IPSIndian Journal of Pharmacology
Home  IPS  Feedback Subscribe Top cited articles Login 
Users Online : 13639 
Small font sizeDefault font sizeIncrease font size
Navigate Here
 » Next article
 » Previous article 
 » Table of Contents
Resource Links
 »  Similar in PUBMED
 »  Search Pubmed for
 »  Search in Google Scholar for
 »Related articles
 »  Article in PDF (124 KB)
 »  Citation Manager
 »  Access Statistics
 »  Reader Comments
 »  Email Alert *
 »  Add to My List *
* Registration required (free)

In This Article
 »  Abstract
 »  Materials and Me...
 »  Results
 »  Discussion
 »  Acknowledgments
 »  References
 »  Article Tables

 Article Access Statistics
    PDF Downloaded868    
    Comments [Add]    
    Cited by others 25    

Recommend this journal


Year : 2007  |  Volume : 39  |  Issue : 6  |  Page : 281-283

Free radical scavenging activity of gossypin and nevadensin: An in-vitro evaluation

1 Pharmacognosy and Phytochemistry Division, College of Pharmaceutical Sciences, Andhra University, Visakhapatnam - 530 003, Andhra Pradesh, India
2 Department of Pharmacology, Hanagal Shri Kumareshwar College of Pharmacy, BVVS Campus, Bagalkot - 587 101, Karnataka, India
3 Center of Biotechnology, Institute of Science and Technology, JNTU, Hyderabad - 500 072, Andhra Pradesh, India

Date of Submission18-Oct-2007
Date of Decision22-Nov-2007
Date of Acceptance08-Jan-2008

Correspondence Address:
S Ganapaty
Pharmacognosy and Phytochemistry Division, College of Pharmaceutical Sciences, Andhra University, Visakhapatnam - 530 003, Andhra Pradesh
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0253-7613.39147

Rights and Permissions

 » Abstract 

Objectives: The antioxidant potential of gossypin and nevadensin, two flavonoid compounds, were evaluated by in vitro methods.
Materials and Methods: Gossypin, nevadensin, and the reference standard, butylated hydroxyl toluene (BHT), were evaluated for DPPH (1, 1-diphenyl-2-picrylhydrazyl), nitric oxide, superoxide, and hydroxyl radical scavenging activity.
Results: Gossypin and BHT showed the potential for significant DPPH radical inhibition of up to 88.52 and 91.45% at 100 µg/ml concentration. With a 100 µg/ml concentration of gossypin, the in vitro nitric oxide, superoxide, and hydroxyl radical scavenging activity was found to be 74.00, 74.22, and 67.15%, respectively; and with 100 µg/ml of BHT the corresponding values were 82.24, 81.76, and 73.03% of inhibition, respectively.
Conclusion: The study results showed that gossypin has significant antioxidant activity.

Keywords: Antioxidant activity, gossypin, nevadensin

How to cite this article:
Ganapaty S, Chandrashekhar V M, Chitme H R, Narsu M L. Free radical scavenging activity of gossypin and nevadensin: An in-vitro evaluation. Indian J Pharmacol 2007;39:281-3

How to cite this URL:
Ganapaty S, Chandrashekhar V M, Chitme H R, Narsu M L. Free radical scavenging activity of gossypin and nevadensin: An in-vitro evaluation. Indian J Pharmacol [serial online] 2007 [cited 2023 Sep 28];39:281-3. Available from: https://www.ijp-online.com/text.asp?2007/39/6/281/39147

Oxygen-derived free radicals such as super oxide anion and hydroxyl radical are cytotoxic and promote tissue injuries. Antioxidants act as a major defense against radical-mediated toxicity by protecting against the damages caused by free radicals. [1] The cellular antioxidant status determines the susceptibility to oxidative damage and is usually altered in response to oxidative stress. [2]

The protection offered by fruits and vegetables against oxidative stress in several diseases has been attributed to various antioxidants and vitamins. Dietary phenolic compounds and flavonoids have generally been considered as non- nutrients and their possible beneficial effect on human health have only recently been recognized. Flavonoids are known to possess anti-inflammatory, antioxidant, antiallergic, hepatoprotective, antithrombotic, neuroprotective, and anticarcinogenic activities. [3],[4] Therefore, the search for natural antioxidants of plant origin has gained momentum in recent years. In this work, two flavonoids, gossypin and nevadensin, are studied for their in vitro antioxidant activities.

