The supernatant was assayed for SOD activity by following the inhibition of epinephrine auto-oxidation. 0.5ml of sample was diluted with 0.5 ml of distilled water, to this 0.25 ml ethanol, 0.5 ml of chloroform (all reagents chilled) was added. The mixture was shaken for 1 min and centrifuged at 2000 rpm for 20 min. The enzymatic activity in supernatant was determined. To 0.05 ml of carbonate buffer (0.05 M, pH 10.2) and 0.5 ml of EDTA (0.49 M) was added.
The absorbance of the mixture was measured at 30s (A1), 5min and 6min intervals. The value A2 was obtained by averaging the values of the 5th and 6th minute. b. Urea 750 ml of urea reagent 1 and 450ml of urea reagent 2 was pipetted into the same 1ml cuvette. Following that, 12ml of patient's sample was transferred into the cuvette. The absorbance of the mixture was measured at 30s (A1), 5min and 6min intervals.
HPLC-grade methanol (15 mL) was added to a quantity of the powdered tablets equivalent to one tablet. This solution is sonicated for 20 min. Filtration through Whatman No. 1 filter paper followed by quantitative transfer of the filtered solution into 25 mL volumetric flask and then dilution was made to volume with methanol. A further dilution was made to reach final concentration of 102.8 µg/mL, 97.2 µg/mL for VAL and SAC respectively.
Twenty tablets were weighed accurately and powdered. An amount of the powder equivalent to 5 mg of amoxicillin trihydrate (content of one tablet) was dissolved in 60 ml of diluent. The solution was stirred for 10 min using a magnetic stirrer and filtered into a 100 ml volumetric flask through 0.45µ nylon membrane filter. The residue was washed 3 times with 10 ml of diluent and then the volume was completed to 100 ml with the same solvent. This solution was diluted with diluents to gae a concentration of 0.1 mg/ml solution each of Amoxicillin trihydrate.
Linoleic acid peroxidation was initiated by the addition of 4 mM FeSO4.7H2O, incubated for 60 min at 37oC and terminated by the addition of 2 mL of ice cold trichloroacetic acid (10% v/v). An amount of 1 mL of thiobarbituric acid (1% w/v in 50 mM NaOH) was added to 1 mL of the reaction mixture, followed by heating at 95oC for 60 min. The reaction sample was read at 532 nm.7 The percentage of linoleic acid peroxidation inhibition activity was calculated using the following equation: % Inhibition = [(AB - AA)/AB] x 100, where AB, absorption of blank sample, AA, absorption of test sample. 2.5.4. 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.
Upon finding the actual concentrations of salicylic acid, concentration of aspirin in the flask at various times can be found using the equation [aspirin]t = [aspirin]0 – [salicylic acid], since at constant volume, number of moles of initial aspirin decrease to form salicylic acid. Initial concentration of aspirin formed as follows: [aspirin]0 = 0.212g / (180.157gmol-1 * 50/1000 L) = 0.0235 mol L-1. Thus using the first test as sample, [aspirin]t = 0.0235 – 9.981*10-4 = 0.0225 mol L-1. To find the rate constant, we will need to log the value of [aspirin]t and plot it against time to find the rate constant. Figure 1 shows the diluted and actual concentrations of salicylic acid, the concentration and log value of aspirin at various times.
The residual biomass was separated by filtration and washed with distilled water. For alginate extraction, the acidified algal biomass was suspended in 3% Na2CO3 solution at different alkali: alga ratio (20, 40, and 60 mL/g). The different extraction temperatures ranged from 25 to 45º C, and lasted for 1 to 3 h. For each experimental run, sodium alginate was collected by filtration and precipitated with absolute ethanol (1:2 v/v). The mixture was maintained at 4º C overnight. The precipitate was collected by vacuum filtration and allowed to dry at room temperature.
The sample was transferred to a 250 ml conical flask kept in water bath for alkali treatment. 75 ml of 17.5% caustic soda was measured using a measuring cylinder at 20°C. 15 ml of 17.5% NaOH was added and fibres were macerated gently with a flattened glass rod for 1 minute. 10 ml more NaOH was added and the solution was mixed for 45 seconds. 10 ml NaOH was again added and mixed for 15 seconds to make lump free slurry.
The 2.5 mL of Albumin Standard (200µg/mL) was distributed among test tubes 0 through 5 (0.00, 0.25, 0.50, 0.75, 1.00, and 0.00 mL respectively). Then, 1.00, 0.75, 0.50, 0.25, 0.00, and 0.00 mL of water was added to test tubes 0 through 5 respectively. One mL of the unknown was added only to test tube #5. Four mL of the biuret reagent was added to each test tube and allowed to stand for 30 minutes at room temperature. Then, the absorbance was determined for each test tube at 545 nm after tube 0 was used to tare the colorimeter to zero absorbance.
Ltd. 4 Melting point Sentwin India 5 NMR Bruker Advance II 400MHz 7 Heating Mantle Inco 6 Structure builder Chem draw Ultra 8.0 4.2 Experimental work: 4.2.1 General procedure for Chalcones: 2’-hydroxy acetophenone or 2’-hydroxy propiophenone (0.2ml) and substituted benzaldehydes (0.5 g) were mixed in the round bottom flask. After that 40% NaOH solution (4g NaOH in 10 ml of distilled water) and ethanol were added in round bottom flask. The reaction mixture was stirred upto 6-48 hours. Completion of reaction was monitored in TLC plate (n-Hexane: Ethyl acetate 9:1). The reaction mixture was poured into ice cold water acidify with 1% HCl and precipitates were collected, filtered and dried and recrystalized with ethanol.