Đặng Xuân Cường và Đtg Tạp chí KHOA HỌC & CÔNG NGHỆ 111(11): 137 - 141

EVALUATION OF PHLOROTANNIN CONTENT WITH ANTIOXIDANT

ACTIVITIES OF BROWN ALGAE SARGASSUM COLLECTED IN
NHA TRANG BAY, VIET NAM

Dang Xuan Cuong1*, Tran Thi Thanh Van1

Vu Ngoc Boi2, Le Nhu Hau1
1
Nha Trang Institute of Technology Application and Researching, VAST
2
Nha Trang University

SUMMARY
Brown algae Sargassum has many bioactive subtances such as: phlorotannin, fucoidan, alginate,
laminaran,…. Bioactive subtances have bioactives such as: antioxidant, antibacterial, antifungal,
anticancer, anti HIV, radioprotective effect, antiallergic effect,…. Phlorotannin content with reducing
power and total antioxidant activity of 16 brown algae species Sargassum collected in Nha Trang bay,
Viet Nam were researched. These species were collected in the growth time. The result showed
phlorotannin/ polyphenol content was the highest in brown algae Sargassum swartzii. Reducing power
and total antioxidant activity were the highest in S. crassifolium species. Phlorotannin content and
antioxidant activities were the lowest in S. baccularia species. Reducing power activity was the stronger
than total antioxidant activity in 16 the researched brown algae species.
Key words: antioxidant, brown algae, Nha Trang, phlorotannin, Sargassum, reducing power

INTRODUCTION* had close relationship with skeleton molecule

Brown algae was natural resources with
abundance and diversification of bioactive
such as antioxidant, antibacterial, antifungal,
anticoagulant and antiradiation UV-B, wound
healing and cell structure regeneration [8].
Many recent research showed that antioxidant
activity from brown algae was one of
bioactives which was focused in the most
research on the world and this activity was
studied by [2]; [8]. It is interesting that most
of the studies on the antioxidant activity were
indicated a close relationship between
contents, structure and antioxidant
mechanisms of polyphenols / phlorotannin in
the brown seaweed.
The before research showed phlorotannins
were phenolic compounds with polymer
nature of phloroglucinol, identified in many
families of brown algae phylum, such as:
Alariaceae, Fucaceae and Sargassaceae.
Many research showed phenolic compounds
in brown algae was phlorotannin [8]; [18].
Strong antioxidant activity of purification
phlorotannin from some brown algae species
* living cell death in a synmatic way. Radicals
Email: cuong_mails@yahoo.com.vn
[2]. Simultaneous, high antioxidant
polyphenols had close relationship with
phenol ring and through electronic trap on
phenol ring to scavenge OH-, ROO-, O2-.
Phlorotannins in brown algae had 8 phenol
ring linked to each other. So, their free radical
scavenging activity was higher than
polyphenol of terrestrial plant, such as
catechins in green tea with 4 phenol ring [5].
Molecular size spectrum of phlorotannin was
very wide, from (400 - 400.000 Da) and
phlorotannins content in each seaweed
species was also different (0.5 - 20% dry
matter) [8].
Move over, free radical was a molecule that
lack of a electron, so that their electrolysis
was usually not equal, they trended to take
electrolysis from other molecule and took
form new free radicals, this thing caused
function disturbance of cell [1]. Free radical
effect on human health popular follow 3
pathway: adaptive with antioxidant system,
cause injury to life cell and kill living cell in
human that showed [3]. Inside, mechanism of
living cell killing could make necrosis or

