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Vol 3 | Issue 1 | Jan – Mar 2024                                                                                     Indian J Pharm Drug Studies | 35  

Original Article 

In-vitro anti-oxidant activity of leaves of Cicer arietinum 

Supriya Mhamane1, Priyanka Adat2 

From, 1Sahyadri College of Pharmacy, Methwade, Sangola, Maharashtra, 2Assistant Professor, Pharmaceutical Chemistry, Sahyadri 

College of Pharmacy, Methwade, Sangola, Maharashtra, India. 

ABSTRACT  

Background: Cicer arietinum also known as chickpea. Chickpea plant belongs to family Fabaceae. Chickpeas are an essential food 

plant in India, Africa and Central and South America. This study concludes the Cicer arietinum leave’s anti-oxidant activity. It shows 

anti-oxidant activity and it is used to reduce oxidative stress. Materials and Methods: This work conceived the anti-oxidant activity 

of leave’s of Cicer arietinum via DPPH scavenging activity and measuring the absorbance at 517nm. Result: The maximum peak of 

free radical scavenging activity by using ethanolic and hydro-alcoholic extract is observed at 400µg/ml. By comparing both the 

extract hydro-alcoholic extract shows maximum percentage inhibition. Conclusion: Thus, the high percentage inhibition of hydro-

alcoholic extract which is 51.31% can be more effective as an antioxidant than the ethanolic extract. 

Keywords: Cicer arietinum, DPPH, Ascorbic acid, Free radical scavenging activity, Oxidative stress. 

ll over the world, traditional medicine has a long 

history of serving people. As ancient as human 

civilization, there is use of natural products with 

therapeutic properties. Main sources of drugs for a long time 

are mineral, plant and animal products. In almost all ancient 

civilizations, there is an evidence of herbs being used in the 

treatment of diseases and for revitalizing body systems. Free 

radicals included in ROS (Reactive Oxygen Species) can 

cause liver cirrhosis, atherosclerosis, cancer, diabetes. To 

protect body from damage due to reactive oxygen species, 

antioxidants are used. In the diabetic state, oxidative stress is 

increased. Oxygen free radical activity can lead to 

peroxidation of lipids, which in turn activates glycation of 

protein, inactivation of enzymes and alterations in the 

structure and function of collagen, basement and other 

membranes and play a role in the long team of diabetes [1,7]. 

MATERIALS AND METHODS 

Extraction method: In soxhlet apparatus, add 50gm of 

powdered drug and pour the solvent system. Pour sovent upto 

which three cycles were complete. Then start the heating 

mentle at 550c and start water supply to condenser. The 

solvent pass from powder and along with constituents present 

in powder it settles in round bottom flask. Keep this process 

upto the solvent comes colourless [2,8]. 
 
 

Access this article online 

Received – 24th Jan 2024 

Initial Review – 02nd Feb 2024 

Accepted – 08th Feb 2024 Quick Response Code 

In-Vitro Anti-Oxidant Activity 

Oxidative stress: The imbalance between formation and 

accumulation of reactive oxygen species (ROS) in the body 

cells and tissues is called as oxidative stress [3]. 

Oxidative stress can lead to: Cancer, cardiovascular disease, 

neurologic disease, respiratory disease, rheumatoid arthritis, 

kidney disease, delayed sexual maturation, etc. [3,5]. 

Antioxidants: 

Exogenous antioxidants: Vitamin E, flavonoid. 

Prooxidant agents: Ascorbic acid, polyphenols, radiation [4]. 

Dpph Free Radical Scavenging Activity: 

Principle: DPPH i.e. 2,2-diphenyl-1-picrylhydrazyl is a deep 

violet colored stable free radical. It has unpaired electron. 

Antioxidant donate electron to DPPH, so non radical of pale 

yellow color form from free radical of deep violet color. Due 

to this, absorbance get decreased which is observed by UV 

spectrophotometer at 517 nm wavelength [1,6]. 

Chemicals used in the assay: DPPH (2,2-diphenyl-1-

picrylhydrazyl), methanol, ascorbic acid, ethanol. 

