Original Article

, Volume: 15( 4)

• View PDF
• Download PDF

Antioxidant Activity of Wood and Leaf Extracts of Achyranthes aspera Using Radical Scavenging Method

*Correspondence:

Bahule BBDepartment of Chemistry, Nowrosjee Wadia College, Savitribai Phule Pune University, Pune, Maharashtra, India, Tel: 09763225085; E-mail: bharatbahule@gmail.com

Abstract

Achyranthes aspera Linn. is an indigenous herb. It is traditionally used as an abortifacient. The aqueous extract is given as a medicine against pneumonia. The aqueous extracts of this plant were screened for their antioxidant activity using DPPH radical scavenging method. The standard used is an ascorbic acid and the phytochemicals exhibited better activity than the standard compound. This extract is comprised of alkaloids and terpenoids as its major constituent and the biological activity can be attributed to the presence these compounds.

Keywords

Achyranthes aspera; Aqueous extract; Phytochemicals; Antioxidant activity

Introduction

The countries like India and China are well known for the various applications of medicinal plants for the treatment of diseases. The polyherbal and herbal therapies are employed combine for effective medication with minimum or no side effect [1]. In the treatment of cancer and diabetics, some of these methods were found to be more useful than the conventional methods. Many medicinal plants show the antioxidant and antidiabetic properties [2]. Most of these plants contain glycosides, alkaloids, terpenoids and flavanoids [3]. Many countries are using different plant species for the prevention and control of diabetics. Naturally derived drugs are having low side effects as compared to synthetic drugs. The root extracts of Achyranthes aspera is known to possess the diuretic property [4]. The dried plant is used for the treatment of gonorrhoea, fever, dysentery, asthama, hypertension and diabetics. The flowering spikes are used for external application in case of reptile and snake bite treatment [5]. The plant is useful in liver complaints, skin diseases [6]. Ash of the plant is used for the treatment of ulcer and warts [7]. Considering the wide range of applications of this plant, we have designed our study to screen the biological activity of the same. For that purpose, we have prepared the aqueous extracts of the plant and stored them in the refrigerator and used whenever required.

Materials and Methods

Collection of plants

Achyranthes aspera was collected from Pune in Maharashtra state, India. Fresh plants were used for extraction and isolation of their phytochemicals. Phytochemicals and extracts were stored in the refrigerator and used whenever needed.

Extraction

Aqueous extraction: The 25 g of dried and crushed plant material was soaked in 25 mL, 50 mL and 100 mL of distilled water respectively for 24 h. The extracts were filtered through the muslin cloth. The final volumes were corrected to viz. 25 mL, 50 mL and 100 mL by washing residue with distilled water and used for all activity.

Procedure for isolation of phytochemicals: The phytochemicals were isolated using known method [8]. Whole fresh plant material weighing 35 g Achyranthes aspera was dried, powdered and homogenized by methanol: water mixture (4:1). Then it was filtered and the filtrate was evaporated. 2MH₂SO₄ was added and the organic compounds were extracted with chloroform to furnish terpenoids. Aqueous layer was basified with NH₄OH and then extracted with chloroform-methanol (3:1). This extract afforded most of the alkaloids whereas remaining aqueous basic layer was evaporated and extracted with methanol yielded quaternary alkaloids. During purification of quaternary alkaloids, tannins were separated.

Phytochemicals: The phytochemicals were analyzed and identified using existing characterization techniques.

Dilution and concentration: Dilution and concentrations were prepared using literature procedure [9].

Dilution of water extracts: 25 mL, 50 mL and 100 mL of these dilutions were prepared by washing residue with distilled water and these dilutions were used for evaluation of all the activity.

Concentration of isolated phytochemicals: 50 μg/mL, 100 μg/mL, 200 μg/mL and 500 μg/mL concentration were prepared using respective solvent for DPPH radical scavenging activity [10,11].

DPPH radical scavenging activity: Free radical scavenging activity of different extracts of leaves and flowers of Achyranthes aspera Linn. Plant was measured by 1, 1-diphenyl-2-picryl hydrazyl (DPPH). In brief, 0.1 mM solution of DPPH in ethanol was prepared. This solution (1 mL) was added to 3 mL. of different extracts in Methanol at different concentration (50, 100, 200, 500 ppm). Here, only those extracts are used which are soluble in methanol and their various concentrations were prepared by dilution. The mixture was shaken vigorously and allowed to stand at room temperature for 30 min; the absorbance was measured at 517 nm by using spectrophotometer.

