In-vitro antioxidant activity of hydroalcoholic extract of leaves of Hydnocarpus pentandrus (Buch. - Ham) Oken

Abstract

Cellular damage can arise from the reactions of free radicals with membrane lipids, nucleic acids, proteins, enzymes, and other micro molecules. Free radical-induced cell damage seems to be a primary factor in the aging process and degenerative diseases, including but not limited to cancer, heart disease, cataracts, liver disorders, diabetes mellitus, inflammation, and renal failure. Naturally, the body produces free radicals, and antioxidants scavenge them to shield the body from harmful consequences. This dynamic equilibrium exists between the two. It's possible that there aren't enough antioxidants in the body under typical physiological circumstances to offset the production of free radicals. Thus, it stands to reason that adding antioxidants to our diet will help shield us from dangerous illnesses. Thus, the creation of "natural antioxidants" from plant material has drawn more attention from the food business and preventative medicine. Given the importance of antioxidant activity, a hydroalcoholic crude extract from the leaves of the Achariaceae family plant Hydnocarpus pentandrus (Buch. - Ham) Oken was made, the 1,1-diphenyl-2-picrylhydrazyl (DPPH) Radical Scavenging Assay, the Ferric Ion Reducing Assay (FRAP), the Nitric Oxide Assay, and the H2O2 Radical Scavenging Assay were used to measure the radical scavenging activity. The antioxidant activity of the hydroalcoholic extract was examined in comparison to the reference; in all four methods, solvent-based findings were observed. Higher antioxidant activity is shown by the FRAP and DPPH assays, which are followed by the nitric oxide, and H2O2 radical assays. DPPH > FRAP > H2O2 > Nitric oxide.

Keywords

Introduction

Hydnocarpus pentandrus (Buch. - Ham) Oken belongs to Family Achariaceae. It is deciduous tropical tree distributed throughout India and few other tropical countries. Traditionally boiled leaves were applied in sprains and bruises, local swellings, pains of body and in elephantiasis [1]. Five dihydroflavonoids were isolated and identified from the leaves of Hydnocarpus pentandrus [2]. The leaves are useful in cuts, wounds, bruises, ulcers, ophthalmia, sprains, diarrhoea and dysentery. The fruit paste of Hydnocarpus pentandrus is therapeutically used for bone fractures by tribes in western ghats of India [3]. The pharmacological properties of extracts of Hydnocarpus pentandrus leaves were screened for antidiabetic activity [4]. In the present study we investigated radical scavenging potential of hydroalcoholic extract of leaves of Hydnocarpus pentandrus.

Oxygen consumption inherent in cell growth leads to the generation of a series of reactive oxygen species (ROS). They are continuously produced by the body’s normal use of oxygen such as respiration and some cell-mediated immune functions. ROS include free radicals such as superoxide anion radicals (O2), hydroxyl radicals (OH) and non - free radical species such as hydrogen peroxide (H₂O₂) and single oxygen [5]. ROS are continuously produced during normal physiologic of membrane lipids, leading to the accumulation of lipid peroxides. ROS are also capable of damaging crucial biomolecules such as nucleic acid, lipids, proteins and carbohydrates and may cause DNA damage that can lead to mutations. If ROS are not effectively scavenged by cellular constituents, they lead to disease conditions. ROS have been implicated in more than 100 diseases [6]. Antioxidants can protect the human body from free radicals and ROS effects. They retard the progress of many chronic disease as well as lipid peroxidation [7]. Hence, a need for identifying alternative natural and safe sources of food antioxidants has been created, and the search for natural antioxidants, especially of plant origin, has notably increased in recent years. Antioxidants have been widely used as food additives to provide protection against oxidative degradation of food [8].

MATERIAL & METHODS

The plant material of Hydnocarpus pentandrus for the present work was collected from Dapoli, Chandra Nagar, Maharashtra, India (17?44’36.3’’N 73?09’39.1’’E) & authenticated from the Blatter Herbarium, St. Xavier’s College, Mumbai (Accession no.- R3060). The leaves were shade dried. Dried leaves were ground to coarse powder, extracted with hydro alcohol. The antioxidant activity of the plant extract and standard were assessed based on radical scavenging effect of the stable DPPH free radical according to method of [9]. The reducing power of the hydroalcoholic extract was determined according to the method of [10]. The ability of the extract to scavenge hydrogen peroxide was determined according to the method of [11]. Nitric oxide was generated from nitroprusside and measured by the Griess reaction [12].

Result & Discussion

DPPH Free radical scavenging activity

DPPH is a free radical compound and has been widely used to test the free radical-scavenging ability of various samples [13]. The assay is based on the increase of DPPH radical in hydroalcoholic extract which causes an absorbance drop at 517 nm. DPPH is known to abstract labile hydrogen [14]. DPPH being a stable free radical can accept an electron or hydrogen radical to become stable diamagnetic molecule. Due to its odd electron, the hydro-alcoholic solution of DPPH shows a strong absorption at 517nm, DPPH radicals react with suitable reducing agents and then electrons become paired off and the solution loses colour stoichiometrically with the number of electrons taken up. Such reactivity has been widely used to test the ability of compounds in plant extracts to acts as a free radical scavenger. Antioxidants molecules can quench DPPH free radicals and convert them to a colourless product. Figure 1. shows the dose responses curve of DPPH radical scavenging activity of hydroalcoholic extract, compared with ascorbic acid, as standard.