 » Materials and Methods Top

The chemicals DPPH (1,1-diphenyl-2-picrylhydrazyl), TBA (thiobarbituric acid), Griess reagent, and NBT (nitroblue tetrazolium) were obtained from Sigma, St. Louis, MO, USA. Trichloroacetic acid (TCA) and potassium superoxides were obtained from Merck, KGOA, Germany. All other chemicals and reagents used were of analytical grade.

DPPH radical scavenging activity

The free radical scavenging activity was measured in terms of hydrogen donating or radical scavenging ability, using the stable radical, DPPH. [5] A 0.1 mM solution of DPPH in methanol was prepared and 1.0 ml of this solution was added to 3.0 ml of control (without the test compound, but with an equivalent amount of methanol), i.e., standard butylated hydroxyl toluene (BHT) at different concentration (25-100 μg/ml), and test solutions at different concentrations (5-100 μg/ml) in different test tubes. Thirty minutes later, the absorbance was measured at 517 nm.

Nitric oxide scavenging activity

Nitric oxide scavenging activity was measured by the spectrophotometry method. [6] Sodium nitroprusside (5 mmol) in phosphate-buffered saline was mixed with a control without the test compound, but with an equivalent amount of methanol. Test solutions at different concentrations (5-100 μg/ml) were dissolved in methanol and incubated at 25°C for 30 min. After 30 min, 1.5 ml of the incubated solution was removed and diluted with 1.5 ml of Griess reagent (1% sulphanilamide, 2% phosphoric acid, and 0.1% naphthyl ethylenediamine dihydrochloride). The absorbance of the chromophore formed during the diazotization of the nitrite with sulphanilamide and the subsequent coupling with naphthylethylene diamine dihydrochloride was measured at 546 nm.

Superoxide scavenging

Superoxide scavenging was carried out using the alkaline dimethyl sulfoxide (DMSO) method. [6] Solid potassium superoxide was allowed to stand in contact with dry DMSO for at least 24 h and the solution was filtered immediately before use; the filtrate (200 μl) was added to 2.8 ml of an aqueous solution containing nitroblue tetrazolium (56 μmol), EDTA (10 μmol), and potassium phosphate buffer (10 mmol, pH 7.4). Test solutions at different concentrations (5-100 μg/ml) were added and absorbance was recorded at 560 nm against the control, in which pure DMSO had been added instead of alkaline DMSO.

Hydroxyl radical scavenging activity

The scavenging capacity for hydroxyl radical was measured according to the modified method. [7] The assay was performed by adding 0.1 ml EDTA, 0.01 ml of FeCl 3 , 0.1 ml H 2 O 2 , 0.36 ml of deoxyriboase, 1.0 ml of test solutions (5-100 μg/ml) dissolved in distilled water, 0.33 ml of phosphate buffer (50 mM, pH 7.4), and 0.1 ml of ascorbic acid in sequence. The mixture was then incubated at 37°C for 1 h. A 1.0 ml portion of the incubated mixture was mixed with 1.0 ml of 10% TCA and 1.0 ml of 0.5% TBA to develop the pink chromogen, which was measured at 532 nm.

Statistical analysis

The results are presented as means ± SEM. All parameters were analyzed using Student's t-test. P < 0.05 was considered significant.

 » Results Top

Inhibition of DPPH radical

The potential decrease in the concentration of DPPH radical due to the scavenging ability of gossypin and BHT (reference standard) showed significant free radical scavenging activity: 91.45% and 88.52% of inhibition, respectively, at 100 μg/ml. The IC 50 (the inhibitory concentration at which there is 50% reduction of free radical) of gossypin was found to be 31 μg/ml. Nevadensin did not show any significant activity [Table - 1].

Nitric oxide scavenging activity

The scavenging of nitric oxide by gossypin and BHT was concentration dependent. There was a moderate inhibition of nitric oxide formation, with the maximum inhibition being 74.00 and 82.24% at 100 μg/ml of gossypin and BHT, respectively. Similar activity was not found in the case of nevadensin [Table - 1].

Superoxide radical scavenging

A moderate inhibition of the superoxide radical was observed with 100 μg/ml each of gossypin and BHT (74.22 and 81.76% respectively). There was no significant inhibition of superoxide radical by nevadensin [Table - 2].

