Question

3b. A student found that in the P60 gel filtration chromatography, there were two absorption peaks at 400 nm, the first peak occurred in fraction 2 while the second occurred in fraction 9. They reasoned that the first was haemoglobin and that the second was vitamin B12. They concluded that gel filtration chromatography separates molecules on the basis of their mass. Haemoglobin had a molecular mass of 63 kDa while vitamin B12 had a molecular mass of 1.4 kDa. Haemoglobin was eluted first since it was heavier than vitamin B12 and thus due to gravity sunk to the bottom of the mobile phase as it moved through the column.

1. Did the student draw the correct conclusion from the chromatography result? Justify your answer.
2. Describe the elution profile of your experiment including a description of the theory behind the separation of molecules by gel filtration chromatography.

Answer 1