Methods
Materials and reagents
The plant material was collected in September 2013
from Manasi，Xinjiang，China． It was identified by Pro-
fessor Yan Ping (School of Life Science)of Shihezi U-
niversity． A voucher specimen was deposited in the la-
boratory of the College of Chemistry and Chemical En-
gineering，Shihezi University，China．
Five bacterial species:Escherichia coli，Soil agrobacte-
ria，Pseudomonas syringaw pv． tomato，Xanthomonas
vesicatoria and Bacillus subtilis，and one fungal strain，
Alternaria solani were used in this study． 1，1-diphenyl-
2-picrylhydrazyl (DPPH)，Ascorbic acid (AA)and
tert-butyl hydroquinone (TBHQ)were purchased from
Sigma Aldrich;2，4，6-tris(2-pyridyl)-s-triazine (TPTZ)
was purchased from Aladdin Technology Corporation;
and streptomycin sulfate was purchased from Hubei
Xingyinhe Chemical Co． Ltd． ． All chemicals were of
analytical grade．
Sample preparation and analysis
Ethanol was used to extract the sample from 10 kg of
dried and powdered plant material，and then the entha-
nol extract was partitioned by liquid-liquid chromatog-
raphy to obtain 4 fractions:PE，CHCl3，EtOAc，and n-
BuOH． 4-hydroxyacetophenone(1)，syringic acid(2) ，
and chrysoeriol (3)were isolated from CHCl3 extract;
vanillic acid (4)，4-hydroxy-3-methoxy benzene etha-
nol (5) ，N-［2-(3，4-dihydroxyphenyl)-2-hydroxyeth-
yl］-3-(4-methoxyphenyl)prop-2-enamide (6) and
isorhamnetin-3-O-rutinoside (7) were isolated from
EtOAc extract． All the compounds were isolated and
purified for the first time by TLC，column chromatogra-
phy on silica gel，Sephadex LH-20，and identified by
NMＲ and ESI-MS［8］．
Antibacterial assay
The antibacterial activity was evaluated by determining
the minimum inhibitory concentration (MIC)and the
median inhibition concentration (IC50)，using MTT［3-
(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium
bromide］ assay［9］． The concentrations of the samples
were set at 50，100，200，300，400，and 500 μg /mL．
Minimum inhibitory concentrations (MIC):The 96 mi-
cro well plates were used，and 90 μL of liquid consis-
ting of bacteria containing 106 colony forming unit
(CFU)microbial cells and 10 μL of tested samples
were added into each well． The cells were incubated for
24 h at 28 ℃，subsequently，10 μL MTT (5 mg /mL)
was added into the wells，and the cells were further in-
cubated at 28 ℃ for 4 h． Sample concentration without
violet sediment was MIC． The values obtained from the
assay were listed in Table 1．
Median inhibition concentration (IC50):Similar to the
above mentioned MIC method，the bacterial cells were
incubated for 28 h at 28 ℃ ． The cells were then centri-
fuged (5，000 rpm，5 min) ，clear liquid was removed，
200 μL of dimethyl sulfoxide (DMSO)was added，os-
cillated until violet substance dissolved completely，and
recentrifuged (5，000 rpm，5 min)． The supernatant
fluid was drained，100 μL was transferred to 5 mL cen-
trifuge tube followed by the addition of 1． 9 mL DMSO．
The contents were shaken well and then monitored by
measuring the absorbance at 590 nm． The percentage
inhibition by monomer compounds was calculated from
mean values by using the following formula (1) :
Percentage inhibition = ［(Am-An)÷ Am］×100%(1)
In which，Am:absorbance values of the solvent com-
pared;An:absorbance values of the sample．
Testing sample concentration logarithm (X)，bacterio-
static rate conversion into biological statistical probabil-
ity values (Y) ，formed a straight line for virulence re-
gression equation (Y = aX + b)and value of IC50 was
calculated． The values were listed in Table 1．
Antioxidant assay
Ferric ion reducing antioxidant power (FＲAP)assay
The FＲAP assay was performed according to the proce-
dure reported by Yin［10］． Standard solution of ferrous
sulfate (FeSO4，0． 1 mL)with different concentrations，
FＲAP liquid (3 mL)and distilled water (0． 3 mL)
were taken in each tube． The reaction mixtures were
252 Nat Prod Ｒes Dev Vol. 27
Table 1 In vitro antibacterial activities of the isolated compounds and positive control (mg/mL)
1 2 3 4 5 6 7 S．
MIC IC50 MIC IC50 MIC IC50 MIC IC50 MIC IC50 MIC IC50 MIC IC50 MIC IC50
A 0． 4 0． 307 0． 3 0． 259 0． 4 0． 321 0． 1 0． 059 0． 05 0． 034 － － － － 0． 1 0． 046
B － － 0． 4 0． 291 0． 4 0． 339 0． 5 0． 359 0． 4 0． 352 － － 0． 4 0． 350 0． 2 0． 095