Hydroxyl radical activity

The effect of gossypin and BHT on hydroxyl radical and iron (II)-dependent deoxyribose damage was protected significantly at all concentrations; the percentage of inhibition of hydroxyl radical being 67.15% and 73.03%, respectively, at 100 μg/ml. There was moderate inhibition of hydroxyl radical activity by nevadensin (62.83%) as compared to BHT, which showed 73.03% inhibition at 100 μg/ml concentration [Table - 2].

 » Discussion Top

Oxidative stress, in which large quantities of reactive oxygen species (ROS) like hydrogen peroxide, superoxide ( * O -2 ), hydrogen radical (OH - ), singlet oxygen, and nitrogen species are generated, is one of the earliest responses to stress. These ROS have a role in disease and aging in animals. [8] The antioxidative system protects the organism against ROS-induced oxidative damage. There are restrictions on the use of synthetic antioxidants, such as BHT, as they are suspected to be carcinogenic. [9] Natural antioxidants, therefore, have gained importance.

DPPH is a stable free radical at room temperature and accepts an electron or hydrogen radical to become a stable diamagnetic molecule. The reduction capability of DPPH radicals was determined by the decrease in its absorbance at 517 nm, which is induced by antioxidants. The significant decrease in the concentration of the DPPH radical is due to the scavenging ability of gossypin.

Nitric oxide was generated from sodium nitroprusside and measured by the Greiss reduction. Sodium nitroprusside in aqueous solution at physiological pH spontaneously generates nitric oxide, which interacts with oxygen to produce nitrate ions that can be estimated by use of Greiss reagent. Scavengers of nitric oxide compete with the oxygen, leading to reduced production of nitric oxide. [10] Significant scavenging activity was observed for gossypin.

The potentially reactive hydroxyl radicals can cause oxidative damage to DNA, lipids, and proteins. The effect of gossypin and BHT on the inhibition of free radical-mediated deoxyribose damage was assessed by means of iron (II)-dependent DNA damage assay, which showed significant results. [11]

The flavonoid gossypin has potent antioxidant and free radical scavenging effects in different in vitro systems, but nevadensin showed no significant effects as compared to standard BHT. Further work is necessary to elucidate the mechanism involved in the antioxidant activity of gossypin.

 » Acknowledgments Top

The authors thank the Principal, H.S.K. College of Pharmacy, Bagalkot, for providing the facilities necessary to carry out the work. We also thank Mr. Giresh. K. Gowda, Research scholar, University of Leads, Leeds (UK), for providing the literature.

 » References Top

1.Mallika J, Shyamala Devi CG. In vitro and in vivo evaluation of free radical scavenging potential of Cissus quadranguloris . Afr J Biomed Res 2005;8:95-9.  Back to cited text no. 1    
2.Bhor VM, Raghuram N, Sivakami S. Oxidative damage and altered antioxidant enzyme activities in the small intestine of Streptozotocin induced diabetic rats. Int J Biochem Cell Biol 2004;36:89-97.  Back to cited text no. 2  [PUBMED]  [FULLTEXT]
3.Araceli S, Camen RM, Guillermo RS, Salvador M, Rosa MG. Assessment of the anti-inflammatory activity and free radical scavenger activity of tiliroside. Eur J Pharmacol 2003;461:53-61.  Back to cited text no. 3    
4.Rajnaryana K, Sripalreddy M, Chalavadi MR, Krishna DR. Bioflavonoids classification, pharmacological, Biochemical effects and therapeutic potential. Indian J Pharmacol 2001;33:2-16.  Back to cited text no. 4    
5.Sreejayan N, Rao MN. Free radical scavenging activity of curcuminoids. Drug Res 1996;46:169-71.  Back to cited text no. 5    
6.Madan MP, Raghavan G, Ajay Kumar Singh R, Palpu P. Free radical scavenging potential of Saussarea costus . Acta Pharm 2005;55:297-304.  Back to cited text no. 6    
7.Rajeshwar Y, Senthil Kumar GP, Malay AG, Mazumder UK. Studies on in vitro antioxidant activities of methanol extract of Mucuna pruriens (Fabaceae) sedds. Eur Bull Drug Res 2005;13:131-8.  Back to cited text no. 7    
8.Melov S. Animal models of oxidative stress, aging and therapeutic antioxidant interventions. Int J Biochem Cell Biol 2002;34:1395-400.  Back to cited text no. 8  [PUBMED]  [FULLTEXT]
9.Singh RP, Chidamara Murthy KN, Jayaprakasha GK. Studies on the antioxidant activity of Pomegranate ( Punica granatum ) Peel and seed extracts using in vitro models. J Agric Food Chem 2002;50:81-6.  Back to cited text no. 9    
10.Govindarajan R, Rastogi S, Vijayakumar M, Shirwaikar A, Rawat AK, Mehrotra S, et al . Studies on antioxidant activities of Desmodium gangeticum . Biol Pharm Bull 2003;26:1424-7.  Back to cited text no. 10  [PUBMED]  [FULLTEXT]
11.Jornot L, Petersen H, Junod AF. Hyrogen peroxide induced DNA damage is independent of nuclear calcium but dependent on redox active ions. Biochem J 1998;335:854-5.  Back to cited text no. 11    


