Evaluation of in vitro antioxidant activity of different extracts of entire plant of Ipomoea pestigridis Linn

Bheemreddy Thrinitha; Murali R; Manichandrika P

doi:https://doi.org/10.26452/ijrps.v11i1.1805

Evaluation of in vitro antioxidant activity of different extracts of entire plant of Ipomoea pestigridis Linn

Bheemreddy Thrinitha ✉ ¹ , Murali R ¹ , Manichandrika P ²

1 Bojjam Narasimhulu Pharmacy college for woman, Vinay Nagar,Champapet, Hyderabad, India, 9704245374

2 Department of Pharmacy, Annamalai University, Annamalai Nagar-608 002, Tamil Nadu, India

Abstract

Ipomoea pestigridis (Linn) (family Convolvulaceae) is commonly known as “Tiger Foot Morning Glory” in English and locally known as ‘Pulichuvadi’ or ‘Pulichuvadu’ in Malayalam. The current study, aerial parts of different concentrates(Pet.ether, ethyl acetate, and methanol) of I.pestigridis, was evaluated for its in-vitro antioxidant potential by nitric oxide activity, total antioxidant activity, iron chelating activity taking ascorbate & Ethylenediamine tetraacetate as the standard correspondingly. An IC₅₀ value was originated that EA concentrates of I.pestigridis more efficient in nitric oxide activity, total antioxidant activity, Iron chelating capacity compared methanolic & PE concentrates. The ethyl acetate concentrates of I.pestigridis & ascorbic acid exhibited antioxidant potential possessing IC₅₀226µg/ml & 66µg/ml (Nitric oxide). 185µg/ml & 60µg/ml (total antioxidant) , 287µg/ml & 65µg/ml (iron-chelating Activity) respectively. The difference in the scavenging potential of the extracts can be due to variation in the percentage of bioactive compounds present in different solvents. Invitro antioxidant studies obviously show EA concentrates of I.pestigridis have better antioxidant activity. These results indicate that aerial parts of methanolic concentrate I.pestigridis could serve as a natural antioxidant, which may be useful in preventing free radical-induced diseases.

Keywords

I.pestigridis, Nitric oxide radical scavenging, total antioxidant, Iron chelating activity

Introduction

Free radicals might be defined as one or more unpaired electrons present in the molecules as it's outmost orbital and are able of autonomous existence. It is regularly represented as hydrogen radical (H^·), which contains one proton and one electron (unpaired) is the simplest free radical. A free radical reaction might be finished by a reaction among two free radicals or by neutralization by substances such as the antioxidants (Fang, Yang, & Wu, 2002). Free radicals may be generated both in vivo and in vitro through one of the subsequent mechanisms. Loss of a single electron from a regular molecule. Addition of an electron to regular molecule (Halliwell & Gutteridge, 1999). Ethnomedical literature contains a huge amount of herbs that may be used for the various diseases, in which ROS and free radical participate vital responsibility. A huge numerical herbs are used for strong antioxidant activity (Badami, Gupta, & Suresh, 2003). Current reports revealed that there is a converse connection among the intake of antioxidant-rich foods and the occurrence of human diseases (Halliwell et al., 1999).

Table 1: Nitric oxide scavenging activity of I. pestigridis PE Extract

S.no	Extract (µg/ml)	% of activity(±SEM)*
		PE extract	Ascorbic acid
1	125	34.65±0.025	48.54 ±0.046
2	250	40.34±0.042	57.31 ±0.034
3	500	45.34±0.052	68.24 ±0.045
4	1000	55.02±0.021	76.34 ±0.012
		IC₅₀ = 750 µg/ml	IC₅₀ = 135 µg/ml

* Every value was articulated as mean ± SEM for 3 experimentation

Ipomoea pestigridis (Linn) (family Convolvulaceae) is commonly known as “Tiger Foot Morning Glory” in English and locally known as ‘Pulichuvadi’ or ‘Pulichuvadu’ in Malayalam (Pawar & Patil, 2004; Sahu & Gupta, 2014). I.pestigridis was used for the treatment of wound healing (Amor-Prats & Harborne, 1993; Austin, 1975). I.pestigridis was used for different diseases like headaches, swellings, poisonous stings, snake bites. I.pestigridis was used for analgesic, antimicrobial, thrombocytic, cytotoxic activity (Pratap, Sudarsanam, Jyothi, Prasad, & David, 2011). Still, no literature are available on the antioxidant activity of the entire plant of I.pestigridis. Thus, the present study to assess the antioxidant activities of the entire plant of I.pestigridis.

