Antioxidant Activity by DPPH Radical Scavenging Method of Ageratum Conyzoides Linn. Leaves

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Antioxidant Activity by DPPH Radical Scavenging Method of Ageratum

conyzoides Linn. Leaves
Article · January 2014
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American Journal of Ethnomedicine, 2014, Vol. 1, No. 4, 244-249
Available online at http://www.ajethno.com
© American Journal of Ethnomedicine
Antioxidant Activity by DPPH Radical

Scavenging Method of Ageratum conyzoides
Linn. Leaves
Tailor Chandra Shekhar*1 and Goyal Anju2
1
Division of Pharmaceutical sciences, Shri guru ram rai institute of technology &
science, Patelnagar, Dehradun (Uttarakhand) India-248001
2
B.N. Institute of Pharmaceutical sciences, Sevashram road, Udaipur (Rajasthan)
India -313001
ABSTRACT
In this study Antioxidant activity was performed by DPPH (1, 1diphenyl-2-picryl

hydrazyl) radical scavenging method for different extracts of aerial parts like
leaves and flowers of Ageratum conyzoides Linn. plant species which showed that
alcoholic extract of leaves of this plant on higher concentration possess better
antioxidant potential when compare to reference standard ascorbic acid. They
exhibited strong antioxidant DPPH radical scavenging activity with IC50 value of
9.3 and 24.8 µg/ml for ascorbic acid and alcoholic leaves extract respectively. The
absorbance for reducing power was found to be 0.0390, 0.0989 for ascorbic acid
and alcoholic leaves extract respectively. The strongest antioxidant activity of
ethanol extract could be due to the presence of flavonoids and phenols.
Keywords- Ageratum conyzoides linn., Antioxidants, DPPH, Flavonoids etc.
INTRODUCTION of compounds with a wide variety of

physical and chemical properties.3 A rapid,
Antioxidants play an important role as simple and inexpensive method to measure
health protecting factor. Scientific evidence antioxidant capacity of food involves the use
suggests that antioxidants reduce the risk for of the free radical, 2, 2-Diphenyl-1-
chronic diseases including cancer and heart picrylhydrazyl (DPPH) which is widely used
disease. Primary sources of naturally to test the ability of compounds to act as free
occurring antioxidants are whole grains, radical scavengers or hydrogen donors and
fruits and vegetables.1 Plant sourced to evaluate antioxidant activity.4
antioxidants like vitamin C, vitamin E, The DPPH assay method is based on
carotenes, phenolic acids etc. have been the reduction of DPPH, a stable free
recognized as having the potential to reduce radical.5 The free radical DPPH with an odd
disease risk.2 Most of the antioxidant electron gives a maximum absorption at 517
compounds in a typical diet are derived from nm (purple colour). When Antioxidants
plant sources and belong to various classes react with DPPH, which is a stable free
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radical becomes paired off in the presence of the residue were dried in a desiccator over
a hydrogen donor (e.g., a free radical- Sodium sulphite. After this, practical yield
scavenging antioxidant) and is reduced to was weighed and calculated as 2.5 gm, 1.5
the DPPHH and as consequence the gm and 3 gm for pet. ether, chloroform and
absorbance’s decreased from the DPPH.6 ethanol extracts respectively.
Radical to the DPPH-H form, results in
decolorization (yellow colour) with respect Evaluation of antioxidant activity by DPPH
to the number of electrons captured.7 More radical scavenging method
the decolorization more is the reducing Free radical scavenging activity of
ability. This test has been the most accepted different extracts of leaves and flowers of
model for evaluating the free radical Ageratum conyzoides Linn. plant were
scavenging activity of any new drug.8 When measured by 1, 1- diphenyl-2-picryl
a solution of DPPH is mixed with that of a hydrazyl (DPPH). In brief, 0.1 mM solution
substance that can donate a hydrogen atom, of DPPH in ethanol was prepared. This
then this gives rise to the reduced form solution (1 ml) was added to 3 ml. of
(Diphenylpicrylhydrazine; non radical) with different extracts in ethanol at different
the loss of this violet colour (although there concentration (5, 10, 15, 20, 25, 30 µg/ml).
would be expected to be a residual pale Here, only those extracts are used which are
yellow colour from the picryl group still Solubilise in ethanol and their various
present).9 concentrations were prepared by dilution
This plant has been reported to method.14 The mixture was shaken
possess antioxidant properties.10 So, this vigorously and allowed to stand at room
study has been undertaken to evaluate temp for 30 min. then, absorbance was
Ageratum conyzoides Linn. plant for their measured at 517 nm. by using
possible potential to antioxidant action by spectrophotometer (UV-VIS Shimadzu).15
DPPH scavenging method.11 Reference standard compound being used
was ascorbic acid and experiment was done
MATERIALS AND METHODS in triplicate.16 The IC 50 value of the
sample, which is the concentration of
Ageratum conyzoides Linn. plant sample required to inhibit 50% of the
species along with leaves and flowers were DPPH free radical, was calculated using Log
collected from the local areas of dehradun dose inhibition curve. Lower absorbance of
along road side. Then, this plant was the reaction mixture indicated higher free
identified and authenticated from Botanical radical activity.17 The percent DPPH
survey of india, Dehradun (Uttarakhand) scavenging effect was calculated by using
under accession no.- 114004. following equation:
DPPH scavenging effect (%) or
Extraction Percent inhibition = A0 - A 1 / A0 × 100.
Shade dried leaves and flowers part Where A0 was the Absorbance of
of this plant were pulverized and about 100 control reaction and A1 was the Absorbance
gms of powdered leaves and flowers were in presence of test or standard sample.18
extracted with increasing order of polarity
solvents series starting from Pet. ether, RESULTS
Chloroform, Ethanol via soxhlet apparatus
by successive hot continuous percolation The leaves and flowers ethanolic
method.13 At last, all extracts were extract of this plant showed better
concentrated in a rotary flash evaporator and antioxidant potential when compare to
Page 244-249
standard ascorbic acid by DPPH scavenging 3. Sravani, T, Paarakh, P.M, “Antioxidant