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 Đặng Xuân Cường và Đtg Tạp chí KHOA HỌC & CÔNG NGHỆ
such as: superoxide, hydroxyl, peroxyl,
alkoxyl, hydroperoxyl, nitric oxide and
nitrogen dioxide were evaluated to be free
radical [3].
So, the paper researched on antioxidant
activities and phlorotannin content evaluation
of 16 brown algae species collected in Khanh
Hoa coast, Viet Nam. The data of this study
will contribute to promote the application of
antioxidants from seaweed into food and
pharmaceutical.
MATERIALS AND METHODS
Sample collection, classification and
preservation
16 brown algae species Sargassum were
collected in Nha Trang bay, Viet Nam. The
samples were classified [12] after collected,
the seaweed was cleaned by sea water, dried
to 19% moisture and preserved at room
temperature.
Sample preparation
Maceration extraction method was used in the
study. Extraction at room temperature, 24-
hour time with the ratio of solvent: material
(25:1). 100g dry weight (19% humidity) was
used for each experiment. The dry sample
was crushed and ground into a powder form
and passed through a 40 mesh sieve. After
extraction, the extract was filtered in the dark.
The collected extract was analysed
phlorotannin with antioxidant activity.
Determination of phlorotannin content
Total phlorotannin content
Total phlorotannin content (TPC) was
determined according to Folin-Ciocalteus’
method as described by [18], phloroglucinol
being the standard substance. 01 ml Folin-
Ciocalteus’ 10% was added to 300 µl extract
and the compound was kept for 5 minutes.
The compound was added with 2 ml Na2CO3
10% mixed and kept for 90 minutes in the
dark and finally the absorbance was measured
at 750 nm by the UV-Vis Spectrophotometer
JenWay 6400/ 6405.
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111(11): 137 - 141
Method of bioactive test
Total antioxidant activity
Total antioxidant (TA) activity was
determined following the method by [13].
900µl distilled water and 3 ml A solvent
(H2SO4 0.6 M, sodium phosphate 28 mM and
ammonium Molybdate 4 mM) were added to
100µl extract. The compound was kept for 90
minutes at 950C. Acid ascorbic was used as
the standard substance and the absorbance
was measured at 695 nm.
Reducing power activity
Reducing power activity was determined
according to the method of [20]. Firstly,
0.5ml phosphate buffer (pH = 7.2) was added
to 500µl extract. Secondly, 0.2 ml
K3[Fe(CN)6] 1% was added to the compound.
The compound was kept at 500C for 20
minutes. Thirdly, 500µl CCl3COOH 10%
with 300µl distilled water and 80µl FeCl3
0.1% were added. Finally, the compound was
measured at 655 nm with the FeSO4 standard
substance.
Data analysis
The experiments have been repeated 3 times
(n = 3), anova and regression were analysed
using Microsoft Excel 2010 software.
RESULTS AND DISCUSSION
Phlorotannin content
Figure 2 showed phlorotannin content
vacillate from 0.0497 ± 0.006 to 6.8940 ±
0.09mg/g dry algae and the average of 3.2014
± 0.03mg/g dry algae. Inside, S. baccularia
species got the lowest phlorotannin content,
phlorotannin content in S. swartzii was the
highest in the researched species.
Phlorotannin content in S. muticum species
was 6% dry algae [1]. Phlorotannin content in
S. polyceratium; S. angustifolium; S.
filipendula; S. cinereum; S. hystrix; S.
siliquosum and S. mcclurei species vacillate
from 0.006 - 0.65g PGE/100g the extract with
the extract ratio of 10:1 (v/w) [16].
Phlorotannin content of S. vulgare got 12.71
± 0.54mg acid gallic/g extract when it was
extracted at room temperature and got 12.66 ±
 Đặng Xuân Cường và Đtg Tạp chí KHOA HỌC & CÔNG NGHỆ
0.65mg acid gallic/g extract when it was
extracted using soxhlet method with the ratio
of 10:1 (v/w) that showed by [11]. These
things were performed clearly in the figure 2.
Fig 2. Phlorotannin content of 16 brown algae
species Sargassum
Sargassum pteropleuron and Sargassum
ramifolium species growth in the tropical
marine of Mexico have the phlorotannin
content of 0.76 ± 0.04 and 0.95 ± 0.10% dry
algae [10]. So phlorotannin content of brown
algae species Sargassum growth in Nha Trang
bay were the lower than some species
Sargassum and black tea in the world.
However, all phlorotannin/ polyphenol of all
plant in the world has phenol rings and the
electron trap on these rings.
Total antioxidant activity
The results showed TA activity of the studied
species vacillate from 0.1105 ± 0.003 to
56.021 ± 0.011mg acid ascorbic/g dry algae,
the average of 8.2213 ± 0.061mg acid
ascorbic/g dry algae. Inside, TA activity was
the lowest in S. baccularia species and
highest in S. crassifolium species. TA activity
of S. pallidum species vacillate from 0.04 ±
0.02 đến 52.08 ± 0.02µmol FeSO4/mg dry
algae [6]. These things were illustrated clearly
in the figure 3.
Fig 3. Total antioxidant activity of 16 brown
algae species Sargassum
111(11): 137 - 141
TA activity of the ethyl acetate fraction from
brown algae S. marginatum growth in India
correspond 39.62 mg ascorbic acid/g extract
that noticed by [4]. So brown algae species
growth in Nha Trang bay had the lower TA
activity than the over species, except S.
crassifolium species with the TA activity of
56.021mg/g dry algae. TA activity was
determined on reaction of extract with
phosphomolybdenum to eliminate Mo6+ to
Mo5+ and form the green solution in the acid
condition.
Reducing power activity
The RP activity of brown algae was showed
in the most researchs [15], [19]. RP activity
was from 0.105 – 0.505 µmol FeSO4/mg that
noticed by [6]. The absorption of RP activity
of five brown seaweed species S. fusiforme, S.
kjellmanianum, S. pallidum, S. thunbergii và
S. horneri correspond with 0.706 ± 0.074;
0.982 ± 0.081; 0.744 ± 0.064; 0.607 ± 0.011
and 0.748 ± 0.071 that noticed [9]. Amphiroa
sp., Halimeda macroloba, S. binderi and T.
conoides species had the RP activity of
69.204 ± 0.71; 14.323 ± 0.051 and 15.597 ±
0.072mg acid gallic/ 1g sample that noticed
[14]. The experiment results were illustrated
more clearly in the figure 4.
Fig 4. Reducing power activity of 16 brown algae
species Sargassum
RP activity of 16 studied brown algae species
vacillate from 0.4736 ± 0.005 to 110.028 ±
0.036mg FeSO4/g dry algae, inside RP
activity was the highest in S. crassifolium
species and the lowest in S. baccularia
species. This thing noticed that reducing
power of other algae species in the world is
very diversiform. The experiment showed RP
activity of extract which contain
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 Đặng Xuân Cường và Đtg Tạp chí KHOA HỌC & CÔNG NGHỆ
phlorotannin/ polyphenol from 16 studied
species was stronger than their TA activity.
Antioxidant activity of phlorotannin may be
due to the structure characteristic with 8
phenol rings to form the electron trap and
reducing power [5].
CONCLUSIONS
The research results of 16 brown algae
species collected in Nha Trang bay, Viet Nam
in the specific condition showed phlorotannin,
total antioxidant and reducing power activity
were the lowest in S. baccularia species,
correspond 0.0497 ± 0.006mg phloroglucinol,
0.1105 ± 0.003mg acid ascorbic and 0.4736 ±
0.005mg FeSO4 dry algae; phlorotannin
content was the highest in S. swartzii species,
corresponds 6.8940 ± 0.09mg
phloroglucinol/g dry algae; total antioxidant
and reducing power activity was the highest
in S. crassifolium species, correspond 56.021
± 0.011mg acid ascorbic, 110.028 ± 0.036mg
FeSO4/g dry algae.
Acknowledgements Support during this work
by the Viet Nam Ministry of Science and
Technology and Viet Nam Academy of
Science and Technology were acknowledged.
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TÓM TẮT
ĐÁNH GIÁ HÀM LƯỢNG PHLOROTANNIN VỚI HOẠT TÍNH CHỐNG OXY
HÓA CỦA RONG NÂU SARGASSUM THU Ở VỊNH NHA TRANG, VIỆT NAM