Materials and Methods

Collection and Identification of Plant

The entire plant of I.pestigridis (family Convolvulaceae ) were gathered form kalakad , Tirunelveli District of Tamilnadu India. Plant identification was made from Botanical investigation of India, Palayamkottai The I.pestigridis were desiccated under shadowy, segregate, crushed through a grinder (Satheeshkumar, KottaiMuthu, & Manavalan, 2011).

Table 2: Nitric oxide scavenging activity of I. pestigridis EA Extract

S.no	Extract (µg/ml)	% of activity(±SEM)*
		EA extract	Ascorbic acid
1	125	43.54 ±0.022	48.54 ±0.046
2	250	55.32 ±0.043	57.31 ±0.034
3	500	64.56 ±0.065	68.24 ±0.045
4	1000	69.34 ±0.054	76.34 ±0.024
		IC₅₀ = 198 µg/ml	IC₅₀ = 135 µg/ml

* Every value was articulated as mean ± SEM for 3 experimentation

Table 3: Nitricoxide scavenging activity of I. pestigridis methanol Extract

S.no	Extract(µg/ml)	% of activity(±SEM)*
		Methanolic extract	Ascorbate
1	125	32.34 ±0.025	48.54 ±0.046
2	250	44.84 ±0.054	57.31 ±0.034
3	500	52.38 ±0.062	68.24 ±0.045
4	1000	66.22 ±0.038	76.34 ±0.024
		IC₅₀= 435 mg/ml	IC₅₀= 135 mg/ml

*Every value was articulated as mean ± SEM for 3 experimentation

Table 4: Total antioxidant activity of I. pestigridis PE Extract

S.no	Extract (µg/ml)	% inhibition(±SEM)*
		PE extract	Ascorbate
1	50	12.23 ±0.043	50.76 ±0.024
2	100	21.32 ±0.012	61.68 ±0.035
3	200	29.22 ±0.076	74.64 ±0.048
4	300	39.55 ±0.046	98.12 ±0.021
		IC₅₀ = 460 µg/ml	IC₅₀ = 57 µg/ml

* Every value was articulated as mean ± SEM for 3 experimentation

Table 5: Total antioxidant activity of I. pestigridis EA Extract

S.no	Extract (µg/ml)	% inhibition(±SEM)*
		EA extract	Ascorbate
1	50	33.45 ±0.043	50.76 ±0.024
2	100	40.65 ±0.054	61.68 ±0.035
3	200	50.85 ±0.062	74.64 ±0.048
4	300	59.46 ±0.014	98.12 ±0.021
		IC₅₀ = 190 µg/ml	IC₅₀ = 57 µg/ml

* Every value was articulated as mean ± SEM for 3 experimentation

Table 6: Total antioxidant activity of I. pestigridis methanol Extract

S.no	Extract (µg/ml)	% inhibition(±SEM)*
		Methanolic extract	Ascorbate
1	50	17.82 ±0.035	50.76 ±0.024
2	100	26.54 ±0.042	61.68 ±0.035
3	200	37.22 ±0.033	74.64 ±0.048
4	300	44.32 ±0.012	98.12 ±0.021
		IC₅₀ = 340 µg/ml	IC₅₀ = 57 µg/ml

* Every value was articulated as mean± SEM for 3 experimentation

Table 7: Iron-binding potential of I.pestigridis PE concentrates

S.no	Extract (µg/ml)	% inhibition (±SEM)*
		PE extract	Ethylenediamine tetraacetate
1	125	29.56 ±0.018	57.52 ±0.014
2	250	37.48 ±0.038	64.76 ±0.022
3	500	45.23 ±0.042	82.12 ±0.045
4	1000	55.48 ±0.023	96.34 ±0.034
		IC₅₀ = 760 µg/ml	IC₅₀ = 65 µg/ml

