Study of in-vitro antioxidant activity of various extracts of aerial parts of Olax scandens
Abstract
Olax scandensRoxb. (family Olacaceae) Available in throughout tropical India. The current study, aerial parts of different concentrates of Olax scandenswas evaluated for its in-vitro antioxidant potential by Diphenylpicrylhydrazyl activity, superoxide scavenging activity, iron chelating activity taking ascorbate, quercetin & Ethylenediamine tetraacetate as the standard correspondingly. An IC50 value was originated that methanolic concentrates of Olax scandensis more efficient in Diphenylpicrylhydrazyl radical, superoxide radical activity, Iron chelating capacity compared EA & PE concentrates. The methanolic concentrates of Olax scandens & ascorbate exhibited antioxidant potential possessing IC50 226µg/ml & 66µg/ml (DPPH). 185µg/ml & 60µg/ml (Superoxide) , 287µg/ml & 65µg/ml (iron-chelating Activity) respectively. Invitro antioxidant studies obviously show methanolic concentrates of Olax scandens have better antioxidant activity. These results indicate that aerial parts of methanolic concentrates Olax scandens could serve as a natural antioxidant, which may be useful in preventing free radical-induced diseases.
Keywords
Olax scandens, DPPH radical scavenging, Superoxide radical, Iron chelating activity
Introduction
A free radical is any species capable of freelance existence that contains a lot of odd unpaired electrons. Electron unpaired being single that is only in an orbital (Halliwell, 1991). The electron unpaired gives convinced characteristic property to the free radical, like as paramagnetism. The chemical reactivity of free radicals is usually more. They may be + charged, - charged, or neutral charge electrically (Slater, 1984). These RO having a major task in the pathogenesis of many oxidative stress associated disorders like carcinogenesis, cardiovascular diseases, arthritis, diabetics, & neurological disorders (Halliwell & Gutteridge, 1990). The generally used synthetic antioxidants at present are BHA, BHT, PG, & TBHQ. However, these drugs are assumed accountable for liver harm and performing as carcinogens in lab animals (Anagnostopoulou, Kefalas, Papageorgiou, Assimopoulou, & Boskou, 2006). The investigate for new products with antioxidative potential & a smaller number of adverse effects is an active domain of research. Thus, the extension and utilization of huge efficient antioxidants of the natural source was popular (Sakagami, Aoki, Simpson, & Tanuma, 1991).
Olax scandens Roxb. (family Olacaceae ) is commonly known as “Parrot Olax, Sprawling olax ” in English & locally known as ‘Kurpodur’ in Telugu. This plants fruits and leaves has been used for therapeutic & food purpose. O. Scandens leaves were used as constipation. Olax scandens(Roxb.), is generally known as Badrul in Odiya, used for cooking & different therapeutic purposes (Sinha & Lakra, 2005). Olax scandens bark decoction is used treatment fever & cough. (Duraipandiyan, Ayyanar, & Ignacimuthu, 2006). O. scandens leaves were used for mouth ulcers. (Kumar, Rao, & Naidu, 2015). O.scandens boiled leaves were applied in the head for the treatment of headache. (Kumar, Naidu, & Raja, 2010). Still, no literature are available on the antioxidant potency of Olax scandens. Thus, the present study to assess the antioxidant activities of Olax scandens.
Materials and Methods
Gathering & Identification of Plant
The aerial parts of O.scandens(family Olacaceae) were gathered from Medak, Telangana state, India. Plant identification was made from the Botanical investigation of India, Telangana regional center, Hyderabad, (BSI/DRC/2019-20/Tech/174). The O.scandens, were desiccated under shadowy, segregate, crushed through the grinder (Satheeshkumar, Muthu, & Manavalan, 2011).
