(EA) belongs to Euphorbiaceae,
(GG) belongs to Fabaceae,
(PK) belongs to Scrophulariaceae,
(TD) belongs to Aizoaceae,
(XM) belongs toMeliaceae. The reason for selection is that, theseplants have a wide range of applications in traditionalmedicines and are involved in the treatment ofvarious skin diseases, liver disorders, cancer, HIV,cough, asthma, diarrohea, dysentery, ulcers andcontrolling of blood pressure etc,. In addition, someof these plants were also reported to be the potentantioxidants. Hence, in the present study, we madean attempt to study the total antioxidant power andradical scavenging capacity of methanolic extractsof these plants. This data will be used to estimatethe phytochemical and antioxidant intake of the localpopulation and to understand the therapeutic usesof these plants.
MATERIAL AND METHODS
Diphenyl picryl hydrazyl (DPPH) wasobtained from Himedia laboratories Pvt. Ltd,Mumbai, India. Ferric Chloride, 2,4, 6-tri pyridyl-s-triazine, Butylated hydroxy toluene (BHT) wereobtained from Sisco research laboratories Pvt. Ltd.,Mumbai, India. All other chemicals used were ofanalytical grade obtained from commercial sources.
Plant collection and preparation of the extraction
The plants are collected from the near byhilly region, Paderu and coastal region ofVisakhapatnam, Andhra Pradesh, India andauthenticated by Dr. M. Venkayya, Associateprofessor, Dept. of Botany, Andhra University,Visakhapatnam, Andhrapradesh.
Plant leaves werecleaned with deionized water and dried at 50ºC for24 hours. The dried plants were ground and thensieved 80mesh. The dried leaves were ground to apowder using a milling machine. The dried powderwas then weighed and extracted with methanol, withreflux on a water bath at 40°C for three consecutivedays. The extracts were filtered and evaporatedunder vacuum to dryness with a rotary evaporatorand then placed in an oven at 60°C until constantweight was obtained and the solutions wereprepared with a concentration 1mg/ml usingmethanol.
Total Antioxidant powerFerric reducing ability of Plasma (FRAP) Assay
The total antioxidant potential of a samplewas determined using the ferric reducing ability ofplasma i.e., FRAP assay as a measure of antioxidantpower by Benzie and Strain
. The assay was basedon the reducing power of a compound (antioxidant).A potential antioxidant will reduce the ferric ion(Fe3+) to the ferrous ion (Fe2+); the latter forms ablue complex (Fe2+/TPTZ), which increases theabsorption at 593 nm. Briefly, the FRAP reagent wasprepared by mixing acetate buffer (300µM, pH 3.6),a solution of 10 µM TPTZ in 40 µM HCl, and 20µMFeCl3 at 10:1:1 (v/v/v). The reagent (300 µM) andsample solutions (10 µL) were added to each welland mixed thoroughly. The absorbance was takenat 593 nm after 10 min. Standard curve was preparedusing different concentrations of FeCl
. All solutionswere used on the day of preparation. The resultswere corrected for dilution (e.g.1000ml) andexpressed as ascorbic acid equivalents (µmoles/ml)or FRAP units. All determinations were performedin triplicates.
Free radical scavenging capacityDi Phenyl Picryl Hydrazyl radical scavengingassay (DPPH Assay)
The DPPH assay was carried out asdescribed by Cuendet
., (11). 5.0 ml of DPPHsolution (0.004%) in methanol was added to 50 µlof plant extract. After 30 min of incubation at 37
C,the absorbance was read against control at 517 nm.Ascorbic acid and BHT were used as positivecontrols. Percentage of inhibition = (absorbance ofcontrol –absorbance of test / absorbance of control)x100. All determinations were performed in triplicate.
RESULTS AND DISCUSSION
Recently much attention has been focusedon reactive oxygen species and free radicals, whichplay an important role in the genesis of variousdiseases such as inflammation, cataract, livercirrhosis and ischemia/reperfusion injury (12).Herbal drugs containing radical scavengers aregaining importance in the prevention and treatmentof such diseases¹³. Hence the present study isfocused on the determination of Total antioxidantpower by FRAP Assay and DPPH Assay.838Rao
Biosci., Biotech. Res. Asia,
(2), 837-840 (2008)