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Der Pharmacia Lettre, 2016, 8 (6):245-248
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ISSN 0975-5071
USA CODEN: DPLEB4
The study of antioxidant activities of an Ayurvedic medicine Kulathadi
Kashayam
N. S. Sharada1, Mudiganti Ram Krishna Rao2*, Minu Priya3, K. Prabbu4, V. S. Kalaivani5,
D. Kumaran6 and Lakshmi Sundaram7
1
Department of Gynecology, SIMS, Vadapalani, Chennai
Professor, Department of Industrial Biotechnology, Bharath University, Chennai-600073
3
Associate Professor, Department of Gynecology, Sree Balajee Medical College and Hospital, Chennai
4
Associate Professor, Department of Anatomy, Sree Balajee Medical College and Hospital, Chennai
5
Professor, Depatment of Biochemistry, Sree Balajee Medical College and Hospital, Chennai
6
Student, Sree Balaji Medical College and Hospital, Bharath University, Chennai
7
Scientist, V Clin Bio, Central Research Facility, Sri Ramachandra University, Chennai
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2
ABSTRACT
The present study deals with the antioxidant assays, namely, DDPH assay, FRAP assay and Hydrogen peroxide
Scavenging Activity assay of one Ayurvedic formulation, Kulathadi Kashayam, which is a known medicine for
gynecological disorders. The report suggests that all the three assays have clearly indicated that Kulathadi
Kashayam has very good antioxidant potential which could be one of the reasons for its action as an effective
medicine for gynecological disorders.
Key words Kulathadi Kashayam, DDPH, FRAP, Hydrogen Peroxide Scavenging Assay.
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INTRODUCTION
The understanding of the mechanism of action of Ayurvedic and Sidha drugs is imperative to bring them to the fore
of medicinal practice all over the world. Although majority of population world over depend on one or the other
form of traditional medicines like, Ayurveda, Sidha, Unani, Chinese medicines etc., there is a lack of scientific
validation towards the efficacy of these drugs. This validation could be to study the parameters like
pharmacological, pharmacokinetic, toxicological and other common tests conducted for validation of medicines.
This knowledge will not only help prove the efficacy of these alternative forms of medicines but also the world will
get cheap, affordable and easily available medicines for all the ailments. Of late some scientific documentation in
this regard is forthcoming, which is a welcome sign. [1-18]
Most of the medicines play a vital role in curing diseases due to their antioxidant capacity. Plants products contain
phytochemicals such as flavonoids, tannins, alkaloids, phenols etc. which are natural antioxidants. Thus the study of
antioxidant activities of Ayurvedic preparations could show some clue to the mechanism of action of these
medicines. Some reports in this regard are available [19-22]
In continuation of our study of finding out the efficacy of on Ayurvedic medicines, the present report deals with the
antioxidant studies of Kulathadi Kashayam. Kulatahadi Kashayam is an Ayurvedic preparation which contains
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Der Pharmacia Lettre, 2016, 8 (6):245-248
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important medicinal plants namely, Dolichochos biflorus (Kulatha, Horse gram), Allium sativum (Lahsuna, garlic),
Ricunus cumminis (castor) and Sesamum indicum (Tila, Sesame). All the four plants have very important medicinal
roles and this Kashayam is used to treat different types of Amenorrhoea and Cryptomenorrhoea. The GC MS
analysis report was published by us earlier indicating the presence of various medicinal biomolecules, which act a
medicine. The present study deals with the antioxidant assays of Kulathadi Kashyam. This knowledge can throw
some light on the mechanism of action of his medicine.
MATERIALS AND METHODS
Kulathadi Kashayam was procured from standard Ayurvedic shop from Chennai. Antioxidant studies, namely,
DPPH assay, FRAP assay, Hydrogen Peroxide scavenging activity assay and were conducted by standard methods.
Antioxidant Study
Antioxidant study was performed by DPPH Assay, FRAP Assay and Hydrogen Peroxide Scavenging Activity
assay.
