Antioxidant activity and phytochemical screening of Ficus benghalensis aerial roots fractions

Zeinab Etratkhah; Seyed Esmaeil Sadat Ebrahimi; Nafiseh Khosravi Dehaghi; Yousef Seifalizadeh

doi:10.4103/jrptps.jrptps_20_18

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ORIGINAL ARTICLE

Year : 2019 | Volume : 8 | Issue : 1 | Page : 24-27

Antioxidant activity and phytochemical screening of Ficus benghalensis aerial roots fractions

Zeinab Etratkhah¹, Seyed Esmaeil Sadat Ebrahimi², Nafiseh Khosravi Dehaghi³, Yousef Seifalizadeh¹
¹ Faculty of Pharmacy, International Campus, Tehran University of Medical Sciences, Tehran, Iran
² Faculty of Pharmacy, International Campus; Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
³ Evidence-Based Phytotherapy and Complementary Medicine Research Center; Department of Pharmacognosy, School of Pharmacy, Alborz University of Medical Sciences, Karaj, Iran

Date of Web Publication

28-Mar-2019

Correspondence Address:
Nafiseh Khosravi Dehaghi
Evidence-Based Phytotherapy and Complementary Medicine Research Center, Alborz University of Medical Sciences, Karaj; Department of Pharmacognosy, School of Pharmacy, Alborz University of Medical Sciences, Karaj
Iran

Source of Support: None, Conflict of Interest: None

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DOI: 10.4103/jrptps.jrptps_20_18

Abstract

Context: Ficus benghalensis (Moraceae) is an evergreen tree found in south and southeast of Iran as wild and cultivated plants. Different parts of this plant have different effects such as antitumor, antipyretic, analgesic and anti-inflammatory. Aims: The aim of this study was investigated the phytochemical screening and antioxidant activities of different fractions of plant roots. Materials and Methods: Phytochemical investigation was done by different methods in references. Antioxidant activity was evaluated by DPPH and FRAP assay. All chemical materials and solvents were prepared from Sigma-Aldrich, Scharlau and Merk. Statistical Analysis: All measurements were carried out in triplicate and the data were expressed as mean ± SD. Statistical analysis was performed using one-way analysis of variance (ANOVA) and tukey test. Results: Phytochemical screening showed steroids, flavonoids, tannins, phenolic compounds, and anthraquinone glycoside are F. benghalensis constituents. This plant had antioxidant activity, but it was lower than the Indian kinds. Conclusion: This study elucidated Ficus benghalensis could be useful plant with antioxidant activity. Further investigation needs for details.

Keywords: 2,2-diphenyl-picrylhydrazyl, antioxidant activity, ferric reducing antioxidant power, Ficus benghalensis, phytochemical screening

How to cite this article:
Etratkhah Z, Ebrahimi SE, Dehaghi NK, Seifalizadeh Y. Antioxidant activity and phytochemical screening of Ficus benghalensis aerial roots fractions. J Rep Pharma Sci 2019;8:24-7

How to cite this URL:
Etratkhah Z, Ebrahimi SE, Dehaghi NK, Seifalizadeh Y. Antioxidant activity and phytochemical screening of Ficus benghalensis aerial roots fractions. J Rep Pharma Sci [serial online] 2019 [cited 2023 Mar 26];8:24-7. Available from: https://www.jrpsjournal.com/text.asp?2019/8/1/24/255049