|
S.no |
Extract (µg/ml) |
% of activity(±SEM)* |
|
|---|---|---|---|
|
PE concentrates |
Ascorbate |
||
|
1 |
100 |
10.29±0.023 |
54.19 ±0.024 |
|
2 |
200 |
19.74±0.045 |
59.24 ±0.032 |
|
3 |
400 |
34.56±0.063 |
65.32 ±0.054 |
|
4 |
800 |
45.08±0.043 |
72.82 ±0.062 |
|
IC50 = 990 µg/ml |
IC50 = 66 µg/ml |
||
*Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% of activity(±SEM)* |
|
|---|---|---|---|
|
(EA concentrates) |
(Ascorbate) |
||
|
1 |
100 |
28.33 ± 0.024 |
54.19 ±0.024 |
|
2 |
200 |
39.75 ± 0.034 |
59.24 ±0.032 |
|
3 |
400 |
47.12 ± 0.023 |
65.32 ±0.054 |
|
4 |
800 |
59.12 ± 0.015 |
72.82 ±0.062 |
|
IC50 = 504µg/ml |
IC50 = 66 µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% of activity(±SEM)* |
|
|---|---|---|---|
|
(Methanolic concentrates) |
Ascorbate |
||
|
1 |
100 |
40.23±0.022 |
54.19 ±0.024 |
|
2 |
200 |
49.34±0.043 |
59.24 ±0.032 |
|
3 |
400 |
57.55±0.048 |
65.32 ±0.054 |
|
4 |
800 |
66.24±0.023 |
72.82 ±0.062 |
|
IC50 = 226µg/ml |
IC50 = 66µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% inhibition (±SEM)* |
|
|---|---|---|---|
|
(PE concentrates) |
(Quercetin) |
||
|
1 |
125 |
19.15 ±0.012 |
72.76 ±0.012 |
|
2 |
250 |
27.09 ±0.014 |
90.29 ±0.014 |
|
3 |
500 |
35.33 ±0.022 |
96.89 ±0.010 |
|
4 |
1000 |
48.65 ±0.026 |
99.12 ±0.018 |
|
IC50 = 1070 µg/ml |
IC50 = 60 µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% of inhibition (±SEM)* |
|
|---|---|---|---|
|
(Ethyl acetate concentrates) |
(Quercetin) |
||
|
1 |
125 |
20.12 ±0.014 |
72.76 ±0.012 |
|
2 |
250 |
44.49 ±0.025 |
90.29 ±0.014 |
|
3 |
500 |
50.23 ±0.033 |
96.89 ±0.010 |
|
4 |
1000 |
64.12 ±0.024 |
99.12 ±0.018 |
|
IC50 = 495 µg/ml |
IC50 = 60 µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% inhibition (±SEM)* |
|
|---|---|---|---|
|
Methanolic extract |
Quercetin |
||
|
1 |
125 |
35.18 ±0.071 |
72.76 ±0.012 |
|
2 |
250 |
54.23 ±0.024 |
90.29 ±0.014 |
|
3 |
500 |
62.34 ±0.044 |
96.89 ±0.010 |
|
4 |
1000 |
70.28 ±0.017 |
99.12 ±0.018 |
|
IC50 = 185 µg/ml |
IC50 = 60 µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% inhibition (±SEM)* |
|
|---|---|---|---|
|
PE concentrates |
Ethylenediamine tetraacetate |
||
|
1 |
125 |
26.90 ±0.012 |
57.52 ±0.014 |
|
2 |
250 |
34.56 ±0.032 |
64.76 ±0.022 |
|
3 |
500 |
42.65 ±0.010 |
82.12 ±0.045 |
|
4 |
1000 |
51.76 ±0.016 |
96.34 ±0.034 |
|
IC50 = 980 µg/ml |
IC50 = 65 µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% inhibition (±SEM)* |
|
|---|---|---|---|
|
EA concentrates |
Ethylenediamine tetraacetate |
||
|
1 |
125 |
32.30 ±0.024 |
57.52 ±0.014 |
|
2 |
250 |
43.38 ±0.013 |
64.76 ±0.022 |
|
3 |
500 |
50.28 ±0.023 |
82.12 ±0.045 |
|
4 |
1000 |
64.70 ±0.026 |
96.34 ±0.034 |
|
IC50 = 495µg/ml |
IC50 = 65µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
|
S.no |
Extract (µg/ml) |
% inhibition(±SEM)* |
|
|---|---|---|---|
|
Methanol concentrates |
EthyleneDiamine tetra acetate |
||
|
1 |
125 |
36.22 ±0.010 |
57.52 ±0.014 |
|
2 |
250 |
47.42 ±0.014 |
64.76 ±0.022 |
|
3 |
500 |
60.56 ±0.016 |
82.12 ±0.045 |
|
4 |
1000 |
68.89 ±0.021 |
96.34 ±0.034 |
|
IC50 = 287µg/ml |
IC50 = 65µg/ml |
||
* Every value was articulated as mean ± SEM for 3 experimentation
Preparation of Concentrates
The pulverized materials were progressively concentrated with PE (40-600C) through hot constant percolation method in Soxhlet equipment (J B Harbrone, 1984) for twenty-four hours. At that moment, the marc was subjected to EA (76-780C) for 24 hrs & then mark was subjected to methanol for 24 hrs. The concentrates were concentrated through the rotational evaporator and subjected to solidify drying in a lyophilizer till dry powder was acquired. (Vasagam, Muthu, & Manavalan, 2010).
