BTEC Applied science unit 2C Chromatography
Each practical was done separately and written up differently so there are 3 assignments
All are distinction level
Apparatus- Justification -
Amino acids solution (the acids used are Required as it’s the essential part of the
aspartame, analine, leucine) practical.
Chromatography strip (about 10-12cm long) Required part of the practical to place the
amino acids on.
Running solvent (Butanol, acetic acid, Required for the practical to help move the
water) solvent up the chromatography paper.
Capillary tubes/ Disposable pipette tips Used to transfer the amino acids onto the
chromatography paper.
Crayon Pencil Used to draw a line about 1cm from the
paper so there is a clear line on where the
amino acid is placed.
Ruler Used to draw a straight line onto the
chromatography paper.
Wooden splint Used to keep the chromatography paper
upright.
Ninhydrin solution in acetone Required for the chromatography paper.
Oven (set at 105oC) Required for the practical.
Hairdryer Used to dry the chromatography paper.
Beaker Used to put the solution and the
chromatography plate in.
Variables- What they are-
Control variable - You use the same amino acids for the
second trial.
Independent variable- We changed the pipette after every amino
acid was transferred
Dependent variable- The Rf value how far the acid travelled up
the chromatography paper.
Risk assessments:
Level of risk - Risk - Preventions -
Low Apparatus such as beakers Plastic apparatus may be
may break due to the fact used so that it doesn’t break
that its glass and may cause causing injuries.
cuts on the person dealing
with them.
Medium Be careful to avoid burns To prevent the burns, I could
when melting the capillary wear gloves, or I could open
tubes. the flame on the Bunsen
burner in a low heat so that I
can be cautious when
handling the capillary tube.
High Butanol is a flammable Make sure there are no
liquid. Some of the solvents open flames and/or heat
will go down the drain near the developing solvent.
, which may cause a chemical Dispose of any left-over
reaction which is not developing solvent
wanted. (butanol/acetic acid/water)
in reclaim container
provided. No solvent should
be allowed to go down the
drain.
Method-
1. Draw a pleasant horizontal line with a pencil approximately 1cm from one cease of the
chromatography paper, at the same time as doing this, take care to keep away
from damaging the floor and do not touch the paper with naked hands. Forceps had been used
to choose up the paper by means of the outer edge and gloves had been worn to avoid getting
sweat or oil at the paper.
2. Sweat on the skin incorporates amino acids and this could confuse the results.
Minimum pressure become carried out when drawing the start line.
3. A Bunsen burner was used to warmth and pull the capillary tube to make a finely pointed
tip.
4. A small quantity of the primary amino acid changed into transferred onto a
marked factor at the pencilled line on the chromatography strip the usage of the tip of the
capillary tube. A hairdryer changed into used dry the pattern. Once dry,
the manner become repeated to give a small concentrated pattern.
5. The other amino acids had been carried out in the same manner to one-of-a-kind,
marked points at the pencilled line taking care to area the extraordinary samples
an identical distance away from every other. 0.5cm3 of solvent turned into poured into the
beaker.
6. The chromatography paper strip became decreased into the solvent making sure that
the pattern spots have been not immersed.
7. The paper being supported by means of a wooden splint to maintain it away from the
rims of the beaker.
8. The chromatogram turned into left for about 1 hour; through this time the solvent the
front had travelled two/three of the manner up the strip. The paper strip changed
into eliminated from the solvent and the solvent the front was carefully marked with a pencil
line.
9. The strip became then left to dry. Once dry, the chromatography strip was moved into
a fume cupboard and sprayed with Ninhydrin. Ninhydrin reacts with unfastened alpha-amino
groups (NH2-C-COOH) that are present in all amino acids. Ninhydrin solution exposes the
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