The document summarizes a study on the antioxidant effects of water and ethanol extracts of Andrographis paniculata leaves. Total phenolic content, total flavonoid content, ferric reducing antioxidant power, and DPPH radical scavenging activity were measured to evaluate the antioxidant activities of the extracts. The ethanol extract contained higher levels of total phenolics and flavonoids than the water extract. Both extracts showed significant antioxidant effects in the ferric reducing and DPPH assays, with the ethanol extract demonstrating stronger activity.

1. An#oxidant
Effect
of
Water
and
Ethanol
(80%)
Extracts
of
Andrographis
paniculata
(Burm.
f.)
Nees.
leaves
Presented
by
Sekar
Galuh
(Indonesia)
Supervised
by
Assoc.
Prof.
Dr.
Jamia
Azdina
Jamal
Kuala
Lumpur,
18th
August
2016

2. 2
INTRODUCTION
Andrographis
paniculata
Ness.
(King
of
BiHer/Brotowali
(indo)/Hempedu
Bumi
(malay)
• Major
compounds
:
• Diterpenoid
(Andrographolide)
• Flavonoid
• Polyphenols
(Chao
and
Lin,
2010)
“Compounds
in
plant
extracts,
herbs,
and
syntheVc
showed
the
an#oxidant
ac#vity”
(Brewer,
2011)
“Flavonoid
showed
many
biological
acVviVes,
such
as
anVallergenic
and
anVinflammatory.
An#oxidant
capacity
of
flavonoid
in
vitro
has
been
studied.”
(PieHa,
2000)
“Polyphenols,
as
secondary
metabolites
of
plants,
is
involved
against
ultraviolet
radia#on”
(Pandey
and
Rizvi,
2009)

3. 3
Objective
To
quanVfy
Phenolic
and
Flavonoid
contents
in
water
extract
and
ethanol
80%
extract
of
Andrographis
paniculata
leaves
The
aim
of
this
research
are
To
evaluate
anVoxidant
acVvity
of
water
extract
and
ethanol
80%
extract
of
Andrographis
paniculata
leaves

4. 4
Methodology
Water
and
Ethanol
80%
Extracts
QuanVfy
Phenolic
and
Flavonoid
Contents
Evaluate
AnVoxidant
AcVviVes
Total
Phenolic
Content
(TPC)
Total
Flavonoid
Content
(TFC)
Ferric
Reducing
AnVoxidant
Power
(FRAP)
Diphenyl-­‐
picrylhidrazil
(DPPH)
Analysis
(using
Microsod
Excel)
mg
Gallic
Acid
Equivalent/g
mg
QuerceVn
Equivalent/g
mg
Trolox
Equivalent/g
RSA;
AAI;
IC50

5. 5
Methodology
Total
Phenolic
Assays
(TPC)
(ISO
14502,2005)
20
μL
sample
(1mg/mL)/
standard
(Gallic
acid)/DMSO
100
μL
Folin-­‐
Ciocalteu
(10%)
IncubaVon
5’
96-­‐well
plates
80
μL
Na2CO3
(75%)
Slightly
shaken
IncubaVon
30min
(darkness
at
RT)
Absorbance
read
at
735nm
using
microplate
reader
Total
Flavonoid
Assays
(TFC)
(Yang
et
al,
2011)
100
μL
sample/
standard
(QuerceVn)/
DMSO
100
μL
AlCl3
(2%)
IncubaVon
15’
96-­‐well
plates
Absorbance
read
at
435nm
using
microplate
reader

6. 6
Methodology
Ferric
Reducing
An#oxidant
Power
Ac#vity
(FRAP)
(Benzie
&
Strain,
1996;
Yang
et
al,
2011)
20
μL
sample
(8-­‐0.0156mg/
mL)/standard
(Trolox;
Ascorbic
Acid)/
DMSO
100
μL
FRAP
reagent
shake
30sec
96-­‐well
plates
IncubaV
on
4
min
37°C
Absorbance
read
at
593
nm
using
microplate
reader
Diphenyl-­‐picrylhidrazyl
Assay
(DPPH)
(Zongo
et
al,
2010)
100
μL
sample
(8-­‐0.0156mg/
mL)/standard
(Ascorbic
Acid)/
DMSO
100
μL
DPPH
SoluVon
reagent
IncubaVon
15min
at
RT
(dark
room)
96-­‐well
plates
Absorbance
read
at
540
nm
using
microplate
reader

7. 7
Results
y
=
0.0049x
+
0.0338
R²
=
0.9955
0.0000
0.1000
0.2000
0.3000
0.4000
0.5000
0.6000
0
20
40
60
80
100
Absorbance
Concentra#on
(ug/mL)
Standard
Curve
TPC
Series1
Linear
(Series1)
No.
Type
of
Extracts
Concentra#on
of
Phenolic
(μg/mL)
Total
Phenolic
Contents
(mg
GAE/g)
1.
Water
extract
12.4097
1979.2175
±
239.19
2.
Ethanol
(80%)
extract
19.9066
3132.4274
±
72.46
TOTAL PHENOLIC CONTENT (TPC)

8. 8
Results
y
=
0.0208x
-­‐
0.1725
R²
=
0.99284
0.0000
0.5000
1.0000
1.5000
2.0000
2.5000
0
20
40
60
80
100
Absorbance
Concentra#on
(ug/mL)
Standard
Curve
TFC
Series1
Linear
(Series1)
TOTAL FLAVONOID CONTENT (TFC)
No.
Type
of
Extracts
Concentra#on
of
Flavonoid
(μg/mL)
Total
Flavonoid
Contents
(mg
QE/g)
1.
Water
extract
13.8880
1269.4681
±
50.36
2.
Ethanol
(80%)
extract
25.0878
7895.4420
±
44.09