  [Table - 1], [Table - 2]

This article has been cited by
1 Prediction Mechanism of Nevadensin as Antibacterial Agent against S. sanguinis: In vitro and In silico Studies
Aldina Amalia Nur Shadrina, Yetty Herdiyati, Ika Wiani, Mieke Hemiawati Satari, Dikdik Kurnia
Combinatorial Chemistry & High Throughput Screening. 2022; 25(9): 1488
[Pubmed] | [DOI]
2 Natural Compounds That Enhance Motor Function in a Mouse Model of Muscle Fatigue
Shuichi Shibuya, Kenji Watanabe, Daiki Sakuraba, Takuya Abe, Takahiko Shimizu
Biomedicines. 2022; 10(12): 3073
[Pubmed] | [DOI]
3 Gossypin'in farkli kanser hücre dizilerinde HSP60 ve HSP70'in gen ekspresyonu üzerindeki etkisi
Ebubekir DIRICAN, Irfan ÇINAR
Cukurova Medical Journal. 2022; 47(2): 497
[Pubmed] | [DOI]
Viswanath K, Binoy C. Varghese, Vijey Aanandhi M
Indian Drugs. 2022; 58(12): 32
[Pubmed] | [DOI]
5 Antimicrobial Activity of Brassica rapa L. Flowers Extract on Gastrointestinal Tract Infections and Antiulcer Potential Against Indomethacin-Induced Gastric Ulcer in Rats Supported by Metabolomics Profiling
Badriyah Alotaibi, Fatma Alzahraa Mokhtar, Thanaa A El-Masry, Engy Elekhnawy, Sally A Mostafa, Dalia H Abdelkader, Mohamed E Elharty, Asmaa Saleh, Walaa A Negm
Journal of Inflammation Research. 2021; Volume 14: 7411
[Pubmed] | [DOI]
6 Modulation of Liver P-Glycoprotien Expression May Contribute to Gossypin Protection against Methotrexate-Induced Hepatotoxicity
Mervat Mohamed, AzzaKamal El Sheikh, HanaaHassanien Mohammed
Indian Journal of Pharmacology. 2021; 53(1): 25
[Pubmed] | [DOI]
7 Antioxidant and anti-inflammatory activities of methanol and aqueous extracts of Sargassum wightii
Sradhasini Rout, Bandana Rath, Subrat Kumar Bhattamisra, Ishani Rath, Anjan Kumar
Journal of Herbmed Pharmacology. 2021; 11(1): 75
[Pubmed] | [DOI]
8 Evaluation of the protective effects of gossypin for ischemia/reperfusion injury in ovary tissue
Busra Dincer, Irfan Cinar, Muhammed Yayla, Erdem Toktay
Journal of Obstetrics and Gynaecology Research. 2021;
[Pubmed] | [DOI]
9 Phytochemical analysis of Ocimum gratissimum by LC-ESI–MS/MS and its antioxidant and anxiolytic effects
M.P. Venuprasad,Hemanth Kumar Kandikattu,Sakina Razack,Farhath Khanum
South African Journal of Botany. 2014; 92: 151
[Pubmed] | [DOI]
10 In Vitro Antioxidant and Anticancer Activity of Flavonoid Fraction from the Aerial Parts of Cissus quadrangularis Linn. against Human Breast Carcinoma Cell Lines
A. Vijayalakshmi,P. R. Kumar,S. Sakthi Priyadarsini,C. Meenaxshi
Journal of Chemistry. 2013; 2013: 1
[Pubmed] | [DOI]
11 Neuroprotective activity of gossypin from Hibiscus vitifolius against global cerebral ischemia model in rats
VM Chandrashekhar,S Ganapaty,A Ramkishan,MLaxmi Narsu
Indian Journal of Pharmacology. 2013; 45(6): 575
[Pubmed] | [DOI]
12 Antioxidant activity free radical-scavenging potential of Pithecellobium dulce Benth seed extracts