DPPH Assay (1,1-diphenyl-2-picrylhydrazyl) (Blios, 1958) [23]
The sample was dissolved in Ethanol in 1mg/ml concentration and used as stock. From the stock, various
concentrations (100, 200, 300, 400mg) were taken for further analysis.
Respective solvents were taken as negative control.
Conc.
= Concentration of the sample
OD
= OD of the sample
Neg. Control
= The Solvent
Activity
= Neg. Control – OD / Neg. Control
% of Activity
= Activity/100
IC50
= 50 – c value / m value
IC50/ml
= IC50/3 (3 ml of DPPH for the assay. To find the activity in 1 ml, the value had been divided by
3).
FRAP Assay (Ferric Reducing/Oxidant Power) (Pulido et al., 2000) [24]
Kulathadi Kashyam was dissolved in Ethanol. Triplicates had been put for all the Processes.
Conc.
= Concentration of the sample
OD
= OD of the sample
Linearity (y)
= mx + c
M
= Slope
C
= The point x crosses y axis
X
= OD – c value / m value
mM Fe/mg
= X value / concentration x 1000
Mean
= Average of mM Fe/mg
STDEV
= Standard Deviation for mM Fe/mg.
Hydrogen Peroxide Scavenging Activity (Ruch et al. 1989) [25]
Kulthadi Kashayam was dissolved in Ethanol.
Triplicates had been put for all the Processes.
Conc.
= Concentration of the sample
OD
= OD of the sample
Neg. control
= The solvent
Activity
= Negative control – OD / Negative control
% of activity
= Activity / 100
Mean
= Average of % of Activity
STDEV
= Standard Deviation of % of Activity
Graph
= (For Mean of % of Inhibition vs samples) Drawn using 2D clustered column.
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RESULTS AND DISCUSSION
Table. No. 1 Indicates the results of DDPH assay with Ethanol Kulathadi Kashayam.
Sl.No
1
2
3
4
Solution
Ethanol
Conc.
100
200
300
400
OD
0.586
0.475
0.322
0.212
Neg.Control
0.989
% Activity
40.74823
51.97169
67.44186
78.56421
m value
0.1838
C value
10.981
IC50
212.2905
IC50/ml
70.76351
From the results it shows that IC50/ml was lowest value (70.76351) indicating highest activity.
FRAP test Results are mentioned in Table No.2
Table No.2 Indicates the FRAP assay patterns of Kulathadi Kashayam in Ethanol solution
Solvent
Ethanol
Conc.
10
10
10
OD
0.486
0.488
0.435
m Value
0.0274
0.0274
0.0274
c Value
0.1086
0.1086
0.1086
X
13.77372
13.84672
11.91241
mM Fe(II)/mg
137.7372263
138.4671533
119.1240876
Mean
STDEV
131.78
10.96
From the Table no. It is clear that ethanol solution of Kulatahadi Kashayam indicated antioxidant activity (131.78).
Hydrogen peroxide scavenging assay results of Kulathadi Kashyam in shown in Table No.3
Table No. 3 Indicates the Hydrogen Peroxide Assay results of Kulathadi Kashayam
.
Solvent
Ethanol
Conc
100
100
100
OD
0.357
0.374
0.312
Neg. Control
0.748
0.748
0.748
% Activity
52.27273
50.00000
58.28877
Mean
STDEV
53.52
4.28
From the results it is clear that the Ethanolic solution of Kulathadi Kashayam indicated antioxidant percent activity
(53.52%) with respect to Hydrogen peroxide scavenging activity.
Reactive oxygen species which are produced naturally in the body can lead to a number of physiological imbalances
leading to various diseases which include physiological diseases and even cancer. The role of photochemical present
in plants as antioxidant agents is well documented. The choice of the plants for the preparation of a certain medicine
by the Ayurvedic proponents to address a particular disease indicate their depth of knowledge of the etiology of the
disease as well as the mechanism of action of the chosen plants or plant parts. This is high time for the scientific
community to understand the hidden secrets of Ayurveda and other traditional medicinal forms for making mankind
healthy and disease free.
CONCLUSION
From the antioxidant assay results it is evident that Kulathadi Kashayam has good antioxidant activity which could
be one of the major reasons for its action as medicine.
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