INTRODUCTION:

The term "antioxidant" is mostly used for two entirely different groups of substances: industrial chemical that are added to products to prevent oxidation, and naturally occurring compounds that are present in foods and tissue. The former, industrial antioxidants, have diverse uses: acting as preservatives in food and cosmetics, and being oxidation-inhibitors in fuels.⁽¹⁾A substance that inhibits oxidation, especially one used to counteract the deterioration of stored food products.Antioxidants are an inhibitor of the process of oxidation, even at relatively smallconcentration and thus have diverse physiological role in the body. Antioxidant constituents of the plant material act as radical scavengers, and helps in converting the radicals to less reactive species. A variety of free radical scavenging antioxidants is found in dietarysources like fruits, vegetables and tea, etc.⁽²⁾Oxidative stress is characterized as an imbalance between the production of reactive species and antioxidant defense activity, and its enhanced state has been associated with many of the chronic diseases such as cancer, diabetes, neurodegenerative and cardiovascular diseases.⁽³⁾

There is our days, an increasing interest in the measurement and use of plant antioxidants for scientific research as well as industrial (dietary, pharmaceutical and cosmetic) purposes. This is mainly due to their strong biological activity, exceeding those of many synthetic antioxidants which have possible activity as promoters of carcinogenesis.⁽⁴⁾

The traditional medicine all over the world is nowadays revalued by an extensive activity of research on different plant species and their therapeutic principles. Experimental evidence suggests that free radicals (FR) and reactive oxygen species (ROS) can be involved in a high number of diseases (Richards and Sharma, 1991, Niwa, 1991). As plants produce a lot of antioxidants to control the oxidative stress caused by sunbeams and oxygen, they can represent a source of new compounds with antioxidant activity. Ayurveda, the Indian traditional health care system (ayus=life, veda=knowledge, meaning science of life), is the oldest medical system in the world and is being revived in its complete form under the name of Maharishi Ayurved (Glaser, 1988). The World Health Organization has approved its efficacy (Zaman, 1974). This system provides an approach to prevention and treatment of different diseases by a large number of medical procedures and pharmaceuticals. One of the clinical specialties’ of Ayurveda is Rasayana. Rasayana is not only a drug therapy but is a specialized procedure practiced in the form of rejuvenating recipes, dietary regimen promoting good habit. The purpose of rasayana is two-fold: prevention of disease and counteraction of aging processes which result from optimization of homeostasis. The meaning of the word Rasayana (rasa=essence, water, ayana=going) essentially refers to nutrition and its acquisition, movement, circulation and perfusion in the body tissues (Singh, 1992). With regard to the Rasayana drug therapy Sharma et al. (1992) reported the strong antioxidant activity of any rasayana: these compounds were found to be 1000 times more potent than ascorbic acid, α-tocopherol, and probucol.⁽⁵⁾Highly reactive free radicals and oxygen species present in biological systems can oxidize nucleic acids, proteins and lipids, initiating degenerative diseases.⁽⁶⁾⁽⁷⁾Antioxidants significantly delay or prevent the oxidation of easily oxidable substrates.

Plants contain high concentrations of numerous redox-active antioxidants, such as polyphenols, carotenoids, tocopherols, glutathione, ascorbic acidand enzymes with antioxidant activity, which fight against hazardous oxidative damage of plant cell components. Plantsourced food antioxidants like vitamin C,vitamin E, carotenes, phenolic acids, phytates and phytoestrogenes have been recognized as having the potential to reduce disease risk.The main characteristic of an antioxidant is its ability to trap free radicals. Antioxidant compounds like phenolic acids, polyphenols and flavonoids scavenge free radicals such as peroxide, hydroperoxide or lipid peroxyl and thus inhibit the oxidative mechanisms that lead to degenerative diseases.⁽⁸⁾During the last few decades there has been an increasing interest in the study of traditional plants and their medicinal value in different parts of the world. The medicinal properties of plants have been investigated due to their potent pharmacological activities, low toxicity and economic viability.⁽⁹⁾This revival of interest in plant-derived drugs is mainly due to the current widespread belief that green medicine is safe and more dependable than the costly synthetic drug, many of which may have adverse side effects.⁽¹⁰⁾

Medicinalplants have played as significant role in various ancient traditional system of medication.Even today, plants provide a cheap source of drugs for majority ofworld’s population. Several pharmacological investigations on the medicinal plants used in traditional antiurolithiatic therapy have revealed their therapeutic potential in thein-vitro or in-vivo models.⁽¹¹⁾Senna occidentalis is an erect foetid annual herb. It belongs to the family Fabaceae and commonly called as coffee Senna and in English; it is called as septic weed. It grows up to60 to 150 cm in height and it is found throughout India up to an altitude of 1500 cm. Senna occidentalis has many traditional to treat typhoid, malaria, dog bites. It has pharmacological activities like antihelmintic antifungal, antimutagenic, antipyretic and antifeedant⁽¹²⁾.

