EVALUATION OF ANTIOXIDANT ACTIVITY OF ETHANOL AND CHLOROFORM EXTRACTS OF WILD AND MICROPROPAGATED CLEOME VISCOSA LINN.HTML Full Text
EVALUATION OF ANTIOXIDANT ACTIVITY OF ETHANOL AND CHLOROFORM EXTRACTS OF WILD AND MICROPROPAGATED CLEOME VISCOSA LINN.
M. Deventhiran, W. John Wyson, P. Saravanan and D. Anand *
Department of Botany, Ramakrishna Mission Vivekananda College (Autonomous), Mylapore, Chennai - 600004, Tamil Nadu, India.
ABSTRACT: Herbal medicines are now creature in modern dosage forms using modern manufacture in and processing techniques in pharmaceutical industry. Antioxidants assuage oxidative stress in cells and thereby help in the prevention and treatment of many diseases of humans. The aim of this study was to assess the antioxidant activity of ethanol and chloroform extracts of wild and micro-propagated Cleome viscosa. Antioxidant activity was performed by DPPH ((2, 2 – diphenyl 1, picryl hydrazyl) radical scavenging method for ethanol and chloroform extracts of wild and micro-propagated C. viscosa. The ethanol and chloroform extracts results were exhibited by strong antioxidant DPPH radical scavenging activity with percentage of antioxidant potential concentration wild and micro-propagated were found to be 77.74%, 76.45% and 63.54%, 55.16% respectively. The percentage was compare to positive standard compounds of ascorbic acid. The results were concluded that ethanol extracts possess effective antioxidant activity than that chloroform extracts.
Antioxidant, Micro-propagated, DPPH, Ascorbic acid, Cleome sp.
INTRODUCTION: Phytomedicine also known as herbal medicine has become a mainstream phenomenon worldwide. Recently, it has been reported that more than 80% of the world population is dependent on herbal medicine 1. Plant besides therapeutic agents is also a big source of information for a wide variety of chemical constituents which could be developed as drugs with precise selectivity. These are the reservoirs of potentially useful chemical compounds which could serve as newer leads and clues for modern drug design 2.
Since, people are aware of organic pesticides and are intended towards using plant extracts to offer alternatives to synthetic insecticides 3. Moreover, the excessive use of medicinal plants for drug formulation increases the need of more biomass of plants which can be met with the biotechnological tools like micro-propagation 4.
Cleome viscosa Linn. commonly known as "wild or dog mustard," is an annual, sticky herb belonging to family Cleomaceae found as a common weed all over the plains of India and throughout the tropics of the world. The whole plant and its parts (leaves, seeds, and roots) are widely used in traditional and folkloric systems of medicine. In Asia and Africa the leaves and seeds used to treat infections, fever, rheumatism and headache. The whole herb is used in treatment of inflammation of the ear pain and applied on wounds and ulcers.
A decoction is used as an expectorant and digestive stimulant and the vapour from a steaming decoction of the whole plant is inhaled to treat headache 5. Traditionally, this plant is used in various disorders such as diarrhoea, fever, inflammation, liver diseases, bronchitis, skin disease and malarial fever 6. Antioxidants act as a defense mechanism that protects oxidative damage, and include compounds to repair damage molecules. It can prevent the oxidation caused by free radicals and sufficient intake of antioxidants is supposed to protect against diseases 7. Antioxidants may be classified according to their mode of action as being free radical terminators, chelators of metal ions involved in catalyzing lipid oxidation or oxygen scavengers that react with oxygen closed system 8.
Free radicals are natural by-products of human metabolism. These are charged molecules which attack cells, breaking cellular membranes, reacting with the nucleic acids, proteins, and enzymes present in the cells. These attacks by free radicals, collectively known as oxidative stress, are capable of causing cells to lose their structure, function and eventually result in cell dysfunction. They are continuously produced by our body's use of oxygen, such as in respiration and some cell mediated immune functions. Free radicals are also generated through environmental pollutants, cigarette smoke, automobile exhaust, radiation, air-pollution, pesticides 9. Hence, in the present study C. viscosa (in vitro and wild plant) extracts was employed to study its antioxidant property.
MATERIALS AND METHODS:
Collection of Plant Material: The healthy plants of Cleome viscosa L. were collected during the month of March from the natural habitats of Kanchipuram district, Tamil Nadu, India. The plant specimen was identified and authenticated by Botanical Survey of India (BSI), Coimbatore, Tamil Nadu, India. Specimens Reference number - BSI/SRC/5/23/2016/Tech/1195.
Sterilization and Inoculation of Explants: The nodal segment of the plant was chosen as explants for the present investigation. Actively growing shoots were selected as the source for explants. The explants were sterilization followed by Muthusamy Govarthanan et al., 2015 10. The nodal segments were cut into 5 mm in size and carefully transferred to the sterile MS basal medium supplemented with 3% sucrose, 0.8% agar and different concentration of PGRs such as 6-Benzylaminopurine (BAP), Kinetin (KIN), Naphthalene-3-acetic acid (NAA). The inoculated cultures were maintained in growth chamber. Data was recorded after 28 days 11.
