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An Investigation using Chromatography to determine the different Amino Acids

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Introduction

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. ...read more.

Middle

acids present in them; I think this because an egg white contains approximately 40 different proteins so obviously when it contains that many proteins it will contain a lot of amino acids as they are what makes up the proteins but in different formations. Diagram Gas Jar Lid Gas Jar Chromatography Paper Solvent Results Chicken egg Rf = Retention Factor Rf = x y x = Distance from starting point to spot y = Distance to solvent front Distance to solvent front/y = 166mm Distance to amino acid/x1 = 86mm Distance to amino acid/x2 = 132mm Calculation for the Rf factor of x1 Rf factor of x1 = 0.158 166mm = 0.158 86mm x1 = Methionine Calculation for the Rf factor of x2 Rf factor of x2 = 0.795 166mm = 0.795 132mm x2 = Leucine Amino Acid Distance Rf Factor Identified Amino Acid x1 86 0.158 Methionine x2 132 0.795 Leucine Duck egg Rf = Retention Factor Rf = x y x = Distance from starting point to spot y = Distance to solvent front Distance to solvent front/y = 220mm Distance to amino acid/x1 = 25mm Distance to amino acid/x2 = 60mm Distance to amino acid/x3 = 97mm Distance to amino acid/x4 = 132mm Calculation for the Rf factor of ...read more.

Conclusion

chromatography where by which a chromatogram is turned by ninety degrees, and placed in a different solvent in the same way as before; some spots separate in the presence of more than one pigment. As before, the value is calculated, and the two pigments are identified. I could have also used a square of chromatography paper and done the experiment turning it 90 degrees each time until four results are shown on the paper. From research, I have found that chicken eggs contain 17 different amino acids, this is shown in the table below and so in my prediciton I do not think I was entirely wrong as it may not have shown all of these on the chromatography paper but I have already discussed a possible reason for this. Amino Acids--g. Alanine 0.348 0.203 0.143 Arginine 0.375 0.191 0.199 Aspartic acid 0.628 0.358 0.272 Cystine 0.145 0.091 0.05 Glutamic acid 0.816 0.467 0.353 Glycine 0.21 0.123 0.086 Histidine 0.148 0.079 0.072 Isoleucine 0.341 0.199 0.141 Leucine 0.534 0.296 0.244 Lysine 0.449 0.239 0.221 Methionine 0.195 0.121 0.069 Phenylalanine 0.332 0.205 0.119 Proline 0.249 0.137 0.116 Serine 0.465 0.242 0.238 Threonine 0.3 0.16 0.148 Tryptophan 0.076 0.043 0.033 Tyrosine 0.255 0.137 0.124 Valine 0.381 0.224 0.155 Table from: http://www.aeb.org/LearnMore/NutrientBreakdown.htm Craig Thompson 12R ...read more.

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