Dnyaneshwar M. Nagmoti,Dharmendra K. Khatri,Parikshit R. Juvekar,Archana R. Juvekar
Free Radicals and Antioxidants. 2012; 2(2): 37
[Pubmed] | [DOI]
13 In Vitro Antioxidant Activity of Cucurbita Maxima Aerial Parts
Prerona Saha,U.K. Mazumder,P.K. Haldar
Free Radicals and Antioxidants. 2011; 1(1): 42
[Pubmed] | [DOI]
14 On-line coupling of dynamic microwave-assisted extraction with high-speed counter-current chromatography for continuous isolation of nevadensin from Lyeicnotus pauciflorus Maxim.
Xing Tong, Xiaohua Xiao, Gongke Li
Journal of Chromatography B. 2011;
[VIEW] | [DOI]
15 Solid state characterization of an antioxidant alkaloid boldine using vibrational spectroscopy and quantum chemical calculations
Anubha Srivastava, Poonam Tandon, A.P. Ayala, Sudha Jain
Vibrational Spectroscopy. 2011; 56(1): 82
[VIEW] | [DOI]
16 On-line coupling of dynamic microwave-assisted extraction with high-speed counter-current chromatography for continuous isolation of nevadensin from Lyeicnotus pauciflorus Maxim
Tong, X., Xiao, X., Li, G.
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences. 2011; 879(24): 2397-2402
17 Free radical scavenging activity of Amaranthus blitum
Saranya, P., Gurusamy, K.
Pharmacologyonline. 2010; 3: 625-636
18 Synthesis of Gossypin derivatives as potential anticancer and anti-inflammatory agents
Kotra, V., Ganapatya, S., Adapa, S.R.
Indian Journal of Chemistry - Section B Organic and Medicinal Chemistry. 2010; 49(11): 1542-1546
19 Nevadensin: Isolation, chemistry and bioactivity
Brahmachari, G.
International Journal of Green Pharmacy. 2010; 4(4): 213-219
20 Synthesis, characterization and biological activity of some α-aminophosphonates
Oniţa, N., Şişu, I., Penescu, M., Purcarea, V.L., Kurunczi, L.
Farmacia. 2010; 58(5): 531-545
21 Evaluation of anti-allergic activity of gossypin and suramin in mast cell-mediated allergy model
Ganapaty, S., Chandrashekhar, V.M., Narsu, M.L.
Indian Journal of Biochemistry and Biophysics. 2010; 47(2): 90-95
22 Antioxidant studies of methanolic extract of Randia Dumetorum Lam
Dharmishtha, A.M., Mishra, S.H., Falguni, P.G.
Pharmacologyonline. 2009; 1: 22-34
23 Free radical scavenging activity of Stereospermum suaveolens DC: An in-vitro evaluation
Chandrashekhar, V.M., Muchandi, A.A., Sudi, S.V., Muchandi, I.S.
Pharmacologyonline. 2009; 1: 50-56
24 Antioxidant studies of aqueous extract of Phyllanthus reticulatus Poir
Abstract + Refs Show Abstract Aswatha Ram, H.N., Shreedhara, C.S., Falguni Gajera, P., Sachin Zanwar, B.
Pharmacologyonline. 2008; 1: 351-364
25 In vitro free radical scavenging potential of methanol extract of entire plant of Phyllanthus reticulatus poir
Aswatha Ram, H.N., Shreedhara, C.S., Gajera, F.P., Zanwar, S.B.
Pharmacologyonline. 2008; 2: 440-451


Print this article  Email this article


Site Map | Home | Contact Us | Feedback | Copyright and Disclaimer | Privacy Notice
Online since 20th July '04
Published by Wolters Kluwer - Medknow