Biochemistry 1b
Biological chemistry 1B Experiment 1: Thin layer chromatography [TLC]
Aim The aim of this experiment is to apply the technique of TLC to first separate out a mixture of coloured dyes and then to do the same for selected range of amino acids. Procedure 2 MMs of each different colour were collected into separate beakers. Approximately 5 drops of water is added to each of the beakers containing the MMs and the tablets were stirred until almost all of the surface colourings were dissolved. A line was drawn [pencil] onto a TLC plate 1cm from the bottom, and 6 dots were drawn separated from each other. Then a spotter was used to apply the dissolved colourings onto each of the dots drawn. This was done for all the beakers. The plate was then placed into a beaker fill to the height of ~0.5cm with a 12:1:1 mix of ethanol:water:ammonia solution. Next the above is repeated on another plate, but instead of MM colouring, a selection of amino acids was applied onto the spots drawn. Also the solution that the plate was placed into is 70:30 ethanol:water instead. Once the solvent front had reached approx. 1cm from the top of the 2 plates, they were taken out to be dried. In addition the second plate was applied with a Ninhydrin spray, before being placed in a heating oven for 15min. Results Part 1 | MM colour | measured distances of [cm] | blue | red | green | yellow | Orange | brown | tartrazine | component 1 | 5.3 | 0 | 4.7 | 1.1 | 2.8 | 2 | 4.7 | component 2 | | 4.6 | 5.1 | 4.6 | 4.4 | 3.2 | | component 3 | | | | | | 4.8 | | component 4 | | | | | | 5.1 | | | Fig 1 | MM colour | Rf value of | blue | red | green | yellow | Orange | brown | tartrazine | component 1 | 0.77 | 0.00 | 0.68 | 0.16 | 0.41 | 0.29 | 0.68 | component 2 | | 0.67 | 0.74 | 0.67 | 0.64 | 0.46 | | component 3 | | | | | | 0.70 | | component 4 | | | | | | 0.74 | |
Aim The aim of this experiment is to apply the technique of TLC to first separate out a mixture of coloured dyes and then to do the same for selected range of amino acids. Procedure 2 MMs of each different colour were collected into separate beakers. Approximately 5 drops of water is added to each of the beakers containing the MMs and the tablets were stirred until almost all of the surface colourings were dissolved. A line was drawn [pencil] onto a TLC plate 1cm from the bottom, and 6 dots were drawn separated from each other. Then a spotter was used to apply the dissolved colourings onto each of the dots drawn. This was done for all the beakers. The plate was then placed into a beaker fill to the height of ~0.5cm with a 12:1:1 mix of ethanol:water:ammonia solution. Next the above is repeated on another plate, but instead of MM colouring, a selection of amino acids was applied onto the spots drawn. Also the solution that the plate was placed into is 70:30 ethanol:water instead. Once the solvent front had reached approx. 1cm from the top of the 2 plates, they were taken out to be dried. In addition the second plate was applied with a Ninhydrin spray, before being placed in a heating oven for 15min. Results Part 1 | MM colour | measured distances of [cm] | blue | red | green | yellow | Orange | brown | tartrazine | component 1 | 5.3 | 0 | 4.7 | 1.1 | 2.8 | 2 | 4.7 | component 2 | | 4.6 | 5.1 | 4.6 | 4.4 | 3.2 | | component 3 | | | | | | 4.8 | | component 4 | | | | | | 5.1 | | | Fig 1 | MM colour | Rf value of | blue | red | green | yellow | Orange | brown | tartrazine | component 1 | 0.77 | 0.00 | 0.68 | 0.16 | 0.41 | 0.29 | 0.68 | component 2 | | 0.67 | 0.74 | 0.67 | 0.64 | 0.46 | | component 3 | | | | | | 0.70 | | component 4 | | | | | | 0.74 | |