According to the Ayurvedic literature Launaea pinnatifida Cass is belong to the class of controversial drug Gohjiva. This plant is well known and valuable herb as per the traditional and Ethnobotanical information. Leaves of this plant has been used since ancient time as herbal remedy for jaundice, diuretic, blood purifier and hepatoprotective action by the tribal people of the Western Ghats. However, the plant remains largely unexplored. The antioxidant capacity of the various fraction of L. Pinnatifida leaves and roots extracts, obtained by successive solvent extraction with various polarity of solvents namely Pet. Ether, Chloroform, Methanol and Water, was evaluated by means of different In vitro methods; DPPH radical scavenging activity (IC50 value for standard ascorbic acid and L. pinnatifida leaves methanol fraction was 165.8 and 220.3 respectively), Ferric ion reducing antioxidant power assay (FRAP) and total antioxidant capacity (TAC) by Phosphomolybdenum method. Those findings means that L. Pinnatifida leaves methanol fraction was most active fraction for free radical scavenging activity compared to all other fractions hence consider as a most potent fraction for the isolation of phytochemical also used in a food industry for protection against oxidative damage. Antioxidant results also support the traditional claim of this plant. The strongest antioxidant activity of methanol fractions of leaves could be due to the presence of flavonoids and phenols.

1. Antioxidant potential of various fractions of
Launaea pinnatifida leaves and roots extracts
by In vitro methods
PRESENTED BY :
HIMANSHU MAKWANA
Ph. D. Scholar,
School of Pharmacy, RK University
Email: himanshu23makwana@gmail.com
1
GUIDE:
DR.DEVANG PANDYA
Dy. Director,
School of Pharmacy, RK University
Email: director.sop@rku.ac.in
NATIONAL CONFERENCE ON
RECENT INNOVATIONS IN SCIENCE NCRIS-2019

2. 2
TABLE
OF
CONTENT
BACKGROUND01
02
03
04
05
06
07
AIM & OBJECTIVES
MATERIAL & METHODS
RESULTS & DISCUSSION
CONCLUSION
FUTURE SCOPE OF WORK
REFERENCES

3. 3
BACKGROUND: SANDIGDHA DRAVAYS
3
As per the Ayurvedic literature, “Sandigdha
dravays” is a word used for the medicinal
plants whose correct botanical identity has
not yet been ascertained.
Why controversy..??????
Use of different species in different part of
country.

4. 44
RATIONALE: GOJIHVA
Mainly 6 plants are considered as Gojihva[1,2]
ELEPHANTOPUS SCABER 1
ONOSMA BRACTEATUM 2
LAUNAEA PINNATIFIDA 3
ANCHUSA STRIGOSA 4
COCCINIA GLAUCA 5
MACROTOMIA BENTHAMI 6

5. 55
अथ गोजिह्वा । तस्या नामानन गुणााँश्चाह
गोजिह्वा गोजिका गोभी दार्विका खरपणिनी । गोजिह्वा वातला शीता ग्राहििी कफर्पत्तनुत् ॥२९७॥
हृद्या प्रमेिकासास्रव्रिज्वरिरी लघुुः । कोमला तुवरा ततक्ता स्वादुपाकरसा स्मृता ॥२९८॥ भा.प्र
Gojihva mainly used in the treatment of cough, cold, fever heart and blood related
problems. Decoction of the plant is useful for the brain disorders and also work as a
memory enhancer.
Name and traditional information about herbs is already given in the Ayurvedic
literature but not as per the botanical source of the plant. Still many a time herbs is
used as per therapeutic indication in the literature.
Now a days it is necessary to find the correct plant as per Ayurvedic literature hence
Modern standardization techniques is required for such work.
RATIONALE: GOJIHVA

6. 6
The present investigation is undertaken by utilizing the plant Launaea pinnatifida with
following objectives.
AIM & OBJECTIVES
To study the Antioxidant potential of various
fractions of Launaea pinnatifida leaves and
roots extracts by means of In vitro methods
2
1
To study the Antioxidant effect
of different fraction of Launaea
pinnatifida
To highlight the Importance of
Gojihva
3
Differentiate Launaea pinnatifida
plant from other species of Gojihva
on the basis of antioxidant study

