Chromatography of amino acids 21/3/2015

Chromatography of amino acids

Jessica Lorenzana
Date:21/3/2015
Supervisor:Mr.Osborne

Chromatography of amino acids 21/3/2015

Introduction

Purpose or setting or background: Amino acids are the building blocks of proteins which are found
in all living beings.
Research question or problem: Can the amino acids which are present in the proteins of raw fruit
juice, be identified?"
Hypothesis: The amino acids found in raw fruit juice can help determine the structure of the
proteins and its Rf values.
Prediction: Through the process of chromatography I believe there will be colour on the paper
that will help us identify the Rf values of the raw fruit juice. This will thus help us determine the types of
amino acids that in the juice.

Method

Materials & Equipment:

Glass jar, with lid and a device to suspend the chromatogram
Chromatography paper this should be cut to a length just a little longer than the height of the gas
jar.
Scissors
Fine capillary tube
Petri dish
Gloves
test-tube of solvent (4 parts Butan-1-ol: 1 part glacial Ethanoic acid: 1 part water)
Small amount of a mixture of known amino acids, including lysine, aspartic acid and leucine
Small amount of fresh juice orange
Ninhydrin
access to the fume cupboard

Risk Assessment & Safety: Ninhydrin stains the hands so it is important that gloves are worn
while handling and the Ninhydrin is used outside. It is also important to note that the solvent place inside
the jar should be covered at all times when it is not being used.
Ethical and environmental considerations: When using the Ninhydrin outside it is important to not
use this chemical on anything but the chromatography paper

Chromatography of amino acids 21/3/2015

Results
Treatment of results
Due to the errors in this investigation there were not any results for the investigation. After
two days there was only appeared to be slight colouring in the chromatography paper. This does not mean
that that the amino acids in the fruit juice cannot be identified, but that the conditions in which the
investigation took place were unstable.
Errors and limitations
the weather- rained constantly while the papers were drying
the orange juice was not concentrated enough with the solvent
we used ripe oranges, all of the amino acids could have been used to make protein
We could have touched the chromatography paper and perhaps taken the amino acids away

EVALUATION
Conclusions: I still believe that the hypothesis is true because there were still hints of colour in the
chromatography paper.
Discussion: There were no results for this investigation, but the hypothesis can still be supported due to
the errors and conditions of the investigation.
Proposals for extension: What could have caused the amino acids not to appear on the chromatography
paper?
Modifications and improvements: The experiment can be done on a clear day in the afternoon where
the paper is more likely to dry faster by receiving more sunlight. We could use unripe oranges so the
solution of amino is less dilute. Finally we could have a more concentrated amount of drops in the
chromatography paper so it will appear when the Ninhydrin is added.