       
           
       
Figure 1: DPPH Free radical scavenging activity of hydroalcoholic extracts

Reducing power assay (FRAP)

Hydrogen peroxide

Nitric oxide scavenging activity

CONCLUSION

The antioxidant activity of the hydroalcoholic extract was examined in comparison to the reference; in all four methods, solvent-based findings were observed. higher antioxidant activity is shown by the FRAP and DPPH assays, which are followed by the nitric oxide, and H₂O₂ radical assays. DPPH > FRAP > H₂O₂ > Nitric oxide. based on results obtained in the present study, it is concluded that hydroalcoholic extract of Hydnocarpus pentandrus leaves, exhibits high antioxidant and free radical scavenging activities. It also chelates iron and has reducing power. These in vitro assays indicate that this plant extract is a significant source of natural antioxidant, which might be helpful in preventing the progress of various oxidative stresses.

ACKNOWLEDGMENT

The authors are grateful to the Department of Botany, Mithibai College (Empowered Autonomous) for providing the required necessary facilities.

REFERENCES

1. Blois MS Antioxidant determinations by the use of stable free radical, Nature Journal of Pharmaceutical (1995),181:1199-1200.
2. Branen AL, Davidson PM & Salminen S Food Additives New York (1990),248:246-252.
3. Gordon MH The mechanism of antioxidant action in vitro. Food antioxidants. Elsevier Applied Science, London (1998), 46:667-672.
4. Govindachari TR, Parthasarathy PC, Desai HK & Mohammed PA: Cluytyl ferulate, a new long chain ester from Gmelina arborea and Lannea grandis. Indian Journal of Chemistry (1999), 9:1027.
5. Gulcin I Antioxidant and antiradical activities of I- Carnitine. Life Sci (2006),78:803-811.
6. Gulcin I, Buyukokurogu ME & Oktay M In vitro antioxidant properties of melatonin. Journal Pineal Res (2002),33:167 -171.
7. Gulcin I, Mshvildadze V & Gepdiremen A Antioxidant activity of a triterpenoid glycoside isolated from the berries of Hedera colchica :3-O-(?-d-glucopyranosyl)-hederagenin, Phytother. Res. (2006),20:130-134.
8. Halliwell B & Gutteridge JMC Role of free radicals and catalytic metal ions in human disease: an overview. Methanol Enzymol (1990), 186: 1-85.
9. Islam MT, Ito T, Sakasai M & Tahara S: Zoosporicidal Activity of Polyflavonoid Tannin Identified in Lannea coromandelica Stem Bark against Phytopathogenic Oomycete. Aphanomyces cochlioides. Journal Agric Food Chem (2002),6(23): 6697-6703.
10. Marcocci L, Maguire JJ, Droy-Lefaix MT & Packer L The nitric oxide scavenging properties of Gingko biloba ECB 761.Biochem. Biophy. Res. Commun. (1994), 15:748-755.
11. Matsubara T, Fuchimoto S & Iwagaki H The possible involvement of free radical scavenging properties in the action of cytokines. Res. Commun. Chem. Pathol. Pharmacol (1991),71(2):239-242.
12. Moncada A, Palmer RM & Higgs EA Nitric oxide physiology, pathophysiology and pharmacology. Pharmacol. Rev. (1991),43(2):109-142.
13. Nair AGR., Subramanian SS & Sridharan K Chemical examinations of flowers of Lannea coromandelica Current Science 3, (1963),4:115-116.
14. Oyaizu M Studies on products of browning reaction: Antioxidative activities of browning reactions products prepared from glucosamine. Eiyogaku Zasshi (1986), 44:307-315.
15. Rahman MS, Begum B, Chowdhury R, Rahman KM & Rashid MA Preminary cytotoxicity screening of some medicinal plants of Bangladesh, Dhaka Univ. Journal Pharm Sci. (2008), 7(1): 47-52.
16. Ratnam KV & Venkata RR Traditional medicine used by the adivasis of eastern Ghats Andra Pradesh -For bone fractures. Ethanobotanical leaflets (2000),7:19-22.
17. Rush RJ, Cheng SJ & Klaunig JE Preventation of cytotoxicity and inhibition of intracellular communication by antioxidant catechins isolated from Chinese green tea. Carcinogenesis (1989),10:1003-1008.
18. Sakanaka S, Tachibana Y & Okada Y Preventation and antioxidant properties of extracts of Japanese persimmon leaf tea (Kakinoha-cha). Food Chemistry (2005), 89:569-575.