Int J Curr Pharm Res, Vol 10, Issue 4, 51-54Original Article


ANTIOXIDANT ACTIVITY OF HYDRO-ALCOHOLIC EXTRACT ON THE ROOTS OF NYCTANTHES ARBORTRISTIS

SUMIT DAS, SHILPA PAUL, AKM ABDULLAH, ABDUL SAMIM

Girijananda Chowdhury Institute of Pharmaceutical Science, Azara, Hatkhowapara, Guwahati 17
Email: shilpanita875@gmail.com

Received: 23 Apr 2018, Revised and Accepted: 10 Jun 2018


ABSTRACT

Objective: To determine the antioxidant activity of Nyctanthes arbortristis (Family-Oleaceae).

Methods: The hydroalcoholic extract of the root of plant Nyctanthes arbortristis was taken into considerations to determine the phytochemicals present in it. The extracts of the roots were evaluated for antioxidant activity by using different in vitro model like Reducing Power Method and DPPH method.

Results: In the current investigation it has been found that the Pet. Ether and Hydroalcoholic extracts showed potent antioxidant activity by reducing power, as the concentration of the extracts increased, the absorbance was also increased correspondingly.

Conclusion: The hydroalcoholic extracts of this plant showed potent antioxidant activity against the standard drug (Kaempferol).

Keywords: Nyctanthes arbortristis, Phytochemistry, Antioxidant Activity


INTRODUCTION

Free radicals are generally highly reactive and unstable compounds that can generate in the body during normal metabolic functions or it can form in the body from the external environmental sources such as environmental pollution and cigarette smoking. Antioxidants are the substances that can protect human bodies from oxidative damage of free radicals [1]. To balance the oxidative state, plants and animals maintain a complex system of overlapping antioxidants such as glutathione and enzymes (e. g. Catalase and superoxide dismutase) produced internally or the dietary antioxidants like vitamin A, vitamin C (ascorbic acid), β-carotene and α-tocopherol (a synthetic vitamin E) etc. Excessive production of free radicals can cause severe damage of cells and these may lead to various chronic diseases like cancer, stroke, heart disease, diabetes etc. Vitamins containing antioxidant are also helpful for the improvement of immune system functions [2]. Now a day’s lot of researches is going on to establish the therapeutic efficacy on the basis of herbs and spices as antioxidant potentiality [3].

Nyctanthes arbortristis is one of the most useful conventional plants in India. The various parts of plant-like fruits, leaves, seeds, flowers, barks and stem have important phytochemicals and have some medicinal importance for treatment and management of different disease states. Phytochemicals such as flavonoids, oleanic acid, carbohydrates,saponins, tannic acid, carotene, lupeol, benzoic acid present in various parts of plant which have significant antiviral, antifungal, antipyretic, antihistamine, anti-malarial, antibacterial, anti-inflammatory, antioxidant activities On the basis of traditionally claim on Nyctanthes arbortristis plants and due to the lack of proper scientific investigation of their potential pharmacological properties, the main aim of this present study has to determine the antioxidant activity of the plant (Nyctanthes arbortristis)by using reducing power (RP), DPPH scavenging methods [4, 5].

Scientific classification

Biological Source: Nyctanthes arbortristis

Family: Oleacea

Kingdom: Plantae

Order: Lamiales

Genus: Nyctanthes

Species: N. arbortristis

Vernacular names: Night-flowering jasmine (English), Harsingar (Hindi), Shefali (Bengali), Sheali (Assamese).

Distribution of the plant

It is widely distributed in south Himalayan regions and southwards to Godavari. Each and every part of the plant has some medicinal value so that it is commercially available. It grows at sea level up to 1500 m altitude, within a range of rainfall patterns. Nyctanthes prefers a scheduled and semi-shady place to grow [6].

Materials and methods

The roots of Nyctanthes arbortristis was collected from the localities in Rangiya, kamrup, Assam during the month of January 2018. N. arbortristis were collected, shade dried, powdered manually and sieve through a no. 60 mesh sieve. About 100 gms of powdered roots are extracted with the solvents (petroleum ether and methanol) in Soxhlet Apparatus and then it was filtered by muslin cloth and evaporated to dryness by using Rotary Evaporator. The percentage yield of the extract was listed below in table 1:

Phytochemical screening of the extract

Phytochemical screening was carried out for petroleum ether and hydro-alcoholic extract of N. arbortristis for the presence of of different phytoconstituents like alkaloid, flavonoid, phenolics, carbohydrate, glycoside and proteins [7].