Plant Collection andAuthentication:

The leaves of the plant Senna occidentalis were collected in the month of August 2017 in Khagazmaddur Village Narsapur Mandal, Medak District, Telangana, India. The plant was authenticated by M.Malla Reddy (M.Sc, M.Phil in Botany),Retired lecturer in Botany, Vikarabad, Telangana.

Materials used:

In the present study1,1diphenyl, 2-picryl–hydrazyl free radical (DPPH) and alcohol used during investigation of antioxidant activity. All the materials were used in the laboratory grade.

Preparation of Plant extract:

The leaves of Senna occidentalis were shade dried and crushed into powder and sieved to get a coarse powder. The powder was subjected to soxhlet extraction using ethanol for 72 hours. The solvent was evaporated using rotary evaporator then the extract was used for the evaluation of Antioxidant activity.

Phytochemical screening

Table 1: Phytochemical screening of leaves extract of Senna occidentalis

Constituents	Ethanolic extract
Alkaloids	+
Tannins	+
Glycosides	+
Saponins	+
Flavonoids	+
Resins	+
Anthraquinones	+
Phenols	+
Reducing sugars	+

Preliminary phytochemical screening of Senna occidentalis indicates the presence of alkaloids, tannins, glycosides, saponins, flavonoids, resins, anthraquinones, phenols and reducing sugars.

Anti oxidant activity:

DPPH Scavenging activity:

The molecule 1,1-diphenyl-2-picrylhydrazyl (a,a-diphenyl-b-picrylhydrazyl;DPPH) is characterized as a stable free radical by virtue of the delocalisation of the spare electron over the molecule as a whole, so that the molecule does not dimerize, as would be the case with most other free radicals. The delocalization of electron also gives rise to the deep violet color, characterized by an absorption band in ethanol solution centered at about 517 nm. When a solution of DPPH is mixed with that of a substrate (AH) that can donate a hydrogen atom, then this gives rise to the reduced form with the loss of this violet color.

Procedure:

The free radical scavenging activity of all the samples was evaluated by 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) according to the previously reported method by Shen et al., 2010. Briefly, a 0.1mM solution of DPPH in ethanol was prepared and 1mL of this solution was added to 3 ml of the solution of all samples in ethanol at different concentration (20, 40, 60, 80&100μg/mL).The mixtures were shaken vigorously and allowed to stand at room temperature for 30 minutes. Then the absorbance was measured at 517 nm using a UV-VIS spectrophotometer. Ascorbic acid was used as the reference. Lower absorbance values of reaction mixture indicate higher free radical scavenging activity. The capability of scavenging the DPPH radical was calculated by using the following formula.

Diphenyl Picryl Hydrazyl Free Radical

DPPH scavenging effect (% inhibition)=

{(A0–A1)/A0)*100}

Where,

A0 is the absorbance of the control reaction

A1 is the absorbance in presence of all of the extract samples and reference.

All the tests were performed in triplicates and the results were averaged¹³.

Determination of IC₅₀:

The IC₅₀value (The concentration of sample required to scavenge 50% of DPPH free radicals) was determined by interpolation from the calibration curve plotted between percentage inhibition and sample concentration and expressed as microgram per ml.

RESULTS AND DISCUSSION:

Antiradical activity assay is based on the reduction of 1, 1-diphenyl-2-picrylhydrazyl (DPPH). Due to the presence of an odd electron it gives a strong absorption maximum at 517 nm. As this electron becomes paired off in the presence of a hydrogen donor, i.e. a free radical scavenging antioxidant, the absorption strength is decreased, and the resulting decolorization is stoichiometric with respect to the number of electrons captured. The decomposition of DPPH free radicals by Senna occidentalis may at least partly result from its antioxidant and free radical scavengingactivity.

Table 2 : DPPH free radical scavenging activity

Concentrations µg/ml	Standard percent scavenging (%)	Ethanol extract Percent scavenging (%)
20	91	84.8
	92	84.2
60	94	85.9
80	96	83.8
100	97	89.4

Figure. free radical scavenging activity of Standard (Ascorbic acid) and Sample (leaves extract of Senna occidentalis)

CONCLUSION:

The results obtained in the present study indicate that Senna occidentalis ethanolic leaf extract exhibit significant free radical scavenging and antioxidant activity. Theoverall antioxidant activity might be attributed to its phytochemical constituents. The findings of the present study suggest that this Senna occidentals could be a potential source of natural antioxidant thatcould have great importance as therapeutic agent in preventing or slowing theprogress of aging and age associated oxidative stress related degenerative diseases.

ACKNOWLEDGMENT:

The authors sincerely thankful to our chairman Shri.K. V. Vishnu Raju Garu and our college Vishnu Institute of Pharmaceutical Education and Research Principal Dr. Ramesh Alluri and staff members for towards our project.

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Received on 20.09.2018 Modified on 15.10.2018

Research J. Pharm. and Tech 2019; 12(2):549-552.

DOI: 10.5958/0974-360X.2019.00097.0