7. 7
LAUNAEA PINNATIFIDA
Launaea Pinnatifida also recognized as Launaea sarmentosa (Willd.) belong to family
Asteraceae (Compositae).It has been traditionally used as a folk herbal remedy for the
many disease[4].
All the extract fractions where prepared using successive solvent extraction technique
with increasing polarity of solvents.
Figure 1. Leaves and Roots of L. pinnatifida Figure 3. Flower of L. pinnatifidaFigure 2. Whole plant
MATERIAL & METHODS

8. 8
The plant Launaea pinnatifida was collected in month of Jan-Feb 2017 from Nagoa beach,
Diu (A costal range of Gujarat). The plant was authenticated by Dr. Hitesh Solanki,
Professor, Department of Botany, School of science, Gujarat university, Ahmedabad. A
voucher specimen (HM1) of the same has been deposited in the museum of the
Department of Botany, School of science, Gujarat University, Ahmedabad, Gujarat, India
for future reference.
Figure 4. Root of L. pinnatifida Figure 5. Freshly preserve
sample of L. pinnatifida
Figure 6. Plant at the collection site of collection
near Nagoa beach (Diu)
SOURCE OF PLANTS
MATERIAL & METHODS

9. 9
FORMATION OF FREE RADICALS

10. 10
ANTIOXIDANTS: WHY ARE THEY IMPORTANT?[1,2,3]
• Antioxidant is a molecule that inhibits the oxidation
of the other molecules.
• Antioxidants may help to protect cells from damage
caused by free radicals.
• Naturally occurring antioxidants include flavonoids,
tannins, phenols and lignans.
• Plant-based foods are the best sources. These include
fruits, vegetables and their juices, whole-grain
products, nuts, seeds, herbs and spices, and even
cocoa.

11. 11
All chemicals and solvents were of analytical grade and were procured from Finar
Chemicals Private Limited, Ahmedabad, India. 1, 1-Diphenyl-2-picryl-hydrazyl (DPPH), was
procured from Sigma Chemicals, USA. The other chemicals used were ascorbic acid, ferric
chloride, Ammonium molybdate tetrahydrate extra pure, sodium phosphate and
Dimethyl sulphoxide(DMSO) were of analytical grade.
REAGENTS AND CHEMICALS
MATERIAL & METHODS

12. 12
RESULTS: IN VITRO ANTIOXIDANT SCREENING
A. Ferric ion reducing antioxidant power assay (FRAP) [5]
y = 0.0363x + 0.1388
R² = 0.9951
0
0.5
1
1.5
2
2.5
0 10 20 30 40 50 60
Absorbance(700nm)
Concentration (µg/ml)
CALIBRATION CURVE FOR
ASCORBIC ACID
Calibration curve
for Ascorbic acid
Linear (Calibration
curve for Ascorbic
acid)
PLANT EXTRACT
FRACTIONS
ABSORBANCE
AT 700 NM
% FREE RADICAL
SCAVENGING
LPL Pet. Ether extract 0.024 0.003
LPL chloroform extract 0.213 0.218
LPL methanol extract 1.195 2.910
LPL water extract 1.295 3.184
LPR Pet. Ether extract 0.390 0.703
LPR chloroform extract 0.309 0.481
LPR methanol extract 0.991 2.350
LPR water extract 0.662 1.440
0.000
0.500
1.000
1.500
2.000
2.500
3.000
3.500
LPL Pet.
ether
extract
LPL
chloroform
extract
LPL
methanol
extract
LPL water
extract
LPR Pet.
ether
extract
LPR
chloroform
extract
LPR
methanol
extract
LPR water
extract
%FREERADICALSCAVENGING
Figure 7. FRAP Assay With Samples
Graph 1: FRAP Assay of L. pinnatifida fractions
Table 1: FRAP Assay of L. pinnatifida leaves and roots fractions

