24 sobiyana et al., Available Online http://www.ijncse.com ISSN Online: 2395-7018 2(6) (2015) 24-30 Antioxidant and Antimicrobial Activities of Tabebuia Rosea ABSTRACT P.Sobiyana *, G.Anburaj, R.Manikandan * P.Sobiyana A.V.V.M Sri Pushpam College, Poondi. G.Anburaj A.V.V.M Sri Pushpam College, Poondi. R.Manikandan Assistant Professor of Poondi The presence of natural antioxidant and antimicrobial in plants is well known. This paper reports the antioxidative and antimicrobial activities of petroleum ether tabebuia rosea extracts. Petroleum ether extracts of tabebuia rosea flowers were evaluated for their free radical scavenging activity using the DPPH radical assay. Reduction of DPPH radicals can be observed by the decrease in the absorbance at λ max517nm. The tabebuia rosea flowers extract and ascorbic acid showed antioxidant activity with different IC50 values. The antimicrobial activity of tabebuia rosea flowers extract was done by using well diffusion method against different species of bacter. Keywords: antioxidant activity, ascorbic acid, DPPH, free radical assay, antimicrobial activity, well diffusion method. INTRODUCTION: Antioxidants have an essential role in body defences system against Reactive Oxygen Species (ROS). Natural antioxidants that are present in the food increase the resistance toward oxidative damages and they may have an essential impact on human health. Therefore, consumption of food that is containing phytochemical with potential antioxidant properties can decrease the danger of human diseases. Chain breaking antioxidants are highly reactive with free radicals and form stable compounds that do not contribute to the oxidative chain reaction 1.
25 sobiyana et al., In other way an oxidation can be defined as a molecule that is capable of slowing or preventing the oxidation of other molecules. Antioxidants are often reducing agents such as thiols or polyphenols. They are believed to play an important role in preventing the developments of such chronic diseases as cancer, heart disease, stroke, Alzheimer, s disease, Rheumatoid arthritis and cataracts 2. Some plant families exhibit antimicrobial activities including antibacterial and antifungal. Helicobacter pylori are susceptible to a wide range of antimicrobial agents, including Acacia nilotica, Daturastramonium, Mangiferaindica and Eucalyptus globulis. It has been found that aglycones inhibit the growth of H pylori, whereas glycosides are inactive. The presence of a methoxyl group at C-4, was also important, and its replacement with a hydroxyl group caused a significant decrease in the activity of the compound 3.The antifungal fatty acids naturally can insert themselves into the lipid bilayer of the fungal membranes and physically disrupt the membrane, leading to enhanced fluidity of the membrane.these elevation in membrane fluidity will cause a generalized disorganization of the cell membrane that lead to conformational changes in membrane proteins, the release of intracellular components, cytoplasmic disorder and eventually cell disintegration 4. METERIALS AND METHODS COLLECTION OF PLANT METERIALS The plant flowers were collected from the tree found in the Sastar University, vallam, thanjavur(dt). The collected flowers were botanically authenticated by Dr.S. John BrittoRapinat Herbarium, st.joseph s college, Trichy. PREPARATION OF POWDER AND EXTRACT The flower (1Kg) was shade dried,crushed by hand and extracted with petroleum ether for 48 hours. The distillation using soxhlet apparatus then the extract was filtered and vaccum dried. The extract was used antioxidant and well diffusion method. ANTIOXIDANT ACTIVITY Each sample stock solution (1.0 mg ml/1) will be diluted to final concentrations of 500, 250, 100, 50 and 10 mg ml/1, in ethanol. A total of 1 ml of a 0.3 mm DPPH ethanol solution will be added to 2.5 ml of sample solution of different concentrations and allowed to react at room temperature. After 30 min, the Ab values will be measured at 518 nm and converted into the percentage antioxidant activity using the following equation:
