In the biomolecule lab, we conducted the lipid chromatography experiment to separate a mixture of biomolecules from lipids. If a positive test is conducted by the end of the experiment, there should be smudges aligned vertically on the chromatography paper, or silica gel strip, indicating the separation of different lipid molecules that were present in the substance. In contrast, if a negative test is conducted there would be no smudges aligned vertically on the chromatography paper.
Substances Tested and Hypothesis In the lipid chromatography experiment, the purpose was to carry out a TLC, thin-layer chromatography, so the substances we wish to separate are absorbed into the thin layer. That being said, the substance that was being tested …show more content…
A silica gel is then obtained and a line is marked across the bottom 1 cm from the bottom. Using a capillary tube, a drop of mixed lipid solution is placed in the middle of the line drawn. This is repeated several times to create multiple coats of the lipid solution. After the final layer of the solution has dried, the silica gel strip is placed in a jar (pigment dot above the solvent line) and observed the results. Next, we removed the chromatography paper as the solvent approached the top of the silica gel strip, and drew a line across the stip. To view the experiment more clearly, Amino black solution was sprayed on the silica gel strip and dried with a blow dryer. Once it had dried, the silica gel strip was placed in a Coplin jar containing iodine crystals for a few minutes (3-5 minutes) (Harris-Haller, Ninth Edition). After the allotted time, the strip was observed and the results were recorded in table 3-2.
Results
The lipid chromatography experiment tested positive by showing three different substances present in the lipid solution: phospholipids, fatty acids, and triglycerides. By calculating the R_f factor ratio we were able to quantitatively exhibit our results. Triglycerides moved approximately 3.5 cm from the origin point of 4.1cm, giving it a R_f value of .854. The fatty acids moved 2.0 cm form the same point of origin, giving it a R_f factor or .488. Last, the phospholipids
Substances Tested and Hypothesis In the lipid chromatography experiment, the purpose was to carry out a TLC, thin-layer chromatography, so the substances we wish to separate are absorbed into the thin layer. That being said, the substance that was being tested …show more content…
A silica gel is then obtained and a line is marked across the bottom 1 cm from the bottom. Using a capillary tube, a drop of mixed lipid solution is placed in the middle of the line drawn. This is repeated several times to create multiple coats of the lipid solution. After the final layer of the solution has dried, the silica gel strip is placed in a jar (pigment dot above the solvent line) and observed the results. Next, we removed the chromatography paper as the solvent approached the top of the silica gel strip, and drew a line across the stip. To view the experiment more clearly, Amino black solution was sprayed on the silica gel strip and dried with a blow dryer. Once it had dried, the silica gel strip was placed in a Coplin jar containing iodine crystals for a few minutes (3-5 minutes) (Harris-Haller, Ninth Edition). After the allotted time, the strip was observed and the results were recorded in table 3-2.
Results
The lipid chromatography experiment tested positive by showing three different substances present in the lipid solution: phospholipids, fatty acids, and triglycerides. By calculating the R_f factor ratio we were able to quantitatively exhibit our results. Triglycerides moved approximately 3.5 cm from the origin point of 4.1cm, giving it a R_f value of .854. The fatty acids moved 2.0 cm form the same point of origin, giving it a R_f factor or .488. Last, the phospholipids