* Every value was articulated as mean ± SEM for 3 experimentation

Table 8: Iron-binding potential of I.pestigridis of EA concentrates

S.no	Extract (µg/ml)	% inhibition (±SEM)*
		EA extract	Ethylenediamine tetraacetate
1	125	38.12 ±0.010	57.52 ±0.014
2	250	53.92 ±0.024	64.76 ±0.022
3	500	61.43 ±0.036	82.12 ±0.045
4	1000	69.87 ±0.065	96.34 ±0.034
		IC₅₀ = 217 µg/ml	IC₅₀ = 65µg/ml

* Every value was articulated as mean ± SEM for 3 experimentation

Table 9: Iron-binding potential of I.pestigridis Methanolic concentrates

S.no	Extract (µg/ml)	% inhibition(±SEM)*
		Methanol extract	EthyleneDiamine tetraacetate
1	125	30.84 ±0.045	57.52 ±0.014
2	250	43.22 ±0.063	64.76 ±0.022
3	500	55.84 ±0.072	82.12 ±0.045
4	1000	67.32 ±0.028	96.34 ±0.034
		IC₅₀ = 385 µg/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 (Harborne, 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. (AlagumaniVasagam, KottaiMuthu, & Manavalan, 2010).

Assessment of Antioxidant potential through invitro methods

The variety of concentrates of I.pestigridis were used assessment of antioxidant activity by (Garrat, 1964) method was adopted for NO radical assay & (Prieto, Pineda, & Aguilar, 1999) method described for total antioxidant activity, and (Benzie & Strain, 1996) method was adopted to determine the Iron chelating activity.

Results and Discussion

Nitric oxide scavenging activity

Nitric oxide scavenging activity of PE concentrates of I. pestigridis appeared in Table 1. The PE concentrates of I. pestigridis exhibit a more Nitric oxide scavenging activity of 55.02% at 1000 µg/ml & ascorbate was recorded 76.34% at 1000 µg/ml. The IC₅₀ of the PE concentrates of I. pestigridis & ascorbic acid were recorded 750µg/ml & 135µg/ml correspondingly.

Nitric oxide scavenging activity of the EA concentrates of I. pestigridis appeared in Table 2. The EA concentrates of I. pestigridis exhibit a more Nitric oxide scavenging activity of 69.34% at 1000 µg/ml & ascorbic acid was recorded at 76.34% at 1000 µg/ml. The IC₅₀ of the EA concentrates of I. pestigridis & ascorbic acid were recorded 198µg/ml & 135µg/ml correspondingly.

Nitric oxide scavenging activity of methanol concentrates of I. pestigridis appeared in Table 3. The methanol concentrates of I. pestigridis exhibit a more Nitric oxide scavenging activity of 66.22% at 1000 µg/ml & ascorbic acid was recorded at 76.34% at 1000 µg/ml. The IC₅₀ of methanol concentrates of I. pestigridis & ascorbic acid were recorded 435µg/ml & 135µg/ml correspondingly.

IC₅₀ values & Nitric oxide scavenging potential revealed that ethyl acetate concentrates of I. pestigridis is better activity in scavenging Nitric oxide scavenging activity when compared methanol & PE extracts.

Phosphomolybdic acid method

Total antioxidant activity of PE concentrates of I. pestigridis appeared in Table 4. The PE concentrates of I. pestigridis exhibit a more total antioxidant activity of 39.55% at 300 µg/ml & ascorbic acid was recorded at 98.12% at 300 µg/ml. The IC₅₀ of the PE concentrates of I. pestigridis & ascorbic acid were recorded 460µg/ml & 57µg/ml correspondingly.

