Harry Sedgwick

8/10/07
Chromatography practical-
write up

Aim: - Was to find out the amino-acids in the mixture using the
Chromatography paper and equipment.

Introduction: - Chromatography is a method of separating a

mixture of compounds by the use of a porous material. Using
chromatography paper I will determine what amino-acids are in the
mixture of substances- these amines being: valine, glylcine, leucine,
alaline. The amines will travel different distances from the origin
therefore showing what the amines are in the mixture. There is
different chromatography to E.G two way chromatography- this is
when a mixture of amines is put on one spot in the corner of a piece
of Chromatography paper therefore it can be checked twice:
(diagram to visualise):

The RF (Retardation factor) can be illustrated easily as well:

Safety:- There are some procedures in this experiment that need to

be carried out with caution e.g. the Ninhydrin in this experiment
(liquid substance in which the paper will be soaked in before heated)
is a cancerous material and also the heating of the paper requires
great safety issues so students cannot deal with these procedures
only teachers can safely take care of this. We also need to wear
overalls and gloves in this experiment to not contaminate our
experiment and ourselves.

Apparatus:- we need some equipment to complete this practical,

this being:-
 • Chromatography paper.
• The right amines (valine, glycine, leucine, alaline) and the
mixture we have to find the contents of.
• 5 different pipettes, 1 for each amine therefore preventing
contamination.
• Gloves and overalls.
• Ruler and pencil for marking points on the paper.
• And the oven chromatography equipment needed by teachers
to complete the experiment : ninhydrin for soaking in a
chromatography jar and an oven set to 100 degrees
Diagrams of equipment needed:-

Method:- Firstly I put my gloves and overalls on and cleaned the

table I was working on ( to prevent contamination). I then marked
the initial points on my Chromatography paper e.g. the origin line
and 5 crosses indicating where to put my amines and the mixture.
After that I started to put my amines on the paper on the right
points marked 1-5 e.g.:
1-
2-
3-
4-
5- Mixture
I had to be careful that I did not apply too much on the crosses
because I did not want the amine to spread. I then had to wait for it
to dry to apply another dot of the same amine on each spot to get a
more concentrated dot. I then gave it to a teacher to process the
rest of the experiment - to put in the chromatography jar and then
put in the oven at 100 degrees.

Conclusion:- I could have carried out this experiment much more

efficiently by using different methods- if I had the time. I could have
used a smaller pipette end:
This would help because it would give a spot that is more defined
therefore a spot of amine that is a lot less likely to merge with other
amines. We could also apply more amine to one spot using the
smaller pipette (remembering to let the amine dry before the next
application). I could have also used more covering on the table and
myself to lower the level of contamination. Lastly I think I should
have made another copy of the experiment and maybe used 2-way
chromatography to get better results.