In vitro antioxidant study

Reducing power methods (RP)

The experiment was performed according to the method given by Oyiazu (1986)[8]. In this method the desired concentration of the extract (different conc.10-100μg/ml) suspended in distilled water, 2.5 ml of 0.2 M-phosphate buffer (pH6.6), and 2.5 ml of K3Fe(CN)6 (1% w/v) were added. The mixture was incubated at 50 °C for 20 min. followed by addition of 2.5 ml of TCA (10% w/v). The mixture was centrifuged at 3000 rpm for 10 min. The upper layer of the solution (2.5 ml) was mixed with distilled water (2.5 ml) and 0.5 ml FeCl3 (0.1% w/v), and the absorbance was measured at 700 nm against blank sample by using Kaempferol as a standard.

Table 1: Percentage yield of various extracts

Extract % yield
Petroleum ether 6.3
methanol 8.9

Free radical scavenging activity measured by 1,1-diphenyl-2-picryl-hydrazil (DPPH)

The experiment was performed according to the method given by Blois [9]. 0.1 mM solution of DPPH· was dissolved in ethanol to be prepared. 1 ml of this solution was added to 3 ml of each extract solution in water at different concentrations (4-500 μg/ml).

The mixture was shaken vigorously and allowed to stand at room temperature for 30 min.

Then the absorbance was measured at 517 nm using a UV-Vis spectrophotometer.

Lower absorbance values of the reaction mixture indicated higher free radical scavenging.

Activity by using Kaempferol as standard [10].

DPPH scavenging effect (%) = [(Ao − A1/Ao) × 100]

Where,

Ao was the absorbance of the control reaction

A1 was the absorbance in the presence of the sample at various concentrations.

Thiobarbituric acid method (TBA)

The experiment was done according to the method given by Chang et. al [11]. Here the same samples were used those were prepared for FTC method. To desired sample solution, 1.0 ml of 20% aqueous trichloroacetic acid (TCA) and 2.0 ml of aqueous thiobarbituric acid (TBA) solution were added. The mixture was placed in a boiling water bath and kept for 10 min. Then we have to cool the reaction mixture. After cooling, it was then centrifuged at 3000 rpm for 20 min. Absorbance of the supernatant layer was measured at 532 nm [12]. Antioxidant activity was recorded based on the absorbance of the final day of the FTC assay. Both methods (FTC and TBA) were used to describe antioxidant activity by percentage inhibition:

RESULTS AND DISCUSSION

Preliminary phytochemical screening

Pet. Ether and Hydroalcholic extract of N. arbortristis roots showed the presence of phenolic, steroids, flavonoids and glycosides (table 2).

In vitro antioxidant effect of Kaempferol, RP, TBA and DPPH scavenging determination of N. arbortristis roots

In the current investigation, it has been found that the Pet. Ether and Hydro alcoholic extracts showed potent antioxidant activity by reducing power, as the concentration of the extracts increased, the absorbance was also increased correspondingly. But the Hydroalcoholic extracts showed more potent activity than the other extracts, as compared with standard substances (Kaempferol) (table 1). The lipid peroxidation of Pet. Ether, Hydroalcoholic extracts and Kaempferol was found to be 29.25, 32.12, 33.05 respectively (table 3). The in vitro free radical scavenging activity of N. arbortristis by Thiobarbituric Acid Method showed the potent percentage inhibition in case of hydroalcoholic extracts as compared to the other extracts. The in vitro free radical scavenging of Pet. Ether, Hydro-alcoholic extract and Kaempferol was found to be 33.05, 8.23, 30.31 respectively (table 2). The free radical scavenging by DPPH method also showed potent result at higher doses for hydroalcoholic extract. From the above findings, it can be concluded that the plant N. arbortristis showed the strong antioxidant property. The IC50 value showed good results for hydroalcoholic extracts of N. arbortristis in reducing power, TBA and DPPH methods (5.93, 6.16 and 14.81 respectively). The results for IC50 values for the standard Kaempferol was found to be 2.48 for the above three methods (table 4). The graphical representation of the above-mentioned activities for the various extracts are given in fig. 1, 2, 3 and 4 respectively.