Assessment of Antioxidant potential through invitro methods
The variety of concentrates of O.scandens were used assessment of antioxidant activity by Mensor et al. (2001) method was adopted for Diphenyl picrylhydrazyl radical assay & (Winterbourn, Hawkins, Brian, & Carrell, 1975) method described for Superoxide radical (O2 -) assay and (Benzie & Strain, 1996)ss method was adopted to determine the Iron chelating activity.
Results and Discussion
DPPH scavenging activity
The DPPH activity of PE concentrates of Olax scandens appeared Table 1. The PE concentrates of Olax Scandens exhibit, a more DPPH activity of 45.08% at 800 µg/ml & ascorbate was recorded 72.82% at 800 µg/ml. The IC50 of the PE concentrates of Olax scandens & ascorbic acid were recorded 90µg/ml & 66µg/ml correspondingly.
DPPH activity of EA concentrates of Olax scandens summarized in Table 2. The EA concentrates of Olax Scandens exhibit more DPPH scavenging potential of 59.12% at 800 µg/ml & ascorbate was recorded 72.82% at 800 µg/ml. The IC50 of the EA concentrates of Olax scandens & ascorbic acid were recorded 504µg/ml & 66µg/m correspondingly.
DPPH potential of methanolic concentrates of O.scandens appeared in Table 3. The methanolic concentrates of O.scandens having more DPPH scavenging potential of 66.24% at 800 µg/ml & ascorbate was recorded 72.82% at 800 µg/ml. The IC50 of the methanolic concentrates of O.scandens & ascorbic acid were recorded 226µg/ml & 66µg/m correspondingly.
The methanolic concentrates of Olax scandens was recorded to more activity than PE & EA concentrates. The IC50 of the methanolic concentrates of O.scandens& ascorbic acid were found to be 226µg/ml & 66µg/ml correspondingly.
Superoxide activity
Superoxide radical potential of PE concentrates of O.scandens appeared Table 4. The more Superoxide radical potential of PE concentrates & standard at 1000 µg/ml was recorded at 48.65% & 98.01 %. IC50 of PE concentrates & standard was recorded as 1070µg/millilitre & 60µg/millilitre correspondingly.
Superoxide radical potential of EA concentrates of O.scandens appeared in Table 5. The more SO scavenging potential of EA concentrates & standard 1000 µg/ml was recorded 64.12% & 99.12% correspondingly. EA concentrates & Quercetin IC50 was recorded as 495µg/ml & 60µg/ml correspondingly.
Superoxide radical scavenging potential of methanolic concentrates of O.scandens appeared in Table 6. Superoxide radical scavenging potential was more in methanolic concentrates & Quercetin (standard) at 1000 µg/ml was recorded 70.28% & 99.12% . Methanolic concentrates & standard IC50 was recorded as 185µg/ml & 60µg/ml correspondingly.
