A flame test and a halide ion test were performed. To start with the flame test, weigh 1 gram of Unknown substance in an analytical balance using a scoopula and mixed it in with 20 mL of water into a 150 mL. Repeat the same step with NaC2H3O2. Repeat the same step for the flame test and record down data. Lastly is the halide ion test.
(ii) Acidify to methyl orange end point with concentrated H2SO4. (iii) Add 5 ml concentrated HNO3. (iv) Add 2 ml 30% H2O2 . (v) Evaporate on a hot plate to 15 to 20 ml. (vi) Transfer the concentrate and any precipitate to a 250 ml conical flask using 5 ml concentrated HNO3.
1.25mL of acetic acid and ferric chloride mixture was added to 2.5mL of the pure extract. Occurrence of blue-green color indicates the presence of glycosides. Test for Saponins. 10mL of distilled water was added to 5 mL of the pure extract the mixture was shaken in a graduated cylinder for 15minutes. Occurrence of layer of foam or bubbles indicates presence of saponins.
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
3) Centrifuged at 2500 rpm for 12 mins. Upper hexane layer (supernatant) was transferred carefully into another test tube. 4) Evaporated the hexane under a stream of grade 1 nitrogen gas and added 100 µl of methanol to the residue left and vortexed for 1 min. 5) Injected 100 µl of extract in HPLC vials and closed properly. Standard curves and calculations- Retinol was quantified from standard curves peak area for each vitamin.
In a 150 mL Erlenmeyer flask with a magnetic stir bar placed 0.25 g of anhydrous sodium hydroxide and dissolved it with 10 mL of methanol. The mix was stirred until all of the sodium hydroxide was dissolved. In a 250 mL beaker placed 5 mL of algal oil. The oil was heated to 60ºC. Added 2 ml of the dissolved sodium hydroxide into the heated oil, immediate the mixture turns cloudy.
Standardization of NaOH solution Accurately weigh out a sample of approximately 0.3-0.4 g of primary standard potassium hydrogen phthalate, KHPh, which has been previously dried at 120°C. Do not use more than 0.4 g. To obtain an accurate mass, weigh the sample on weighing paper, slide it into a clean (but not necessarily dry) 250 mL Erlenmeyer flask and reweigh the paper to account for any KHPh that may remain on it. Dissolve the KHPh sample in about 50 mL of CO2-free water and add 2-3 drops of 0.1% phenolphthalein indicator. Begin adding the approximately 0.1 M sodium hydroxide solution from the buret while continuously swirling the flask contents. Do not open the stopcock completely.
3.2.1 Ferric Chloride Test The sample extraction will be dissolved in 2 ml ethanol. A few drops of 10% ferric chloride solution will be added in test tube F. A green-blue coloration indicates the presence of a phenolic hydroxyl group (Bhatt et. al., 2011) 3.2.2 Sodium Hydroxide Test Few drops of 10% aqueous sodium hydroxide solution will be added in a test tube E with 2-3 ml of the extract. Formation of intense yellow color that became colorless on the addition of few drops of diluted HCl indicates the presence of flavonoids (Bello et. al.,
For TLC profiling, 4 TLC plates were prepared for the testing of each solvent. As shown in Figure 1, the green food dye was placed at the bottom center, specifically 0.5 cm away from the bottom of the plate, with the use of a capillary tube. Each one of the silica plates were then vertically placed in a small beaker with its inside surrounded by a filter paper saturated with the solvent to be tested and a small amount of the same solvent at the bottom. The TLC plate was then taken out when the rising solvent was about to reach the top of plate. The ammonia: 1-butanol solvent was tested 7 times due to some personal
Analytical indices related to fats and oils can be distinguished as structure or quality indices. Structure indices are the iodine value (IV), a measure of total unsaturation of an oil or fat; the saponification value (SV), an indicator of average M.W. ; and the hydroxyl value (HV), which is applicable to fatty compounds (or their mixtures) containing (Knothe, 2002). Saponification is the hydrolysis of fats or oils under basic conditions to afford glycerol and the salt of the corresponding fatty acid (Chalmers and Bathe, 1978). The saponification number is the number of milligrams of potassium hydroxide required to neutralize the fatty acids resulting from the complete hydrolysis of 1g of fat.