9. 9
Results
FERRIC REDUCING ANTIOXIDANT ASSAYS (FRAP)
No.
Serial
Dilu#on
(mg/mL)
Equivalent
Trolox
(mg
TE/g)
Water
extract
Ethanol
(80%)
Extract
1.
0.5
3293.0087
3686.2979
2.
1
8795.7385
7259.0896
3.
2
15174.6569
12936.0615
4.
3
33478.3691
25521.3156
5.
8
63305.2887
49297.8875
y
=
0.0106x
-­‐
0.0459
R²
=
0.99987
0.0000
0.2000
0.4000
0.6000
0.8000
1.0000
1.2000
0
50
100
150
Absorbance
Concentra#on
(ug/mL)
Standard
Curve
Trolox
Series1
Linear
(Series1)

10. 10
Results
DIPHENYL-PICRYLHIDRAZYL (DPPH) ASSAY
No
Serial
Dilu#on
(mg/mL)
Radical
Scavenging
Ac#vity
(RSA)
An#oxidant
Ac#vity
Index
(AAI)
IC50
Water
extract
Ethanolic
Extract
Water
extract
Ethanolic
Extract
Water
extract
Ethanolic
Extract
1.
0.0625
44.9251
45.3432
34.13481
40.04599
2.9287
±0.08
2.4967
±0.013
2.
0.125
46.1005
45.5141
3.
0.25
46.1400
46.2083
4.
0.5
46.4871
46.1321
5.
1
47.7020
46.9420
6.
2
49.5138
49.7952
7.
3
53.0980
52.7456
8.
8
55.8564
59.9350
Standard
Curve
Ascorbic
Acid
y
=
-­‐0.0044x
+
1.0858
R²
=
0.99841

11. 11
Discussions
• Ethanol
(80%)
extract
contains
more
phenolic
and
flavonoid
compounds
than
water
extract
• Its
an#oxidant
ac#vity
is
also
higher
than
water
extract
• There
is
a
linear
relaVonship
between
natural
compound
and
anVoxidant
acVvity
• The
higher
phenolic
and
flavonoid
exhibited
in
plants,
the
higher
its
anVoxidant
acVvity
• Based
on
(Qader,
2011),
Aqueos
extract
of
Andrographis
paniculata
has
more
phenolic
and
higher
anVoxidant
capacity
• They
might
use
different
solvent
• Phenolic
and
Flavonoid
more
soluble
in
ethanol
because
its
polarity
lower
than
water

12. 12
Conclusions
Ethanol
(80%)
extract
of
Andrographis
paniculata
leaves
contains
higher
phenolic
and
flavonoid
Ethanol
(80%)
extract
of
Andrographis
paniculata
leaves
has
stronger
anVoxidant
acVvity

13. 13
Acknowledgement
1. Eldiza
Puji
Rahmi
2. Ms.
KarVniwaV
Muhammad
3. Nor-­‐Ashila
Aladdin
4. Zalita
Yaacob
5. Faculty
of
Pharmacy
UKM
6. SEP
CommiHee
MyPSA

14. 14
References
Benzie,
I.F.F.
&
Strain,
J.J.
1996.
The
ferric
reducing
ability
of
plasma
(FRAP)
as
a
measure
of
“anVoxidant
power”:
the
FRAP
assay.
AnalyVcal
Biochemistry
239:
70-­‐76.
Brewer,
MS.,
2011,
Natural
AnVoxidants:
Sources,
Compounds,
Mechanisms
of
AcVon,
and
PotenVal
ApplicaVons,
Comprehensive
Reviews
in
Food
Science
and
Food
Safety,
10
(4):221-­‐247
Chao,
Wen-­‐Wan.,
Lin,
Bi-­‐Fong.,
2010,
IsolaVon
and
idenVficaVon
of
bioacVve
compounds
in
Andrographis
paniculata
(Chuanxinlian),
Chinese
Medicine,
5:17
Pandey
KB.,
Rizvi
SI.,
2009,
Plant
polyphenols
as
dietary
anVoxidants
in
human
health
and
disease,
Oxid
Med
Cell
Longev,
2(5):
270-­‐278
PieHa,
PG.,
2000,
Flavonoid
as
AnVoxidants,
Journal
Natural
Products,
63(7):1035-­‐42
Qader,
SW.,
Abdulla,
MA.,
Chua,
LS.,
Najim,
N.,
Zain,
MM.,
Hamdan,
S.,
2011,
AnVoxidant,
Total
Phenolic
Content
and
Cytotoxicity
EvaluaVon
of
Selected
Malaysian
Plants,
Molecule,
16:3433-­‐3443
Yang,
H.,
Dong,
Y.,
Du,
H.,
Shi,
H.,
Peng,
Y.
&
Li,
X.
2011.
AnVoxidant
compounds
from
propolis
collected
in
Anhui,
China.
Molecules
16:
3444-­‐3455.
Zongo,
C.,
Savadogo,
A.,
OuaHara,
L.,
Bassole,
I.H.N.,
OuaHara,
C.A.T.,
2010.
Polyphenols
content,
anVoxidant,
and
anVmicrobial
acVviVes
of
Ampelocissus
granVi
(Baker)
Planch.
(Vitaceae):
a
medicinal
plant
from
Burkina
Faso.
InternaVonal
Journal
of
Pharmacology
6(6),
880-­‐887.

15. 15
THANK YOU
COPYRIGHT
©
2016