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*Corresponding author: E-mail: xjg71@163.com; 
 
 
 

Asian Journal of Immunology 
 
3(1): 160-165, 2020; Article no.AJI.56639 
 

 
 

 

 

Polyphenol and Flavonoids Content and  
Antioxidant Activity of Different Solvent  

Extracts from Artemisia argyi 
 

Dong-Qin Wang1,2, Jing Yang2 and Jianguo Xu1,2* 
 

1
Modern College of Humanities and Sciences, Shanxi Normal University, Linfen 041004, China. 

2
School of Food Sciences, Shanxi Normal University, Linfen 041004, China. 

 
Authors’ contributions  

 
This work was carried out in collaboration among all authors. Author DQW designed the study, 
performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript. 

Authors JY and JX managed the analyses of the study. Author JX managed the literature searches. 
All authors read and approved the final manuscript. 

 
Article Information 

 
Editor(s): 

(1) Dr. Wagner Loyola, Brazilian Agricultural Research Corporation (Embrapa), Brazil. 
Reviewers: 

(1) Douati Togba Etienne, Université Felix Houphouet Boigny, Côte d’Ivoire. 
(2) Yves Latune, University of Abidjan, Côte d’Ivoire. 

Complete Peer review History: http://www.sdiarticle4.com/review-history/56639 

 
 
 
 

Received 27 February 2020 
Accepted 04 May 2020 

Published 11 May 2020 

 
 

ABSTRACT 
 

The effects of extraction solvents (methanol, ethanol, acetone, petroleum ether and n-hexane) on 
the content of total polyphenols and flavonoids, as well as antioxidant activities of Artemisia argyi 
were investigated. The results showed that, the ethanol extract had the highest total polyphenols 
and flavonoids content, and it exhibited stronger antioxidant activities, followed by methanol and 
acetone extracts. Correlation analysis revealed that the content of phytochemicals was well 
correlated with antioxidant activities of extracts from Artemisia argyi, which indicates that different 
solvents had a great influence on the level of total polyphenols, flavonoids and antioxidant activities 
of extracts. Therefore, selective extraction from Artemisia argyi, by an appropriate solvent, is 
important for obtaining fractions with high antioxidant activity, which will be useful for the 
developing and application of Artemisia argyi. 
 

 
Keywords: Artemisia argyi; extraction solvent; polyphenols; flavonoid; antioxidant activity. 

Short Communication 



 
 
 
 

Wang et al.; AJI, 3(1): 160-165, 2020; Article no.AJI.56639 
 

 

 
161 

 

1. INTRODUCTION  
 
Artemisia argyi (A. argyi) is known in Chinese as 
“Aicao” and in Japanese as “Gaiyou” and 
belongs to the Arteraceae family that comprises 
over 500 species [1], which is mainly distributed 
in Asia, Europe and North America [2]. As a 
traditional medicinal and edible plant, ancient 
Chinese commonly pick the buds and leafs of A. 
argyi consumed as a food supplement or a 
flavoring and colorant for the Chinese dish 
Qingtuan [3]. A. argyi also contains many active 
compounds such as volatile oil [1], flavonoids [4], 
phenols, organic acids [4], polysaccharides [5], 
terpenoids [6] and glycosides [4,7-9]. Several 
Studies have reported that A. argyi extracts 
possess various biological activ ities, such as 
antioxidant [8], anti-inflammatory [10], 
antibacterial, anti-tumor [11], hemostatic, anti-
hypertensive effects, immunoregulatory, anti-
asthmatic, oncogene inhibitory and anti-
osteoporotic activities [7,8,12,13]. Extract 
solvents is one of the most important factors 
affecting the chemical composition and biological 
activity of plant extracts [14,15]. Therefore, the 
objective of this work was to investigate the 
content of total polyphenols, flavonoids and 
antioxidant activity of different solvent extracts 
(methanol, ethanol, acetone, petroleum ether, n-
hexane) from Artemisia argyi. 
 

2. MATERIALS AND METHODS 
 

2.1 Preparation of Extracts 
 
Fresh leaves of artemisiae argyi were collected 
around the fifth day of the fifth lunar month in 
2019 from Linfen (Shanxi Province) and were 
dried at 40°C. The dried leaves (100 g) were 
grounded and were extracted with 250 mL 
solvents (methanol, ethanol, acetone, petroleum 
ether, n-hexane) and kept in a shaker at 25°C for 
1 h. And then the mixture was centrifuged at 
4000 rpm/min for 15 min at 4°C. The precipitation 
was extracted with 250 mL solvent once again 
and mixed supernatants. The extracts were dried 
under vacuum and stored at 4°C until analysis.  
 

2.2 Determination of Polyphenolic 
Content 

 

Polyphenolic content of extracts was determined 
according to method described by Bettaieb et al. 
with some modifications [14]. Briefly, an aliquot 
(0.1 mL) extract was mixed with 2.8 mL of 
distilled water and 0.1 mL of Folin-Ciocalteu 
reagent (1.0 mol/L), and then the solution was 

mixed and incubated at room temperature (25°C) 
for eight minutes. Following that, 2 mL of 7.5% 
sodium carbonate (Na2CO3) solution was added 
and shaken thoroughly. The mixture was 
incubated for 2 h in the dark at room temperature 
(25°C) and the absorbance was determined at 
765 nm. Gallic acid was used for calibration of 
the standard curve and total phenolic content 
was expressed as milligram gallic acid equivalent 
per gram dried weight. 
 

