Unit 2 Assignment C__ Chromatography
Unit 2 Undertake chromatographic
techniques to identify components
in mixtures Assignment C
Contents
Chromatography __ Amino Acids..........................................................................................................1
Title....................................................................................................................................................2
Introduction.......................................................................................................................................2
Method..............................................................................................................................................2
Results...............................................................................................................................................3
Analysis..............................................................................................................................................4
Conclusion.........................................................................................................................................6
Evaluation..........................................................................................................................................7
Suggest improvements......................................................................................................................9
Chromatography __ thin layer chromatography (TLC)........................................................................10
Title..................................................................................................................................................10
Introduction.....................................................................................................................................10
Method............................................................................................................................................10
Results.............................................................................................................................................11
Analysis............................................................................................................................................13
Conclusion.......................................................................................................................................15
Evaluation........................................................................................................................................17
Suggests improvements...................................................................................................................17
Glossary...........................................................................................................................................18
References.......................................................................................................................................19
Chromatography __ Amino Acids
1
, Unit 2 Assignment C__ Chromatography
Title
Use chromatography paper to separate a mixture of amino acids,
identify and compare them to be able to find the Rf value.
Introduction
I will find which amino acids are present in the mixture using 5 known samples of
amino acids and another mixture using paper chromatography.
Method
1. Draw a pencil line 2cm from the bottom of the chromatography paper.
2. Add 6 crosses in pencil on the line equally spaced out. Write your name or initials
in one of the upper corners in pencil.
3. Label the crosses of the amino acids Tyrosine (T), Lysine (L), Glutamic acid (Glu),
Glycine (Gly), Valine (V), and the mixture (M).
4. Use a capillary tube to spot the samples and mixture onto the crosses. Use as
small a spot as possible.
5. Add 1cm of solvent to the beaker (must be below origin line when paper is
added).
6. Solvent is ethanol, water, and concentrated ammonia at a ratio of 8:1:1.
7. Make sure the paper is not touching the sides of the beaker or curled.
8. Leave the solvent to travel up the paper until it is a few cm from the top.
9. Remove the paper before the solvent reaches the top and mark the solvent front.
10. Amino acids are invisible so to show the distance they have travelled they need to
be sprayed with Ninhydrin to become visible.
11. Leave the paper to dry.
12. Once dried circle the area covered by the several types of amino acids and pencil
a cross in the centre.
13. Calculate the Rf values using the formula distance by amino acids/distance of
solvent.
14. Compare the Rf values in the mixture to the known amino acids to identify them.
2
, Unit 2 Assignment C__ Chromatography
Results
Amino Distance in mm Rf values
Acids travelled
Solvent Spot
Glycine 99 41 0.41
Glutamic 99 22 0.23
Valine 100 61 0.61
Lysine 99 31 0.31
Tyrosine 99 35 0.35
Mixture 99 1. 19 1. 0.19
2. 0.67
2. 66
3
Unit 2 Undertake chromatographic
techniques to identify components
in mixtures Assignment C
Contents
Chromatography __ Amino Acids..........................................................................................................1
Title....................................................................................................................................................2
Introduction.......................................................................................................................................2
Method..............................................................................................................................................2
Results...............................................................................................................................................3
Analysis..............................................................................................................................................4
Conclusion.........................................................................................................................................6
Evaluation..........................................................................................................................................7
Suggest improvements......................................................................................................................9
Chromatography __ thin layer chromatography (TLC)........................................................................10
Title..................................................................................................................................................10
Introduction.....................................................................................................................................10
Method............................................................................................................................................10
Results.............................................................................................................................................11
Analysis............................................................................................................................................13
Conclusion.......................................................................................................................................15
Evaluation........................................................................................................................................17
Suggests improvements...................................................................................................................17
Glossary...........................................................................................................................................18
References.......................................................................................................................................19
Chromatography __ Amino Acids
1
, Unit 2 Assignment C__ Chromatography
Title
Use chromatography paper to separate a mixture of amino acids,
identify and compare them to be able to find the Rf value.
Introduction
I will find which amino acids are present in the mixture using 5 known samples of
amino acids and another mixture using paper chromatography.
Method
1. Draw a pencil line 2cm from the bottom of the chromatography paper.
2. Add 6 crosses in pencil on the line equally spaced out. Write your name or initials
in one of the upper corners in pencil.
3. Label the crosses of the amino acids Tyrosine (T), Lysine (L), Glutamic acid (Glu),
Glycine (Gly), Valine (V), and the mixture (M).
4. Use a capillary tube to spot the samples and mixture onto the crosses. Use as
small a spot as possible.
5. Add 1cm of solvent to the beaker (must be below origin line when paper is
added).
6. Solvent is ethanol, water, and concentrated ammonia at a ratio of 8:1:1.
7. Make sure the paper is not touching the sides of the beaker or curled.
8. Leave the solvent to travel up the paper until it is a few cm from the top.
9. Remove the paper before the solvent reaches the top and mark the solvent front.
10. Amino acids are invisible so to show the distance they have travelled they need to
be sprayed with Ninhydrin to become visible.
11. Leave the paper to dry.
12. Once dried circle the area covered by the several types of amino acids and pencil
a cross in the centre.
13. Calculate the Rf values using the formula distance by amino acids/distance of
solvent.
14. Compare the Rf values in the mixture to the known amino acids to identify them.
2
, Unit 2 Assignment C__ Chromatography
Results
Amino Distance in mm Rf values
Acids travelled
Solvent Spot
Glycine 99 41 0.41
Glutamic 99 22 0.23
Valine 100 61 0.61
Lysine 99 31 0.31
Tyrosine 99 35 0.35
Mixture 99 1. 19 1. 0.19
2. 0.67
2. 66
3
