An Investigation using Chromatography to determine the different Amino Acids

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An Investigation using Chromatography to determine the different Amino Acids, which are contained in both a Chicken and Duck egg and to establish the difference between them.

I am going to do an experiment to find out which amino acids are contained in both a chicken egg and a duck egg.  I will do this using chromatography, I will then use a equation which is relevant to my experiment, to calculate how far the amino acids have moved towards the solvent front, thus letting me compare with a table to see which amino acids are present.

Equation

Rf = x

        y

Equipment

Chicken egg                          Gas Jar                                    Ninhydrin

Chromatography Paper         Gas Jar Lid                              Fume cupboard

Pipette                                   Double Sided Sellotape

Hair Dryer                              A Solvent

Butaniol – Glacial Ethanoic Acid – Distilled H2O

With a ratio of 4:1:1

Method

I will first need to prepare the egg white by separating it from the egg yolk and then adding the protease mixture to the egg white.  This will then be left over night in an incubator so that the enzymes have time to react with the proteins in the egg whites, and break down the bonds between the amino acids, so that they are more accessible and easier to identify.  After the egg/protease mixture has been left over night I will then get a strip of chromatography paper and draw a line approximately 2cm from the end of the strip I will then get the egg/protease mixture and using the pipette slowly and carefully drop the mixture into the middle of the line that has just been drawn, when the paper looks saturated I will take the strip over to the hair dryer which I will be using to accelerate the drying process, once the paper has been fully dried using the hair dryer I will take the strip back over to the bench and repeat the process of saturating the paper and then drying it.  I repeated this process of saturating and drying 4 times in total; I did this because the more of the mixture that was present on the chromatography paper then logically the more possibility of more and different amino acids being present.

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I then placed the double-sided sellotape onto the Gas Jar Lid and folded the other end of the chromatography paper and stuck it to the sellotape so that the saturated end of the chromatography paper was dangling just above the bottom of the Gas Jar, this is so that when I put the solvent into the Gas Jar it was just submerged into the solvent.  So as I just mentioned I then put the solvent into the bottom of the Gas Jar, so that it could rise up the chromatography paper, put then lid on so that the vapours from ...

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