Reference standard compound used was ascorbic acid and experiment was done in triplicate. The value of the sample, which is the concentration of samples required to inhibit 50% of the DPPH free radical, was calculated using long dose inhibition curve. Lower absorbance of the reaction mixture indicated higher free radical activity (TABLES 1 to 3). The percent DPPH scavenging effect was calculated by using following equation:

TABLE 1. Absorbance of different isolated phytochemicals with standard ascorbic acid at 517 nm.

TABLE 2. % Inhibition of different isolated phytochemicals with standard ascorbic acid.

TABLE 3. Absorbance of different aqueous extracts at 517 nm.

Where A0 was the absorbance of control reaction and A1 was the absorbance in presence of test or standard sample. Control absorbance=0.2492.

Results and Discussion

This study determined that isolated phytochemicals and aqueous extract of Achyranthes aspera species showed better antioxidant potential by DPPH radical scavenging method compare to standard ascorbic acid (TABLE 4). The absorbance at 517 nm by UV visible spectrophotometer were found to be as compared to ascorbic acid more % of inhibition showed at 25% means it is nothing but 100% aqueous extract. In isolated phytochemicals 50 ppm terpenoid showed high percentage of inhibition as compared to other phytochemicals. In 100 ppm, 200 ppm and 500 ppm quaternary alkaloid showed high % of inhibition as compared another isolated phytochemical.

TABLE 4. % Inhibition of different aqueous extract.

Conclusion

By DPPH radical scavenging activity aqueous and isolated phytochemicals are exhibited better activity or better % of inhibition with standard ascorbic acid. The method used for activity is simple, fast and economical.

Acknowledgement

The authors are thankful to the Principal Nowrosjee Wadia College Pune for providing the necessary infrastructure and constant encouragement.

References

1. Rashid ANM, Hossain MS, Hassan N, et al. A review on medicinal plant with anti-diabetic activity. J Pharmacognosy and Phytochem. 2014;3(4):149-59.
2. Malviya N, Jain S, Malviya S. Antidiabetic potential of medicinal plants. Acta Poloniaepharmaceutica: Drug Res. 2010;67(2):113-8.
3. Nwaoguikpe RN. The effect of extract of bitter leaf (Vernonia amygdalina) on blood glucose levels of diabetic rats. Indian J Biochem. 2010;4(3):721-9.
4. Ebong PE, Atangwho J, Eyong EU, et al. The Antidiabetic Efficacy of combined extracts two continental plants: Azadirachta indica and Vernonia amygdalina. Am J Biochem and Biotechnol. 2008;4(3):239-44.
5. Srivastava S, Singh P, Mishra G. Achyranthes aspera: An important medicinal plant: A review. J Nat Prod Plant Resour. 2011;1(1):1-14.
6. Shibeshi W, Makonnen E, Zerihun L, et al. Effect of Achyranthes aspera on fetal abortion, uterine and pituitary weights. Serum lipids and hormones. African Health Sci. 2006;6(2):108-11.
7. Gomes A, Das R, Sarkel S, et al. Herbs and herbal constituents active against snake bite. Indian J Exp Biol. 2010;48:865-78.
8. Verma S. Medicinal plants used in cure of skin diseases. Adv in App Sci Res. 2016;7(3):65-7.
9. Arumugam G, Manjula P, Pari N. A review: Antidiabetic medicinal plant used for diabetes mellitus. J Acute Dis. 2013;pp:196-200.
10. Shimpi SR, Chaudhari LS, Bharambe SM, et al. Evaluation of antimicrobial activity of organic extract of leaves of Aristolochia bracteata. Pesticide Res J. 2005;17(1):16-8.
11. Rita MP, Shailendra BM, Pramod PM. Evaluation of in vitro antimicrobial activity of phytochemicals and extract of Enicostemma littorale. J Pure and App Microbiol. 2010;4(1):379-85.