Procedure: Methanolic solution of DPPH was used to test 

free radical scavenging action of Cicer arietinum leaves 

__________________________________________________ 

Correspondence to: Supriya Mhamane, Korwali, Tal. Mohol, 

Dist. Solapur, Maharashtra, India. Email: 

supriyamhamane99@gmail.com. Tel.: +91 8080244609. 

A 

mailto:supriyamhamane99@gmail.com


Mhamane & Adat                An investigational study towards use of leaves of cicer arietinum as an anti-oxidant 

Vol 3 | Issue 1 | Jan – Mar 2024                                                                                     Indian J Pharm Drug Studies | 36  

extract. In this assay as a free radical, DPPH is used. 

Antioxidants reduce DPPH which is observed at 517 nm 

wavelength. In this assay, as a standard antioxidant, ascorbic 

acid is used. In this assay, reaction mixture is used, which 

contain 0.1 ml leaves extract of 6 different concentrations in 6 

different test tubes and add 3.9 ml methanolic DPPH solution 

in each test tube as shown in (Figure 1). As a standard, 0.1 ml 

ascorbic acid was used and as a control 0.1 ml methanol was

  used instead of extract. At a room temperature for 30 

minutes it is incubated. Observe the absorbance at 517 nm 

wavelength [1,6].  

 
Figure 1: Dilution of Anti-oxidant Activity  

Calculation: Percentage inhibition were calculated by using 

following formula: 

 
RESULTS  

In-vitro anti-oxidant activity of standard (Ascorbic acid), 

having percentage inhibition was found to be 96.83%. 

Table: In-vitro antioxidant activity of Cicer arietinum 

Conc. 

(µg/ml) 

Percentage Inhibition 

(Ethanolic) (%) 

Percentage Inhibition 

(Hydro-alc.) (%) 

25 5.26 5.17 

50 7.09 6.5 

100 7.27 8.01 

150 7.61 10.57 

200 9.62 12.99 

400 22.92 51.31 

DISCUSSION 

Cicer arietinum plant includes plant constituents like 

alkaloids, glycosides, steroids, carbohydrates, amino acids, 

inorganic elements. These constituents are responsible for 

showing anti-oxidant activity. In this study, the antioxidant 

activity was determined using DPPH free radical scavenging 

activity in which the purple color of DPPH gets reduced to 

yellow color due to transfer of electrons. Due to this, decrease 

in value of absorbance depicts that constituents of leaves of 

Cicer arietinum are able to reduce free radicals. 

Due to free radicals of oxygen, the number of reactive 

oxygen species (ROS) increases, so that the oxidative stress 

increases this can lead to various diseases. To overcome it, the 

antioxidants are used. Among two samples ethanolic and 

hydro-alcoholic extract, the highest percentage inhibition was 

observed in hydro-alcoholic extract at 400µg/ml which was 

51.31%. The percentage inhibition increases as concentration 

increases, which is as shown in table and graph. Ethanolic 

extract shows high percentage inhibition at 400µg/ml i.e. 

22.92%. Thus it could be concluded that hydro-alcoholic 

extract shows good antioxidant property. 

CONCLUSION 

The conclusion of this study shows that, the ethanolic and 

hydro-alcoholic extract of leaves of Cicer arietinum shows 

anti-oxidant activity by DPPH free radical scavenging 

activity. The hydro-alcoholic extract shows high percentage 

inhibition as compared with ethanolic extract. Hydro-

alcoholic extract at 400µg/ml shows high percentage 

inhibition i.e. 51.31%. It was observed as the concentration of 

extract increases from 25µg/ml to 400µg/ml The percentage 

inhibition also increases. From given Table, it can be 

concluded that hydro-alcoholic extract has the highest 

antioxidant activity or good scavenging activity. 

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5. Gabriele Pizzino, Natasha Irrera, Mariapaola Cucinotta, et al. 

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How to cite this article: Supriya Mhamane, Priyanka Adat. 

In-vitro anti-oxidant activity of leaves of Cicer arietinum. 

Indian J Pharm Drug Studies. 2024; 3(1):35-36. 

Funding: None;                   Conflicts of Interest: None Stated 

 

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