Antioxidant and Antimicrobial Activity of Canavalia gladiata
Sai Koteswar Sarma1*, D. Umamaheswari2, B. R. Balakrishnan3
1Research Scholar, Vinayaka Mission’s Research Foundation, Salem, Tamilnadu, India - 636008.
2Professor, Department of Pharmaceutical Chemistry, Vinayaka Missions College of Pharmacy,
Vinayaka Mission’s Research Foundation, Salem, Tamilnadu.
3Professor and HOD, Department of Pharmacognosy, Vinayaka Missions College of Pharmacy,
Vinayaka Mission’s Research Foundation, Salem, Tamilnadu, India - 636008.
*Corresponding Author E-mail: dsksharma@gmail.com
ABSTRACT:
The present study deals with the antioxidant and antimicrobial activities of Canavalia gladiata. Antioxidant activity by nitric oxide and DPPH methods reveals that methanol extract of Canavalia gladiata shows good results when compared to the aqueous extract. It indicates that methanol extract of Canavalia gladiata shows maximum percentage of inhibition when compared to the standard drug (Ascorbic acid).Antioxidants help to neutralize free radicals, which are unstable molecules that are linked to the development of a number of degenerative diseases such as cancer, cardiovascular disesase, cognitive impairment immune dysfunction, cataract and macular degeneration. Antibacterial activity by Agar well diffusion method reveals that methanol extract of Canavalia gladiata shows better results when compared to the aqueous extract. It indicates that methanol extract of Canavalia gladiata shows maximum zone of inhibition when compared to the the standard drug (chloramphenicol).
KEYWORDS: Canavalia gladiata nitric oxide method, DPPH method, agar well diffusion method,Antimicrobial and Antioxidant activities.
INTRODUCTION:
canavalia gladiata is a popular plant commonly called as sword bean and belongs to the family Fabaceae and is used for various ailments1,2. It is commonly cultivated in almost Asian countries and European countries such as in India, Myanmar or adjacent China3-5. Sword Bean is a twining, nearly erect annual herb, some cultivars may also be semi-erect and the root system is deep, growing up to 6 ft. Stems are sparsely pubescent or glabrous and Leaf blades are elliptic or ovate-elliptic, base cuneate, apex acute or acuminate; stipules deciduous. Leaves6-8 are shiny, trifoliate. Flowers are pink and white in colour .Flowers are bisexual, papilionaceous, all joined; ovary superior, style slender, curved, stigma is small. Pods are 30 cm long and 5cm wide. Seeds are elliptical. Seeds are red or red-brown, rarely black, pink or white.
Fruits9-11 are linear-oblong pod, slightly compressed, sometimes curved. The main chemical constituents are carbohydrates, alkaloids Phenolic compounds, flavonoids and amino acids such as cystin, tyrosin, and tryptophan. Seeds contain three crystalline globulins canavalin, concanavalin A and B etc.Six compounds were isolated and their structures were identified Gallic acid, Methyl gallate, 1,6-di-o- galloyl-β-D- glucopyranoside, Β- sitosterolm, Lupeol and δ-tocopherol1-15. All parts of the plant have been used as crude drug for the treatment of vomiting, abdominal dropsy, kidney-related lumbago, asthma, obesity, stomach-ache, dysentery, coughs, headache, intercostal neuralgia, epilepsy, schizophrenia, inflammatory diseases and swellings12-16.
MATERIALS AND METHODS:
The leaves of Canavalia gladiata were collected from the local area of Nellore, SPSR Nellore district (India) . The plant material (PARC/2020/4331-Voucher Specimen) was identified and authenticated by the botanist Prof.P.Jayaraman M.Sc, Ph.D. (PARC), Director Institute of Herbal Botany, Tambaram Chennai.
A) Methanol extract of Canavalia gladiate:
About 650 grams of the dry powder extracted with 2 litres of 95% ethanol by continous hot percolation using soxhlet apparatus. After completion of extraction it was filtered and concentrated to dry mass by vaccum distillation. A pale green colour residue was obtained. The extract was then stored in a dessicator.
B) Aqueous extract of Canavalia gladiate:
About 650 grams of the dry powder extracted with 2 litres of distilled water by continuous hot percolation method using soxhlet apparatus. After completion extraction, the aqueous extract was filtered and concentrated to dry mass by vaccum distillation .A green colour residue was obtained.
Antioxidant Activity:
Nitric oxide method17-28
The reaction mixture (3ml) containing sodium nitroprusside (10mM, 2ml), phosphate buffer saline (0.5ml) and extract or standard solution (0.5ml) will be incubated at 25 degrees Centigrade for 150 min. After incubation, 0.5 ml of the reaction mixture containing nitrate will be pipetted and mixed with 1 ml of sulphanilic acid reagent (0.33%i in 20% glacial acetic acid ) and will be allowed to stand for 5 min for completing diazotization.Then 1 ml of naphthylethylene diamine dihydrochloride (1%) will be added, mixed and will be allowed to stand for 30 min.The absorbance of these solutions will be measured at 540 nm. against the blank solution The percentage inhibition will be calculated by comparing the absorbance values of control and test by using the formula.
