Metal chelating activity Briefly, 2 mM FeCl2 was added to different concentrations of test sample and reaction was initiated by the addition of 5 mM ferrozine. The mixture was vigorously shaken and left to stand at room temperature for 10 min. Absorbance was measured at 562 nm after 10 min.8 % Inhibition = [(AB - AA)/AB] x 100, where AB, absorption of blank sample, AA, absorption of test sample. 2.6. Antibacterial
Preparation of chitosan microspheres49 In this method, the polymer chitosan was initially dissolved 1% acetic acid, by magnetic stirring for about 2-3 hrs, to the prepared chitosan solution, diltiazem hydrochloride was added, stirred for 1hr, the resulted polymer solution was added to liquid paraffin containing span 60 as surfactant. The resultant emulsion was stirred for one hr further cross linking agent gluteraldehyde was added stirred over night for rigidisation of coating. Microsphers were filtered, washed with petroleum ether, and air dried. Formulation code Diltiazem hydrochloride (mg) Chitosan Gluteraldehyde (ml) Surfactant Conc. FCD1 90 90 3 1 FCD2 90 180 3 2 FCD3 90 240 3 3 FCD4 90 360 3 4 FCD5 90 450 3 5 FCD6 90 90 3 5
Characterization of bacteriocins 6.1. Effect of temperature The effect of temperature on bacteriocin stability was determined by exposing 5 ml aliquots of CFS at 40C, Room temperature, 370C, 630C and 1000C for 30 minutes. Agar well diffusion method was performed to determine the residual activity in the heat-treated CFS17. 6.2. Effect of pH To check the pH stability of the bacteriocins, 5 ml aliquots of CFS was taken in sterile tubes and were adjusted to pH values of 2, 4, 7 and 9 busing 1N HCl or 1 N NaOH.
Determination of antioxidant activity Scavenging DPPH radicals DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) radical is used free radical method is an in antioxidant assay forwas used to evaluate measured the free radical scavenging activity of the lichen extract . Two millilitreers of 0 .05 mg/mL methanol solution of DPPH radical in the concentration of (0 .05 mg/mL) and 1 mL of the lichen extract (1 mg/mL) were placed in cuvettes. The mixture is storewas stored stand at room temperature for 30 min. Then, the absorbance was measured at 517 nm in a spectrophotometer (Jenway, UK). Ascorbic acid was used as a positive control.
The peel powder was soaked overnight (or for desired period) at room temperature (30-32°C) for extraction with intermittent shaking. After extraction, it was centrifuged at 2000 rpm for 5 min and filtered through Whatman No. 2 filter paper. The effective volume obtained after centrifugation was noted. 2.3 Determination of radical scavenging activity (RSA) Determination of RSA was carried out according to Murthy, Jayaprakasha, & Singh (2002) using DPPH as stable free radical and butylatedhydroxyanisole (BHA) as standard.
 Appropriate concentrations of SBP hydrolysate were mixed 1: 4 (v/v) with 200 M DPPH solution in anhydrous methanol for 30 min in the dark. The absorbance of the samples was recorded spectrophotometrically on a microplate reader (Multiskan Go, Thermo Fisher Scientific) at 517 nm. A control group containing DPPH solution without sample was also prepared. Ascorbic acid was used as a positive control. ABTS radical scavenging
Care is taken to maintain the temperature between 160 °C to 170 °C. The contents are gradually stirred for about 55 minutes. The modified bitumen is cooled to room temperature and suitably stored for testing. 4.2 Common tests on the modified bitumen Penetration test and Softening point tests on both the plain and modified CRMB are performed and the results are analyzed for further study. 4.3 Preparation of Bituminous mix For the present study Bituminous concrete mix gradation was used following specifications stated by MORT & H table
2.3. Preparation procedure for DS loaded PC-SA combined beads The ionotropic gelation method was used for the preparation of DS loaded PC-SA bead. The gum (PC) was dissolved in distilled water and initially boiled for 10 minutes, then cool and keep stirring for 24 hours at 400 RPM. Then filtration
This reaction is refluxed in a waterbath for two hours and their volume were reduced to 70% of it’s original volume and residue was obtained. The coloured product obtained was filtered under suction, washed with ethanol. The product were recrystallized from ethanol. Their yields ranges from 50-55%, the product obtained were pinkish colour and melting point was 2020C. Step-III: Synthesis of Cr(II) Complexes: The Schiff's base complexes were synthesized by mixing the Schiff's base (1.5 g) in ethanolic solution of Chromium chloride [CrCl2].
Four solvents were used for the extraction of the plant material. They were as follows i) N hexane ii) Petroleum ether iii) Methanol iv) Chloroform The powdered plant material (50 gm) was successively extracted in a Soxhlet extractor with an elevated temperature using 250 ml of n-hexane, followed by petroleum ether, methanol and chloroform, according to increasing solubility. All extracts were filtered individually and poured on petri dishes to evaporate the solvents from the extracted material. After drying, crude