Table 2: Results of preliminary phytochemical screening

Chemical test Pet. ether Hydroalcoholic
Alkaloid -ve -ve
Tannins +ve +ve
Saponins +ve +ve
Glycoside +ve +ve
Carbohydrates -ve -ve
Flavonoids +ve +ve
Proteins and amino acid +ve +ve
Phenolics +ve +ve

*(+ve) and (-ve) symbol indicates the presence and absence of respective plant constituents.

Table 1: In vitro Antioxidant activity of Kaempferol (Stand.), Pet. Ether, Hydro-alcoholic extract of N. arbortristis by RP method

  • Reducing the power method

S. No. Extracts Activity 20 μg/ml 40 μg/ml 60 μg/ml 80 μg/ml 100 μg/ml
01 Kaempferol(Stnd.)

Reducing

Power

33.05±0.001 45.57±0.002 57.02±0.001 67.30±0.05 73.87±0.01
O2 Pet. Ether Extract 12.44±0.020 21.10±0.018 28.02±0.022 35.15±0.012 43.02±0.039
03 Hydro alcholic Extract 17.47±0.019 26.34±0.020 34.13±0.025 46.36±0.010 57.12±0.030

Fig. 1: Reducing power activity of different extracts of N. arbortristis

Table 2: In vitro antioxidant activity of Kaempferol (Stand.), Pet. Ether, Hydro-alcoholic extract of N. arbortristis by DPPH scavenging method

  • DPPH assay

S. No. Extracts Activity 20 μg/ml 40 μg/ml 60 μg/ml 80 μg/ml 100 μg/ml
01 Kaempferol(Stnd.) DPPH 33.05±0.001 45.57±0.002 57.02±0.001 67.30±0.05 73.87±0.01
O2 Pet. Ether Extract 08.23±0.12 14.49±0.012 15.06±0.003 18.01±0.032 21.12±0.43
03 Hydro alcholic Extract 30.31±0.32 35.47±0.017 42.06±0.014 47.51±0.032 54.32±0.43

Fig. 2: DPPH scavenging activity of (% inhibition) of N. arbortristis

Fig. 3: TBA scavenging power activity (% inhibition) of N. arbortristis

Table 3: In vitro antioxidant activity of Kaempferol (Stand.), Pet. Ether, Hydro-alcoholic extract of N. arbortristis by TBA scavenging method

  • TBA method

S. No. Extracts Activity 20 μg/ml 40 μg/ml 60 μg/ml 80 μg/ml 100 μg/ml
01 Kaempferol(Stnd.) TBA 33.05±0.001 45.57±0.002 57.02±0.001 67.30±0.05 73.87±0.01
O2 Pet. Ether Extract 29.25±0.011 31.24±0.002 38.40±0.101 39.67±0.04 45.82±0.011
03 Hydro alcholic Extract 32.12±0.101 37.43±0.001 43.14±0.005 45.34±0.05 50.54±0.02

Table 4: IC50 value from reducing power, DPPH, TBA Scavenging activity of roots of N. arbortristis

S. No. Activity Extract IC50
01 Reducing Power Standard 2.48
Pet. Ether 4.38
Hydroalcholic 5.93
02 DPPH Standard 2.48
Pet. Ether 4.34
Hydroalcholic 14.81
03 TBA Standard 2.48
Pet. Ether 4.85
Hydroalcholic 6.16

AUTHORS CONTRIBUTIONS

All the author have contributed equally

CONFLICT OF INTERESTS

There was no conflict of interest

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About this article

Title

ANTIOXIDANT ACTIVITY OF HYDRO-ALCOHOLIC EXTRACT ON THE ROOTS OF NYCTANTHES ARBORTRISTIS

Keywords

Nyctanthes arbortristis, Phytochemistry, Antioxidant Activity

DOI

10.22159/ijcpr.2018v10i4.28463

Date

16-07-2018

Additional Links

Manuscript Submission

Journal

International Journal of Current Pharmaceutical Research
Vol 10, Issue 4 (July-Aug), 2018 Page: 51-54

Online ISSN

0975-7066

Authors & Affiliations

Sumit Das
Girijananda Chowdhury Institute of Pharmaceutical Science, Azara, Hatkhowapara, Guwahati 17

Shilpa Paul
Girijananda Chowdhury Institute of Pharmaceutical Science, Azara, Hatkhowapara, Guwahati 17

Akm Abdullah
Girijananda Chowdhury Institute of Pharmaceutical Science, Azara, Hatkhowapara, Guwahati 17

Abdul Samim
Girijananda Chowdhury Institute of Pharmaceutical Science, Azara, Hatkhowapara, Guwahati 17


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