Introduction

Ficus includes 800 species that is found in most tropical zones all over the world.^[1] Ficus benghalensis (Moraceae) is one of them and it is a large evergreen tree 23–34 m tall ^[2] that is found in south and southeast of Iran as wild and cultivated plant. Various studies show that different parts of this plant can be effective as traditional medicine because of the presence of different chemical constituents such as triterpenoids, flavonoids, tannin, different glucosides, steroids, resin, albumin, and malic acid.^[2],[3] Methanolic extracts of root, bark, and leaf of F. benghalensis were studied for antimicrobial effects, and all three parts of plant extracts inhibited the growth of Gram-positive bacteria as well as Gram-negative bacteria. Among the tested microbial strains, bacteria were found to be more sensitive than fungi. The methanolic extracts of root and leaf are potent against Candida albicans.^[2] This plant has antipyretic activities, analgesic effects,^[4] antitumor activities,^[5] and anti-inflammatory ^[6] activities. The bark of this plant has anti-inflammatory and analgesic properties in animal models,^[7] and it is useful for burning sensation, ulcers, and painful skin diseases.^[8] The aqueous and methanolic extracts of this plant kill the earthworm and it was comparable by antihelminthic drugs.^[9] Antidiabetic activity of the aqueous extract of F. benghalensis at a dose of 500 mg/kg/day was shown with histological studies in normal and streptozotocin-induced diabetic rats.^[10] Indian researchers study various biological activities of different parts of F. benghalensis. This plant is one of the Iranian traditional medicine that is used in different traditional formulations There is not enough study about Iranian kind besides useful uses of it. Therefore, the aim of this study was preliminary phytochemical screening and investigation of antioxidant activity of Iranian F. benghalensis root as a traditional medicine for comparing with Indian kind to further pharmaceutical investigations.

Materials and Methods

Plants materials

The roots of F. benghalensis were purchased from botanical market of Tehran. The dried roots (300 g) were powdered and extracted by ethanol 80% in percolator at room temperature (3 × 48 h). The extract was concentrated using a rotary evaporator at 37°C (190 g). Twenty-nine grams of dried extract was fractionated based on increasing polarity by chloroform, ethyl acetate, and methanol at room temperature by silica gel (mesh 35–70) and filtration set. The different fractions were concentrated using a rotary evaporator at 37°C. All fractions, i.e., chloroform (8.3 g), ethyl acetate (5.5 g), and methanol (7.1 g), were stored at 4°C until use.

Chemicals

2,2-diphenyl-picrylhydrazyl (DPPH), butylated hydroxyanisole (BHA), and 2, 4, 6-tripyridyl-striazine (TPTZ) were bought from Sigma-Aldrich (Germany). Sodium carbonate, sodium acetate, ferrous sulfate, and FeCl3 were prepared from Scharlau, Spain. Other chemicals and all solvents were purchased from Merck (Germany).

Phytochemical screening

Standard phytochemical methods were used to evaluate the presence of different constituents such as saponins, alkaloids, tannins, anthraquinones, cardiac glycosides, cyanogenic glycosides, amino acid, and protein and flavonoids.^{[11],[12],[13],[14],[15]}

Antioxidant activity

Free radical-scavenging assay 2,2-diphenyl-1-picrylhydrazyl assay

The DPPH assay is a popular method in natural product antioxidant studies. This assay is based on the theory that a hydrogen donor is an antioxidant. It measures compounds that are radical scavengers.^[16] The assay was carried out according to Sarker et al.^[17],[18] The stock solution of DPPH was prepared at the concentration of 8.0 × 10⁻² mg/mL in methanol. The extract dilutions were made in methanol to get the concentrations of 5.0 × 10⁻¹, 2.5 × 10⁻¹, 1.25 × 10⁻¹, 6.25 × 10⁻², 3.13 × 10⁻², and 1.6 × 10⁻² mg/mL. The prepared solutions of extracts (2.0 mL each) were mixed with DPPH solution (2.0 mL). After 30 min, ultraviolet absorbances of the solutions were recorded at 517 nm. BHA, a synthetic antioxidant, and Vitamin C, a natural antioxidant, were used as the positive control. All experiments were carried out at least three times. Inhibition of DPPH-free radical was calculated as:

Inhibition % = 100 – ([Sample absorption − control absorption]/Blank absorption) × 100.