The total antioxidant activity of the EA concentrates of I. pestigridis appeared in Table 5. The EA concentrates of I. pestigridis exhibit a more total antioxidant activity of 59.46% at 300 µg/ml & ascorbic acid was recorded at 98.12% at 300 µg/ml. The IC₅₀ of the EA concentrates of I. pestigridis & ascorbic acid were recorded 190µg/ml & 57µg/ml correspondingly.

The total antioxidant activity of methanol concentrates of I. pestigridis appeared in Table 6. The methanol concentrates of I. pestigridis exhibit a more total antioxidant activity of 44.32% in both table (Table 6) and text. The IC₅₀ of the methanol concentrates of I. pestigridis & ascorbic acid were recorded 460µg/ml & 57µg/ml correspondingly.

IC₅₀ values & total antioxidant potential revealed that ethyl acetate concentrates of I. pestigridis is better activity in scavenging total antioxidant potential when compared methanol & PE extracts

Iron chelating potential

The iron complex potential of PE concentrates I.pestigridis & Ethylenediamine tetraacetate were appeared in Table 7. The more iron-binding potential of PE concentrates & Ethylenediamine tetraacetate 1000 µg/ml were recorded, 55.48% & 96.34 %. The IC₅₀ of PE concentrates of I.pestigridis & Ethylenediamine tetraacetate were found as 760µg/ml & 65µg/ml correspondingly.

The iron complex potential of EA concentrates of I.pestigridis & Ethylenediamine tetraacetate was were presented in Table 8. The more iron-binding capacity of EA concentrates & Ethylenediamine tetraacetate 1000 µg/ml was recorded at 69.87% & 96.34 %. The IC₅₀ value of ethyl acetate concentrates of I.pestigridis & Ethylenediamine tetraacetate were found 217µg/ml & 65µg/ml correspondingly.

The iron complex potential of methanolic concentrates of I.pestigridis & Ethylenediamine tetraacetate were presented in Table 9. The more iron-binding potential of methanolic concentrates & Ethylenediamine tetraacetate 1000 µg/ml were recorded, 67.32% & 96.34 %. The IC₅₀ value of methanol concentrates of I.pestigridis & Ethylenediamine tetraacetate was recorded as 385µg/ml & 65µg/ml correspondingly.

IC₅₀ values & iron-binding potential revealed that ethyl acetate concentrates of I.pestigridis is a huge activity in iron-chelating potential when compared methanol & petroleum ether concentrates. But when compared to all the three concentrates, the ethyl acetate concentrates of the I.pestigridis showed the better result.

Assessment of antioxidant activity, so many in vitro methods have been used a variety of concentrates of I.pestigridis. The results of antioxidant activity by NO radical activity, total antioxidant activity & iron-chelating potential were expressed in terms of % inhibition of generated free radicals, respectively, with respect to various concentrations. NO is produced from L-arginine through vascular endothelial cells, phagocytes, and certain cells of the brain. NO is a free radical since its unpaired electron and displays significant reactivity with confident types of proteins and other free radicals. The toxicity of Nitric oxide becomes a side effect when it reacts with superoxide radical, forming a highly reactive peroxynitrite anion (ONOO−) (Nagmoti, Khatri, Juvekar, & Juvekar, 2012). NO produced from sodium nitroprusside reacts with oxygen to form nitrite. The nitrite ions diazotize with sulphanilamide acid and couple with naphthyl ethylenediamine, producing pink coloured, which absorbs at 546 nm (Balakrishnan, Panda, Raj, Shrivastava, & Prathani, 2009). Among the three different plant concentrates tested, interestingly, in the NO radical activity of the EA extract of I.pestigridis exhibited more NO radical potential comparable with that of ascorbic acid.

The iron-chelating potential of all the concentrates was measured by Fe-ferrozine complex formation. Ferrozine-Fe complex is producing red coloured, 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). The iron-chelating potential of ethyl acetate extract of I.pestigridis exhibited a higher ability in scavenging compared to standard Ethylenediamine tetraacetate.

Conclusions

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 extract ethyl acetate extracts of I.pestigridis exhibited higher potency of antioxidant activity. These results indicate that ethyl acetate extract of I.pestigridis might serve as a natural antioxidant, which may be useful in preventing free radical-induced diseases.

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