2.3 Determination of Flavonoid Content 
 
An aliquot (1 mL) extract was mixed with 0.3 mL 
of 3% NaNO3 solution and incubated for 6 min. 
After that, 0.3 mL of 10% Al(NO3)3 was added 
and the solution was kept at room temperature 
(25°C) for 6 min. Finally, mix the solution with 4 
mL of 4% NaOH solution and add water to 10 mL. 
After 20 minute of incubation, the absorbance of 
the mixture at 510 nm was measured. Rutin was 
used for calibration of the standard curve, and 
flavonoid content was expressed as milligram 
rutin equivalents per gram dried weight. 

 
2.4 DPPH Radical Scavenging Assay 
 
DPPH radical scavenging assay was measured 
the method described by Xu et al. with some 
modifications [16]. Extract was serially diluted to 
different concentrations and 0.5 mL of diluted 
extract mixed with 2.5 mL of 60 μmol/L DPPH 
solution dissolving in methanol. The mixture was 
shaken thoroughly and incubated in the dark at 
room temperature (25°C) for 30 min, and the 
absorbance was measured at 517 nm. Trolox 
was used for calibration of the standard curve 
and the DPPH radical scavenging activity of 
sample was expressed as milligram Trolox 
equivalent per gram dried weight (mg Trolox/g). 
 

2.5 ABTS Radical Scavenging Assay 
 
ABTS radical scavenging assay was determined 
according to the method described by Xu et al 
[16]. Briefly, a certain quality of ABTS and 
potassium persulfate was dissolved in water to 
keep the final concentrations of the two 
substances to be 7 mmol/L and 2.45 mmol/L 
respectively. The mixture was kept in the dark for 
16~24 h to make the ABTS radical working 
solution and its absorbance at 734 nm was 
adjusted to 0.700±0.050. The ABTS radical 
scavenging ability was measured by adding 50 
μL of diluted extract to 1.9 mL of ABTS radical 
working solution and the absorbance at 734 nm 
after 6 min was recorded. Trolox was used for 

app:ds:rutin
app:ds:rutin
app:ds:potassium
app:ds:persulfate


 
 
 
 

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162 

 

calibration of the standard curve and the ABTS 
radical scavenging activity of sample was 
expressed as milligram Trolox equivalent per 
gram dried weight (mg Trolox/g). 
 

2.6 Ferric Reducing Antioxidant Power 
(FRAP) Assay 

 

The reducing ability was determined as 
described by Xu et al [16]. Briefly, the FRAP 
reagent was freshly prepared from 300 mM 
sodium acetate buffer (pH 3.6), 10 mM 2,4,6-
Tri(2-pyridyl)-s-triazine (TPTZ) solution in 40 mM 
HCl and 20 mM FeCl3 solution in proportions of 
10:1:1 (v/v), respectively. The FRAP reagent was 
prepared fresh daily and was warmed to 37

o
C in 

a water bath prior to use. Then 0.1 mL sample 
solution was mixed with 1.8 mL FRAP reagent 
and 3.1 mL ultra-pure water. The absorption of 
the reaction mixture was measured at 593 nm 
after incubation for 30 min at 37

o
C. The standard 

curve was constructed using FeSO4 solution 
(100-1000 μM), and FRAP value was expressed 
as micromoles Fe(II) per gram DW.  
 

3. RESULTS AND DISCUSSION 
 

3.1 Contents of Total Polyphenols and 
Flavonoids 

 

The contents of phytochemicals of different 
solvent extracts were showed in Fig. 1. 
Depending on the solvent used, the total 
polyphenol content ranged from 6.58 to 25.6 mg 
GAE/g. Ethanol extract had the highest 
polyphenol content, followed by methanol, 
acetone, the lowest for petroleum ether and n-
hexane. However, no significant difference in 
total phenolic content was found between 
Petroleum ether and n-hexane extracts. The 
range of flavonoid content in different solvent 
extracts was 12.6-40.2 mg Re/g and their order 
corresponded to the same order in the 
polyphenol content. Generally speaking, 
differences in the content of total polyphenols 
and flavonoids from different solvent extracts 
may be come from differences in the polarity of 
solvents. However, there was not a one-to-one 
relationship between the polarity of solvent and 
the content of phytochemicals. These differences 
in the content of total phenolics and flavonoids 
from different extracts may be come from 
differences in the polarity, chemical structure, 
dielectric constant of solvents [17], which can 
influence the extraction efficiency and 
compositions of extracts. 