Đặng Xuân Cường1*, Vũ Ngọc Bội2

Trần Thị Thanh Vân1, Lê Như Hậu1
1
Viện Nghiên cứu và Ứng dụng Công nghệ Nha Trang, VAST
2
Đại học Nha Trang

Rong nâu Sargassum chứa rất nhiều hoạt chất sinh học như phlorotannin, fucoidan, alginate,
laminaran,…. Những hoạt chất này có hoạt tính sinh học như chống oxy hóa, kháng khuẩn, kháng
nấm, kháng ung thư, kháng HIV, chống dị ứng, kháng viêm…. Hàm lượng phlorotannin với hoạt
tính chống oxy hóa tổng và khử sắt của 16 loài rong nâu Sargassum thu ở vịnh Nha Trang, Việt
Nam đã được nghiên cứu. Những loài rong này được thu mẫu trong thời gian sinh trưởng. Kết quả
nghiên cứu cho thấy hàm lượng phlorotannin/ polyphenol trong rong nâu S. swartzii là cao nhất.
Hoạt tính khử sắt và chống oxy hóa tổng cao nhất ở loài S. crassifolium. Loài S. baccularia có hàm
lượng phlorotannin và hoạt tính chống oxy hóa là thấp nhất. Hoạt tính khử sắt mạnh mẽ hơn hoạt
tính chống oxy hóa tổng trong 16 loài được nghiên cứu.
Từ khóa. Chống oxy hóa, Nha Trang, khử sắt, phlorotannin, rong nâu, Sargassum.

Ngày nhận bài: 30/10/2013; Ngày phản biện: 08/11/2013; Ngày duyệt đăng: 15/11/2013
Phản biện khoa học: GS.TS Từ Quang Hiển – Đại học Thái Nguyên

*
Email: cuong_mails@yahoo.com.vn

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