Preparation of Plant and Callus Extracts: The wild and micro-propagated plants were washed thoroughly in sterile distilled water. The plants were shade dried and ground to fine powder using mortar and pestle. One gram (dry weight) of powdered extract was soaked in 10 ml of ethanol for 3 hours and sonicated in an Ultrasonic Sonicator at 20 pulses for 20 min. The extract was centrifuged at 10,000 rpm for 10 min and the supernatant was freeze-dried and stored at 4 °C until further use 12.
Antioxidant Activity by in vitro Techniques: The free radical scavenging ability of the crude extracts of Cleome viscosa was evaluated by DPPH free radicals scavenging assay.
DPPH Radical Scavenging Assay: The effect of ethanol and chloroform extracts of Cleome viscosa on DPPH (2, 2 - diphenyl 1, picryl hydrazyl) radical was estimated according to the procedure described by Von Gadow et al., 1997 13. 2 ml of 6 ×10-5 M methonolic solution of DPPH were added to 50 μl of an ethonolic solution (1 mg/ml) of the sample. Absorbance measurements commenced immediately. The decrease of absorbance at 515 nm was continuously recorded in a spectro-photometer for 16 min at room temperature. Methanolic solutions of pure compound (ascorbic acid) were tested at 1 mg/ml concentration. The scavenging effect (decrease of absorbance at 515 nm) was plotted against the time and the percentage of DPPH radical scavenging ability of the sample was calculated from the absorbance value at the end of 16 m in duration as follows: The percentage inhibition of the DPPH radical by the samples was calculated according to the formula of Yen and Duh (1994) 14.
IP = [(AC (0) - AA (t) / AC (0))] × 100
Where AC (0) is the absorbance of the control at t = 0 min; and AA (t) is the absorbance of the antioxidants at t = 16 min.
RESULTS AND DISCUSSION: C. viscosa was efficiently regenerated from nodal explants from field grown young plants on MS medium on supplemented with different concentration of cytokinins and auxins BAP, KIN and NAA. The callus was observed in 15 days old culture on media tested and it was found the number of shoots developed on nodal explants (Fig. 1A to Fig. 1C).
A B C
FIG. 1: IN VITRO SHOOT PROLIFERATION OF C. VISCOSA ON MS MEDIUM WITH DIFFERENT CONCENTRATION OF CYTOKININS AND AUXINS
Scavenging activity for free radicals of 2, 2-diphenyl-2-picrylhydrazyl (DPPH) has been widely used to evaluate the antioxidant activity of natural products from plant and microbial sources. The antioxidant potential of the ethanol and chloroform extracts of wild and micro-propagated C. viscosa was shown 77.74%, 76.45% and 63.54%, 55.16% respectively. The positive standard of ascorbic acid compound showed 94.83% of antioxidant potential (Table 1 and Fig. 2).
TABLE 1: DPPH RADICAL SCAVENGING ACTIVITY OF ETHANOL AND CHLOROFORM EXTRACTS OF WILD AND MICROPROPAGATED C. VISCOSA
Name of the Plants
|O.D. at 515 nm||Antioxidant Potential concentration (%)|
|Initial O.D. (0 min)||Final O.D. (16 min)|
|1||C. viscosa ethanol (Wild)||0.620||0.138||77.74|
|2||C. viscosa ethanol (callus)||0.620||0.146||76.45|
|3||C. viscosa chloroform (wild)||0.620||0.226||63.54|
|4||C. viscosa chloroform (callus)||0.620||0.278||55.16|
FIG. 2: EFFECT OF WILD AND CALLUS EXTRACTS IN DPPH RADICAL SCAVENGING ASSAY WITH COMPARED STANDARD COMPOUND ASCORBIC ACID
In response to the increased popularity and greater demand for medicinal plants, a number of conservation groups are recommending that wild medicinal plants be brought into cultivation.
A large number of medicinal plants and their purified constituents have shown beneficial therapeutic potentials. Various herbs and spices have been reported to exhibit antioxidant activity. The majority of the antioxidant activity is due to the flavones, isoflavones, flavonoids, anthocyanin, coumarin lignans, catechins and isocatechins 15. Previous authors analyzed the efficacy of free radical scavenging by using DPPH radical scavenging assay with ethanol crude extracts of C. viscosa showed value of IC50 of leaf, stem, pod and root were 8.32, 12.26, 21.62 and 35.99 mg/ml, respectively 16. The present study was ethanol and chloroform extracts of the wild and callus exhibited significant antioxidant activities determined by DPPH assay. Cleome viscosa showed higher activities in antioxidant assay.
Ethanol and chloroform extracts wild and callus is results was observed in 77.74%, 76.45% and 63.54%, 55.16% respectively at the concentration of 1 mg/ml. The DPPH radical scavenging activity methanolic extracts of C. viscosa has the lowest free radical scavenging potential. The maximum highest antioxidant potential percentage value was reported to be 50% 17. In the present study, wild and callus ethanol extracts of C. viscosa was effective DPPH radical scavenging activity compared with chloroform extracts.
