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Analysis of amino acids by paper chromatography

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Introduction

Sumudu D L L6B Monday, October 21, 2002 Analysis of amino acids by paper chromatography Introduction- Proteins may be thought of natural polymers of amino acids, as the composition of proteins is of amino acids. The technique known as paper chromatography is used to separate amino acids for analysis. In this technique small spots of amino acids are introduced to a piece of porous filter paper. The bottom of the paper is then placed in a small bath of an appropriate solvent. The solvent is allowed to rise up the paper. The various amino acids are attracted to the paper to different degrees due to their differences in polarity. Various amino acids travel at different rates. The location of the amino acids can be determined by spraying the paper with a 2% solution of Ninhydrin in ethanol. The Ninhydrin forms a purple complex with an amino acid which is readily identified. Materials- Solutions of amino acids used are - Amino acid mixture 'X' Aspartic Acid Leucine Proline Asparagine Phenylalanine Solvent (ammonia and propan-2-ol in the ratio 2:1) ...read more.

Middle

All in all about six spots are sufficient. To speed up time a hair dryer can be used to dry the amino acids quicker. Do not use the same capillary for each--you will contaminate each spot with the previous substance. Use a different capillary for each solution (or use the capillary that was present in the solution). 4. The Paper rolled up and stapled is lowered into the jar inside the fume cupboard itself . The penciled end is lowered first so that the paper is upright in the jar. The bottom end should be 5mm into the solvent, thus keeping the line of origins above the solvent and the paper must not be touching the sides of the jar as much as possible. 5. The top of the jar must be replaced and the apparatus is left in the fume cupboard until the solvent has run up the chromatography paper. The amino acids will move up the paper by capillary action at different rates depending on their relative solubilities in the developing solvent. 6. The strip is best left overnight and then removed. ...read more.

Conclusion

Phenylaline - 0.68 The figures obtained and the official figures are almost the same. Anomalies are however quite obvious here due to experimental error. The largest error probably came from the measurement of the stains as they were not spots but elongated stains. The amino acids were also dabbed on the origins with a large drop at times which probably added to the inaccuracy of the corresponding stains. Some stains were longer than usual, probably because the amino acids mixed. Other sources of error include the fact that the amino acid solutions may have been impure. Blow drying the amino acids may have interfered with the amino acids. The chromatography paper may have been a little The mixture X is found to contain Phenylalinine, proline, and aspartic acid. We know that X had Proline as there was a yellow section. Only Proline stains yellow. There was a similar stain(in shape and height) in x to that of aspartic acid therefore we know that aspartic acid was in X. There was also another stain in X of the same height and shape as Phenylaline so obviously this amino acid was also present in X. ...read more.

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