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Unit 19 A3 All Answers for Pass , Merit and Distinction
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unit 19 A3 with all answer to question achieved distinction in this assignment .
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Task 11.You will use paper to investigate the range of amino acids in a sample of orange
juice
(or grapefruit or tomato, or similar).
Lenten Paper Chromatography
Aim: To use lantern chromatography paper to find 2 unknown amino acids and
amino acids present in the orange juice.
Equipment:
Chromatography paper (stationary phase)
Coffee filters or blotting paper or paper towels may also work
Chromatography chamber
Paper clip
Skewer (or pencil)
Water soluble black marker(s)
Beaker
Ethanol
Water
Ammonia
Ruler
Ninhydrin
Solvents (mobile phase)
Amino Acids (from a sample of orange juice): Glutamic acid, leucine, Phenylalanine,
alanine, glycine, proline, tyrosine,unknown x and y.
Method:
Method for the Lenten paper chromatography for identifying the amino acids
present in the 2 unknown samples and in fruit juice
1.Firstly draw a line on the chromatography paper of 1.50cm height with a
pencil.Then the line which we drew is called the baseline.
2.Secondly acquire a piece of tissue and place it on the table and place the
chromatography paper on top of the tissue.
3.Thirdly label the paper with each amino
acid(Tyrosine,alanine,leucine,phenylalanine,aspartic acid,X,X and orange juice)
4.Fourthly now place 5 drops of each amino acid on the baseline.
5. Sixthly after you have placed all five drops get a hair dryer and dry the dots
completely.
6. Seventhly when all the dots have dried up, clip the two ends of the paper to
, make a cylinder shape.
7.Eightly fill a beaker with ethanol water ammonia of a height of 0.5cm and place in
a fume cupboard( the solvent should not touch the baseline).Place the Lenten
paper inside the beaker.
8.Then ensure the beaker is covered with cling film and allow it to absorb
everything for 5 mins.
9.Next leave aside, until the solvent has absorbed and risen to a maximum level
ensuring it is not overflowing.Then take it out from the beaker and unclip it.
10.Lastly proceed the chromatogram in the oven for 5 mins and then spray
ninhydrin to see how the chromatogram develops.
Result:
Lenten paper chromatography to find the 2 unknown amino acids and the amino acid
present in the orange juice.
Sample of Distance of Distance of Rf value Confirmation of
amino acid the amino acid the solvent unknown
from the from the
baseline baseline
Tyrosine 4.7 cm 8.3 cm 4.7/8.3=0.56
Proline 4.6 cm 8.3cm 4.6/8.3=0.55
Leucine 6.5cm 8.3cm 6.5/8.3=0.78
Alanine 4.5cm 8.3cm 4.5/8.3=0.54
Phenylalanine 5.7cm 8.3cm 5.7/8.3=0.68
Aspartic Acid 1cm 8.3cm 1/8.3=0.12
X 2.9cm 8.3cm 2.9/8.3=0.34 Proline and
alanine
Y 6.6cm 8.3cm 6.6/8.3=0.79 Leucine
Orange juice 4.4cm 8.3cm 4.4/8.3=0.53 Proline and
alanine
In Conclusion:
I can confirm that unknown x is proline and alanine and unknown y is leucine.This
was deduced by comparing RF indexes, unknown x measured 0.34 and proline
and alanine was the closest match on the table with 0.55 for proline and alanine
0.54. Unknown y measured 0.79 and leucine was the closest match on the table
with 0.78. For orange juice measured 0.53 and proline and alanine was the closest
match on the table with 0.55 for proline and alanine 0.54.
, 2. You will use thin-layer chromatography to investigate the range of plant
pigments in a sample of freshly grown spinach leaves (or nettle leaves, or similar).
Thin-layer Chromatography
Aim: To use thin-layer chromatography to investigate the range of plant
pigments in a sample of freshly grown spinach leaves (or nettle leaves, or
similar).
Equipment:
TLC Glass silica gel plate
Pencil
Solvents (clear liquid acetone)
Sand
Hairdryer
Carrot and spinach leaves
Hexane
Acetone
Method:
1. Place some leaves in a pestle and mortar and mash them to extract pigment. Add
some drops of the sample extracted pigments.
2. A horizontal line is drawn near one end of the silica plate (about 1 cm from the
bottom edge) of the slide.
3. Place a small drop of one of the leaf juices on the line on the paper using a
capillary tube. Labelling the drop by a pencil mark with a letter (either G or S) to
identify the compound later. The drops are then soaked on the paper x 7 times and
air dried in between.
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