CONCLUSION: In conclusion the current study describes the in vitro antioxidant activity of ethanol and chloroform extracts of wild and micro-propagated Cleome viscosa. The extracts showed efficient free radical scavenging activity compared with standard compound ascorbic acid. Further the phytochemical screening of C. viscosa showed the presence of various chemical constituents of important pharmacological actions and needed to find the compounds which are responsible for treating radical-related pathological damage.
ACKNOWLEDGEMENT: The authors are thankful to the Secretary and Principal, Ramakrishna Mission Vivekananda College (Autonomous), Mylapore, Chennai, India for providing all facilities.
CONFLICTS OF INTEREST: Nil
- Prabhakar VK, Jaidka A and Singh R: In vitro study on alpha-amylase inhibitory activity and phytochemical screening of few indian medicinal plants having anti-diabetic properties. International Journal of Scientific and research publication 2013; 8: 1-6.
- Vijyalakshmi R and Ravindran R: Preliminary comparative phytochemical screening of root extracts of Diospyrus ferrea (Wild.) Bakh and Arva lanata (L.) Juss. Ex Schultes. Asian Journal of Plant Science and Research 2012; 2: 581-587.
- Ladhari A, Laarif A, Omezzine F and Haouala R: Effect of the extracts of the spider flower, Cleome arabica, on feeding and survival of larvae of the cotton leafworm, Spodoptera littoralis. Journal of Insect Science 2013; 13: 1-14.
- Bhatia A, Anand M, Singla R and Sharma A: Antioxidant activity of native and micro-propagated Tylophora indica leaves extract: A comparative study. Journal of Natural Product and Plant Resources 2013; 3(1): 1-7.
- CSIR, The wealth of India: A dictionary of Indian raw materials and industrial products. Council of Scientific and Industrial Research, New Delhi, India, 1950; 2: 427.
- Chandak RR, Bharat NK, Devdhe SJ and Majmudar HF: In vitro evaluation of Anthelmintic potential of leaves Cleome viscosa International Journal of Pharmaceutical Sciences Review and Research 2010; 5(3): 77-79.
- Celiktar OY, Girgin G, Orhan H, Nichers HJ, Bedir E and Sukan FV: Screening of free radical scavenging capacity and antioxidant activities of Rosmarinus officinalis extracts with focus on location and harvesting times. European Food Research and Technology 2007; 24: 443-451.
- Shahidi F, Janitha PK and Wanasundra PD: Phenolic antioxidants. Critical Reviews in Food Science and Nutrition 1992; 32: 67.
- Li Y and Trush MA: Reactive oxygen dependent DNA damage resulting from the oxidation of phenolic compounds by a copper redox cycle, Cancer Research 1994; 54: 1895 - 1898.
- Govarthanan M, Rajinikanth R, Kamala-Kannan S and Selvankumar T: A comparative study on bioactive constituents between wild and in vitro propagated Centella asiatica. Journal of Genetic Engineering and Biotechnology 2015; 13(1): 25-29
- Sharma A, Bhansali S and Kumar A: Micro-propagation of Eclipta alba (l.) hassk. An important medicinal plant of traditional medicine. International journal of life science and pharma research 2013; 3 (2): 47-51.
- Singh J and Tiwari KN: In vitro plant regeneration from decapitated embryonic axes of Clitoria ternatea An important medicinal plant. Industrial Crops and Products 2012; 35: 224-229.
- Von Gadow A, Joubert E and Hansmann CF. Comparison of antioxidant activity of aspalathin with that of other plant phenols of Rooibosed tea (Aspalathon linearis), α- tocopherol, BHT and BHA. Journal of Agricultural and Food Chemistry 1997; 45: 632-638.
- Yen GC and Duh PD: Scavenging effect of methanolic extracts of peanut hulls on free radical and active oxygen species. Journal of Agricultural and Food Chemistry 1994; 42: 629-632.
- Aqil F, Ahmed I and Mehmood Z: Antioxidant and free radical scavenging properties of twelve traditionally used Indian medicinal plants. Turkish Journal of Biology 2006; 30: 177-183.
- Sriwatcharakul S: Antimicrobial, Antioxidant and Cytotoxic Activities of Cleoma viscosa Crude Extracts. International Journal of Biological, Biomolecular, Agricultural, Food and Biotechnological Engineering. 2016; 10(7): 425- 428.
- Aparadh VT, Naik VV and karadge BA: Antioxidative properties (TPC, DPPH, FRAP, Metal chelating ability, Reducing power and TAC) within some Cleome Annali di Botanica (Roma) 2012; 2: 49-56.
How to cite this article:
Deventhiran M, Wyson WJ, Saravanan P and Anand D: Evaluation of antioxidant activity of ethanol and chloroform extracts of wild and micropropagated Cleome viscosa linn. Int J Pharmacognosy 2017; 4(8):269-72:.doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.4(8).269-72.
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M. Deventhiran, W. J. Wyson, P. Saravanan and D. Anand*
Department of Botany, Ramakrishna Mission Vivekananda College (Autonomous), Mylapore, Chennai, Tamil Nadu, India.
01 April, 2017
23 May, 2017
30 May, 2017
01 August, 2017