C 0． 4 0． 202 0． 2 0． 155 0． 4 0． 227 0． 4 0． 296 － － 0． 4 0． 264 0． 2 0． 133 0． 2 －
D 0． 4 0． 348 － － 0． 2 0． 164 0． 2 0． 164 － － 0． 3 0． 209 0． 2 0． 105 0． 2 0． 159
E 0． 4 0． 375 0． 05 0． 022 0． 3 0． 271 － － － － － － － － 0． 1 0． 082
F 0． 5 0． 519 0． 2 0． 071 － － － － － － 0． 2 0． 110 0． 1 0． 055 0． 2 0． 185
Note:‘A’Escherichia coli;‘B’Soil agrobacteria;‘C’Pseudomonas syringaw pv． tomato;‘D’Xanthomonas vesicatoria;‘E’Bacillus subtilis;‘F’Alternaria
solani;‘S．’Streptomycin sulfate;‘-’no inhibitory activity in the respective concentration．
then incubated at 37 ℃ for 30 min，and monitored by
measuring the absorbance at 593 nm． The linear regres-
sion equation:y = 0． 702 + 0． 084x，Ｒ2 = 0． 9942 was
used to obtain the results． Value of IC50 was calculated
based on the sample concentration and scavenging
rate． AA and TBHQ were used as positive control． The
corresponding values were listed in Table 2．
Table 2 The FＲAP (mmol /g)of the isolated compounds
1 2 3 4 5 6 7 AA TBHQ
FＲAP 0． 282 4． 610 1． 309 1． 596 9． 402 4． 925 1． 059 7． 945 17． 464
DPPH free radical scavenging assay
Scavenging effect of seven compounds on DPPH radi-
cals was analyzed by a modified method utilized by
Wu［11］． In vitro experiments were performed． Samples
with different concentration (0． 1 mL)and DPPH (3
mL，0． 1 mmol /L)were mixed together and allowed to
react for 30 min;the progress was monitored by meas-
uring absorbance at 517 nm． AA and TBHQ were used
as positive control． The scavenging ability was calculat-
ed by using the formula (1)． The values were listed in
Table 3．
Table 3 DPPH radical scavenging activities of the isolated compounds (mg/mL)
1 2 3 4 5 6 7 AA TBHQ
IC50 7． 2266 0． 2927 1． 0005 1． 1263 0． 2964 0． 2452 2． 5776 0． 0896 0． 0660
Ｒesults and Discussion
In the antibacterial test，most of the isolated compounds
strongly inhibited the growth of microorganisms． The
values corresponding to the in vitro antibacterial activity
of the isolated compounds were listed in Table 1． 4-
Hydroxyacetophenone，syringic acid and chrysoeriol ex-
hibited inhibitory activity on most of selected bacterial
strains． Syringic acid exhibited the strongest inhibitory
effect on B． subtilis，with MIC value of 0． 05 mg /mL，
and IC50 value of 0． 022 mg /mL;It was found to be
more effective than the positive control with the value
of MIC as 0． 1 mg /mL，and IC50 as 0． 082 mg /mL． The
above mentioned compounds strongly inhibited E． coli．
N-［2-(3， 4-dihydroxyphenyl )-2-hydroxylethyl］-3-
(4methoxyphenyl)prop-2-enamide and isorhamnetin-3-
O-rutinoside did not exhibit significant activity． Moreo-
ver，they were not active against all the bacterial strains
employed in this study (Table 1)． However，N-［2-(3，
4-dihydroxyphenyl)-2-hydroxethyl］-3-(4-methoxyphe-
nyl)prop-2-enamide exhibited effective inhibition a-
gainst the fungal strain A． solani，for which the value of
MIC was 0． 2 mg /mL and IC50 was 0． 110 mg /mL．
Isorhamnetin-3-O-rutinoside exhibited the best inhibito-
ry activity against P． syringaw pv． tomato，X． vesicatoria
and A． solani． The MIC values were 0． 2，0． 2 and 0． 1
mg /mL，respectively and the IC50 were 0． 133，0． 105，
and 0． 055 mg /mL，respectively，which were stronger
352Vol. 27 WANG Ying，et al:Antibacterial and Antioxidant Properties of Compounds Extracted from Petrosimonia sibirica L．
than that of Streptomycin sulphate (0． 159 and 0． 185
mg /mL，respectively)．
In the antioxidant test，most of the compounds exhibited
powerful antioxidant potential． The values obtained by
the two testing methods were listed in Table 2 and Ta-
ble 3，respectively． 4-hydroxy-3-methoxy benzene etha-
nol was relatively more active than standard AA in
FＲAP assay，with FＲAP value of 9． 402 mmol /g． N-
［2-(3，4-dihydroxyphenyl)-2-hydroxyethyl］-3-(4-
meth oxyphenyl)prop-2-enamide was found to be the
most effective in DPPH method，with IC50 value of
0. 2452 mg /mL．
Conclusions
This study indicated that the compounds isolated from
P． sibirica can be exploited as ideal，inexpensive and
environmentally acceptable agrochemicals for future
plant disease management program． Moreover，these
compounds can also be used in the treatment of various
diseases caused by E． coli． The experimental presented
in this study provided a scientific support for the further
study of P． sibirica．
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