13. 13
RESULTS: IN VITRO ANTIOXIDANT SCREENING
B. Evaluation of total antioxidant capacity (TAC) by Phosphomolybdenum method[6]
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
%Scavenging
SR. NO. PLANT EXTRACT FRACTIONS
ABSORBANCE AT 695
NM
% FREE RADICAL SCAVENGING
1 LPL Pet. Ether extract 0.008 1.48
2 LPL chloroform extract 0.15 2.51
3 LPL methanol extract 0.119 3.78
4 LPL water extract 0.154 4.61
5 LPR Pet. Ether extract 0.034 1.76
6 LPR chloroform extract 0.118 3.75
7 LPR methanol extract 0.129 4.01
8 LPR water extract 0.124 3.89
Graph 2: Total antioxidant capacity (TAC) of L. pinnatifida fractions
Figure 8. : Total antioxidant capacity (TAC) with
samples
Table 2: TAC assay of Launaea pinnatifida leaves and roots most active fractions along with standard drug L-ascorbic acid.

14. 14
RESULTS: IN VITRO ANTIOXIDANT SCREENING
C. DPPH radical scavenging activity [7]
Principle:
The antioxidant reacts with stable free radical, DPPH and converts it to 1, 1- diphenyl-2-
picryl hydrazine. The ability to scavenge the free radical, DPPH was measured in the
absorbance at 517 nm.
SR NO. SAMPLE IC50 VALUE
1 STANDARD 165.8
2 LPLM 220.3
3 LPLW 306.6
4 LPRM 328.5
5 LPRW 348.6
Table 3: IC50 value of Launaea pinnatifida leaves and roots most active
fractions (LPLM, LPLW, LPRM AND LPRW) along with standard drug L-
ascorbic acid.
Figure 8. : Principle of DPPH assay

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RESULTS: IN VITRO ANTIOXIDANT SCREENING
y = 0.2433x + 40.806
R² = 0.5654
y = 0.1879x + 41.716
R² = 0.5472
y = 0.1322x + 40.986
R² = 0.4967
y = 0.1488x + 49.132
R² = 0.6161
y = 0.1398x + 48.963
R² = 0.6088
0.00
20.00
40.00
60.00
80.00
100.00
120.00
140.00
0 100 200 300 400
%SCAVENGING
CONCENTRATION (µg/ml)
In vitro antioxidant studies
STANDARD
LPLM
LPLW
LPRM
LPRW
Linear
(STANDARD)
Linear (LPLM)
Linear (LPLW)
Linear (LPRM)
Linear (LPRW)
Graph 3: % scavenging of Launaea pinnatifida leaves and
roots most active fractions (LPLM, LPLW, LPRM AND
LPRW) along with standard drug L-ascorbic acid.
SR NO. SAMPLE ABSORBANCE
%
SCHAVENGING
1 CONTROL 0.51 -------
2 STANDARD 0.024 95.29
3 LPLPE 0.388 23.92
4 LPLC 0.442 13.33
5 LPLM 0.087 82.94
6 LPLW 0.154 69.80
7 LPRPE 0.401 21.37
8 LPRC 0.269 47.25
9 LPRM 0.095 81.37
10 LPRW 0.064 87.45
Table 4: % scavenging of Launaea pinnatifida leaves
and roots fractions along with standard drug L-
Ascorbic acid.

16. 16
CONCLUSION
The strongest antioxidant activity of methanol fractions of leaves could be
due to the presence of flavonoids and phenols.
Among all the fractions of leaves and roots of L. pinnatifida methanol and
water fraction were found to be more active.
Antioxidant results also support the traditional claim of this plant.
To conclude, studies have shown that, Launaea pinnatifida leaves methanol
fraction (LPLM) possess marked antioxidant activity thus have a supportive role
in the treatment of liver disease.

17. 17
This includes:
Bioactivity-guided fractionation & isolation of bioactive phytoconstituents
by means of Spectrometric & Chromatographic evaluation using UV, IR,
NMR, GC-MS/LC-MS, HPTLC
FUTURE SCOPE OF WORK
Explore L. pinnatifida as well as investigate the scientific evidence for
traditional claim
Generate standardization data for comparison study with other species of
Gojihva.

18. 18
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20076428
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http://plants.jstor.org/flora/ftea004644. [Last accessed on 2018 Nov 20].
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269:337-341.
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REFERENCES

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