The concentration that caused 50% decrease in the initial DPPH radical concentration was defined as IC₅₀. The experiments were repeated three times, and the IC₅₀ values were expressed as mean ± standard deviation.^[17],[18]

Ferric reducing antioxidant power assay

The antioxidant activity of the plant fractions was evaluated according to the method of Benzie and Strain.^[12] The ferric reducing antioxidant power (FRAP) reagent included 10 mM TPTZ solution in 40 mM HCl, 20 mM FeCl3 solution, and 0.3 M acetate buffer (pH 3.6) in proportions of 1:1:10 (v/v). Fifty microliters of each diluted extract was mixed with 3 mL of freshly prepared FRAP reagent, and the reaction mixtures were incubated at 37°C for 30 min. The absorbance was determined at 593 nm against distilled water as blank. Aqueous solutions of ferrous sulfate (100–2000 μM) were used for calibration. Triplicate measurements were taken, and the FRAP values were expressed as μmol of Fe (II)/g dry weight of plant powder.^[19],[20]

Results

Phytochemical screening of Ficus benghalensis aerial roots

The total extract and different fractions of the plant were used for preliminary phytochemical screening that is shown in [Table 1].

Table 1: Phytochemical screening of total extract and different fractions of Ficus benghalensis aerial root

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2,2-diphenyl-1-picrylhydrazyl radical scavenging activity

All results are reported in [Table 2]. IC₅₀ value is the concentration of sample required to scavenge 50% of DPPH-free radical and was calculated by plotting a graph of concentration and % inhibition. The calibration curve for the quantification of DPPH assay was linear over the range of standard concentrations of 500–2000 μg/ml with correlation coefficient of R² = 0.957 about the validity, accuracy, and precision of the method being also in the acceptable range. Methanolic fraction was more effective than the others.

Table 2: Antioxidant activity of different fractions of Ficus benghalensis aerial roots by 2,2-diphenyl-picrylhydrazyl assay

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Ferric reducing antioxidant power radical scavenging activity

All results are reported in [Table 3]. Calibration curve of standard solution of sulfate iron is shown in [Figure 1]. The calibration curve for the quantification of FRAP assay was linear over the range of standard concentrations of 100–1000 μM/L with correlation coefficient of R² = 0.987 about the validity, accuracy, and precision of the method being also in the acceptable range. Ethyl acetate fraction was more effective than the others.

Table 3: Ferric reducing antioxidant power radical scavenging activity of Ficus benghalensis aerial roots

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Figure 1: Calibration curve and regression equation calculated standard solution of sulfate iron

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Discussion

Ficus species are rich source of natural antioxidants.^[21] There are different researches about antioxidant activity of different parts of this genus. F. benghalensis is one of the species that Indian researches worked on it. One study elucidated that the aqueous extract of F. benghalensis Linn. root had maximum scavenging of DPPH radical (96.07%) at 250 μg mL ⁻¹ concentration and hydrogen peroxide (69.23%) at 1000 μg mL ⁻¹ concentration.^[22] The methanolic extract of roots of this plant showed good antioxidant activity by reducing power assay and hydrogen peroxide assay with IC₅₀ value 27.5 and 25 μg/ml, respectively. One study evaluated that methanolic extract of F. benghalensis aerial roots had immune stimulatory activity with specific and nonspecific mechanisms, which may be due to the presence of prominent amount of flavonoids, phenols, and tannins.^[23] The antioxidant and immunomodulatory activities of F. benghalensis leaves fractions were evaluated, and this article indicated that the radical scavenging effects of fractions on DPPH were less than reference antioxidants and the n-hexane fraction showed the best activity.^[24] In this study, the different fractions of Iranian aerial root showed antioxidant activity, but this effect is less than the Indian kinds. Preliminary phytochemical screening showed that the roots of this plant have different constituent such as steroids, tannins, phenolic compounds, and flavonoid glycosides as the Indian kind. The methanolic fraction has the best antioxidant activity by DPPH assay, and ethyl acetate fraction has the best ferric reducing power. This ability happens because of the presence of different constituents such as phenolic compounds, flavonoids, and tannins in this plant. Another study will need for accurate elucidation of its chemical constituents.

Conclusion

The present studies indicated that Ficus benghalensis root could be useful as antioxidant constituents in pharmaceutical industries.

Acknowledgment

This study was supported by International Campus, Tehran University of Medical Sciences (Thesis Code: 8923120022).

Financial support and sponsorship

This study was supported by International Campus, Tehran University of Medical Sciences (Thesis Code: 8923120022).

Conflicts of interest

There are no conflicts of interest.

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Figures

[Figure 1]

Tables

[Table 1], [Table 2], [Table 3]

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