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Amino Acid Lab Report

Name: Shanya Shayal Lab Group: Thursday(8-12pm)

ID: S11157053

Pre-labs
Question 1

leucine phenylamine

Threonine glutamine

Question 2
Leucine phenylalanine

threonine glutamine
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Question 3
Dissociation equations (reactions) of lysine

pl lysine= (pk2+ pk3)/2

=(8.98+10.53)/2
=9.76
Question 4

The structure of the dipeptide Leu-Ile in the neutral (unionized) form.

Results and Data

AMINO ACIDS SOLVENT A (pH = 9.8-10) SOLVENT B (pH = 3-4)

Rf Color Rf Color
Glycine 0.89 purple 0.44 Pink
Alanine 0.91 purple 0.52 pink
Serine 0.88 purple 0.41 pink
Cysteine 0.91 purple 0.83 pink
Aspartic acid 0.86 purple 0.37 pink
Asparagine 0.83 Light brown 0.35 Light brown
Glutamic acid 0.9 Purple 0.45 pink
Valine 0.95 purple 0.7 pink
Leucine 0.97 purple 0.82 pink
Unknown 1 0.9 Purple 0.33 pink
0.51
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Unknown 2 0.9 Purple 0.58 pink

Calculation, Analysis and Discussion

1. Show clearly how you calculated the Rf values for serine, unknown 1 and unknown 2.
Rf value for serine = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑐𝑜𝑚𝑝𝑜𝑢𝑛𝑑 (𝑐𝑚)
𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 (𝑐𝑚)
=8.8/10
=0.88
Solution A

Rf value for unknown 1 = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑐𝑜𝑚𝑝𝑜𝑢𝑛𝑑 (𝑐𝑚)

𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 (𝑐𝑚)
=9/10
=0.9

Rf value for unknow 2 = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑐𝑜𝑚𝑝𝑜𝑢𝑛𝑑 (𝑐𝑚)

𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 (𝑐𝑚)
=0.9/10
=0.9
Solution B

Rf value for unknown 1 = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑐𝑜𝑚𝑝𝑜𝑢𝑛𝑑 (𝑐𝑚)

𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 (𝑐𝑚)
=3.3/10
=0.33
Rf value for unknown 2 = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑐𝑜𝑚𝑝𝑜𝑢𝑛𝑑 (𝑐𝑚)
𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 (𝑐𝑚)
=5.1/10
= 0.51

Rf value for unknow 2 = 𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑐𝑜𝑚𝑝𝑜𝑢𝑛𝑑 (𝑐𝑚)

𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 (𝑐𝑚)
=5.8/10
=0.58

2.
(a) Alanine because the rf values for unknown 1 (rf for solvent a=0.9 and solvent b rf = 0.51) is
similar to that of alanine with rf value for solvent a=0.9 and solvent rf=0.52, and has similar
colours of purple for solvent a and pink for solvent b.
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(b) Unknown 2 is alanine because the rf value for alaine in solvent a is same as unknown 2 in
solvent a with color purple and rf value for alanine in 0.52 in solvent b which is same to 0.58
of unknown 2 in solvent b which is also same colour pink.

3. A.)

(a) Unknown 1 in solution a=Alanine at pH=10

(b.) unknown 1 solvent B (pH ~ 4).

(c.) unknown 2 in solvent A (pH ~10).

(d.) the main form of unknown 2 in solvent B (pH ~4).

Post-lab questions

1.) Handle the chromatography paper only at its edges or with the gloves to prevent getting oils from
your fingers on the paper or wetting the paper .

2.) Pencil lead is almost 100% graphite and won't be separated by the solvent. Ink, however, is a
mixture of dyes and/or pigments and solvents or other liquids. It will be separated by the
chromatography solvent.
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3.) Rf is only useful if the solvent front distance is known.the solvent starts to evaporate at the TLC
chamber is open. Letting the solvent to evaporate in equilibrium with the chamber’s vapor.

4.) Ninhydrin is the most well known spray reagent for identification of amino acids. Spring with
Ninhydrin as a non-specific reagent is well-known and widely used for its remarkable high sensitivity.
And amino acids are colorless compounds and they can be detected on the chromatogram by using
Ninhydrin reagent.

5.) They all have a stationary phase (a solid, or a liquid supported on a solid) and a mobile phase (a
liquid or a gas). The mobile phase flows through the stationary phase and carries the components of
the mixture with it.The stationary phase is a liquid or solid component that is fixed in a place for the
procedure.