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.
Session 1-8 Development and Impact of SIM-MOD (Strategic Intervention MAterial and Module COmbined) On Students' Academic Achievement in Mathematics at The 8th Grade Leve PDF