(A control-A test)
(%) scavenged = ---------------------------- X 100
A control
A control is the absorbance of the control reaction mixture. A test is the absorbance of sample of the extracts at different concentrations.
DPPH free radical scavenging activity17-28
The free radical scavenging activity will be measured in vitro by 1, 1-diphenyl-2-picryl-hydrazyl assay. About 0.3 mM solution of DPPH in 100% ethanol will be prepared and 1ml of this solution will be added to 3ml of the extract dissolved in ethanol at different concentrations (5-80mcg/ml).The mixture has to be shaken and allowed to stand at room temperature for 30 min .Absorbance will be measured at 517 nm using a spectrophotometer.The capability to scavenge the DPPH radicals will be calculated using the formula.
(A control-A test)
(%) scavenged = -----------------------------X 100
A control
A control is the absorbance of the control reaction mixture. A test is the absorbance of sample of the extracts at different concentrations
Determination of Antimicrobial Activity:
Fresh aqueous and methanolic extracts of Canavalia gladiata were used for the determination of antimicrobial activity.
Microorganism used:
Salmonella typhi
Method:
Agar well diffusion method18-28
From the above mentioned organisms, inoculums was prepared by inoculating the organisms in 10 ml of nutrient broth and incubated at 37 degrees centigrade for 18 hrs. Nutrient agar medium was poured in to each sterilized petridish and organism was inoculated. Wells were made in to the medium by using sterile cork borer and each sample of the extracts (100ul) was filled in to the wells of agar plates directly by using a micro liter syringe .Then the plates were incubated at 37 degrees centigrade for 24 hr.After incubation, the zone of inhibition was observed and measured in mm.
RESULTS:
Table 1: DPPH method
|
S. No |
Type of extract |
Concentration (ug/ml) |
Absorbance |
% Inhibition |
IC50 (ug/ml) |
|
1 |
Methanol extract
|
100 |
1.0953 |
34.37 |
|
|
200 |
0.8776 |
47.42 |
|
||
|
300 |
0.7788 |
53.34 |
291 |
||
|
400 |
0.5628 |
66.28 |
|
||
|
500 |
0.5151 |
69.14 |
|
||
|
2 |
Aqueous extract
|
100 |
1.3932 |
16.52 |
|
|
200 |
1.2596 |
24.53 |
|
||
|
300 |
1.1291 |
32.35 |
487 |
||
|
400 |
0.9630 |
42.30 |
|
||
|
500 |
0.7634 |
54.26 |
|
||
|
3 |
Ascorbic acid
|
100 |
0.9640 |
42.24 |
|
|
200 |
0.7624 |
54.32 |
|
||
|
300 |
0.6079 |
63.58 |
263 |
||
|
400 |
0.3609 |
78.38 |
|
||
|
500 |
0.2978 |
82.16 |
|
Table 2: Nitric oxide method
|
S.No |
Type of extract |
Concentration (ug/ml) |
Absorbance |
% Inhibition |
IC50 (ug/ml) |
|
1 |
Methanol extract
|
100 |
0.7473 |
32.87 |
|
|
200 |
0.6150 |
44.76 |
|
||
|
300 |
0.5420 |
51.32 |
290 |
||
|
400 |
0.3948 |
64.54 |
|
||
|
500 |
0.3635 |
67.35 |
|
||
|
2 |
Aqueous extract
|
100 |
0.8859 |
20.42 |
|
|
200 |
0.8179 |
26.53 |
|
||
|
300 |
0.7152 |
35.76 |
510 |
||
|
400 |
0.6208 |
44.24 |
|
||
|
500 |
0.4881 |
56.16 |
|
||
|
3 |
Ascorbic acid
|
100 |
0.6202 |
44.29 |
|
|
200 |
0.5171 |
53.55 |
|
||
|
300 |
0.4300 |
61.38 |
211 |
||
|
400 |
0.2645 |
76.24 |
|
||
|
500 |
0.2191 |
80.32 |
|
Table 3 :Antimicrobial activity of Canavalia gladiata by Agar well diffusion method against S.typhi
|
S.No |
Name of the Extract |
Concentration (μg/ml) |
Zone of inhibition (mm) |
|
1 |
Methanol extract |
100 |
16 |
|
2 |
Aqueous extract |
100 |
0 |
|
3 |
Std (Chloramphenical) |
100 |
26 |
DISCUSSION:
Antioxidant activity by nitric oxide method and DPPH method states that methanol extract of Canavalia gladiata shows good antioxidant activity when compared to aqueous extract. In both cases as concentration increases, % of scavenging activity also increases for methanol extracts of Canavalia gladiate When compared to aqueous extracts. Antimicrobial activity by agar well diffusion method by using microorganism salmonella typhi states that methanol extract of Canavalia gladiata shows the better antagonist effect against the microorganism when compared to the standard drug chloramphenicol.
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Received on 05.08.2020 Modified on 27.12.2020
Accepted on 03.02.2021 © RJPT All Right Reserved
Research J. Pharm.and Tech 2021; 14(12):6507-6510.
DOI: 10.52711/0974-360X.2021.01125