assay method. The absorbance at 517 nm activity of Hedychium spicatum Buch. Ham
by UV visible spectrophotometer were Rhizomes” Indian journal of natural
found to be as 0.0390 and 0.0989 for products & resources, Sep.2012, 3(3), 354-
358.
standard ascorbic acid and alcoholic extract
4. Kirtikar, K.R, Basu, B.D, Indian medicinal
respectively and IC 50 value obtained were plants, International book distributors,
as 9.3 and 24.8 µg/ml. for same ascorbic Dehradun, 2006, 993-994.
acid and alcoholic extract respectively. It 5. Warrier, P.K, Nambier, VPK, Raman Kutty
means alcoholic extract of plant at higher C, Indian medicinal plants- A compendium
concentration captured more free radicals of 500 species, Orient longman Ltd, Madras,
formed by DPPH resulting into decrease in 1994, Vol-I, 95-97.
absorbance and increase in IC 50 value. 6. Harborne, J.B, Phytochemical methods- A
guide to modern techniques of plant
DISCUSSION analysis, 3rd Edn, Springer (India) Pvt. Ltd,
New delhi,1998, 5-32.
This study determined that Ethanolic 7. Ghosh, M.N, Fundamentals of Experimental
extract of leaves & flowers of Ageratum Pharmacology, 2nd Edn., Scientific Book
plant species showed better antioxidant Agency, Calcutta,1998, 174-179.
potential by DPPH radical scavenging 8. Wagner, H, Bladet, S, et.al, Plant Drug
Analysis-A TLC Atlas, 1st Edn, Springer
method when compare to standard ascorbic
verlag Berlin, Heidel berg, New York, 1996,
acid19 and IC 50 value found to be as 9.3 195-214.
and 24.8 µg/ml for ascorbic acid and 9. Handa, SS, Vasisht, K, et.al, Compendium
alcoholic extract respectively.20 So, we can of Medicinal and Aromatic Plants-Asia, II,
say this plant is having antioxidant activity. ICS-UNIDO, AREA Science Park,
Padriciano, Trieste, Italy, 2006, 79-83.
ACKNOWLEDGEMENTS 10. http://www.tropilab.com/billy-goat.html
accessed on March, 2012.
The authors are thankful to the 11. http://www.mmhmms.com/downloads/mp03
Management and Staff members of SGRR ageratumconyzoides.pdf accessed on March,
institute of technology & science, patel 2012.
nagar, Dehradun for their support and 12. http://www.wikiherb.info/2011/01/ageratum
Mahant Shri Devendra Das Ji Maharaj, -conyzoidesl.html accessed on March, 2012.
Chairman, SGRR Education Mission, 13. Okunade, A.L, Review- Ageratum
conyzoides L. (Asteraceae), Fitoterapia,
Dehradun for providing me necessary
2002, 73, 1-16.
facilities also Rahul mishra, Nidhi 14. Vaidyaratnam, Varier, PS, Indian Medicinal
Choudhary and Bhawna Bisht for supporting Plants- A Compendium of 500 species, I,
me at SGRR Institute of Technology & Orient longman publishing house,
Science, Dehradun. Kottakkal-India, 2002, 146.
15. Ahmed, M, Saeed, F, et al, “Evaluation of
REFERENCES Insecticidal and Antioxidant activity of
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Table 1. Absorbance of different extract of Ageratum conyzoides. linn with standard ascorbic
acid at 517 nm by uv visible spectorphotometer (dpph scavenging assay method)
Concentration Ascorbic acid
Pet ether (Abs) AlcE (Abs)
(µg/ml) (Abs)
5 0.2380 0.2421 0.2440
10 0.1719 0.2428 0.2420
15 0.0469 0.2440 0.2180
20 0.0415 0.2379 0.1619
25 0.0410 0.2218 0.1420
30 0.0390 0.2386 0.0989
Control: 0.2444
Table 2. % inhibition of different extract of Ageratum conyzoides Linn. with ascorbic acid
Concentration Ascorbic acid Pet. ether Alc. E
(µg/ml) (% Inhibition) (% Inhibition) (% Inhibition)
5 2.61% 0.99% 0.16%
10 29.66% 0.65% 0.98%
15 80.8% 0.16% 10.80%
20 83.01% 2.65% 33.75%
25 83.22% 9.24% 41.89%
30 84.02% 2.37% 59.53%
Page 244-249
Figure 1. Ageratum conyzoides- full plant species12
% ℎ = ( control – Abs test) / Abscontrol × 100
Graph 1. Absorbance in different concentration
Page 244-249
Graph 2. % inhibition in different concentration
Page 244-249
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