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To determine which solvent system gives the best resolution of separation of the individual amino acids.

Extracts from this document...

Introduction

Ratio of volumes (Methanol: Chloroform: Water) Ds* /cm Alanine Aspartic Acid Leucine Glycine Alone In Mixture Alone In Mixture Alone In Mixture Alone In Mixture Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm 8:1:1 14.30 9.10 0.63 8.80 0.62 6.10 0.43 5.70 0.40 11.40 0.80 11.30 0.79 6.20 0.43 5.70 0.40 1:8:1 1:1:8 6:2:2 12.40 8.20 0.66 7.80 0.63 4.80 0.39 4.70 0.38 11.00 0.89 10.80 0.87 5.50 0.44 4.70 0.38 2:6:2 2:2:6 4:3:3 8.80 7.50 0.85 7.60 0.86 7.30 0.83 7.50 0.85 8.30 0.94 8.00 0.91 6.60 0.75 6.40 0.73 3:4:3 3:3:4 Key: Ds represents the distance moved by the solvent up the chromatography paper Da represents the distance moved by the individual amino acid up the chromatography paper Rf represents the Rf value of the individual amino acids, that is, (Rf = Da / Ds) The asterix, * indicates the uncertainty in the values calculated, which is, + 0.05cm. Ratio of volumes (Methanol: Ammonia: Water) Ds* /cm Alanine Aspartic Acid Leucine Glycine Alone In Mixture Alone In Mixture Alone In Mixture Alone In Mixture Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* ...read more.

Middle

The Rf values calculated for the individual amino acids alone and in the mixture, were the most distinct compared to the results obtained for the other proportions. In addition, the spots obtained in the mixture section could easily be identified with the individual spots of each amino acids as their Rf were approximately equal. 3) The proportion which gave the best resolution of separation of amino acids in the solvent system, methanol: ammonia: water, was the proportion, 8:1:1. This was adjudged the best for the following reasons: a. The clarity of the spots of the individual amino acids obtained alone in the proportion, 8:1:1, were relatively the best compared to the other proportions of the same solvent system. However, for all the proportions, including the best proportion, the separation of the individual amino acids in the mixture sections, were not visible at all. In relatively better situations, it was still very difficult to identify the individual amino acids in the mixture. As such, this criterion was not taken into consideration in determining the best proportion of this particular solvent system. b. The Rf values obtained for the four amino acids were the most distinct in the proportion 8:1:1 compared to the other proportions of the same solvent system. ...read more.

Conclusion

70:15:15 f. 60:20:20 g. 50:25:25 Ratio of volumes (Methanol: Chloroform: Water) Ds* /cm Alanine Aspartic Acid Leucine Glycine Alone In Mixture Alone In Mixture Alone In Mixture Alone In Mixture Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm Da* /cm Rf* /cm 90:5:5 15.50 11.70 0.75 - - 10.10 0.65 - - 13.40 0.86 13.30 0.86 9.60 0.62 - - 85:10:5 15.60 8.90 0.57 8.70 0.56 5.40 0.35 5.00 0.32 12.10 0.78 12.00 0.77 5.30 0.34 5.00 0.32 85:5:10 15.30 9.80 0.64 9.80 0.64 6.30 0.41 6.30 0.41 12.30 0.80 12.60 0.82 6.50 0.42 6.30 0.41 80:10:10 14.30 9.20 0.64 8.80 0.62 6.10 0.43 5.90 0.41 11.40 0.80 11.20 0.78 6.20 0.43 5.90 0.41 70:15:15 13.00 8.00 0.62 7.60 0.58 5.00 0.38 4.70 0.36 10.20 0.78 10.30 0.79 4.80 0.37 4.70 0.36 60:20:20 14.00 11.20 0.80 10.90 0.78 10.00 0.71 9.60 0.69 12.50 0.89 12.60 0.90 9.10 0.65 9.40 0.67 50:25:25 14.90 12.40 0.83 12.10 0.81 11.00 0.74 11.30 0.76 13.30 0.89 13.30 0.89 11.00 0.74 11.30 0.76 Key: Ds represents the distance moved by the solvent up the chromatography paper Da represents the distance moved by the individual amino acid up the chromatography paper Rf represents the Rf value of the individual amino acids, that is, (Rf = Da / Ds) The asterix, * indicates the uncertainty in the values calculated, which is, + 0.05cm. Observations: ...read more.

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