IC50 values & Superoxide radical potential revealed that methanol concentrates of Olax scandens is better activity in scavenging superoxide radical when compared EA & PE extracts.
Iron chelating potential
The iron complex potential of PE concentrates O.scandens & Ethylenediamine tetraacetate were appeared in Table 7. The more iron-binding potential of PE concentrates & Ethylenediamine tetraacetate 1000 µg/ml were recorded, 51.76% & 96.34 %. The IC50 of PE concentrates of O.scandens& Ethylenediamine tetraacetate were found as 980µg/ml & 65µg/ml correspondingly.
The iron complex potential of EA concentrates of Olax scandens & Ethylenediamine tetraacetate were presented in Table 8. The more iron-binding capacity of EA concentrates & Ethylenediamine tetraacetate 1000 µg/ml was recorded at 64.70% & 96.34 %. The IC50 value of ethyl acetate concentrates of Olax scandens & Ethylenediamine tetraacetate were found 495µg/ml & 65µg/ml correspondingly.
The iron complex potential of methanolic concentrates of Olax scandens & Ethylenediamine tetraacetate were presented in Table 9. The more iron-binding potential of methanolic concentrates & Ethylenediamine tetraacetate 1000 µg/ml were recorded, 68.89% & 96.34 %. The IC50 value of methanol concentrates of Olax scandens & Ethylenediamine tetraacetate was recorded as 287µg/ml & 65µg/ml correspondingly.
IC50 values & iron binding potential revealed that methanol concentrates of Olax scandens is huge activity in iron chelating potential when compared ethyl acetate & petroleum ether concentrates. But when compared to the all the three concentrates, the methanol concentrates of the Olax scandens showed the better result.
Assessment of antioxidant activity, so many in vitro methods have been used a variety of concentrates of Olax scandens. Diphenylpicrylhydrazyl is a stable N2-centered free radical generally utilized for testing the antioxidant potential of herbal concentrates. When the stable Diphenylpicrylhydrazyl radical accepts an electron from the antioxidant compound, the violet colour of the Diphenylpicrylhydrazyl as reduced to yellow colored diphenylpicrylhydrazine radical which was measured colorimetrically. Substances which are able to perform this reaction can be considered as antioxidants & therefore radical scavengers (Dehpour, Ebrahimzadeh, Fazel, & Mohammad, 2009). The results of antioxidant activity by Diphenylpicrylhydrazyl radical activity, superoxide radical activity & iron-chelating potential were expressed in terms of % inhibition of generated free radicals respectively with respect to various concentrations. Among the three different plant concentrates tested, interestingly, in the DPPH radical activity of the methanolic of Olax Scandens exhibited more Diphenylpicrylhydrazyl radical potential comparable with that of ascorbic acid.
Superoxides could be produced in huge amounts by various biological processes. It is known to be more injurious to cellular components as an originator of the most ROS, contributing to tissue damage & many disorders (G B J M Halliwell, 1999). The methanolic concentrates of Olax scandens exhibited higher ability in scavenging superoxide anion radical when compared to the strand quercetin.
The iron-chelating potential of all the concentrates was measured by Fe-ferrozine complex formation. Ferrozine-Fe complex is producing red-colored, which absorbs at 562nm (Yamaguchi, Ariga, Yoshimura, & Nakazawa, 2000). It was revealed that Ethylenediamine tetraacetate, which forms σ bond with iron, are efficient as secondary antioxidants, for the basis that they decrease the redox potential, thereby stabilizing the oxidized form of the iron ion (Duh, Du, & Yen, 1999). Iron chelating potential of methanolic concentrates of Olax Scandens exhibited higher ability in scavenging compared to standard Ethylenediamine tetraacetate.
Conclusion
The current trends, antioxidative activity of the herbs having more interest due to their possible use as natural additives to substitute synthetic ones. Among the three various concentratess methanolic concentrates of Olax scandens exhibited higher potency of antioxidant activity. These results indicate that aerial parts of methanolic concentrates Olax Scandens could serve as a natural antioxidant, which may be useful in preventing free radical-induced diseases.