26 sobiyana et al., %inhibition= *100 Ethanol (1.0 ml) test solution (2.5 ml) was used as a blank, while DPPH solution plus ethanol was used as a negative control. The positive controls were DPPH solution plus each 1 mm test. AGAR WELL DIFFUSION METHOD Find the extracts from the plants for studying their antibacterial activity. A loop full of bacterial strain was inoculated in of Nutrient broth in a conical flask and incubated for 72 hrs. After solidification 0.25 ml of test strains were inoculated in the media separately and cared for proper homogenization. The experiment was performed under strict aseptic conditions. After the medium solidified, a well was made in the plates with sterile borer (5 mm). Then extract compound was introduced into the well and plates were incubated at 37 C for about overnight. All samples were tested in triplicates. Microbial growth was determined by measuring the diameter of zone of inhibition. RESULTS AND DISCUSSION The scavenging the stable DPPH radical is a widely used method to evaluate the free radical scavenging ability of various samples, including plant extracts. The measured DPPH radical scavenging activity is shown in table 1and figure 1. The Tabebuia rosea flower extract scavenging antioxidant activity was significantly increased with increasing concentration of the extract. The effect of antioxidants on DPPH is thought to be due to their hydrogen donating ability. Although the DPPH radical scavenging abilities of the extracts were significantly lower than those of ascorbic acid, it was evident that the extracts did show some proton-donating ability and could serve as free radical inhibitors or scavengers, acting possibly as primary antioxidants. The quality of the antioxidants in the extracts was determined by the percent inhibition values shown in table 1 and figure 1. Although the precent inhibition values of Tabebuia rosea flower extract were not much greater than the reference antioxidant. Lack of hydrogen donor bioactive constituents in the extract, slow rate of the reaction between DPPH and the substrate molecules resulting in low readings for antioxidant activity probably might explain the low DPPH antioxidant activity of the Tabebuia rosea flower extract [5-8]. Method for DPPH radical scavenging assay Radical scavenging activity of plant extracts was determined using DPPH assay. DPPH reacts with an antioxidant compound, which can donate hydrogen, and reduce DPPH. The change in
27 sobiyana et al., colour (from deep violet to light yellow) was measured at 517 nm on a UV visible light spectrophotometer. 1ml of 0.1mM DPPH solution in methanol was mixed with 1ml of compound A and B (PP and HP) at different concentrations (50, 100, 150, 200 and 250 μg/ml). The samples were kept in the dark for 30 minutes at room temperature and the decrease in absorbance was measured. Table1. DPPH radical scavenging activity of crude extract of Tabebuia rosea S.No Concentration(µg/ml) Crude extract Ascorbic acid (Standard) 1 20 12.55±8.63 34.10± 1.21 2 40 31.90 ± 4.56 40.66 ± 0.64 3 50 45.64 ± 3.04 56.49 ± 1.00 4 80 52.43 ± 4.14 66.28± 0.55 5 100 71.63 ± 4.6 88.15 ± 0.41
28 sobiyana et al., Fig 1: Comparison of % scavenging of DPPH by ascorbic acid vs crude extract at different concentrations Antimicrobial activity Antimicrobial activity was determined by using agar well diffusion assay. Our results indicated a potent antioxidant and antimicrobial activity of Petroleum ether extracts of Tabebuia rosea. In the present study Petroleum ether extracts of plant have been tested against resistant bacteria. The antimicrobial activity of the extracts and their potency was quantitatively assessed by the presence or absence of inhibition zone and zone diameter (Table 2). The results of antimicrobial activity of Tabebuia rosea was encouraging and that the plant extract showed significant antimicrobial activity against different bacterial strains. Antimicrobial activity of Petroleum ether extracts of Tabebuia rosea against five bacterial strain Bacillus subtillus, E.coli, shigellaflexinari, klebsillapeneumonia, p.sudomonas at different concentration (20,40,60,80,100 µg/ml) was found in the following decreasing order Bacillus subtillus>e.coli>shigellaflexinari>klebsillapeneumonia>p.sudomonas (Table 2). Determination of antibacterial activity by agar well diffusion assay showed that Petroleum ether extracts of Tabebuia rosea flower exhibited the antibacterial effect against pathogenic as well as non-pathogenic test bacteria. Fig.1. Zone inhibitions of Antibacterial activity of Tabebuia rosea flower against different pathogens at different concentrations
29 sobiyana et al., Bacillus subtillus E.coli Shigellaflexinari Klebsillapneumonia Pseudomonas Table 2 Antibacterial activity of Tabebuia rosea Pathogens 20µL 40µL 60µL 80µL 100µL Chloramphenicol (mm) (mm) (mm) (mm) mm) (standard) Bacillus subtillus 8 9 11 16 20 29 E.coli - - 7 10 17 27 Shigellaflexinari - - - 7 9 28 Klebsillapneumonia - 7 9 8 8 31 Pseudomonas - - - - 7 29 CONCLUSION In conclusion, Tabebuia species are rich source of naturally occurring antioxidant and antimicrobial activity. Its compound play a vital role in preventing innumerable health disorders related to oxidative stress including cardiovascular diseases, neurodegenerative diseases and cancer. Our results indicated a significant increase in antioxidant activity of
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