 

3.2 DPPH and ABTS Radicals Scavenging 
Abilities of Different Extracts 

 
Due to the differences in the compositions and 
contents of the phytochemicals, the extracts 
showed the different antioxidant abilities. Table 2 
showed that there were significant differences of 
DPPH radical scavenging abilities of different 
solvent extracts. Consistent with the results in 
phytochemical test, ethanol extract owned the 
highest DPPH radical scavenging ability, 
followed by methanol and acetone extracts. 
Petroleum ether and n-hexane extracts had 
lowest DPPH radical scavenging ability, of which 
were 0.63 and 0.86 mg Trolox/g respectively. But 
there was no significant difference between 
petroleum ether and n-hexane extracts. The 
results of ABTS radical scavenging abilities were 
similar to that of DPPH assay. The difference 
was that there was no significant differences 
between methanol and ethanol extracts. 

 
3.3 Ferric Reducing Antioxidant Power 

(FRAP) 
 
Table 1 also showed that different extracts from 
Artemisia argyi exhibited different reducing 
power. There was a significant difference (p < 
0.05) in the reducing power among different 
extracts, and ranged from 16.7 to 55.6 μmol 
Fe(II)/g DW for n-hexane, petroleum ether, 
acetone, ethanol and methanol extracts, 
respectively. As observed in the DPPH and 
ABTS, the reducing power of methanol extracts 
was the highest, followed by ethanol, acetone, 
and petroleum ether extracts, the lowest for 
hexane extracts. The results suggested that 
extracts from Artemisia argyi had a potency to 
donate electron to reactive free radicals, 
converting them into more stable non-reactive 
species and terminating the free radical chain 
reaction [16]. 
 

3.4 Correlation among Total Phenolics, 
Flavonoids and Antioxidant Activities 

 
To further investigate their interrelationship, the 
correlation between the active compounds and 
biological activities were established, and 
correlation coefficients (R) are shown in Table 2. 
In this study, the DPPH and ABTS were highly 
and positively correlated to the content of both 
total phenolics and flavonoids (R ≥ 0.893), 
indicating total phenolics and flavonoids are the 
main constituents contributing to the antioxidant



 
 
 
 

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163 

 

 
 

Fig. 1. The contents of total phenolics and flavonoids of extracts from Artemisia argyi 
 

Table 1. The antioxidant activities of different extracts from Artemisia argyi 
 

 DPPH (mg Trolox/g) ABTS (mg Trolox /g) FRAP (μmol Fe(II)/g) 

Petroleum ether 0.63±0.06d 2.88±0.63c 18.6±1.3d 

n-Hexane 0.86±0.16d 2.55±0.52c 16.7±1.4 d 

Acetone 2.76±0.42c 6.84±0.73 b 31.5±1.9 c 

Ethanol 6.93±0.44a 14.25±1.42a 48.5±2.2 b 

Methanol 4.35±0.52b 12.72±1.57 a 55.6±3.1 a 

 
Table 2. Correlation analysis of polyphenols, flavonoids and antioxidant activities 

 

 Phenolics Flavonoids Antioxidant activities 

 DPPH  ABTS  FRAP 

Phenolics 1     

Flavonoids 0.964
*
 1    

DPPH 0.992
**
 0.988

*
 1   

ABTS 0.930 0.893 0.935 1  

FRAP 0.557 0.501 0.568 0.821 1 
Values are correlation coefficient R. 

a
 Significantly different: 

**
 p < 0.01,

 *
 p < 0.05 

 
and antibacterial activities of extracts, which was 
supported by previous reports studied on some 
cereals or plants [18,19]. However, FRAP was 
moderately correlated to the content of phenolics 
and flavonoids. 
 

4. CONCLUSIONS 
 
Plant polyphenols and flavonoids have been 
considered as one of the most widely distributed 
secondary metabolites which contains various 
complicated compounds. Based on the principle 
of the dissolution in the similar material structure, 
different solvent extracts differ in its active 
constituents as well as their contents due to the 

different polarity and solubility of extracting 
solvents. Our findings indicated that ethanol 
extract had the highest content of polyphenol and 
flavonoid among all of five solvent extracts. 
Consistent with results described above, ethanol 
extract showed the highest antioxidant abilities in 
DPPH, ABTS radicals scavenging and FRAP 
assays. These results indicated that selective 
extraction from Artemisia argyi, by an appropriate 
solvent, is important for the development and 
utilization of Artemisia argyi. 
 

CONSENT 
 

It is not applicable. 

d 

d 

c 

b 

a 

b 

d 

c d 

a 

0 

10 

20 

30 

40 

50 

0 

10 

20 

30 

40 

50 

petroleum ether hexane acetone ethanol methanol 

F
la

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o
n

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id

s 
(m

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 R

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) 

P
o
ly

p
h

en
o

ls
 (

m
g
 G

A
E

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g
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W
) 

Extracts 

polyphenols 

flavonoids 



 
 
 
 

Wang et al.; AJI, 3(1): 160-165, 2020; Article no.AJI.56639 
 

 

 
164 

 

ETHICAL APPROVAL 
 

It is not applicable. 
 

ACKNOWLEDGEMENTS 
 

This work was financially supported by a project 
of the Natural Science Foundation of Modern 
College of Humanities and Sciences of Shanxi 
Normal University, China (project no. 
2019JCYJ18). 
 

COMPETING INTERESTS 
 

Authors have declared that no competing 
interests exist. 
 

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